Alteration of Notch signaling and functionality of adipose tissue derived mesenchymal stem cells in heart failure.

AIM: Circulating mesenchymal cells increase in heart failure (HF) patients and could be used therapeutically. Our aim was to investigate whether HF affects adipose tissue derived mesenchymal cell (adMSC) isolation, functional characteristics and Notch pathway. METHODS AND RESULTS: We compared 25 patients with different degrees of HF (11 NYHA classes I and II and 14 NYHA III and IV) with 10 age and gender matched controls. 100% adMSC cultures were obtained from controls, while only 72.7% and 35.7% from patients with mild or severe HF (p<0.0001). adMSC from HF patients showed higher markers of senescence (p16 positive cells: 14±2.3% in controls and 35.6±5.6% (p<0.05) and 69±14.7% (p<0.01) in mild or severe HF; γ-H2AX positive cells: 3.7±1.2%, 19.4±4.1% (p<0.05) and 23.7±3.4% (p<0.05) respectively), lower proliferation index (Ki67 positive cells: 21.5±4.9%, 13.2±2.8% and 13.7±3.2%, respectively), reduced pluripotency-associated genes (Oct4 positive cells: 86.7±4.9%, 55±12% (p<0.05) and 43.3±8.7% (p<0.05), respectively; NANOG positive cells: 89.8±3.7%, 39.6±14.4% (p<0.01) and 47±8.1%, respectively), and decreased differentiation markers (α-sarcomeric actin positive cells: 79.8±4.6%, 49±18.1% and 47±12.1% (p<0.05) and CD31-positive endothelial cells: 24.5±2.9%, 0.5±0.5% (p<0.001) and 2.3±2.3% (p<0.001), respectively). AdMSC from HF patients also showed reduced Notch transcriptional activity (lowered expression of Hey 1 and Hey 2 mRNAs). Stimulation with TNF-α of adMSC isolated from controls affected the transcription of several components of the Notch pathway (reduction of Notch 4 and Hes 1 mRNAs and increase of Notch 2 and Hey 1 mRNAs). CONCLUSIONS: In HF yield and functionality of adMSC are impaired and their Notch signaling is downregulated.

[Learning motivational interviewing to help patients change their health-related behaviors: medical students confirm it]. / Se former à l'entretien motivationnel pour aider ses patients à changer: Les étudiants en médecine le confirment.

At the Lausanne University, 5th year medical students were trained in Motivational interviewing (MI). Eight hours of training improved their competence in the use of this approach. This experience supports the implementation of MI training in medical schools. Motivational interviewing allows the health professional to actively involve the patient in this behavior change process (drinking, smoking, diet, exercise, medication adherence, etc.), by encouraging reflection and reinforcing personal motivation and resources.

Immunological modifications during treatment with thymosin alpha1 plus antiviral therapy in chronic hepatitis C.

The current standard therapy for the treatment of chronic hepatitis C virus (HCV) is the combination of peginterferon and ribavirin, although many patients fail to clear the virus and their retreatment options are still unsatisfactory. Thymosin alpha1 (Talpha1) is an immunomodulating agent that has been proposed as complementary therapy for chronic HCV, especially in the setting of difficult-to-treat patients. The aim of this study was to evaluate, in patients nonresponsive to previous Peg-based therapy, the effect of standard antiviral therapy with or without Talpha1 on peripheral lymphocyte subsets. Twenty-four patients, 12 receiving Talpha1 and 12 standard therapy, were enrolled. Peripheral subpopulations were analyzed by flow cytometry. Although the addition of Talpha1 did not seem to significantly modify the T-lymphocyte subpopulations, as comparable behaviors were observed in the CD4 and CD8 longitudinal evaluation, Talpha1 produced an earlier increase of natural killer cells. An accurate selection of HCV patients who can benefit from immunomodulation is needed.

Inhibition of serine-peptidase activity enhances the generation of a survivin-derived HLA-A2-presented CTL epitope in colon-carcinoma cells.

Cytotoxic T lymphocytes eliminate tumor cells expressing antigenic peptides in the context of MHC-I molecules. Peptides are generated during protein degradation by the proteasome and resulting products, surviving cytosolic amino-peptidases activity, may be presented by MHC-I molecules. The MHC-I processing pathway is altered in a large number of malignancies and modulation of antigen generation is one strategy employed by cells to evade immune control. In this study we analyzed the generation and presentation of a survivin-derived CTL epitope in HLA-A2-positive colon-carcinoma cells. Although all cell lines expressed the anti-apoptotic protein survivin, some tumors were poorly recognized by ELTLGEFLKL (ELT)-specific CTL cultures. The expression of MHC-I or TAP molecules was similar in all cell lines suggesting that tumors not recognized by CTLs may present defects in the generation of the ELT-epitope which could be due either to lack of generation or to subsequent degradation of the epitope. The cells were analyzed for the expression and the activity of extra-proteasomal peptidases. A significant overexpression and higher activity of TPPII was observed in colon-carcinoma cells which are not killed by ELT-specific CTLs, suggesting a possible role of TPPII in the degradation of the ELT-epitope. To confirm the role of TPPII in the degradation of the ELT-peptide, we showed that treatment of colon-carcinoma cells with a TPPII inhibitor resulted in a dose-dependent increased sensitivity to ELT-specific CTLs. These results suggest that TPPII is involved in degradation of the ELT-peptide, and its overexpression may contribute to the immune escape of colon-carcinoma cells.

[Motivational interviewing: some theoretical aspects and some practical exercises]. / L'entretien motivationnel: quelques repères thèoriques et quelques exercices pratiques.

Heath behavior change is an important task for caregivers. Motivational Interviewing (MI) is an effective evidence-based approach to overcoming the ambivalence that keeps many people from desired changes in their lives. In a warm and empathic atmosphere and in a basic trust in the patient self efficacy for achieving the change, MI is a patient-centered approach to enhance patient motivation to commit them in the change decision. Among critical conditions of change, MI practicing emphasizes on expressing empathy, supporting self-efficacy, rolling with resistance and developing discrepancy. Regular practicing with helping and non judgmental teachers is a necessary condition of a fruitful learning.

Red blood cell-mediated delivery of recombinant HIV-1 Tat protein in mice induces anti-Tat neutralizing antibodies and CTL.

The immunotherapeutic potential of biologically active HIV-1 Tat protein coupled to autologous red blood cells (RBCs) was evaluated in a mouse model. HIV-1 Tat expressed in Escherichia coli and purified to homogeneity was found to be active in viral trans activation and efficiently internalised by monocyte-derived dendritic cells (MDDCs). The product of HIV-Tat biotinylation and coupling to RBCs by means of a biotin-avidin-biotin bridge, (RBC-Tat), showed no trans activation activity and was still efficiently internalized by MDDCs as compared to uncoupled Tat.Balb/c mice were then immunized with 10 microg of soluble Tat in complete Freund's adjuvant or with 40 ng of Tat coupled on RBCs surface and boosted at week 3, 6 and 25 with 5 microg soluble Tat in incomplete Freund's adjuvant or with 20 ng of RBC-coupled Tat, respectively. Anti-Tat antibody response was similar in both groups; however, 2/6 animals immunized with soluble Tat and 6/6 animals immunized with RBC-Tat developed anti-Tat neutralizing antibodies. In addition, at week 28 cytolytic anti-Tat CTLs were detected in all animals although they were slightly higher in mice immunized with RBC-Tat. These results indicate that RBC-mediated delivery of HIV-1 Tat, in amounts 250 times lower than soluble Tat, is safe and induces specific CTL responses and neutralizing antibodies.

Molecular analysis of G6PD variants in northern Italy: a study on the population from the Ferrara district.

In the Ferrara district, an area south of the Po delta, four different variants of glucose-6-phosphate dehydrogenase (G6PD;E.C.1.1.49) have been described as a result of biochemical characterization of the enzyme protein: one was G6PD Mediterranean (G6PD Med) and three were local variants named Ferrara I, II, and III. The Ferrara I variant was recently analysed at the DNA level and shown to correspond to G6PD A376G/202A, while the mutations causing the variants II and III, still remain unknown. We analysed the G6PD coding region of 18 apparently unrelated G6PD deficient subjects, whose families have lived in the Ferrara district for at least three generations: 12 subjects had G6PD Med563T/1311T, 3, G6PD Santamaria376G/542T and 2, G6PD A-376G/202A. In one subject we found a new mutation, a G-->A transition at nucleotide 242 causing an Arg-->His amino-acid replacement at position 81. We named this new variant G6PD Lagosanto242 A. Phenotypically the enzyme has nearly normal kinetic properties and appears different from the variants Ferrara II and III.

Phase II trial of recombinant alpha-2b interferon in the treatment of metastatic skin melanoma.

A pilot study was undertaken to evaluate toxicity and activity of recombinant alpha-2b interferon in patients with metastatic malignant melanoma. Interferon was administered at the dosage of 10 x 10(6) IU/m2, 3 times a week i.m. 21 patients entered the study, 17 pretreated with chemotherapy and/or immunotherapy and 6 untreated. We obtained 3 partial responses (14.3%; 95% CL, 3.0-36.3%); 9 patients had stable disease. All patients experienced flue-like symptoms and fever; most fatigue and worsening of performance status. Recombinant interferon alpha-2b at the dosage and schedule used has limited but definite activity in metastatic malignant melanoma; the substantial subjective toxicity must be taken into consideration. Further trials testing recombinant alpha interferon in combination with chemotherapeutic agents, like DTIC, are warranted.

Neonatal periventricular leukomalacia: diagnosis and evolution evaluated by real-time ultrasound.

Recent advances in imaging techniques have provided the opportunity to obtain prompt diagnosis and to study the natural evolution of periventricular leukomalacia (PVL). Three premature neonates were followed up by brain sonograms from birth to six, four and three months of age, respectively. Sequential ultrasound examinations confirmed previous observations which identified four stages of PVL; 1. increased echogenicity in the periventricular white matter, 2. apparent normalization, 3. cystic cavitation, 4. resolution of cysts and development of ventriculomegaly. Decreased perfusion of the periventricular end-arterial zone is responsible for the development of PVL; this selective hypoperfusion has been documented in infants with a significant history of cardiorespiratory disturbances and group-B streptococcal sepsis. Two of our patients and other cases described in the literature, however, did not exhibit these clinical features. The present study suggests the involvement of chronic hypoxia and toxic insults in the pathogenesis of this condition and confirms the value and accuracy of sequential sonography in the diagnosis of PVL.

Criteria for discontinuing neonatal seizure therapy: a long-term appraisal.

In order to evaluate the criteria for discontinuing neonatal anticonvulsant treatment, 55 newborns with seizures have been studied. Clinical and EEG serial examinations were performed: soon after the first seizure, throughout the hospital course, and during the follow-up every 3 months until a year, and every 6 months later on. Anticonvulsant treatment with phenobarbital was discontinued (at 4 days-19 months; mean 104 days) on the basis of the following variables: type and number of seizures, time taken for their control, type and persistence of EEG abnormalities, initial neurological features, and seizure etiology. At the follow-up (12 months-8 years; mean 36 months) only 4 children had relapsed, 3 of them with a single short seizure without EEG abnormalities. The results obtained by means of the correlation between the length of anticonvulsant treatment and the clinical and EEG variables provide evidence of the value of the criteria employed. Of these, the duration of persistence of EEG abnormalities was the most important for planning the maintenance of anticonvulsant treatment and its discontinuation.

[Problems of the therapy of neonatal convulsions]. / Problemi di terapia nelle convulsioni neonatali.

The newborn with seizures should be treated urgently, because of the high risk of consequent brain damage. In addition to general management in order to correct metabolic and functional unbalancement, associated with the fits, specific causes of neonatal seizures (hypoglicemia, hypocalcemia, hypomagnesiemia, hypo-hypernatremia, pyridoxine deficiency) should be immediately removed. If neonatal seizures depend on other non specific causes (anoxia, cerebral hemorrhagy, malformation, infection or other, a symptomatic anticonvulsant treatment should be carried out without delay. Useful drugs for the newborn are phenobarbital, phenytoin and benzodiazepine e.v. or e.m. After fits have been controlled, an oral maintenance therapy has to be started with phenobarbital or phenytoin in order to avoid seizure-relaps. Clinical EEG and hematological data should be monitored to detect side effects, as well as plasma drugs levels to achieve adequate maintenance doses. Criteria for discontinuing the neonatal seizures treatment have not been well established. On the bases of the data collected through a longitudinal study of 54 newborns who developed seizures in the first day of life, clinical and EEG criteria for discontinuing anticonvulsant therapy are discussed. If the fits are rare, short, immediately controlled and EEG is mildly abnormal, we attempt to discontinuing treatment within 15 days. If fits are unfrequent, varying in length, their therapeutical control is reached within 3 days and the EEG is markedly abnormal but recovered within 1 month, treatment is discontinuing between 15 days and 3 months.(ABSTRACT TRUNCATED AT 250 WORDS)