A bioavailability study on microbeads and nanoliposomes fabricated by dense carbon dioxide technologies using human-primary monocytes and flow cytometry assay.

Supercritical Emulsion Extraction (SEE) and Supercritical assisted Liposome formation (SuperLip), use dense gases such as carbon dioxide (dCO2) to fabricate advanced micro/nanocarriers. SEE uses dCO2 to extract solvent from the oily phase of an emulsion and obtain biopolymer microbead; For this study, poly-Lactic Acid (PLA) microbeads of 1 ±â€¯0.2 µm in mean size loaded at 1 µg/mgPLA with Rhodamine B (ROD) were prepared by SEE; the beads showed a solvent residue lower than 10 ppm and encapsulated the fluorochrome with an efficiency of 90%. SuperLip uses dCO2 to enhance lipid/ethanol/water mixing and to promote the ethanol extraction from liposome suspension. In this case, phosphatidyl-choline (PC) vesicles with a mean size of 0.2 ±â€¯0.05 µm and loaded with Fluorescein Iso-ThioCyanate (FITC) at 8 µg/mgPC were prepared; small unilamellar structure was observed for all the vesicles with FITC encapsulation efficiency of 80%. Ethanol residue of 50 ppm was measured in all the liposome suspensions. The bioavailability of microbeads and nanoliposomes was assessed through incubation with human monocytes previously isolated from healthy donors' blood. A specifically optimized protocol that allowed their quenching on the cell surface was developed to monitor by flow cytometer assay only the cell population that effectively internalized the carriers. When microbeads were tested, the percentage of alive internalizing monocytes was of about 30%. An internalization of 96.1 ±â€¯21% was, instead, obtained at dosage of 0.1 mg/mL for nanoliposomes. In this last case, monocytes showed a vitality of almost 100% after vesicles internalization at all the concentrations studied; on the other hand, cell apoptosis progressively increased in a dose/response manner, after polymer microbeads phagocytosis. The proposed data suggested that dCO2 technologies can be reliably used to fabricate intracellular carriers.

Impact of HLA-G polymorphism on the outcome of allogeneic hematopoietic stem cell transplantation for metastatic renal cell carcinoma.

Renal cell carcinoma (RCC) is particularly sensitive to immune intervention. HLA-G, a non-classical HLA class I molecule with immunomodulatory properties, has been studied with regard to outcome after hematopoietic stem cell transplantation (HSCT), in particular the 14 bp insertion/deletion polymorphism in the 3' untranslated region. Here we analyzed n=56 patients affected by metastatic RCC who received an allogeneic HSCT between 1998 and 2006 in Milano, Marseille, Clermont-Ferrand and Stockholm. The 14 bp polymorphism was analyzed in correlation with overall survival (OS), PFS, acute and chronic GvHD. With a median follow-up of 13 years, a trend towards better outcome was observed when homozygosity for the 14bp-del allele was present: multivariate hazard ratio was 0.50 (95% confidence interval (CI): 0.23-1.13; P=0.10) and 0.57 (95% CI: 0.26-1.26; P=0.17) for OS and PFS, respectively, when 14bp-del/del was compared with 14bp-ins/X. Further exploratory analysis revealed a significant association between T/C at p3003 and improved OS (P=0.05) and PFS (P=0.006) compared with T/T. To our knowledge this is the first study on HLA-G and outcome after HSCT for a solid malignancy. After a coordinated multicenter study, we found that the more tolerogenic polymorphisms (14bp-del/del) is associated with better PFS and OS. The finding on p3003 deserves further investigation.

New insights into HLA-G mediated tolerance.

Human Leukocyte Antigen G (HLA-G) is a nonclassical HLA class I molecule with well-characterized immunomodulatory activities. HLA-G was first described as a regulatory molecule that allows the fetus to elude the maternal immune response. In the last decade it has become evident that HLA-G is involved in modulating both innate and adaptive immune responses, in maintaining tolerance in autoimmune and inflammatory diseases and after transplantation, and in promoting immune escape in cancer and infectious diseases. HLA-G exerts its modulatory/regulatory functions directly by interacting with specific inhibitory receptors. The expression of HLA-G is finely tuned by genetic variations in the noncoding region of the locus. The recent discovery of dendritic cells-10 (DC-10) as naturally occurring HLA-G-expressing dendritic cells opens new perspectives in the identification of the molecular and cellular mechanisms underlying HLA-G-mediated tolerance. An overview on the HLA-G-mediated inhibition of innate and adaptive immune cells, on the genetic influence on HLA-G expression, and on HLA-G-expressing DC-10 is presented. Moreover, we discuss the central and critical role of DC-10 in the HLA-G-mediated tolerance.

The use of first generation versus second generation antipsychotics as add-on or as switch treatment and its effect on QTC interval: the Italian experience in a real-world setting.

Some psychotropic drugs are connected with prolongation of the QT interval, torsade de pointes and sudden death. Recent data suggest that with regard to this adverse effect, the atypical antipsychotic drugs are no safer than the older drugs. The purpose of this study is to evaluate the different use of first generation versus second generation antipsychotics as add-on (Group I) or switch treatment (Group II) and its effect on QTc interval in a sample of schizophrenic and bipolar inpatients without medical illness. All patients had been evaluated twice by using ECG: on admission and after two weeks of hospitalization. Exclusions criteria were: abnormalities in levels of potassium, magnesium and calcium, cardiovascular and metabolic diseases, alcohol or drug abuse. We found a significant (p < 0.01) greater use of first generation antipsychotic in Group I (73.80%) than in the Group II (33.33%). Also Group I showed a significant increase (p < 0.0001) in total chlorpromazine equivalent (476. 78 ± 448.80 mg/day vs 845.48 ± 491.64 mg/day) and in QTc interval (369.14 ± 33.75 ms vs 387.09 ± 31.97 ms), while we did not find any statistical difference in Group II during hospitalization. Our results, in spite of the small sample size, indicate that antipsychotic add-on can increase QTc interval more than switching to other antipsychotic in psychiatric patients without other risk factors.

Health promotion in minority adolescents: a Healthy People 2000 pilot study.

PURPOSE: The purpose was to evaluate the effects of a health promotion curriculum on health knowledge, behavior, cardiovascular fitness, and cardiovascular risk factors. METHODS: A multi-ethnic, multi-cultural sample (n = 54) of 10th grade males and females participated in a study of cardiovascular health promotion and coronary risk factor reduction. The sample was comprised of Asian-Americans (39%), blacks (33%), Hispanics (11%), whites (2%), and others (15%). Intervention consisted of a 10-week health promotion curriculum of classroom education modules in physical activity, nutrition, smoking cessation, stress management and personal problem solving, and an exercise program of walking and running. A nonintervention control group served as a basis for comparison. Classroom and exercise sessions met on alternate days. RESULTS: Following intervention, a significant treatment effect (P = .007) was observed in lowered total cholesterol, and significant within group improvements (P < .01) were observed in diet habits, percent body fat, and cardiovascular health knowledge. Comparisons of knowledge and social effects revealed higher cardiovascular health knowledge (P < .05) in subjects of nonsmoking compared to smoking parents, higher self-perception of health (P < .01) in more active vs less active subjects and better dietary habits (P < .07) in children whose parents were college educated compared to parents who did not attend college. CONCLUSIONS: Preliminary findings suggest that a health promotion curriculum consisting of health education, behavior modification, and regular aerobic exercise lowers cholesterol, improves health behavior and increases health knowledge.

[Acute abdominal pain and appendicitis in childhood]. / Il dolore addominale acuto e l'appendicite nell'infanzia.

The diagnosis of acute appendicitis is still difficult to ascertain in children. However, a complete anamnesis, an accurate physical examination as well as a careful evaluation of other medical and surgical possibilities causing abdominal pain allow to arrive to a correct diagnosis in 80% of cases. Laboratory findings may be helpful but usually don't add further information. Each patient suspected to have appendicitis should be admitted to the hospital and kept under observation; if no improvement is registered during the following hours then a surgical exploration is needed. The surgeon, however, must be acquainted with the different medical affections causing abdominal pain in order to decide whether a laparotomy is required. The Authors report their experience in 426 patients submitted to appendectomy and stress the correlation between abdominal pain and intraoperative finding.

Clinical use of transoesophageal atrial stimulation in terminating ventricular tachycardia.

This study was performed to evaluate whether transoesophageal atrial pacing could also stop ventricular tachycardias with low rates and no haemodynamic impairment. Prior to resorting to ventricular endocardial pacing, seven male patients, aged between 15 and 73 years, were treated by transoesophageal atrial pacing for 10 spontaneous episodes of sustained ventricular tachycardia at rates between 105 and 160 beats per minute, without haemodynamic impairment. When atrial pacing did not allow ventricular capture, atropine sulphate was administered. Transoesophageal atrial pacing led to ventricular capture in seven episodes, which made overdriving possible, and blocked six episodes of ventricular tachycardia. In no case did transoesophageal atrial pacing lead to an acceleration of ventricular tachycardia or to degeneration into ventricular fibrillation. Transoesophageal atrial pacing can block low-rate sustained ventricular tachycardias (less than or equal to 150 beats per minute). For low-rate sustained ventricular tachycardias without haemodynamic impairment, transoesophageal atrial pacing can thus be used as the method of choice thanks to its good ratio of risk to efficiency.

Programmed stimulation in patients with minor forms of right ventricular dysplasia.

Programmed electrical stimulation (PES) was performed in 17 patients, mean age 31 +/- 8 years, with minor forms of right ventricular dysplasia diagnosed because of (1) premature ventricular beats (PVBs) of left bundle branch block (LBBB) morphology; (2) no clinical or non-invasive evidence of cardiac abnormalities; (3) angiographic evidence of right ventricular wall motion abnormalities and bioptic findings of fibro-adipose infiltration. Fifteen patients had frequent and complex PVBs while two had sustained ventricular tachycardia (VT). During PES, sustained VT was induced in 2/2 patients with spontaneous sustained VT; ventricular repetitive responses were induced in 2/15 cases (13%) with complex and frequent PVBs. In conclusion, in minor forms of right ventricular dysplasia, PES induces VT only in patients with clinical VT; on the contrary, in patients with PVBs it is only possible to induce repetitive ventricular responses in a small proportion of cases; it is therefore not possible to select patients at high risk of developing severe ventricular arrhythmias.

Electrophysiologic evaluation of intravenous nicardipine in man.

The acute electrophysiological effects of intravenous nicardipine (0.014 mg/kg per min for 5 minutes) were studied in 12 subjects with estimated normal sinus node functions and atrioventricular conduction parameters. The most important effects were sinus cycle length shortening, increase of corrected sinus node recovery time and reduction of effective and functional refractory period of the atrioventricular node. Sinuatrial conduction time, atrial refractory periods, intranodal conduction, bundle branch refractoriness and ventricular refractoriness were unchanged. Systolic and diastolic blood pressure was reduced. The clinical implications of these properties of the drug are discussed and compared with those of verapamil, diltiazem and nifedipine.