The functional polymorphism rs73598374:G>A (p.Asp8Asn) of the ADA gene is associated with telomerase activity and leukocyte telomere length

Recent evidence demonstrated a relevant role of adenosine deaminase (ADA) in replicative senescence of T cells through its capacity to modulate telomerase activity (TA). Herein, we tested the impact of the functional polymorphism ADA rs73598374:G>A (c.22G>A, p.Asp8Asn) on telomere biology, by measuring TA and leukocyte telomere length (LTL) in healthy subjects selected according to rs73598374 genotype. rs73598374-A carriers showed lower TA (P=0.019) and shorter LTL (P=0.003), respectively, compared to G/G carriers. rs73598374-A carriers showed a stronger cross-sectional age reduction of LTL (r=-0.314, P=0.005) compared to G/G carriers (r=-0.243, P=0.022). The reduced ADA activity associated to rs73598374-A variant predisposes those carriers to display higher levels of adenosine compared to G/G carriers. Consequently, it may lead to an accelerated process of replicative senescence, causing a stronger reduction of TA and in turn shorter LTL. In conclusion, the crucial role played by replicative senescence of the immune system in several human diseases and in the aging process underscores the relevance of the present findings and also spurs interest into the possible involvement of rs73598374 in shaping the susceptibility to several age-related diseases

The functional VNTR MNS16A of the TERT gene is associated with human longevity in a population of Central Italy.

Telomerase, encoded by TERT, is the ribonucleoprotein polymerase that maintains telomere ends and it plays a crucial role in cellular senescence. TERT single nucleotide polymorphisms (SNPs) have been associated both with various malignancies and telomere length (TL). The association of TERT SNPs with longevity remains uncertain and varies with ethnicity. The aim of this study was to investigate whether the functional variable number of tandem repeat (VNTR) MNS16A of TERT is associated with longevity. METHODS: MNS16A genotypes have been determined for 1072 unrelated healthy individuals from Central Italy (18-106 years old) divided into three gender-specific age classes defined according to demographic information and accounting for the different survivals between sexes: for men (women), the first class consists of individuals 88 years old (>91 years old). TL was assessed using genomic DNA from whole blood of 72 selected individuals by a multiplex real-time PCR assay. RESULTS: MNS16A appears associated to longevity, showing significant associations in Comparison 2 (Age Class 3 vs. Age Class 2) under both additive (odds ratio [O.R.] 0.749; p=0.019) and dominant (O.R. 0.579; p=0.011) models. The MNS16A*L allele is significantly underrepresented in Age Class 3 (O.R. 0.759; p=0.020) compared to Age Class 2. A significant telomere attrition is reported along the three age classes (p=0.0001), that remains significant only in L*/L* genotype carriers (p=0.002) when the analysis was conducted according to MNS16A genotype. CONCLUSIONS: The TERT MNS16A*L allele appears negatively associated with longevity. The concomitant significant telomere cross sectional attrition rate observed for L*/L* genotype suggests that this polymorphism could influence human longevity by affecting TL

Further steps in the role of autophagy in methamphetamine toxicity

Methamphetamine (METH) abuse is known to cause a variety of disorders Incluing depression and psychosis. METH induces nigrostriatal damage in animal models and in humans consisting of intracellular alterations in nigral DA cell bodies, degeneration of DA terminals and decreased striatal DA levels. Following METH exposure, the number of nigral cell bodies is quite preserved but autophagy-like vacuoles and cytoplamic accumulation of misfolded proteins are observed. The DA-containing PC12 cell lines represent a simple model of METH toxicity and are commonly used in vitro to understand the pathophysiology of DA neurons. We analyzed at morphological level the effects of plasmid-dependent autophagy modulation on METH toxicity in PC12 cell line. We profited from the high number of authophagic like vacuoles induced by low doses of METH in order to isolate cell fraction in which to study their origin, dynamic structure and molecular composition. We found that authophagy-like vacuoles are positive both for autophagy and proteasome markers. The modulation of autophagy via a p62 containing plasmid protected from METH toxicity, while the inhibition of autophagy machinery worsened METH neurotoxicity. The present data substantiate the protective role of the autophagy machinery in METH-induced DA toxicity where the pro-autophagy protein p62 possesses a key role

Approximation of Fourier Integral Operators by Gabor multipliers

A general principle says that the matrix of a Fourier integral operator with respect to wave packets is concentrated near the curve of propagation. We prove a precise version of this principle for Fourier integral operators with a smooth phase and a symbol in the Sjoestrand class and use Gabor frames as wave packets. The almost diagonalization of such Fourier integral operators suggests a specific approximation by (a sum of) elementary operators, namely modified Gabor multipliers. We derive error estimates for such approximations. The methods are taken from time-frequency analysis.Comment: 22. page

Aggregating Proteins Affect Immunity and Bone Homeostasis in Vivo

Background: DNA vaccines provide high tolerability and safety but commonly suffer from suboptimal immunogenicity. We previously reported that a plasmid vector (pATRex), encoding for the extracellular domain of the Tumor\ud Endothelial Marker-8, when given in combination with plasmid-encoded tumor antigens acted as a powerful molecular adjuvant enhancing immunity against various tumors. Aims: the present study has been developed to determine whether the adjuvant action of pATRex can also extend to vaccines against infectious diseases such as malaria, to identify which is the mechanism of action that underlies pATRex adjuvancy and, finally, to deeply investigate the potential side effects. Results: here we show that co-administration of pATRex potentiates antibody production elicited by an intramuscular injection of plasmid encoding for malaria plasmodium antigens. Cells were transfected with pATRex and this resulted in formation of insoluble intracellular aggregates (as confirmed by 3D molecular modeling) and apoptotic cell death, thus explaining that adjuvancy mechanism is mediated by immunogenic cell death. Besides, histological examination of pATRex-injected mice organs, we have uncovered that proteotoxic aggregates profoundly disrupt the bone and bone marrow architecture and influenced the molecular network and the cytokine interchanges through the skeletal and hematopoietic marrow components. We also discovered that a similar scenario occurs when mice were injected with a\ud plasmid DNA encoding for the mutant Huntington, a well-known aggregating-protein responsible for Huntington's disease in humans. Finally, we provided the first evidence that the impaired bone deposition and the enfeeble bone\ud homeostasis is also present in the transgenic mouse model for human Spinocerebellar Ataxia 1 carrying the mutant aggregating protein Ataxin 1.\ud Conclusions and Perspectives: in the present work we found that plasmids encoding aggregating proteins behave as adjuvants for DNA vaccines but they also controvert the delicate steady-state bone and bone marrow physiology\ud ultimately driving to impaired bone formation in conjunction with bone marrow niche disruption. These findings drew the attention of further investigation in\ud reference to the side effects associated with the use of aggregating proteins as adjuvants. We have also unveiled, for the first time, that the osteoporotic phenotype, which is often present in the neurodegenerative diseases, is a\ud coherent fall-out of the protein aggregates observed in human and animal carriers of the mentioned disease

Recombinant DNA Technology for Melanoma Immunotherapy: Anti-Id DNA Vaccines Targeting High Molecular Weight Melanoma-Associated Antigen

Anti-idiotypic MK2-23 monoclonal antibody (anti-Id MK2-23 mAb), which mimics the high molecular weight melanoma-associated antigen (HMW-MAA), has been used to implement active immunotherapy against melanoma. However, due to safety and standardization issues, this approach never entered extensive clinical trials. In the present study, we investigated the usage of DNA vaccines as an alternative to MK2-23 mAb immunization. MK2-23 DNA plasmids coding for single chain (scFv) MK2-23 antibody were constructed via the insertion of variable heavy (VH) and light (VL) chains of MK2-23 into the pVAC-1mcs plasmids. Two alternative MK2-23 plasmids format VH/VL, and VL/VH were assembled. We demonstrate that both polypeptides expressed by scFv plasmids in vitro retained the ability to mimic HMW-MAA antigen, and to elicit specific anti-HMW-MAA humoral and cellular immunoresponses in immunized mice. Notably, MK2-23 scFv DNA vaccines impaired the onset and growth of transplantable B16 melanoma cells not engineered to express HMW-MAA. This pilot study suggests that optimized MK2-23 scFv DNA vaccines could potentially provide a safer and cost-effective alternative to anti-Id antibody immunization, for melanoma immunotherapy

The extent of pelvic lymph node dissection correlates with the biochemical recurrence rate in patients with intermediate- and high-risk prostate cancer.

OBJECTIVE: • To assess the impact of pelvic lymph node dissection (PLND) and of the number of lymph nodes (LNs) retrieved during radical prostatectomy (RP) on biochemical relapse (BCR) in pNX/0/1 patients with prostate cancer according to the clinical risk of lymph node invasion (LNI). PATIENTS AND METHODS: • We evaluated 872 pT2-4 NX/0/1 consecutive patients submitted to RP between October 1995 and June 2009, with the following inclusion criteria: (i) a follow-up period ≥12 months; (ii) the avoidance of neoadjuvant hormonal therapy or adjuvant hormonal and/or adjuvant radiotherapy; (iii) the availability of complete follow-up data; (iv) no pathological T0 disease; (v) complete data regarding the clinical stage and Gleason score (Gs), the preoperative prostate-specific antigen (PSA) level and the pathological stage. • The patients were stratified as having low risk (cT1a-T2a and cGs ≤6 and PSA level < 10 ng/mL), intermediate risk (cT2b-T2c or cGs = 7 or PSA level = 10-19.9) or high risk of LNI (cT3 or cGs = 8-10 or PSA level ≥ 20). • The 872 patients were divided into two LN groups according to the number of LNs retrieved: group 1 had no LN or one to nine LNs removed; group 2 had 10 or more LNs. • The variables analysed were LN group, age, PSA level, clinical and pathological stage and Gs, surgical margin status, LN status and number of LN metastases; the primary endpoint was the BCR-free survival. RESULTS: • The mean follow-up was 55.8 months. • Of all the patients, 305 (35%) were pNx and 567 (65.0%) were pN0/1. • Of the 567 patients submitted to PLND, the mean number of LNs obtained was 10.9, and 49 (8.6%) were pN1. • In the 402 patients at low risk of LNI, LN group was not a significant predictor of BCR at univariate analysis, while in the 470 patients at intermediate and high risk of LNI, patients with ≥ 10 LNs removed had a significantly lower BCR-free survival at univariate and multivariate analysis. CONCLUSION: • In our study population, a more extensive PLND positively affects the BCR-free survival regardless of the nodal status in intermediate- and high-risk prostate cancer

Accuracy of [11C] Choline positron emission tomography/computed tomography in preoperative staging in patients with bladder cancer referred to radical cystectomy (RC): comparison with conventional computed tomography (CT)

.Accuracy of [11C] Choline positron emission tomography/computed tomography in preoperative staging in patients with bladder cancer referred to radical cystectomy (RC): comparison with conventional computed tomography (CT)

NEFRECTOMIA PARZIALE LAPAROSCOPICA VS OPEN:IMPATTO DELLA CURVA DI APPRENDIMENTO SUI RISULTATI ONCOLOGICI E FUNZIONALI

NEFRECTOMIA PARZIALE LAPAROSCOPICA VS OPEN:IMPATTO DELLA CURVA DI APPRENDIMENTO SUI RISULTATI ONCOLOGICI E FUNZIONAL