Early diagnosis of bladder cancer through the detection of urinary tyrosine-phosphorylated proteins

Background:

A noninvasive, highly sensitive and specific urine test is needed for bladder cancer (BC) diagnosis and surveillance in addition to the invasive cystoscopy. We previously described the diagnostic effectiveness of urinary tyrosine-phosphorylated proteins (UPY) and a new assay (UPY-A) for their measurement in a pilot study. The aim of this work was to evaluate the performances of the UPY-A using an independent cohort of 262 subjects.

Methods:

Urinary tyrosine-phosphorylated proteins were measured by UPY-A test. The area under ROC curve, cutoff, sensitivity, specificity and predictive values of UPY-A were determined. The association of UPY levels with tumour staging, grading, recurrence and progression risk was analysed by Kruskal–Wallis and Wilcoxon''s test. To test the probability to be a case if positive at the UPY-A, a logistic test adjusted for possible confounding factor was used.

Results:

Results showed a significant difference of UPY levels between patients with BC vs healthy controls. For the best cutoff value, 261.26 Standard Units (SU), the sensitivity of the assay was 80.43% and the specificity was 78.82%. A statistically significant difference was found in the levels of UPY at different BC stages and grades between Ta and T1 and with different risk of recurrence and progression. A statistically significant increased risk for BC at UPY-A ⩾261.26 SU was observed.

Conclusions:

The present study supplies important information on the diagnostic characteristics of UPY-A revealing remarkable performances for early stages and allowing its potential use for different applications encompassing the screening of high-risk subjects, primary diagnosis and posttreatment surveillance.     

Dietary acrylamide and cancer risk: An updated meta‐analysis

The debate on the potential carcinogenic effect of dietary acrylamide is open. In consideration of the recent findings from large prospective investigations, we conducted an updated meta‐analysis on acrylamide intake and the risk of cancer at several sites. Up to July 2014, we identified 32 publications. We performed meta‐analyses to calculate the summary relative risk (RR) of each cancer site for the highest versus lowest level of intake and for an increment of 10 µg/day of dietary acrylamide, through fixed‐effects or random‐effects models, depending on the heterogeneity test. Fourteen cancer sites could be examined. No meaningful associations were found for most cancers considered. The summary RRs for high versus low acrylamide intake were 0.87 for oral and pharyngeal, 1.14 for esophageal, 1.03 for stomach, 0.94 for colorectal, 0.93 for pancreatic, 1.10 for laryngeal, 0.88 for lung, 0.96 for breast, 1.06 for endometrial, 1.12 for ovarian, 1.00 for prostate, 0.93 for bladder and 1.13 for lymphoid malignancies. The RR was of borderline significance only for kidney cancer (RR = 1.20; 95% confidence interval, CI, 1.00–1.45). All the corresponding continuous estimates ranged between 0.95 and 1.03, and none of them was significant. Among never‐smokers, borderline associations with dietary acrylamide emerged for endometrial (RR = 1.23; 95% CI, 1.00–1.51) and ovarian (RR = 1.39; 95% CI, 0.97–2.00) cancers. This systematic review and meta‐analysis of epidemiological studies indicates that dietary acrylamide is not related to the risk of most common cancers. A modest association for kidney cancer, and for endometrial and ovarian cancers in never smokers only, cannot be excluded.     

Natural vitamin C intake and the risk of head and neck cancer: A pooled analysis in the International Head and Neck Cancer Epidemiology Consortium

Evidence of associations between single nutrients and head and neck cancer (HNC) is still more limited and less consistent than that for fruit and vegetables. However, clarification of the protective mechanisms of fruit and vegetables is important to our understanding of HNC etiology. We investigated the association between vitamin C intake from natural sources and cancer of the oral cavity/pharynx and larynx using individual‐level pooled data from ten case‐control studies (5,959 cases and 12,248 controls) participating in the International Head and Neck Cancer Epidemiology (INHANCE) consortium. After harmonization of study‐specific exposure information via the residual method, adjusted odds ratios (ORs) and corresponding 95% confidence intervals (CIs) were estimated using unconditional multiple logistic regression models on quintile categories of 'non‐alcohol energy‐adjusted' vitamin C intake. In the presence of heterogeneity of the estimated ORs among studies, we derived those estimates from generalized linear mixed models. Higher intakes of vitamin C were inversely related to oral and pharyngeal (OR = 0.54, 95% CI: 0.45–0.65, for the fifth quintile category versus the first one, p for trend<0.001) and laryngeal cancers (OR = 0.52, 95% CI: 0.40–0.68, p for trend = 0.006), although in the presence of heterogeneity among studies for both sites. Inverse associations were consistently observed for the anatomical subsites of oral and pharyngeal cancer, and across strata of age, sex, education, body mass index, tobacco, and alcohol, for both cancer sites. The inverse association of vitamin C intake from foods with HNC may reflect a protective effect on these cancers; however, we cannot rule out other explanations.     

Antipsychotic drugs counteract autophagy and mitophagy in multiple sclerosis

Multiple sclerosis (MS) is a neuroinflammatory and neurodegenerative disease characterized by myelin damage followed by axonal and ultimately neuronal loss. The etiology and physiopathology of MS are still elusive, and no fully effective therapy is yet available. We investigated the role in MS of autophagy (physiologically, a controlled intracellular pathway regulating the degradation of cellular components) and of mitophagy (a specific form of autophagy that removes dysfunctional mitochondria). We found that the levels of autophagy and mitophagy markers are significantly increased in the biofluids of MS patients during the active phase of the disease, indicating activation of these processes. In keeping with this idea, in vitro and in vivo MS models (induced by proinflammatory cytokines, lysolecithin, and cuprizone) are associated with strongly impaired mitochondrial activity, inducing a lactic acid metabolism and prompting an increase in the autophagic flux and in mitophagy. Multiple structurally and mechanistically unrelated inhibitors of autophagy improved myelin production and normalized axonal myelination, and two such inhibitors, the widely used antipsychotic drugs haloperidol and clozapine, also significantly improved cuprizone-induced motor impairment. These data suggest that autophagy has a causal role in MS; its inhibition strongly attenuates behavioral signs in an experimental model of the disease. Therefore, haloperidol and clozapine may represent additional therapeutic tools against MS.

Multiple sclerosis (MS) is a neuroinflammatory disease, characterized by progressive neurological damage consisting in crumbling of the axonal myelin sheath in the central nervous system (CNS), followed by strong impairment in axonal conductance, axonal transection, and death of oligodendrocytes and neurons (1). MS onset is subacute or chronic, typically in a relapsing–remitting fashion, with phases of neuroinflammation corresponding to clinically evident disease followed by periods of oligodendrocyte proliferation, partial reconstitution of the myelin sheath, and corresponding clinical recovery. MS worsens over time, evolving into a progressive form, characterized by the disappearance of the relapsing–remitting phases and by permanent chronic lesions.Inflammation plays a central and fundamental role in MS (2). The initial phase of MS is characterized by signs of an autoimmune response including disruption of the blood–brain barrier, invasion of the CNS by immune cells, and presence of antibodies against myelin. These are believed to play a causative role in the onset of the disease. Immunological aggression is sustained by a macrophage- or microglia-driven secretion of proinflammatory cytokines, such as tumor necrosis factor alpha (TNF-α) (3, 4) or interleukin-1 beta (IL-1-β) (5, 6), which have been shown to promote neuronal, oligodendrocyte, and vascular damage in models of MS.Autophagy may also play a fundamental pathophysiological role in MS, but this has been only marginally explored thus far (1, 7). Autophagy represents a key intracellular function, which allows the cell to face periods of nutrient deprivation, meanwhile eliminating damaged and potentially harmful molecules, organelles, or invading microorganisms (8). These activities are grounded on the capacity of the cell to organize a double-membrane–lined autophagosome that, after engulfing the unwanted material, brings it to the lysosomal compartment, which in turn ensures degradation of proteins and recycling of nutrients.Many neurodegenerative diseases are characterized by pathological accumulation of misfolded or damaged proteins, suggesting an important role of autophagy in their pathophysiology (9, 10). In Alzheimer’s disease, for example, impairment of the autophagic flux due to reduced vesicle clearance has been associated to accumulation of an aggregated form of hyperphosphorylated tau protein and to extracellular amyloid-β (A-β) plaques (11). Similarly, in Parkinson’s disease, dysfunctional lysosomes and accumulation of autophagosomes are associated with intracellular inclusions of α-synuclein and other polyubiquitinated proteins.Several reports have demonstrated an increase in autophagic markers in human samples of MS patients (12). In particular, our and other groups have shown that autophagic and mitophagic markers are increased in serum and cerebrospinal fluid (CSF) of MS-affected individuals (13–15). However, the role of autophagy in MS remains still controversial. (16, 17).In this study, we investigated in detail the involvement of autophagy in MS. We used in vitro, ex vivo, and in vivo models to test the effect of proinflammatory cytokines, lysolecithin (LPC), and cuprizone (CPZ) on myelin production, axonal myelination, and motor performance. In particular, we investigated the occurrence of autophagy in the different models and examined the effect of autophagy inhibitors and of two antipsychotic drugs, clozapine and haloperidol, on autophagy and behavior, revealing a significant improvement in motor function in MS animals.The rationale for employing these antipsychotic drugs stems from neuroimaging and postmortem investigations on the brain of schizophrenia patients that revealed loss of white matter (18, 19) and down-regulation of genes involved in oligodendrocyte functioning and myelination (20, 21). Antipsychotic drugs, in particular haloperidol and clozapine, proved effective in improving cortical myelination as well as spatial working memory and locomotor activity in animal models (22–25). The molecular mechanism(s) underlying these effects are still obscure, but it has been recently suggested that these antipsychotic drugs may interfere with the autophagic process (26, 27). Our findings explore this mechanism of action of haloperidol and clozapine and disclose the opportunity of repurposing these drugs that are currently employed in other clinical settings for the treatment of MS.     

Mapping and kinetics of microglia/neuron cell‐to‐cell contacts in the 6‐OHDA murine model of Parkinson's disease

As neuroinflammatory processes are involved in the pathogenesis of Parkinson's disease (PD), we provide several key data describing the time‐course of microglial accumulation in relation with behavioral alterations and neurodegeneration in a murine model of PD induced by intrastriatal injection of 6‐hydroxydopamine (6‐OHDA). Our study argues for a major role of microglia which accumulation is somehow early and transient in spite of the neuronal loss progression. Moreover, we observed less 6‐OHDA‐induced neurodegeneration associated with less inflammatory reaction in DAP‐12 Knock‐In mice. The direct cell‐to‐cell contacts that may support physical interactions between microglia and altered dopaminergic neurons are ill‐defined, while it is currently hypothesized that microglia support an immune‐mediated amplification of neurodegeneration by establishing a molecular cross talk with neurons. Indeed, we sought to map microglia/neuron appositions in substantia nigra (SN) of 6‐OHDA injected C57Bl/6 mice and CX3CR1/^GFP/+ mice. Confocal immunofluorescence analyses followed by 3D reconstitutions reveal close appositions between the soma of TH+ neurons and microglial cell bodies and ramifications. Interestingly, some microglial ramifications penetrated TH⁺ somas and about 40% of GFP⁺ microglial cells in the injured SN harbored TH⁺ intracytoplasmic inclusions. These results suggest a direct cross talk between neurons and microglia that may exert a microphagocytic activity toward TH+ neurons. Altogether, these results obtained in a murine PD model may participate in the understanding of microglial cells' function in neurodegenerative diseases. GLIA 2013;61:1645–1658     

On the relationship between retrospective childhood ADHD symptoms and adult BPD features: The mediating role of action-oriented personality traits

A number of studies have reported data suggestive of a significant association between ADHD and BPD, nevertheless, the nature of this relation has not been fully understood yet. In our study, we tried to evaluate if the relationship between retrospectively assessed ADHD symptoms and adult BPD features could mediated by selected temperament/personality traits. Four hundred forty-seven in- and outpatients consecutively admitted to the Clinical Psychology and Psychotherapy Unit of the Scientific Institute H San Raffaele of Milan, Italy, were administered the Italian versions of the following instruments: Structured Clinical Interview for DSM-IV Axis II Personality Disorders, Version 2.0 (SCID-II), Wender Utah Rating Scale (WURS), Temperament and Character Inventory-Revised (TCI-R), Barratt Impulsiveness Scale-11 (BIS-11), and Aggression Questionnaire (AQ). Our mediation analyses showed that the combination of impulsivity, aggression, novelty seeking, and juvenile conduct problems completely mediate the relationship between retrospectively assessed ADHD symptoms and current BPD features.     

Progranulin gene (GRN) promoter methylation is increased in patients with sporadic frontotemporal lobar degeneration

Mutations in progranulin gene (GRN) are the most common cause of autosomal dominant familial frontotemporal lobar degeneration (FTLD). In addition, GRN variability influences the risk to develop the disease in non-carriers (sporadic FTLD). We evaluated progranulin gene (GRN) promoter methylation levels in peripheral blood mononuclear cells isolated from 38 patients with sporadic FTLD compared with 38 controls, and correlate them with GRN mRNA expression rate. The percentage of methylation of the GRN promoter was increased in patients with FTLD compared with controls (61.5 vs. 46.3 %, P < 0.001). A trend towards decreased GRN relative expression was observed in patients compared with controls (threefold decrease over controls, P > 0.05), together with a negative correlation between the degree of GRN promoter methylation and mRNA GRN levels (ρ = ?0.1, P > 0.05). GRN promoter methylation was not correlated with age. In conclusion, the degree of methylation of the GRN promoter is increased in patients with FTLD as compared with controls, likely leading to a decreased expression of GRN.