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The Central Nervous System (CNS) function was shown to be fueled exclusively by oxidative phosphorylation (OXPHOS). This is in line with the sensitivity of brain to hypoxia, but less with the scarcity of the mitochondria in CNS. Consistently with the ectopic expression of FoF1-ATP synthase and the electron transfer chain in myelin, we have reported data demonstrating that isolated myelin vesicles (IMV) conduct OXPHOS. It may suggest that myelin sheath could be a site for the whole aerobic degradation of glucose. In this paper, we assayed the functionality of glycolysis and of TCA cycle enzymes in IMV purified from bovine forebrain. We found the presence and activity of all of the glycolytic and TCA cycle enzymes, comparable to those in mitochondria-enriched fractions, in the same experimental conditions. IMV also contain consistent carbonic anhydrase activity. These data suggest that myelin may be a contributor in energy supply for the axon, performing an extra-mitochondrial aerobic OXPHOS. The vision of myelin as the site of aerobic metabolism may shed a new light on many demyelinating pathologies, that cause an a yet unresolved axonal degeneration and whose clinical onset coincides with myelin development completion., (Copyright © 2013 Elsevier Masson SAS. All rights reserved.)

Fanconi anemia (FA) is a rare and complex inherited blood disorder of the child. At least 15 genes are associated with the disease. The highest frequency of mutations belongs to groups A, C and G. Genetic instability and cytokine hypersensitivity support the selection of leukemic over non-leukemic stem cells. FA cellular phenotype is characterized by alterations in red-ox state, mitochondrial functionality and energy metabolism as reported in the past however a clear picture of the altered biochemical phenotype in FA is still elusive and the final biochemical defect(s) still unknown. Here we report an analysis of the respiratory fluxes in FANCA primary fibroblasts, lymphocytes and lymphoblasts. FANCA mutants show defective respiration through Complex I, diminished ATP production and metabolic sufferance with an increased AMP/ATP ratio. Respiration in FANCC mutants is normal. Treatment with N-acetyl-cysteine (NAC) restores oxygen consumption to normal level. Defective respiration in FANCA mutants appear correlated with the FA pro-oxidative phenotype which is consistent with the altered morphology of FANCA mitochondria. Electron microscopy measures indeed show profound alterations in mitochondrial ultrastructure and shape., (Copyright © 2013 Elsevier Masson SAS. All rights reserved.)

Fanconi anemia (FA) is a rare, complex disorder that manifests in childhood. Children with FA suffer bone marrow failure, leukemias, or solid tumors. FA-associated mutations are found in 15 proteins that are involved in DNA repair. Some of these proteins have extranuclear activities involving redox balance, apoptosis, and energy metabolism; and recent data demonstrate respiratory impairment in FA cells, suggesting that altered mitochondrial function is a factor in this disease., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Myelin sheath is the proteolipid membrane wrapping the axons of CNS and PNS. We have shown data suggesting that CNS myelin conducts oxidative phosphorylation (OXPHOS), challenging its role in limiting the axonal energy expenditure. Here, we focused on PNS myelin. Samples were: (i) isolated myelin vesicles (IMV) from sciatic nerves, (ii) mitochondria from primary Schwann cell cultures, and (iii) sciatic nerve sections, from wild type or Charcot-Marie-Tooth type 1A (CMT1A) rats. The latter used as a model of dys-demyelination. O₂ consumption and activity of OXPHOS proteins from wild type (Wt) or CMT1A sciatic nerves showed some differences. In particular, O₂ consumption by IMV from Wt and CMT1A 1-month-old rats was comparable, while it was severely impaired in IMV from adult affected animals. Mitochondria extracted from CMT1A Schwann cell did not show any dysfunction. Transmission electron microscopy studies demonstrated an increased mitochondrial density in dys-demyelinated axons, as to compensate for the loss of respiration by myelin. Confocal immunohistochemistry showed the expression of OXPHOS proteins in the myelin sheath, both in Wt and dys-demyelinated nerves. These revealed an abnormal morphology. Taken together these results support the idea that also PNS myelin conducts OXPHOS to sustain axonal function., (© 2013 International Society for Neurochemistry.)

Background: Spiral low-dose computed tomography (LDCT) permits to measure whole-lung volume and density in a single breath-hold., Objective: To evaluate the agreement between static lung volumes measured with LDCT and pulmonary function test (PFT) and the correlation between the LDCT volumes and lung density in restrictive lung disease., Design: Patients with Systemic Sclerosis (SSc) with (n = 24) and without (n = 16) pulmonary involvement on sequential thin-section CT and patients with chronic obstructive pulmonary disease (COPD)(n = 29) underwent spirometrically-gated LDCT at 90% and 10% of vital capacity to measure inspiratory and expiratory lung volumes and mean lung attenuation (MLA). Total lung capacity and residual volume were measured the same day of CT., Results: Inspiratory [95% limits of agreement (95% LoA)--43.8% and 39.2%] and expiratory (95% LoA -45.8% and 37.1%) lung volumes measured on LDCT and PFT showed poor agreement in SSc patients with pulmonary involvement, whereas they were in substantial agreement (inspiratory 95% LoA -14.1% and 16.1%; expiratory 95% LoA -13.5% and 23%) in SSc patients without pulmonary involvement and in inspiratory scans only (95% LoA -23.1% and 20.9%) of COPD patients. Inspiratory and expiratory LDCT volumes, MLA and their deltas differentiated both SSc patients with or without pulmonary involvement from COPD patients. LDCT lung volumes and density were not correlated in SSc patients with pulmonary involvement, whereas they did correlate in SSc without pulmonary involvement and in COPD patients., Conclusions: In restrictive lung disease due to SSc there is poor agreement between static lung volumes measured using LDCT and PFT and the relationship between volume and density values on CT is altered.

Study Objective: We investigate the prognostic value of plasma lactate levels in patients with acute pulmonary embolism., Methods: We studied adult patients with symptomatic, objectively confirmed pulmonary embolism presenting to a single emergency department. Plasma lactate and troponin I levels were tested at presentation. We considered lactate values greater than or equal to 2 mmol/L and troponin I values greater than or equal to 0.10 ng/mL to be abnormal. Right-sided ventricular dysfunction was assessed by echocardiography. Primary endpoint was all-cause death occurring on or before 30 days after presentation. Secondary endpoints were the composite of all-cause death and clinical deterioration (defined as progression to shock, mechanical ventilation, or cardiopulmonary resuscitation) and death caused by pulmonary embolism. We tested the association between lactate level greater than or equal to 2 mmol/L and the endpoints using Cox proportional hazards regression analysis., Results: Of the 270 patients included in the study, the mean age was 73 years (SD 12.7 years) and 151 (55.9%) were women. Twelve patients (4.4%) showed shock or hypotension (shock or systolic arterial pressure <100 mm Hg) at presentation, 109 (40.4%) had right-sided ventricular dysfunction, 93 (34.4%) showed troponin I level greater than or equal to 0.10 ng/mL, and 81 (30%) showed lactate level greater than or equal to 2 mmol/L. Seventeen patients (6.3%) died, 12 (4.4%) because of pulmonary embolism, and 37 (13.7%) reached the composite endpoint. Patients with lactate level greater than or equal to 2 mmol/L showed higher mortality (17.3%; 95% confidence interval [CI] 11.9% to 20%) than patients with a lower level (1.6%; 95% CI 0.8% to 1.9%). Plasma lactate level was associated with all-cause death (hazard ratio 11.67; 95% CI 3.32 to 41.03) and the composite endpoint (hazard ratio 8.14; 95% CI 3.83 to 17.34) independent of shock or hypotension, right-sided ventricular dysfunction, or elevation of troponin I values., Conclusion: Patients with pulmonary embolism and elevated plasma lactate level are at high risk of death and adverse outcome, independent of shock or hypotension, or right-sided ventricular dysfunction or injury markers., (Copyright © 2012. Published by Mosby, Inc.)

Myelin sheath is a lipid-rich membrane, consisting of 70% lipid and 30% proteins, that is involved in physiological and pathological processes. For this reason its protein composition has been often investigated, principally by two-dimensional electrophoresis; however, the consistent lipid content makes it difficult to obtain good proteins separation. To improve the resolution of myelin proteins in a denaturing monodimensional gel electrophoresis, we examined several mixtures for the denaturation of the sample, utilizing different detergents and reducing agents. The definition of the protein pattern was analyzed by both "Blue Silver" Coomassie staining and Western Blot analysis against myelin basic protein, one of the most represented myelin proteins. The best resolution is observed when the sample was incubated with a mixture containing 1.25% dithiothreitol, 4 M urea, and 1% dodecyl maltoside or 1 % 3-[(3-cholamidopropyl)-dimethylammonio]-1-propanesulfonate, prior to addition of denaturing agents. In conclusion, this work describes a novel method to improve the separation of myelin proteins in a monodimensional gel electrophoresis. It may be also useful for investigating other lipid-rich samples.

COPD is a heterogeneous disorder whose assessment is going to be increasingly multidimensional. Grading systems such as BODE (Body-Mass Index, Obstruction, Dyspnea, Exercise), mBODE (BODE modified in grading of walked distance), ADO (Age, Dyspnea, Obstruction) are proposed to assess COPD severity and outcome. Computed tomography (CT) is deemed to reflect COPD lung pathologic changes. We studied the relationship of multidimensional grading systems (MGS) with clinically determined COPD phenotypes and CT lung density. Seventy-two patients underwent clinical and chest x-ray evaluation, pulmonary function tests (PFT), 6-minute walking test (6MWT) to derive: predominant COPD clinical phenotype, BODE, mBODE, ADO. Inspiratory and expiratory CT was performed to calculate mean lung attenuation (MLA), relative area with density below-950 HU at inspiration (RAI(-950)), and below -910 HU at expiration (RAE(-910)). MGS, PFT, and CT data were compared between bronchial versus emphysematous COPD phenotype. MGS were correlated with CT data. The prediction of CT density by means of MGS was investigated by direct and stepwise multivariate regression. MGS did not differ in clinically determined COPD phenotypes. BODE was more closely related and better predicted CT findings than mBODE and ADO; the better predictive model was obtained for CT expiratory data; stepwise regression models of CT data did not include 6MWT distance; the dyspnea score MRC was included only to predict RA-950 and RA-910 which quantify emphysema extent. BODE reflect COPD severity better than other MGS, but not its clinical heterogeneity. 6MWT does not significantly increase BODE predictivity of CT lung density changes.

Practicing a visual task commonly results in improved performance. Often the improvement does not transfer well to a new retinal location, suggesting that it is mediated by changes occurring in early visual cortex, and indeed neuroimaging and neurophysiological studies both demonstrate that perceptual learning is associated with altered activity in visual cortex. Theoretical treatments tend to invoke neuroplasticity that refines early sensory processing. An alternative possibility is that performance is improved because of an altered attentional strategy and that the changes in early visual areas reflect locally altered top-down attentional modulation. To test this idea, we have used functional MRI to examine changes in attentional modulation in visual cortex while participants learn an orientation discrimination task. By examining activity in visual cortex during the preparatory period when the participant has been cued to attend to an upcoming stimulus, we isolated the top-down modulatory signal received by the visual cortex. We show that this signal changes as learning progresses, possibly reflecting gradual automation of the task. By manipulating task difficulty, we show that the change mirrors performance, occurring most quickly for easier stimuli. The effects were seen only at the retinal locus of the stimulus, ruling out a generalized change in alertness. The results suggest that spatial attention changes during perceptual learning and that this may account for some of the concomitant changes seen in visual cortex., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

Background: To ascertain if analysis of lung density histograms in thin-section CT was more reproducible than visual assessment of lung changes in systemic sclerosis (SSc), and if such density histogram parameters as mean lung attenuation (MLA), skewness, and kurtosis could more closely reflect pulmonary function as well as exercise and quality of life impairment., Methods: The intraoperator and interoperator reproducibility of visual and densitometric lung CT analysis in 48 SSc patients examined with CT were evaluated by means of weighted kappa statistics. Univariate and multivariate regression analyses were applied to evaluate the relationship of visual and densitometric CT measurements with functional parameters including functional residual capacity (FRC), FVC, FEV(1), diffusion capacity of the lung for carbon monoxide (Dlco), 6-min walking testing (6MWT), and health-related quality of life questionnaire (QLQ) parameters., Results: The intraoperator and interoperator reproducibility of MLA (intraobserver weighted kappa = 0.97; interobserver weighted kappa = 0.96), skewness (intraobserver weighted kappa = 0.89; interobserver weighted kappa = 0.88), and kurtosis (intraobserver weighted kappa = 0.89; interobserver weighted kappa = 0.88) were higher than those of visual assessment (intraobserver weighted kappa = 0.71; interobserver weighted kappa = 0.69). In univariate analysis, only densitometric measurements were correlated with some exercise and QLQ parameters. In multivariate analysis, MLA (square regression coefficient corrected [R(2)c] = 0.70), skewness (R(2)c = 0.78), and kurtosis (R(2)c = 0.77) were predicted by FRC, FVC, Dlco, 6MWT, and QLQ parameters, while visual assessment was associated only with FRC and FVC (R(2)c = 0.40)., Conclusions: In SSc, densitometric analysis is more reproducible than visual assessment of lung changes in thin-section CT and more closely correlated to pulmonary function testing, 6MWT, and QLQ. Density histogram parameters may be useful for cross-sectional and longitudinal studies of lung involvement in SSc.

Objective: To correlate lung density measurements with the results of visual assessment of thin-section computed tomography (CT) and of pulmonary function tests (PFT) in Systemic Sclerosis (SSc)., Methods: Thirty-nine SSc patients underwent sequential thin-section CT and spiral low-dose whole-lung acquisitions. The thin-section CT scans were evaluated with a dedicated visual scale. Mean lung density, skewness, and kurtosis were calculated from the lung density histogram. In addition from the spiral low-dose acquisition, the lung volume was computed. The visual score, the densitometric parameters, and the lung volume were correlated with the PFT., Results: Mean lung density, skewness, and kurtosis computed from thin-section (R = 0,66; R = -0,74; R = -0,75) and low-dose volumetric (R = 0,72; R = -0,71; R = -0,71) CT and the lung volume (R = -0.54) correlated with the visual score. Densitometric values and lung volume consistently better correlated with PFT than the visual score., Conclusions: In SSc the histogram results are more closely correlated to PFT than the visual score. The low-dose spiral CT seems ideal for longitudinal studies.

Bowel herniation through the foramen of Winslow is among the rarest of internal hernias, accounting for less than 0.8%. In its origin, a pivotal role is played by some anatomic variations, or anomalies such as the increased mobility of the right transverse colon, and maybe the exceedingly large bore of the foramen itself. The first case of hernia through the foramen of Winslow was reported by Blandin in 1834. Since then, no more that 200 new cases have been described. Diagnosis usually is established during surgery while treating a bowel obstruction. Only in an exceedingly small group of patients is diagnosis achieved preoperatively on the basis of radiological findings. We describe a preoperatively diagnosed case of transverse colon herniation through the foramen of Winslow, showing a portal vein narrowing and periportal lymphedema at computed tomography (CT). To the best of our knowledge, only a few cases of preoperative CT diagnosis of Winslow foramen hernia have been described in the past. None had the above-mentioned CT findings.

Study Objectives: The aim of this study was to investigate the relationship between high-resolution CT (HRCT) lung attenuation measurements, acquired under spirometric control of inspiratory and expiratory lung volume, and pulmonary dysfunction as well as dyspnea severity in patients with COPD., Patients and Design: In 51 patients with COPD, we compared by linear regression, univariate and multivariate logistic regression airflow limitation (FEV(1)/vital capacity [VC]), hyperinflation (percentage of predicted residual volume [RV%]), parenchymal loss (percentage of predicted diffusing capacity of the lung for carbon monoxide [Dlco%]), and Medical Research Council (MRC) dyspnea scale with relative area with attenuation values < - 950 HU at 90% of VC [RAI(950)] and < - 910 HU at 10% of VC, respectively, and with mean lung attenuation measured at the same levels of VC (mean CT lung density at 10% of VC, and mean CT lung density at 90% of VC [MeanCTEXP])., Results: All HRCT attenuation measurements were significantly related with functional abnormalities and dyspnea severity. In multivariate logistic models, with 1 indicating worse changes in dichotomous outcome variables, MeanCTEXP independently predicted FEV(1)/VC (odds ratio [OR], 0.24; 95% confidence interval [CI], 0.11 to 0.56), RV% (OR, 0.57; 95% CI, 0.42 to 0.77), and MRC dyspnea scale (OR, 0.63; 95% CI, 0.48 to 0.82), while RAI(950) independently predicted Dlco% (OR, 1.90; 95% CI, 1.37 to 2.65)., Conclusions: Spirometrically gated measurements of HRCT lung attenuation reflect differently functional changes and dyspnea perception in COPD. Inspiratory measurements assess the extent of emphysematous tissue loss, and expiratory measurements may reflect airflow limitation and lung hyperinflation with attendant dyspnea perception. Pulmonary dysfunction in COPD cannot be assessed by a single modality of lung attenuation measurement.

Purpose: To prospectively evaluate airway wall thickness and lung attenuation at spirometrically gated thin-section computed tomography (CT) in patients with chronic obstructive pulmonary disease (COPD) and to correlate gated CT findings with pulmonary function test (PFT) results., Materials and Methods: The ethical committee approved the study, and all patients gave informed consent. Forty-two consecutive patients with COPD (20 with and 22 without chronic bronchitis [CB]) underwent gated thin-section CT and PFTs on the same day. The percentage wall area (PWA) and the thickness-to-diameter ratio (TDR) for all depicted bronchi that were round and larger than 2 mm in diameter, the mean lung attenuation (MLA), and the pixel index (PI) at -950 HU were determined. The reproducibility of the airway measurements was preliminarily tested by performing a five-trial examination in a patient with COPD and in a control patient. Differences in airway and lung attenuation measurements between the patients with and those without CB were evaluated at Mann-Whitney U testing. Simple and multiple regression analyses were used to assess the correlation between thin-section CT and PFT measurements., Results: The mean intraoperator coefficient of variation for airway measurements was 7.8% (range, 3.8%-13.4%). An average of nine bronchi per patient were assessed. Patients with CB had significantly higher PWAs, TDRs, and MLAs and significantly lower PIs than patients without CB (P < .05 for all values). The combination of PWA, TDR, and PWA normalized to body weight correlated significantly (P < .05) with the forced expiratory volume in 1 second-to-slow vital capacity ratio and the diffusing capacity of the lung for carbon monoxide in patients with but not in patients without CB. PFT results correlated better with MLA and PI in patients without CB., Conclusion: Bronchial wall measurements differ between patients who have COPD with CB and those who have COPD without CB. The correlation between airway dimensions and indexes of airway obstruction in patients with COPD and CB indicates that the bronchial tree is the site of anatomic-functional alterations in this patient group., ((c) RSNA, 2005.)

Purpose: To report the results of a three-year observational pilot study of lung cancer screening with low dose computed tomography (CT) and to present the study design of a randomised clinical trial named as ''Italung-CT''., Materials and Methods: Sixty (47 males and 13 females, mean age 64+/-4.5 years) heavy smokers (at least 20 packs-year) underwent three low-dose spiral CT screening tests one year apart on a single slice or multislice CT scanner. Indeterminate nodules were managed according to the recommendations of the Early Lung Cancer Action Project., Results: and Indeterminate nodules were observed in 33 (55%) of the subjects (60% at the baseline screening test, 24% at the first annual test and 16% at the second annual test). The size of the largest indeterminate nodule was <5 mm in diameter in 20 subjects, 10 of whom showed the nodule at the baseline test. Forty-five subjects (75%) completed the first annual test and 42 (70%) the second annual test. One (1.6%) prevalent lung cancer (adenosquamous carcinoma) and one (2.2%) incident lung cancer (small cell cancer at the first annual examination) were observed, as well as a pulmonary localisation of Hodgkin's lymphoma (at the second annual test). In addition, one subject underwent lung surgery for a chondromatous hamartoma., Conclusions: The results of the pilot study are substantially in line with those of other observational studies of greater sample size. This justifies optimism about the reliability of the results in the screened arm of the ''Italung CT'' trial which has just begun.

Objective: To compare the quantitative assessment of pulmonary emphysema with spirometric-gated computed tomography (gated CT) using 3 different acquisition techniques and to determine if low-current spiral CT could be used effectively to quantitate emphysema., Methods: Eleven patients with chronic obstructive pulmonary disease (COPD) underwent gated CT and pulmonary function tests (PFTs). Spiral whole-lung 10-mm collimation acquisitions at standard (146 mAs) and low (43 mAs) current and sequential 3-slice 1-mm collimation high-resolution computed tomography (HRCT) acquisitions at standard current were obtained at 90% of the patient's vital capacity. The mean lung density (MLD) and the pixel index (PI) derived from the 3 data sets were compared using one-way analysis of variance and correlated with PFTs using linear regression. Moreover, the radiation dose associated with each technique was measured., Results: The MLDs were not significantly different. The PIs calculated from the standard- and low-current spiral acquisitions were similar, and both were significantly different from that of HRCT. The MLDs correlated with the PFTs in standard-current spiral and HRCT but not in low-current spiral acquisitions, whereas the PIs correlated with the PFTs in all 3 techniques. High-resolution computed tomography implied the lowest dose (0.08 mSv) compared with low-current (1.2 mSv) and standard-current (4 mSv) spiral techniques., Conclusions: Low- and standard-dose spiral CT provides similar lung density data in COPD. The combination of low-dose whole-lung spiral CT and 3-slice HRCT represents the best compromise between the amount of information provided and radiation exposure to the patient and could be substituted for standard-dose spiral CT for quantitative evaluation of COPD.

Background/aims: To assess the value of hepatic-arterial-phase computed tomography (HAP-CT) versus ultrasound (US) plus alpha-fetoprotein (AFP) in the surveillance of cirrhotic patients with previously treated hepatocellular carcinoma (HCC)., Methods: Thirty-six cirrhotic patients, treated for single nodular HCC <4cm with complete response and no evidence of other focal lesions, were enrolled in a prospective study and underwent simultaneous AFP/US/spiral-CT follow-up every 6 months. Focal lesions were considered recurrences when they appeared as globular enhancement areas (EA) at HAP-CT and increased in size during the follow-up., Results: Fifteen of 36 patients showed at least one focal lesion for a total of 43 EA: 38/43 increased in size, four did not change and one disappeared. EA were first observed after a follow-up of 9+/-4 (range 6-18) months. At the same time, no patient had either nodular lesion at US examination or diagnostic levels of AFP. In 22 matched lesions, diagnosis by CT was 8.2+/-3.5 months earlier than by US. In 13 patients, one evolved EA was submitted to US-guided biopsy and histological examination showed HCC in all cases., Conclusions: Periodical spiral-CT examination is more effective than US-AFP in early detection of HCC recurrence in cirrhotic patients successfully treated for HCC.

Purpose: The purpose of this work was to evaluate feasibility of spirometric-gated high-resolution computed tomography (HRCT) in patients with chronic obstructive pulmonary disease (COPD) and to compare the lung density CT measurements obtained with and without spirometric control of lung volume., Method: Twenty-nine patients with COPD underwent pulmonary function tests and spirometric-gated (3 slices at 10% and 90% of vital capacity) and -ungated (12 slices at maximum expiration and inspiration) HRCT in the same day. Four lung density measurements (inspiratory pixel index, expiratory pixel index, inspiratory and expiratory mean lung density) derived from gated and ungated acquisitions were compared using the nonparametric Wilcoxon test, the line of equality, and the Bland and Altman test., Results: The vital capacity measured at pulmonary function tests and on the CT table showed a substantial agreement. All but one patient completed the gated and ungated examination, but only 8 (28%) of 28 patients reached the expiratory and inspiratory gating level for CT acquisitions at the first attempt. Only the inspiratory mean lung density derived from the 3 gated and 12 ungated slices showed borderline agreement. Other CT measurements, and notably all those from the 3 gated and ungated scans, acquired at the same anatomic level, did not agree., Conclusions: Although the procedure can be difficult for individual patients, spirometric gating significantly influences the lung density CT measurements and might improve standardization of CT evaluation of COPD.

The objective of this case report is to demonstrate that a benign tumour arising solely from the olfactory bulb can act as a primary epileptic focus and to illustrate the difficulty in making a preoperative diagnosis of pathological lesions in this area. The case of a 29-year-old male with intractable epilepsy is presented. Radiological imaging demonstrated a conspicuous, calcified lesion in the mesial inferior frontal lobe. Electroclinical findings confirmed this to be the epileptogenic zone. Surgery and subsequent histology surprisingly revealed this focal lesion to be a benign hamartoma arising solely from the olfactory bulb. Resection resulted in seizure resolution. Tumours of the olfactory apparatus are extremely rare, but can present with epilepsy., (Copyright 2002 S. Karger AG, Basel)

Purpose: To assess the feasibility of spirometrically gated HRCT in patients with chronic obstructive pulmonary disease and to correlate the density measures with the results of pulmonary function tests., Material and Methods: Pulmonary function tests and spirometrically gated HRCT were performed, on the same day, in 20 patients with chronic obstructive pulmonary disease. Spirometrically gated HRCT scans at 90 and 10% of the patient's vital capacity were acquired, at three anatomical levels (carena, carena plus 5 cm, carena minus 5 cm). In each section, lung parenchyma was isolated from the other thoracic structures and from the main bronchi and a densitometric evaluation was applied to the regions of interest. A histogram showing the distribution of the pixel frequencies versus lung densities, the mean CT number and the pixel index at -910 HU (for scans acquired at 10% of the patient's vital capacity) and at -950 HU (for scans obtained at 90% of the patient's vital capacity) were obtained. Average acquisition time for the spirometrically gated HRCt was 15-20 minutes, average data processing time was 20 minutes., Results: The mean CT number and the pixel index at -910 HU obtained at 10% of the patient's vital capacity correlated with the Tiffeneau index (r = 0.85; R = 0.78 respectively). The pixel index at -910 HU correlated also with the ratio of residual volume to total lung capacity (r = 0.65). The mean CT number and the pixel index obtained at 90% of the vital capacity correlated with the lung diffusion of carbon monoxide (r = 0.64 and r = 0.79 respectively)., Conclusions: In our study, we found good correlation between the densitometric values obtained with spirometrically gated HRCT and the respiratory function tests routinely employed to assess the presence and the gravity of obstructive pulmonary disease. A good reproducibility of the data is guaranteed by the spirometical gating which enables standardization of the lung acquisition volume, and by the semiautomatic contour tracing program combined with an automated densitometric evaluation. The main disadvantages of the technique are the relatively long acquisition and evaluation times. Spirometrically gated HRCT densitometry is a promising tool for longitudinal studies of diffuse lung disease (pulmonary emphysema and fibrosis).

Background: Adjuvant chemotherapy for colorectal carcinoma was found to improve survival of patients with American Joint Committee on Cancer/International Union Against Cancer Stage III disease. The usefulness of chemotherapy in patients with Stage II disease continues to be debated, and it is likely that only those patients with a poor prognosis should receive adjuvant chemotherapy. Biologic prognostic factors may allow further insight into the optimal treatment strategy for patients with Stage II or earlier disease. In this study the prognostic role of S-phase fraction (SPF) determined by flow cytometry was assessed in patients with Stage I-II colorectal carcinoma., Methods: Specimens of surgically resected colorectal carcinoma were examined for SPF by flow cytometric DNA analysis. Consecutive patients referred to the study institution were considered eligible for this study. The main inclusion criteria were a Stage I-II tumor together with sufficient tumor material and adequate follow-up information. For each stage of disease, SPF data were associated with the recurrence rate and the disease free survival (DFS)., Results: Analysis was performed on 167 patients (65 with Stage I disease and 102 with Stage II disease). Among Stage I patients, SPF was high in 20 patients and low in 45 patients. In Stage II patients, there were 36 patients with low SPF and 66 patients with high SPF. In both stages, the recurrence rate and DFS were significantly worse for the subgroups of patients with high SPF., Conclusions: SPF has revealed prognostic differences among patients with surgically resected Stage I-II colorectal carcinoma. These data should be considered for planning future trials in the adjuvant setting because patients with high SPF may benefit from adjuvant chemotherapy.

The effectiveness and safety of specific immunotherapy (SIT) in allergic diseases such as asthma have increasingly come under question. Some authors advocate eliminating SIT as a therapeutic option for allergic patients, since the risks associated with this form of asthma may outweigh its positive effects. However, in a review of twenty-nine controlled studies in 2077 children and an equal number of controls, 27 (93.1%) have shown the effectiveness of SIT in pediatric age for the treatment of asthma due to inhalant allergens (p < 0.0001). The scope of this study was to ascertain whether this form of therapy is safe and effective in pediatrics. 300 children (median age 4.4 years) with asthma due to pollen or house dust mite were prospectively followed for three years. They were randomly divided into two groups: the study group and control group, being almost equal in number of children and clinical characteristics such as sex and age. No child suffered severe reactions due to SIT. Children receiving SIT had significantly greater reductions in days (p = 0.0001) and nights (p = 0.0005) without asthma and drug usage (p = 0.0003), compared with drug-treated children. In addition, the number of asthma attacks (p = 0.0001), and the quality of life were significantly improved in the study group (p = 0.0001). These findings suggest that if suitable allergen extracts are used with close observation of therapeutic indications, and children are followed by their doctors as frequently as required, SIT is effective in the treatment of pediatric asthma, with few adverse effects.

The proportion of T lymphocytes, mainly CD4 positive, co-expressing the CD40 ligand (CD40-L) was significantly greater (P = 0.001) in the colostrum of 10 breast-feeding mothers than in either autologous or heterologous blood. This surface glycoprotein is a T cell molecule involved in B cell isotype switching and immunoglobulin production with its natural counter-receptor, CD40, expressed by both adult and infant B lymphocytes. As the T cells of newborn infants fail to express the CD40-L when stimulated in vitro, the in vivo upregulation on milk T lymphocytes may be one of the mechanisms through which the mother transfers immune protection to the suckling infant.

Flowcytometric analysis on T-cell subpopulations in the blood and cerebrospinal fluid (CSF) of 10 children with mumps meningitis (MM) revealed that the proportion of lymphocytes which bore the gamma delta T-cell receptor for antigen was significantly higher in CSF than in autologous and heterologous blood samples. gamma delta T-cell values in CSF of MM patients were also significantly higher than those calculated in CSF specimens from nine children with non-inflammatory neurological disorders. Whether and how the expanded CSF gamma delta T-lymphocyte subset, either CD8+ or double-negative (CD4-/CD8-), contributes to the eradication of mumps virus infection from the central nervous system represents a central focus of our ongoing research.

A total of 1292 sera, provided by different USLs (Local Health Centers) in the Marche Region (Italy), was tested by ELISA and Western Blot for the detection of anti-HTLV I/II ad HIV-1 antibodies. It was decided not to follow any particular criterion when collecting the sera, so no protocol of admission was established in advance. Of the 1292 sera tested, 12 were found antibody positive to HTLV I/II, 18 to HIV-1 and 4 to both viruses. Of the seropositives, for HTLV I/II and HIV-1, respectively, 2 and 3 were prisoners; 2 and 4 were IVDAs, 3 and 7 were hematological patients, 3 and 2 were hospitalized subjects (not for hematological disorders), and 2 and 2 were positives belonging to a healthy population group. These data suggest that the viruses may also be spreading among groups other than the high-risk ones (homosexuals, IVDAs, prisoners, blood-transfusion recipients, promiscuous heterosexuals).

In our program to produce monoclonal antibodies against hemopoietic functional antigens we have obtained an hybridoma cell line EA.3 reacting with human transferrin receptor. This monoclonal antibody is an IgM that displays "in vitro" antiproliferative activity against leukemic cells due to its competition with transferrin on the receptor. Cell cytotoxicity was demonstrated either with the blockage of BrdU incorporation in HL-60 cells and with leukemic CFU inhibition. In contrast, K562 cells were insensitive to the blockage of the transferrin receptor induced by this MAb.

Cell to cell interaction play a major role in the induction of a immune response. NK cells represent a special lymphoid subset which displays its cytotoxic function without the engagement of MHC system. In order to investigate the role of different adhesion molecules in the mechanism of binding of the NK cell to the classic tumor target K562 cell, we have employed different unclustered mAbs of the Adhesion session (5th "CD" Workshop, Boston 1993) mostly of the CAM (cell adhesion molecule) subpanel. After their reactivity characterization both on lymphocytes and K562 cells, those that demonstrate reactivity against the tumor target were tested in the binding assay. The target was pretreated with the monoclonal in order to block a possible reacting molecule for the effector. Then, after the incubation of lymphocytes with PE-labelled anti CD16 mAb, their ability to bind to the target was tested in flow. While the majority of the mAbs did not induce any significant change in the binding capacity of the NK subset, few of them did, and precisely anti-CD58 (LFA-3) and anti-CD54 (ICAM-1) which showed different level of inhibition, particularly drastic with S002, S083 and S100. Other mAbs, such as the S011 (anti-CD59), due to the presence of the PI-linked glycoprotein recognized on both target and effector membranes, and to its capacity to stimulate NK activity, produced a total binding of the NK population. The coincubation of targets with anti-CD54 and anti-CD58 allowed to reduce at the lowest level this function. This data seem to support the hypothesis of specific surface molecules involved in the binding process of the NK cell, after recognition of the target.

After successful bone-marrow transplantation (BMT) in thalassaemia, the individual acquires the pattern of globin synthesis of the donor. We call such an individual "ex-thalassaemic after BMT", a term that underscores the cure of the genetic defect but maintenance of residual signs of organ damage due to iron overload and dysfunction acquired during the pretransplant years. We have analysed the extent and fate of tissue iron overload in 151 ex-thalassaemic patients after BMT, according to the risk factors of hepatomegaly, hepatic portal fibrosis, and inadequate chelation therapy. Serum ferritin concentrations decreased and unbound iron binding capacity (UIBC) increased slowly during the years after the transplant. When analysed according to risk group (assigned at the time of the transplant), ferritin and UIBC returned within the normal ranges in only the low-risk group (without hepatomegaly or portal fibrosis, and with adequate chelation pre-BMT). Ferritin and UIBC were still abnormal 7 years after the transplant in the moderate-risk group (those with one or two risk factors) and highly abnormal in the high-risk group (all three risk factors) indicating persistence of, respectively, moderate and severe iron overload at the time of transplant. In ex-thalassaemic patients who were studied before and yearly after the transplant the extent of haemosiderosis, as judged by staining of liver biopsy samples, decreased during the years after transplant. The degree of iron deposition and rate of post-BMT linear growth seem to influence rate of post-BMT decrease in tissue iron overload in different risk groups at the time of BMT.

Interferon (IFN) has been shown to enhance the cytotoxic effects of 5-fluorouracil (5FUra) in colorectal cancer, and clinical trials with this combination resulted in higher response rate with respect to 5FUra alone. IFN is generally administered s.c. three times a week. This prolonged exposure could determine a block of tumor cells in the G0-G1 phase of the cell cycle, thus rendering tumor cells insensitive to 5FUra, an S-phase specific agent. In order to verify the presence of this block, 21 operable colorectal cancer patients were treated with IFN-alpha 2b at the dose of 3 megaunits every other day in the week before operation, while another 22 represented the control group. Samples of tumor tissue were taken at endoscopy and operation. [3H]Thymidine labeling index and flow cytometry were used to assess the S-phase fraction. In IFN treated patients, we found a significant statistical difference between the mean percentage of S-phase fractions evaluated either by labeling index (P = 0.00001) or by flow cytometry (P < 0.001). On the contrary, this difference was not present in the control group: labeling index, P = 0.06; flow cytometry, P = 0.08. Furthermore a significant increase in the G0-G1 phase of the cell cycle was found after IFN administration (P < 0.001) but not in the control group. Our results suggest that IFN reduces the S-phase fraction in colorectal cancer tumors. This action should be considered in the design of the 5FUra/IFN combination because it could decrease 5FUra activity, leading to a loss or a decrease in the advantage of 5FUra modulation by IFN.

A new biphasic system (MB Check, Roche) for isolation of mycobacteria from clinical specimens was evaluated by eight different microbiological laboratories in comparison with methods routinely used in the respective laboratories. Altogether 1125 clinical specimens were processed; pretreatment, if performed, was by a variety of methods. Mycobacteria were recovered from 167 specimens with the biphasic system and 165 specimens with the other methods. The average time required for isolation of Mycobacterium tuberculosis was 22.6 days with the biphasic system and 24.7 days with egg-based media; for other mycobacterial species it was 23.5 versus 20.8 days. The inclusion of a chocolate agar section in the biphasic system facilitated the early detection of contaminants, while the NAP-containing section appeared unable to differentiate Mycobacterium tuberculosis from other mycobacterial species. The biphasic system, which enables recovery of mycobacteria in small laboratories without specialized equipment, is more practical than conventional methods and at least as sensitive.

The antibacterial activity in vitro of Imipenem (N-formimidoyl-thienamycin) was studied on 237 recently-isolated nosocomial bacterial strains and compared with Aztreonam, Cefotaxime, Ceftazidime, Ceftriaxone, Piperacillin, Amikacin and Netilmicin through the determination of susceptibility by the Kirby-Bauer method. Taking into account that 65% bacterial strains, out of the examined ones, came from antibiotic high-pressure departments (45% from bone-marrow transplanted patients in the Hematological Department and 20% from Intensive Care Unit patients), the results obtained in vitro show an activity of Imipenem on Gram-negative bacteria as much as a 92.7% susceptibility and a 75.7% on Gram-positive bacteria; when compared with the activity of the other antibiotics, Imipenem shows a really excellent activity.

One of the most important problems, of very difficult solution, in organizing an A/E department is to co-ordinate the emergency in the field. This peculiar task is carried on by an operative central station, the core of an A/E Dept. The organizer in Arezzo try to find out the efficiency and a cost benefit ratio of this central station and try to jump over an historical hindrance to operate by mutual consent with voluntary service. This seems to be possible with the technological development of telephone system and broadcast system. Staff, technical qualifications, connections, reception, modulation, sorting out of different calls are discussed in conjunction with problems of health education concerning the operative central station that coordinates A/E Dept in Arezzo.

While organizing the Catchment area of A/E Department in Arezzo, a recording of intervention time of prehospital care system (primary transport) was made by land. The analysis recorded that one third of the field on the Catchment area of the A/E Dept was in a red dangerous zone reachable in 15 minutes and more. Such analysis confirmed the importance of local operation bonds all for a correct redistribution of ambulances in the Catchment area and a rational choice of vans and staff suitable for an advanced life support only where and when really necessary.