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Perfusion culture systems are widely used in tissue engineering applications for enhancing cell culture viability in the core of three-dimensional scaffolds. In this work, we present a multichamber confined-flow perfusion system, designed to provide a straightforward platform for three-dimensional dynamic cell cultures. The device comprises 6 culture chambers allowing independent and simultaneous experiments in controlled conditions. Each chamber consists of three parts: a housing, a deformable scaffold-holder cartridge, and a 7 mL reservoir, which couples water-tightly with the housing compressing the cartridge. Short-term dynamic cell seeding experiments were carried out with MC3T3-E1 cells seeded into polycaprolactone porous scaffolds. Preliminary results revealed that the application of flow perfusion through the scaffold favored the penetration of the cells to its interior, producing a more homogeneous distribution of cells with respect to dropwise or injection seeding methods. The culture chamber layout was conceived with the aim of simplifying the user operations under laminar flow hood and minimizing the risks for contamination during handling and operation. Furthermore, a compact size, a small number of components, and the use of bayonet couplings ensured a simple, fast, and sterility-promoting assembling. Finally, preliminary in vitro tests proved the efficacy of the system in enhancing cell seeding efficiency, opening the way for further studies addressing long-term scaffold colonization.

Background: The bone marrow (BM) cytokine milieu might substantially affect T-lymphocyte homeostasis in HIV-positive individuals. Interleukin-7 (IL-7) is a bone marrow-derived cytokine regulating T-cell homeostasis through a CD4+-driven feedback loop. CD4+ T-lymphopenia is associated with increased free IL-7 levels and reduced IL-7R expression/function, which are only partially reverted by highly active antiretroviral therapy (HAART). We investigated the BM production, peripheral expression and signaling (pStat5+ and Bcl-2+ CD4+/CD8+ T cells) of IL-7/IL-7Rα in 30 HAART-treated HIV-positive patients who did not experience CD4+ recovery (CD4+ ≤200/µl) and who had different levels of HIV viremia; these patients included 18 immunological nonresponders (INRs; HIV-RNA≤50), 12 complete failures (CFs; HIV-RNA>1000), and 23 HIV-seronegative subjects., Methods: We studied plasma IL-7 levels, IL-7Rα+CD4+/CD8+ T-cell proportions, IL-7Rα mRNA expression in PBMCs, spontaneous IL-7 production by BM mononuclear cells (BMMCs), and IL-7 mRNA/IL-7Rα mRNA in BMMC-derived stromal cells (SCs). We also studied T-cell responsiveness to IL-7 by measuring the proportions of pStat5+ and Bcl-2+ CD4+/CD8+ T cells., Results: Compared to HIV-seronegative controls, CFs and INRs presented elevated plasma IL-7 levels and lower IL-7Rα CD4+/CD8+ cell-surface expression and peripheral blood production, confirming the most relevant IL-7/IL-7R disruption. Interestingly, BM investigation revealed a trend of higher spontaneous IL-7 production in INRs (p = .09 vs. CFs) with a nonsignificant trend toward higher IL-7-Rα mRNA levels in BMMC-derived stromal cells. However, upon IL-7 stimulation, the proportion of pStat5+CD4+ T cells did not increase in INRs despite higher constitutive levels (p = .06); INRs also displayed lower Bcl-2+CD8+ T-cell proportions than controls (p = .04)., Conclusions: Despite severe CD4+ T-lymphopenia and a disrupted IL-7/IL-7R profile in the periphery, INRs display elevated BM IL-7/IL-7Rα expression but impaired T-cell responsiveness to IL-7, suggesting the activity of a central compensatory pathway targeted to replenish the CD4+ compartment, which is nevertheless inappropriate to compensate the dysfunctional signaling through IL-7 receptor.

Background: Treatment of HIV-infected patients with interleukin-2 (IL-2) produces significant increases in CD4 T cell counts; however an associated qualitative improvement in cells function has yet to be conclusively demonstrated. By measuring mycobacterial killing activity, we evaluated IL-2-mediated functional immune enhancement ex vivo in immunological non-responders (INRs)., Methods and Findings: PBMC from 12 immunological non-responders (INRs) (CD4+<200/µl, HIV-RNA<50 cp/ml) on combination antiretroviral treatment (cART) were collected at baseline, and after 3 IL-2 cycles. Eight INRs receiving only cART were studied as controls. After 21 days of PBMC incubation with a virulent M. avium suspension, counts of residual colony forming units (CFUs) and concentrations of TNF-α, IL-10 and IFN-γ were determined. In IL-2 treated patients, a significant reduction in mean residual CFUs of PBMC cultures was observed (p<0.01). Moreover, following IL-2 treatment, significant increases in PBMC's IFNγ production (p = 0.02) and substantial reductions in IL-10 levels were observed., Conclusions: IL-2 therapy restores the ability of the lympho-monocyte system in eliciting an effective response against mycobacterial infections. Our data indicate the possibility of a clinical role held by IL-2 in enhancing the immune function of subjects unable to achieve immune competence through cART alone.

Mechanisms by which HIV affects the thymus are multiple and only partially known, and the role of thymic dysfunction in HIV/AIDS immunopathogenesis remains poorly understood. To evaluate the effects of HIV infection on intra-thymic precursors of T cells in HIV-infected adults, we conducted a detailed immunophenotypic study of thymic tissue isolated from 7 HIV-infected and 10 HIV-negative adults who were to undergo heart surgery. We found that thymuses of HIV-infected individuals were characterized by a relative depletion of CD4+ single positive T cells and a corresponding enrichment of CD8+ single positive T cells. In addition, thymocytes derived from HIV-infected subjects showed increased levels of activated and proliferating cells. Our analysis also revealed a decreased expression of interleukin-7 receptor in early thymocytes from HIV-infected individuals, along with an increase in this same expression in mature double- and single-positive cells. Frequency of regulatory T cells (CD25+FoxP3+) was significantly increased in HIV-infected thymuses, particularly in priorly-committed CD4 single positive cells. Our data suggest that HIV infection is associated with a complex set of changes in the immunophenotype of thymocytes, including a reduction of intrathymic CD4+ T cell precursors, increased expression of activation markers, changes in the expression pattern of IL-7R and enrichment of T regulatory cells generation.

Vascular access failure causes 20% of all hospitalizations of dialysis patients. Native arteriovenous fistulas, the best type of dialysis vascular access, have a 1-year primary patency rate that is extremely variable, ranging 40-80%. Neointimal hyperplasia is the most important cause of arteriovenous fistula late primary dysfunction. In recent years the arteriovenous fistula late primary patency rate has not improved because of the increase of old uremic patients with a high number of comorbidities and the lack of new therapeutic interventions. Therefore, we performed a long-term case-control study to analyze which factors or drugs may affect native arteriovenous fistula late primary patency rate in 60 incident hemodialysis patients. The arteriovenous fistula late primary patency rate was 75.1% after 12 months, 58.5% after 24 months, and 50% after 987 days. Homocysteine levels during follow-up had a significant direct association with vascular access failure (event vs. event-free 28.5+/-1.9 vs. 22.3+/-1.2 micromol/L, p<0.01). Folate values had a trend toward an inverse relationship with arteriovenous fistula failure (event vs. event-free 11.5+/-1.2 vs. 14.6 vs. 1.1 ng/mL, p=0.06). Patients treated with folic acid and/or statin had an arteriovenous fistula late primary patency rate significantly higher than patients without folic acid and statin therapy, respectively, 81.7% vs. 66% after 1 year and 71.5% vs. 39.1% after 2 years (p=0.02). Many other factors were not associated with vascular access failure. Statin and homocysteine-lowering folic acid therapy is associated with prolonged arteriovenous fistula survival. It is important to perform randomized trials to verify our observation.

The Arabic MS Sprenger 1908 (Staatsbibliothek, Berlin) is a handbook of medieval alchemy. Among the works it preserves, we can find the only extant witness to the Arabic original of the well-known Liber de aluminibus et salibus. In this paper, I focus on a detailed alchemical dictionary preserved in this manuscript (fols. 3r-6r) whose explicit aim is to clarify the meaning of the secret language used by the alchemists to conceal the names of substances and operations. Other versions of the same alchemical lexicon are found in Syriac and karsuni in MSS Oriental 1593 and Egerton 709, both preserved in the British Library. After describing these manuscripts, I analyse the contents of this dictionary, its structure, its different versions, and the features of the alchemical language that it attests to, providing some examples to show how this kind of dictionary is still a useful tool for the contemporary researcher.

Background: Dialysis patients have higher cardiovascular events rate than patients with normal renal function. Hyperhomocysteinemia, a risk factor for cardiovascular disease, is frequently detected in dialysis patients. Vitamin B supplementation lowers hyperhomocysteinemia, but it is unknown whether it reduces cardiovascular events rate. We planned a long-term study to analyze the effects of homocysteine-lowering vitamin B therapy on cardiovascular disease in hemodialysis patients., Methods: We performed a single center open prospective trial. Patients, just on folate therapy at enrolment, were left out from randomization and maintained folate therapy according to study's protocol (group A). Patients, untreated with folic acid at recruitment, were randomly assigned to other 2 groups: patients submitted to folate supplementation according to study's protocol (group B), and untreated ones (group C). We instructed patients to take 5 mg oral daily folic acid or 5 mg every other day whether serum folate levels were up the normal high limit. We measured homocysteine, folate and vitamin B12 plasma levels at baseline and every 4 months. We chose the appearance of fatal and nonfatal cardiovascular events as end-points., Results: We analyzed data of 114 patients for a median follow-up time of 871 days. Stepwise regression analysis demonstrated that baseline homocysteine levels were related to folate (coefficient: -1.02; F: 64.5), creatinine (coefficient: 0.98; F: 11.3), and C reactive protein (coefficient: -0.64; F: 4.3). Patients ended the study for the following reasons: cardiovascular morbidity (n = 44), death (n = 25), renal transplant (n = 9), moved away (n = 4). Cardiovascular events occurred in 58 of 114 patients (51%), in 26 of 63 (41%) treated patients (both group A and group B) and in 32 of 51 (63%; chi2 = 6.0; p = 0.05) untreated patients (group C). Kaplan-Meier survival analysis showed that cardiovascular events were less frequent in treated patients with low homocysteine levels (chi2 24.1; p < 0.0001). Cox regression analysis showed that cardiovascular events were explained by homocysteine, dialysis vintage, past cardiovascular accidents, and age. We noticed not only lower homocysteine levels, but also higher protein catabolic rate values in events-free patients as compared with patients with nonfatal cardiovascular events. After having divided patients into 4 subgroups according to high and low, split at median, Hcy and protein catabolic rate values, we observed in Kaplan-Meier survival curves for cardiovascular events by these subgroups that patients with low Hcy and high protein catabolic rate values showed a significant lower hazard rate than patients with high Hcy and low protein catabolic rate levels (chi2 = 21.7; p < 0.0001)., Conclusions: This trial shows for the first time that homocysteine-lowering folate therapy decreases cardiovascular events in dialysis patients. It is necessary to perform large prospective studies to confirm our results., (Copyright 2006 S. Karger AG, Basel.)

We evaluated the clinical usefulness of spoligotyping, a polymerase chain reaction-based method for simultaneous detection and typing of Mycobacterium tuberculosis strains, with acid-fast bacilli-positive slides from clinical specimens or mycobacterial cultures. Overall sensitivity and specificity were 97% and 95% for the detection of M. tuberculosis and 98% and 96% when used with clinical specimens. Laboratory turnaround time of spoligotyping was less than that for culture identification by a median of 20 days. In comparison with IS6110-based restriction fragment length polymorphism typing, spoligotyping overestimated the number of isolates with identical DNA fingerprints by approximately 50%, but showed a 100% negative predictive value. Spoligotyping resulted in the modification of ongoing antimycobacterial treatment in 40 cases and appropriate therapy in the absence of cultures in 11 cases. The rapidity of this method in detection and typing could make it useful in the management of tuberculosis in a clinical setting.

Spoligotyping was compared with RFLP fingerprinting analysis in the identification of Mycobacterium tuberculosis strains. Spoligotyping sensitivity was 97.6% with a specificity of 47%. The global probability for two strains clustered with spoligotyping to be clustered also with RFLP analysis was 33%; the probability for two strains clustered with RFLP analysis to be clustered also with spoligotyping analysis was 95%. However, comparing the two methods in five outbreak episodes, full concordance was evidenced between spoligotyping and RFLP. Moreover, we evaluated the presence of our 17 largest spoligotyping clusters in spoligotyping databases from Caribbean countries, London and Cuba. Only five out of 17 patterns were present in all the cohorts. The conditional probability comparing spoligotyping and RFLP methods related to these patterns resulted in very low concordance (range from 2 to 38%). In conclusion, we confirm that spoligotyping when used alone overestimates the number of recent transmission and does not represent a suitable method for wide clinical practice application. However, it allows to get a first good picture of strain identity in a new setting and in more localized or confined settings, the probability of reaching the same result compared to RFLP was 100% confirming the usefulness of spoligotyping in the management of epidemic events, especially in hospitals, prisons and close communities.

Purpose: Erythropoietin (EPO) deficiency, erythropoiesis inhibition and reduction in red blood cell survival are the main causes of anemia in endstage renal disease (ESRD). Hemodiafiltration (HDF) with on-line endogenous reinfusion eliminates backfiltration and uses an ultrapure dialysate and reinfusate. This technique should improve anemia correction by increasing uremic toxin removal and reducing inflammatory cytokine production. In this study, we evaluated the effects of HDF with on-line endogenous reinfusion on anemia correction., Methods: This was a single-center, prospective and non-randomized study. We selected 12 patients on EPO therapy with steady haemoglobin (Hb) values; eight patients were treated with bicarbonate dialysis and four patients with on-line HDF. We switched them to HDF with on-line endogenous reinfusion for 6 months, changing the EPO dose when Hb levels were not within the 11-12 g/dL range. Biochemical data were measured every month. Results are expressed as means +/- m.s.e. Data were analyzed using the Student's t-test and analysis of variance for repeated measurements. A value p<0.05 was considered statistically significant., Results: Patients maintained the same Hb, ferritin and dialytic efficiency throughout the study. The EPO supplementation was significantly higher in patients treated with bicarbonate dialysis (108 +/- 8 UI/kg/week as compared with patients treated with on-line HDF (36 +/- 5 UI/kg/week) during the 6 months before treatment by HDF with on-line endogenous reinfusion. In the last 6 months, we detected a reduction in EPO consumption, but not statistically significant, in the patients switched from bicarbonate dialysis to HDF with on-line endogenous reinfusion (83 +/- 6 UI/kg/week)., Conclusions: The change from bicarbonate dialysis to HDF with on-line endogenous reinfusion caused a reduction in EPO supplementation and, consequently, a reduction in the cost of anemia therapy. It is necessary to increase the size of the study group and to prolong the observation period to possibly obtain statistical significances.

Background: Strategies to prevent cardiovascular diseases are still not widely applied. It is vital to involve people for effective prevention, and knowledge of their own coronary risk is a first step. Little is known about how people perceive their coronary risk. For this reason, we examined this perception in a sample of the general population and compared it with a reference risk chart., Methods: We invited the population of two small towns to come for a free check of their blood pressure and blood cholesterol levels. People were first asked to complete a self-administered questionnaire grading their perceived risk of developing a coronary event. Then, basic data were collected for calculating their risk according to a reference chart., Results: A total of 877 individuals (382 males, 495 females, age 30-80 years) without ischemic heart disease completed the questionnaire and had their blood pressure and cholesterol checked. Many important risk factors are still not taken into account by patients, particularly male sex, older age and diabetes. One out of 4 subjects underestimates his level of coronary risk and 9 out of 10 high-risk subjects are not even aware of it., Conclusions: Our study shows that large numbers of persons are still unaware of their level of coronary risk and of its major determinants. Better counseling by general practitioners is needed since a realistic perception of risk is essential for reducing it.

Background: The presence of aortic valve sclerosis accounts for a higher rate of ischemic events and increased cardiovascular mortality. It may reflect coronary artery disease (CAD) because of a shared pathologic background., Hypothesis: We aimed to analyze whether the presence of aortic valve sclerosis might help in identifying patients with coronary atherosclerosis among those with severe nonischemic mitral regurgitation (MR), who undergo coronary angiography before surgery for screening, and not because of suspected ischemic heart disease., Methods: In all, 84 patients (mean age 64 +/- 9 years; 71% men) with mitral valve prolapse and severe regurgitation underwent echocardiography and coronary angiography. Aortic valve sclerosis was defined as focal areas of increased echogenicity and thickening of the leaflets without restriction of leaflet motion on echocardiography. Coronary artery disease was defined by the presence/absence of atherosclerotic plaques, independent of the degree of stenosis., Results: Coronary artery disease was diagnosed in 47.6% of patients with and 15.8% of those without aortic valve sclerosis (p = 0.008). On logistic regression analysis, the presence of aortic valve sclerosis predicted CAD (odds ratio 3.3, 95% confidence interval 1.03-10.5; p = 0.04) independent of age. In female patients, the risk ratio for CAD in the presence of aortic valve sclerosis was 9., Conclusions: Coronary artery atherosclerosis and aortic valve sclerosis are closely associated in patients with severe nonischemic MR.

Objectives: chronic obstructive pulmonary disease and asthma are major causes of hospitalisation and mortality among older patients but respiratory diseases are often under- or misdiagnosed because spirometry is not extensively used at this age., Design: we examined 715 elderly subjects with respiratory symptoms; all underwent a spirometric test and were administered the Mini Mental State Examination, Activities of Daily Living, Instrumental Activities of Daily Living and Geriatric Depression Scale questionnaires for cognitive, functional and effective evaluation. Their educational level and Body Mass Index were also taken into consideration., Results: a total of 585 patients (81.8%) were able to perform spirometry according to ATS'94 criteria while 130 (18.2%) were unable to do it. As regards educational level, Mini Mental State Examination, Activities of Daily Living and Instrumental Activities of Daily Living scores showed a significant difference (P<0.001) between the groups of patients with high-quality spirometries and those with inadequate ones. There was no difference between the two groups in terms of age, Body Mass Index or Geriatric Depression Scale score., Conclusions: the majority of elderly subjects can perform spirometry according to international guidelines; age itself cannot be considered a risk factor for a bad spirometric performance, but it becomes influential if it is associated with cognitive and functional impairment.

Background: Many trials have proved the benefits of early systematic treatment with angiotensin-converting enzyme inhibitors in patients with acute myocardial infarction (AMI). Pathophysiological studies, however, suggest potential harm in excessive reduction of blood pressure (BP) in hypertensive patients with ischemic heart disease., Methods: We analyzed data from the GISSI-3 (Gruppo Italiano per lo Studio della Sopravvivenza nell'Infarto miocardico) trial to assess the effects of early treatment with angiotensin-converting enzyme inhibitors during AMI in patients with a history of hypertension compared with normotensive patients. The trial randomly assigned 19,394 patients to 6 weeks of lisinopril treatment or control, starting treatment within 24 hours of AMI onset., Results: In the 10,661 normotensive patients, lisinopril significantly reduced lethal events, but in the 7362 hypertensive patients, a higher rate of lethal events was reported the first day of treatment, and the benefits only appeared subsequently. These results may be attributable to the subgroup of 1165 hypertensive patients with low baseline systolic BP (lower quintile, BP <120 mm Hg), in whom critical hypotension was more prone to develop after lisinopril treatment. In fact, these patients showed a higher mortality rate as the result of an excess of cardiogenic shock during the first day of lisinopril treatment (odds ratio 3.07, 95% CI 1.39-6.77) and a persistent, unfavorable death trend after 6 weeks., Conclusions: These data suggest that caution should be exercised when using lisinopril in the acute phase of a myocardial infarction in patients with a history of hypertension but low systolic BP at presentation.

A case of sudden death in an old female carrier of Duchenne muscular dystrophy is reported. Typical electrocardiographic features were registered without other signs or symptoms of heart involvement. In particular, cardiomyopathy was excluded by echocardiographic evaluation. We believe that in case of a typical electrocardiogram of Duchenne muscular dystrophy, a careful family medical history should be undertaken to exclude a carrier condition.

In geographical areas with a low incidence of tuberculosis, recurrent tuberculosis is generally due to reactivation of the disease. However, the relative contribution of tuberculosis reinfection increases in parallel with the incidence of disease and is likely to depend on the epidemiological context: factors such as the spread of multidrug resistance, human immunodeficiency virus (HIV) infection, and immigration from developing countries could modify disease transmission in areas at low risk for tuberculosis. A molecular epidemiology study was performed in Lombardy, Northern Italy, where the incidence of tuberculosis is 17.5 cases per 100,000 persons. A total of 2,452 cases of culture-confirmed tuberculosis in 2,127 patients were studied. A group of 32 patients (1.5%), each of whom had two episodes of tuberculosis with cure as the outcome of the first episode and with more than 6 months between the two episodes, were studied by means of restriction fragment length polymorphism DNA fingerprinting analysis. For 5 of the 32 patients (16%), the DNA fingerprinting patterns of Mycobacterium tuberculosis strains responsible for the second episode did not match those of the corresponding isolates of the first episode, indicating exogenous reinfection. Two of these patients developed multidrug-resistant tuberculosis during the second episode, and in three cases the isolates belonged to clusters of M. tuberculosis strains spreading in the community. A fourfold-increased risk for reinfection was observed in immigrant patients compared to Italian subjects. In contrast, a higher risk of relapse rather than reinfection was evidenced in HIV-positive subjects and in patients infected with multidrug-resistant tuberculosis. Episodes of tuberculosis reinfection in areas with a low incidence of tuberculosis are rare compared to those in high-incidence geographical regions. In populations that have immigrated from high-risk areas, reinfection may represent a considerable contributor to the rate of recurrent tuberculosis. This finding emphasizes the importance of containing the spread of epidemic strains in close communities, in order to prevent changes in global tuberculosis trends for developed countries.

We evaluated the sensitivity of a DNA amplification test for the detection of Mycobacterium avium in blood samples using different blood components and different DNA extraction methods. M. avium-inoculated blood samples were processed to obtain separate blood components: peripheral blood mononuclear cells (PBMCs), polymorphonuclear cells (PMNCs), and whole-blood sodium dodecyl sulfate (SDS)-lysate pellets. The sensitivity for the detection of the lowest mycobacterial load (1 CFU/ml) was significantly greater (P < 0.01) with DNA extracted from SDS-lysate pellets than with DNA extracted from PBMCs or PMNCs. Subsequently, DNA extraction methods based on guanidine NaOH, and proteinase were compared. The sensitivity of the guanidine-based method was significantly greater (P < 0.01) than those of the others.

Background: Atrial fibrillation (AF) is the most frequent complication following cardiac surgery. It occurs mostly between the second and fourth postoperative days and often recurs within 30 days after surgery. While postoperative AF has been well analyzed, post-discharge recurrences of AF have not been clearly described even if they are reported as a frequent cause of re-hospitalization., Methods: We followed up 185 patients for 10 +/- 5 months with the aim of characterizing the post-discharge recurrences of AF. All patients had recently undergone cardiac surgery complicated by AF and were in sinus rhythm at the time of admission to our Center. We also compared the efficacy of the main prophylactic regimens adopted in the referral Centers (amiodarone, beta-blockers, amiodarone plus beta-blockers) during the first postoperative month., Results: In the first postoperative month AF recurred after discharge in 60 patients. The event rate was not different in patients treated with amiodarone and controls (47 vs 50%, p = NS), while it was significantly lower in patients taking beta-blockers either alone or associated with amiodarone (10 and 9% respectively, p = 0.002). At the end of follow up (10 +/- 5 months), AF persisted in 3 out of 176 study patients (1.7%)., Conclusions: In patients undergoing cardiac surgery, post-discharge recurrences of AF are frequent during the first postoperative month and have a clinical relevance. Beta-blockers (not amiodarone) seem to be an effective prophylactic measure. The phenomenon tends to vanish in the long term, and a chronic prophylaxis is not justified.

Differentiation between Mycobacterium tuberculosis and M. avium is essential for the treatment of mycobacterial infections. We have developed an easy and rapid detection assay for the diagnosis of mycobacterial diseases. This is a PCR-hybridization assay based on selective amplification of a 16S rRNA gene sequence using pan-Mycobacterium primers followed by hybridization of the amplification products to biotinylated M. tuberculosis and M. avium-specific probes. A total of 55 mycobacterial isolates were tested. For all isolates, results concordant with those of conventional identification methods were obtained. Moreover, we developed a method for extraction of DNA from Ziehl-Neelsen-positive smears which allows the recovery of intact target DNA in our PCR-hybridization assay. Our method was able to confirm all culture results for 59 Ziehl-Neelsen-positive smears from clinical specimens (35 sputum, 11 lymph node biopsy, 6 stool, 4 pus, 2 urine, and 1 pericardial fluid specimens). These data suggest that our PCR-hybridization assay, which is simple to perform and less expensive than commercial probe methods, may be suitable for the identification of M. tuberculosis and M. avium. It could become a valuable alternative approach for the diagnosis of mycobacterial infections when applied directly to DNA extracted from Ziehl-Neelsen-positive smears as well.

The aim of this pilot study was twofold. The first was to show a method for having an objective and dynamic analysis of body posture, evaluating weight distribution and its connections with different mandibular positions. The second was to verify if a neuromuscularly stimulated occlusal position, called myocentric occlusal position, is associated with a positive or negative postural charge. For the second aim a group of 20 subjects (including both males and females), was chosen. Posture of each subject was analyzed in three different conditions: centric occlusion, rest position and myocentric position. To evaluate the dynamic of posture a platform capable of measuring the weight on the feet supporting points and the related variations during time of observation and the swinging of body barycenter was used. The data showed that there is an improvement in the position of the barycenter from the centric occlusion to the myocentric position. Such an improvement can't be observed from the intercuspal position to the rest position. The results of this pilot study are discussed.

The recombinant cytokines interferon (IFN)-gamma and interleukin (IL)-12 stimulate several macrophage-mediated functions that are important in host defense. An experimental pertussis model showed that intraperitoneal (i.p.) administration of 10,000 U of recombinant murine (rm) IFN-gamma to mice at the time of Bordetella pertussis infection caused a marked and significant reduction in the number of colony-forming units of bacteria in the lungs. Administration i.p. of 1 microgram of rmIL-12 or 1 microgram of rmIL-12 at the time of and for 5 consecutive days after B. pertussis challenge also induced a significant reduction in the number. However, i.p. administration of 1 microgram of rmIL-12 with 10,000 U of IFN-gamma at the time of B. pertussis challenge did not provide protection. These findings indicate that exogenous administration of rmIL-12 and rmIFN-gamma enhances resistance of mice to B. pertussis infection.

Several studies have demonstrated that Bordetella pertussis has the ability to enter and survive intracellularly within human polymorphonuclear leukocytes (PMNL) and human monocytes/macrophages. The effects of human recombinant gamma interferon (IFN-gamma) on the survival of B. pertussis in PMNL and human monocytes, and on the oxidative burst activity of PMNL and human monocytes in response to B. pertussis were assessed in this study. IFN-gamma partially increased intracellular killing of phagocytosed B. pertussis in human monocytes, as determined by an orange acridine-crystal violet assay. In contrast, IFN-gamma did not enhance intracellular killing of B. pertussis in PMNL. No significant increase of superoxide production was noted in human monocytes in response to B. pertussis when stimulated with various concentrations of IFN-gamma. The partial increase of B. pertussis killing by IFN-gamma within monocytes, together with poor production of superoxide may explain how B. pertussis can survive within human phagocytic cells, and thus cause a more prolonged course of the disease.

We found a significant increase in fibronectin receptor (FNR) levels in the sera of adult human immunodeficiency virus type 1 (HIV-1)-infected patients, especially in those with AIDS (1,026.9 +/- 583.9 ng/ml; P < 0.0001). In contrast, AIDS patients with neurologic disorders and HIV-1-seropositive patients showed normal levels of FNR in serum. In addition, HIV-1-infected children showed increased levels of FNR in serum (824.4 +/- 333.5 ng/ml; P = 0.03). We suggest that an increase of FNR levels in AIDS patients is related to enhanced expression of FNR on HIV-1-infected cells.

Serum levels of interleukin-1 alpha (IL-1 alpha), interleukin-1 beta (IL-1 beta), interleukin-6 (IL-6), and tumor necrosis factor-alpha (TNF-alpha) in patients with acute viral hepatitis were investigated. Twelve patients suffering from acute viral hepatitis were studied; 8 patients presented with acute hepatitis B, 2 patients with acute hepatitis A, and 2 patients with acute hepatitis C. Serum levels of IL-1 alpha, IL-1 beta, IL-6, and TNF-alpha were significantly increased in all patients with acute viral hepatitis. Decreased serum levels of all cytokines were noted in four patients with acute hepatitis B during the recovery phase of infection. In addition, IL-1 alpha, IL-1 beta, IL-6, and TNF-alpha were undetectable at the end of a follow-up period of 6 months. Our study shows that increased levels of IL-1 alpha, IL-1 beta, IL-6, and TNF-alpha are probably related to hepatitis activity and thus may have some role in hepatocytic injury.

Fibronectin (FN) is present in soluble and matrix forms in various body fluids and tissues, and has been shown to bind to several pathogens, including viruses. The interaction of FN with viral proteins of human immunodeficiency virus (HIV-1) was investigated by immunofluorescence technique using a cell line chronically infected with HIV-1 (H9-V). The results of this study showed that FN binds to HIV-1 infected cells, especially at FN concentration of 5 micrograms/ml. In addition, FN-pentapeptide has shown the ability to bind to HIV-1 infected cells. On the other hand, preincubation with antibodies against FN abolished the binding of FN to HIV-1 infected cells. Finally, FN has shown to bind to HIV-1 glycoproteins, including gp41 and gp120. In contrast, no binding to HIV-1 core proteins, including p15 and p24, was noted. We suggest that FN, in binding HIV-1 particles, may reduce viremia and thus may be involved in the clearance of viral proteins from the cells.

We describe a case of a man with acute myocardial infarction of the inferior wall complicated by cardiogenic shock due to acute mitral regurgitation. Transesophageal echocardiography showed the rupture of the posteromedial papillary muscle. The examination (otherwise difficult) was facilitated by the mechanical ventilation by means of an endotracheal intubation.

Objective: To determine serum levels of acute-phase proteins and interleukin 1B, interleukin 6, tumor necrosis factor alpha, and interleukin 8 in children with pertussis., Design: Cross-sectional study., Setting: Divisions of Infectious Diseases, Regional Hospital, and Pediatrics, University of Pavia, Varese, Italy., Participants: Eight children with pertussis, six with acute febrile infections, and eight healthy control children matched for sex, age, and time presentation over a 32-month study period., Interventions: None., Measurements/main Results: An immunoenzymatic assay was used to detect serum levels of all cytokines. Normal values of C-reactive protein, alpha 1-acid glycoprotein, and erythrocyte sedimentation rate were observed in the serum of patients with pertussis. The mean (+/- SD) detectable levels of tumor necrosis factor alpha (65.0 +/- 50.4 pg/mL) and interleukin 6 (32.3 +/- 17.8 pg/mL) were observed in the serum of patients with pertussis. In contrast, a nonsignificant increment of interleukin 1B levels (66.5 +/- 83.7 pg/mL) and interleukin 8 levels (12.7 +/- 17.8 pg/mL) was noted in the serum of the same patients. Increased and significant levels of all four cytokines were noted in most of the serum samples of patients with acute febrile infections., Conclusions: Acute-phase response is absent in patients with pertussis, whereas detectable and significant serum levels of tumor necrosis factor alpha and interleukin 6 were observed in some such patients.

We investigated the role of lymphocyte function-associated antigen 1 (LFA-1) in human T cell polarization and migration assay by using monoclonal antibody specific to beta chain (CD18) and alpha chain (CD11a). T cell polarization in response to fetal calf serum (FCS) and colchicine was suppressed by the addition of CD18 and CD11a antibodies. Furthermore, T cell migration in response to lymphocyte chemotactic factor (LCF) and casein was markedly depressed by the addition of CD18 and CD11a antibodies. Additional studies to evaluate effects of interleukin 8 (IL-8) on polarization and migration of T cells preincubated with CD18 or CD11a antibody showed that IL-8 restored the capability of migration of T cells, whereas did not restore polarization activity of such cells. These studies indicate that LFA-1 plays a role in the polarization and migration of T cells and that IL-8 may positively interfer with LFA-1-adhesion molecules.

The present study was undertaken to assess the locomotor capacity of lymphocytes from patients with human immunodeficiency virus (HIV) infection by a polarization assay. In addition, the capping phenomenon of lymphocytes from HIV-infected patients or in vitro preincubation of lymphocytes with HIV-envelope glycoproteins, gp41 and gp120 was evaluated. A significant decrease in polarized lymphocytes from patients with acquired immunodeficiency syndrome (AIDS) was observed. On the other hand, the capping phenomenon either of lymphocytes from AIDS patients or of lymphocytes from healthy donors preincubated with gp41 and gp120 was reduced but not significantly.

To prevent delayed cardiomyopathy induced by adriamycin and subsequent severe congestive heart failure, a monitoring schedule with noninvasive techniques to measure cardiac performance (CPK-MB, electrocardiographic, systolic time intervals, echocardiography and quantitative radionuclide angiography) has been used. 15 patients, 33 to 65 years old, with metastatic breast carcinoma, previously treated with a polychemotherapy protocol not including adriamycin have been studied. Monitoring at 0 time allowed to exclude one patient with previous asymptomatic cardiomyopathy and to treat patients at risk with cardiovascular pathology, up to reach and even exceed the dosage of 550 mg/m2. Four of the treated patients showed positive findings of cardiomyopathy at different adriamycin dosage levels: 2 asymptomatic dilatative cardiomyopathies, 1 symptomatic and 1 with congestive heart failure remitted with medical therapy. No patient died because of cardiovascular complications. The results obtained show that of all noninvasive techniques used, only quantitative radionuclide angiocardiography allowed, when employed following an adequate monitoring schedule, to detect reliable findings of moderate and reversible cardiomyopathy thus indicating the appropriate time for drug discontinuation.

The origin of multinucleated giant cells in myeloma kidney is not yet defined. These cells have long been thought to represent tubular epithelial cells transformed in syncytia, but, more recently, they have been considered as histiocytic cells arising from the interstitial tissue. We have studied by electron microscopy the distal tubules of kidney biopsies of 5 patients affected with myeloma kidney: our ultrastructural findings do not seem to clarify the problem. In fact, in the same patient, we have observed multinucleated giant cells probably originating from epithelial tubular cells and others, probably originating from histiocytes.

The association between the presence of areas of myocardial fibrosis and the appearance of unexpected DEVIATIONS OF THE QRS loop-outline on the vectorcardiographic tracing (bites) has been reported. In order to re-evaluate the correlation between the presence of bites and the possible existence of scars we have studied 511 patients. On the basis of clinical data, laboratory data, rest and exercise electrocardiograms, the subjects have been divided into four groups; 195 normal subjects; 267 with ischemic heart disease, 16 with arterial hypertension and 33 with diabetes mellitus (the last two groups as representative of subjects with higher risk for myocardial ischemia). Bites in at least one plane were detected in 9.7% of normal subjects, 27.8% of ischemic patients, 56.3% of hypertensive patients and 18.2% of diabetics (less than 0.001). The genesis of bites and reliability of the diagnostic criteria are discussed. With more restrictive criteria the frequency in the normal subjects falls to 1%, while in the other three groups it remains much higher (10.1%-12.5%-6.1%).

The ECG in Ostium Secundum Atrial Septal Defect (O.S. ASD) shows typically a pattern of incomplete right bundle branch block with right axis deviation and a 12 year old boy suspected of having O.S. ASD on the basis of x-ray and echocardiographic findings showed an ECG pattern of incomplete left bundle branch block. Physical examination and a phonocardiogram revealed a single second heart sound. An ECG recorded at age five showed a typical pattern consistent with O.S. ASD. The angiographic examination confirmed the suggested diagnosis. This is, to our knowledge, the first case of O.S. ASD with incomplete left bundle branch block. The conduction disorder may have abolished the fixed splitting of the second heart sound.