Antiretroviral compounds affect the granule-dependent mechanisms of lysis in CD8 T cells

Cytotoxic T-lymphocytes (CTLs) are essential for suppression of viral replication and, in particular, they have a pivotal role in control of progression of HIV infection. It has been demonstrated that HIV-specific CTL responses are defective in HIV-infected patients undergoing highly active antiretroviral therapy (HAART). In this study we investigated the effects of antiretroviral compounds on the granule-dependent mechanisms of lysis in peripheral blood mononuclear cells (PBMCs). PBMCs of 10 HCs were incubated with 3 different antiretroviral drugs combinations: combination A: AZT (NRTI) + 3TC (NRTI) + IDV (PI); combination B: d4T (NRTI) + ddI (NRTI) + NFV (PI); combination C: 3TC (NRTI) + EFV (NNRTI) + TDF (NRTI). To evaluate the CTLs function we measured: production and release of granule-dependent effector molecules (perforin, granzyme B); production and release of granule-independent effector molecules (IFN-gamma, TNF-alpha); degranulation markers (LAMP1 and LAMP2). To assest the immunomodulant effects of IL-15, PBMCs were also incubate in presence of this cytokyne. Antiretroviral compounds reduce the granzyme B and perforin production (while they don't affect the IFN-gamma and TNF-alpha production). Moreover, one of the 3 tested combinations of antiretroviral compounds (combination B) reduces the granzyme B release and affects the degranulation in CTLs. IL-15 increases the levels of granzyme B and perforin. Antiretroviral compounds mainly affect the expression of genes encoding for Pfp and GranzB, and deteriorate the mechanism of degranulation in CD8+ T cells. L-15 restores both the granule-dependent and the granule-independent cytotoxic mechanisms. Basd on these data, IL-15 seems to be useful in overcoming the negative effects of antiretroviral compounds on the cytotoxic function

Zeolites for Fine Chemical Production State of Art and Perspectives

The paper analyses the role of catalysis and that of renewable resources in the frame of a sustainable development. The possible uses of natural feedstocks for chemical production and the application of catalytic methods to their transformations are reviewed, with emphasis on carbohydrates and vegetable oils and on zeolite catalysts, respectively. The problems arising from the embedment of active sites on the catalyst surface are discussed, with the aid of specific examples taken from oxidation and acid catalysed reactions

A novel approach to cancer therapy using melanin-containing patches

In recent decades, the incidence of melanoma has been steadily raising, even if it is still considered to be a rare cancer. In contrast with what is observed in other solid tumors, melanomas principally affect young and middle-aged individuals who are often still in the prime of their carriers and are involved in raising a family. Surgery is the first-line treatment, in particular for early-stage melanomas; unfortunately, patients often develop metastasis, and the presence of a first tumor increases the chance to develop a second melanoma (1)

High-Dimensional ICA Analysis Detects Within-Network Functional Connectivity Damage of Default-Mode and Sensory-Motor Networks in Alzheimer’s Disease

High-dimensional independent component analysis (ICA), compared to low-dimensional ICA, allows to conduct a detailed parcellation of the resting-state networks. The purpose of this study was to give further insight into functional connectivity (FC) in Alzheimer's disease (AD) using high-dimensional ICA. For this reason, we performed both low- and high-dimensional ICA analyses of resting-state fMRI data of 20 healthy controls and 21 patients with AD, focusing on the primarily altered default-mode network (DMN) and exploring the sensory-motor network. As expected, results obtained at low dimensionality were in line with previous literature. Moreover, high-dimensional results allowed us to observe either the presence of within-network disconnections and FC damage confined to some of the resting-state subnetworks. Due to the higher sensitivity of the high-dimensional ICA analysis, our results suggest that high-dimensional decomposition in subnetworks is very promising to better localize FC alterations in AD and that FC damage is not confined to the DMN

Serum miRNAs Expression and SNAP-25 Genotype in Alzheimer’s Disease

MicroRNAs (miRNAs) are small non-coding RNAs that control gene expression by binding their 3′ untranslated region (3′UTR) region; these molecules play a fundamental role in several pathologies, including Alzheimer’s disease (AD). Synaptosomal-associated protein of 25 kDa (SNAP-25) is a vesicular protein of soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) involved in neural plasticity and in the exocytosis of neurotransmitters, processes that are altered in AD. Recent results showed that a reduction of SNAP-25 is associated with dementia, and that the rs363050 SNAP-25 polymorphism correlates with cognitive decline and brain atrophy, as well as with the outcome of multistructured rehabilitation in AD patients. We verified the presence of possible correlations between the serum concentration of miRNAs that bind the SNAP-25 3′UTR region and AD. Six different microRNAs (miR-181a-5p, miR-361-3p, miR-23a-3p, miR-15b-3p, 130a-3p and miR-27b-3p) that bind the SNAP-25 3′UTR region were measured by qPCR in serum of AD patients (n = 22), mild cognitive impairment (MCI) subjects (n = 22) and age- and sex-matched controls (n = 22); analysis of results was done stratified for the rs363050 SNAP-25 genotype. Results showed that miR-27b-3p, miR-23a-3p and miR181a-5p serum concentration was significantly reduced in rs363050 SNAP-25 GG homozygous AD patients. Notably, concentration of these miRNAs was comparable in rs363050 AA homozygous AD patients, MCI and healthy controls (HCs). Data herein suggest that miRNAs that bind the SNAP-25 3′UTR region interact with SNAP-25 polymorphisms to influence the neural plasticity typical of AD brains, possibly as a consequence of modulatory activity on SNAP-25 mRNA and/or protein

Evolutionary analysis provides insight into the origin and adaptation of HCV

Hepatitis C virus (HCV) belongs to the Hepacivirus genus and is genetically heterogeneous, with seven major genotypes further divided into several recognized subtypes. HCV origin was previously dated in a range between ~200 and 1000 years ago. Hepaciviruses have been identified in several domestic and wild mammals, the largest viral diversity being observed in bats and rodents. The closest relatives of HCV were found in horses/donkeys (equine hepaciviruses, EHV). However, the origin of HCV as a human pathogen is still an unsolved puzzle. Using a selection-informed evolutionary model, we show that the common ancestor of extant HCV genotypes existed at least 3000 years ago (CI: 3192-5221 years ago), with the oldest genotypes being endemic to Asia. EHV originated around 1100 CE (CI: 291-1640 CE). These time estimates exclude that EHV transmission was mainly sustained by widespread veterinary practices and suggest that HCV originated from a single zoonotic event with subsequent diversification in human populations. We also describe a number of biologically important sites in the major HCV genotypes that have been positively selected and indicate that drug resistance-associated variants are significantly enriched at positively selected sites. HCV exploits several cell-surface molecules for cell entry, but only two of these (CD81 and OCLN) determine the species-specificity of infection. Herein evolutionary analyses do not support a long-standing association between primates and hepaciviruses, and signals of positive selection at CD81 were only observed in Chiroptera. No evidence of selection was detected for OCLN in any mammalian order. These results shed light on the origin of HCV and provide a catalog of candidate genetic modulators of HCV phenotypic diversity

Monosodium urate crystals activate the inflammasome in primary progressive multiple sclerosis

Inflammasome-driven inflammation is postulated to play a role in multiple sclerosis (MS), but there is no direct evidence that the nod-like receptor protein 3 (NLRP3) inflammasome is involved in MS pathogenesis. Uric acid was shown to be one of the "danger" signals involved in the activation of NLRP3 inflammasome; notably, the concentration of uric acid is increased in the serum and in the cerebrospinal fluid of MS individuals. To better investigate the role of the NLRP3 inflammasome in MS-associated inflammation, we primed with lipopolysaccharide and stimulated with monosodium urate crystals PBMCs of 41 MS patients with different disease phenotypes. Eleven individuals with primary progressive MS (PPMS), 10 individuals with stable relapsing-remitting MS (SMS), 10 individuals with acute relapsing-remitting MS (AMS), 10 individuals with benign MS were analyzed; 10 healthy controls were enrolled as well in the study. The expression of the NLRP3, apoptosis-associated speck-like protein containing CARD (ASC), caspase-1, caspase-8, IL-1\u3b2, and IL-18 inflammasome genes was evaluated by RT-PCR. NLRP3 and ASC-speck protein expression was analyzed by FlowSight AMNIS, whereas production of the pro-inflammatory cytokines IL-1\u3b2 and IL-18 and of caspase-1 and caspase-8 was measured by ELISA in supernatants. Results showed that uric acid serum concentration was significantly increased in PPMS; in these and in AMS patients, mRNA for NLRP3, ASC, and IL-18 was upregulated as well, but caspase-8 mRNA was upregulated only in PPMS. Expression of NLRP3 and ASC-speck protein was significantly increased in PPMS, SMS, and AMS patients, but IL-18 and caspase-8 production was significantly increased only in PPMS, in whom a direct correlation between hyperuricemia and caspase-8 was detected. The NLRP3/caspase-8 inflammasome pathway is activated in PPMS, possibly as a consequence of hyperuricemia. Therapeutic strategies reducing NLRP3 activation and/or lowering hyperuricemia could be useful in the therapy of PPMS

Effective artifact removal in resting state fMRI data improves detection of DMN functional connectivity alteration in Alzheimer's disease

Artifact removal from resting state fMRI data is an essential step for a better identification of the resting state networks and the evaluation of their functional connectivity (FC), especially in pathological conditions. There is growing interest in the development of cleaning procedures, especially those not requiring external recordings (data-driven), which are able to remove multiple sources of artifacts. It is important that only inter-subject variability due to the artifacts is removed, preserving the between-subject variability of interest\u2014crucial in clinical applications using clinical scanners to discriminate different pathologies and monitor their staging. In Alzheimer's disease (AD) patients, decreased FC is usually observed in the posterior cingulate cortex within the default mode network (DMN), and this is becoming a possible biomarker for AD. The aim of this study was to compare four different data-driven cleaning procedures (regression of motion parameters; regression of motion parameters, mean white matter and cerebrospinal fluid signal; FMRIB's ICA-based Xnoiseifier\u2014FIX\u2014cleanup with soft and aggressive options) on data acquired at 1.5 T. The approaches were compared using data from 20 elderly healthy subjects and 21 AD patients in a mild stage, in terms of their impact on within-group consistency in FC and ability to detect the typical FC alteration of the DMN in AD patients. Despite an increased within-group consistency across subjects after applying any of the cleaning approaches, only after cleaning with FIX the expected DMN FC alteration in AD was detectable. Our study validates the efficacy of artifact removal even in a relatively small clinical population, and supports the importance of cleaning fMRI data for sensitive detection of FC alterations in a clinical environment

Oligomeric Alpha-Synuclein and STX-1A from Neural-Derived Extracellular Vesicles (NDEVs) as Possible Biomarkers of REM Sleep Behavior Disorder in Parkinson's Disease: A Preliminary Cohort Study

REM sleep behavior disorder (RBD) has a tighter link with synucleinopathies than other neurodegenerative disorders. Parkinson's Disease (PD) patients with RBD have a more severe motor and cognitive impairment; biomarkers for RBD are currently unavailable. Synaptic accumulation of α-Syn oligomers and their interaction with SNARE proteins is responsible for synaptic dysfunction in PD. We verified whether oligomeric α-Syn and SNARE components in neural-derived extracellular vesicles (NDEVs) in serum could be biomarkers for RBD. Forty-seven PD patients were enrolled, and the RBD Screening Questionnaire (RBDSQ) was compiled. A cut-off score > 6 to define probable RBD (p-RBD) and probable non-RBD (p non-RBD) was used. NDEVs were isolated from serum by immunocapture, and oligomeric α-Syn and SNARE complex components VAMP-2 and STX-1 were measured by ELISA. NDEVs' STX-1A resulted in being decreased in p-RBD compared to p non-RBD PD patients. A positive correlation between NDEVs' oligomeric α-Syn and RBDSQ total score was found (p = 0.032). Regression analysis confirmed a significant association between NDEVs' oligomeric α-Syn concentration and RBD symptoms (p = 0.033) independent from age, disease duration, and motor impairment severity. Our findings suggest that synuclein-mediated neurodegeneration in PD-RBD is more diffuse. NDEVs' oligomeric α-Syn and SNARE complex components' serum concentrations could be regarded as reliable biomarkers for the RBD-specific PD endophenotype