Sirt3 and metabolic reprogramming mediate the antiproliferative effects of whey in human colon cancer cells

Emerging strategies to improve healthy aging include dietary interventions as a tool to promote health benefits and reduce the incidence of aging-related comorbidities. The health benefits of milk are also linked to its richness in betaines and short-chain acylcarnitines, which act synergisti-cally in conferring anticancer, anti-inflammatory, and antioxidant properties. Whey, despite being a dairy by-product, still has a considerable content of bioactive betaines and acylcarnitines. Here, we investigated the anticancer properties of whey from Mediterranean water buffalo (Bubalus bubalis) milk by testing its antiproliferative effects in colorectal cancer (CRC) cells HT-29, HCT 116, LoVo and SW480. Results indicated that treatment with whey for 72 h inhibited cell proliferation (p < 0.001), induced cell cycle arrest, and apoptosis via caspase-3 activation, and modulated cell metabolism by limiting glucose uptake and interfering with mitochondrial energy metabolism with the highest effects observed in HT-29 and HCT 116 cells. At molecular level, these effects were accompanied by upregulation of sirtuin 3 (SIRT3) (p < 0.01) and peroxisome proliferator-activated receptor (PPAR)-γ expression (p < 0.001), and downregulation of lactate dehydrogenase A (LDHA) (p < 0.01), sterol regulatory-element binding protein 1 (SREBP1) (p < 0.05), and PPAR-α (p < 0.01). Transient SIRT3 gene silencing blocked the effects of whey on the LDHA, PPAR-γ, and PPAR-α protein expressions (p < 0.01) suggesting that the whey capacity of perturbating the metabolic homeostasis in CRC cell lines is mediated by SIRT3

SIRT3 Modulates Endothelial Mitochondrial Redox State during Insulin Resistance

Emerging evidence indicates that defects in sirtuin signaling contribute to impaired glucose and lipid metabolism, resulting in insulin resistance (IR) and endothelial dysfunction. Here, we examined the effects of palmitic acid (PA) treatment on mitochondrial sirtuins (SIRT2, SIRT3, SIRT4, and SIRT5) and oxidative homeostasis in human endothelial cells (TeloHAEC). Results showed that treatment for 48 h with PA (0.5 mM) impaired cell viability, induced loss of insulin signaling, imbalanced the oxidative status (p &lt; 0.001), and caused negative modulation of sirtuin protein and mRNA expression, with a predominant effect on SIRT3 (p &lt; 0.001). Restoration of SIRT3 levels by mimic transfection (SIRT3+) suppressed the PA-induced autophagy (mimic NC+PA) (p &lt; 0.01), inflammation, and pyroptosis (p &lt; 0.01) mediated by the NLRP3/caspase-1 axis. Moreover, the unbalanced endothelial redox state induced by PA was counteracted by the antioxidant δ-valerobetaine (δVB), which was able to upregulate protein and mRNA expression of sirtuins, reduce reactive oxygen species (ROS) accumulation, and decrease cell death. Overall, results support the central role of SIRT3 in maintaining the endothelial redox homeostasis under IR and unveil the potential of the antioxidant δVB in enhancing the defense against IR-related injuries

Psychometric properties and diagnostic accuracy of the short form of the geriatric anxiety scale (GAS-10)

Background: Anxious symptoms have a negative impact on different aspects of the elderly\u2019s quality of life, ranging from the adoption of unhealthy lifestyle behaviours to an increased functional impairment and a greater physical disability. Different brief assessment instruments have been developed as efficacy measures of geriatric anxiety in order to overcome psychometric weaknesses of its long form. Among these, the 10-item Geriatric Anxiety Scale (GAS-10) showed strong psychometric properties in community-dwelling samples. However, its diagnostic accuracy is still unexplored, as well as its discriminative power in clinical samples. Methods: In the present study, we explored the psychometric performance of the GAS-10 in the elderly through Item Response Theory in a sample of 1200 Italian community-dwelling middle-aged and elderly adults (53.8% males, mean age = 65.21 \ub1 9.19 years). Concurrent validity, as well as diagnostic accuracy, was examined in a non-clinical sample (N = 229; 46.72% males) and clinical sample composed of 35 elderly outpatients (74.28% females) with Generalized Anxiety Disorder (GAD). Results: The GAS-10 displayed good internal construct validity, with unidimensional structure and no local dependency, good accuracy, and no signs of Differential Item Functioning (DIF) or measurement bias due to gender, but negligible due to the age. Differences in concurrent validity and diagnostic accuracy among the long form version of the GAS and the GAS-10 were not found significant. The GAS-10 may be more useful than the longer versions in many clinical and research applications, when time constraints or fatigue are issues. Conclusion: Using the ROC curve, the GAS-10 showed good discriminant validity in categorizing outpatients with GAD disorder, and high anxiety symptoms as measured by the GAS-SF cut-off. The stable cut-off point provided could enhance the clinical usefulness of the GAS-10, which seems to be a promising valid and reliable tool for maximize diagnostic accuracy of geriatric anxiety symptoms

Inhibition of NF-κB activation sensitizes U937 cells to 3′-azido-3′-deoxythymidine induced apoptosis

In this study, we investigated molecular mechanisms underlying low susceptibility to apoptosis induced by the nucleoside analog azidothymidine (AZT) and the role of nuclear factor-κB (NF-κB) activation in these phenomena. A preliminary screening in different cell lines indicated U937 monocytic cell line as suitable to this purpose. Treatment of U937 cells even with suprapharmacological concentrations of AZT induced only moderate levels of apoptosis. Surprisingly, SuperArray analysis showed that AZT induced the transcriptional activity of both pro- and anti-apoptotic genes. Interestingly, moreover, several genes upregulated by AZT were NF-κB related. In fact, AZT, after an initial inhibition of NF-κB activation with respect to control, induced a transient, but consistent, increase in NF-κB-binding activity. Inhibition of NF-κB activation in U937 cells, stably transfected with a dominant-negative IκBα or by pharmacological treatment, sensitized them to apoptosis induced by AZT and impaired the upregulation of anti-apoptotic genes in response to AZT treatment, with respect to control cells. These results indicate that NF-κB activation by AZT has a role in protecting target cells from apoptotic cell death, improving our understanding of the toxicology and the therapeutic usage of this drug

Insight from imaging on plaque vulnerability: similarities and differences between coronary and carotid arteries—implications for systemic therapies

Nowadays it is widely accepted that the rupture of the atherosclerotic plaque in coronary and carotid arteries plays a fundamental role in the development of acute myocardial infarctions or cerebrovascular events. In recent years, imaging techniques have explored, with a new level of detail, the atherosclerotic disease generating new evidences that some plaque characteristics are significantly associated to the risk of rupture and subsequent thrombosis or embolization. Moreover, the recent evidence of the anti-atherosclerotic effects determined by lipid-lowering and anti-inflammatory therapies poses a challenge for the choice of therapeutic approaches (best/optimal medical therapy vs. revascularization), maximized by the evidence that coronary and carotid atherosclerosis share common patterns but also differ regarding some important features. In this Review, we discuss the similarities and differences between coronary and carotid artery vulnerable plaque from the imaging point of view and the potential implications for systemic therapies according to the emerging evidence

Inhibition of IκBα phosphorylation potentiates regulated cell death induced by azidothymidine in HTLV-1 infected cells

Adult T cell leukemia/lymphoma (ATL) can be susceptible, at least transiently, to treatments with azidothymidine (AZT) plus IFNα and/or arsenic trioxide. However, the real role of AZT in this effect is still unclear. In fact, while reverse transcriptase (RT) inhibition could explain reduction of clonal expansion and of renewal of HTLV-1 infected cells during ATL progression, this effect alone seems insufficient to justify the evident and prompt decrease of the pro-viral load in treated patients. We have previously demonstrated that AZT is endowed with an intrinsic pro-apoptotic potential towards both peripheral blood mononuclear cells from healthy donors or some tumor cell lines, but this cytotoxic potential cannot be fully achieved unless IκBα phosphorylation is inhibited. Since the constitutive activation of NF-kappa B (NF-κB) appears a common biological basis of HTLV-1-infected cells, a pharmacological inhibition of IκBα phosphorylation seems a potential strategy for treating and preventing HTLV-1 related pathologies. In this study, we have demonstrated that a combination treatment with the IκBα phosphorylation inhibitor Bay 11-7085 and AZT induced increased levels of regulated cell death (RCD) by apoptosis compared to the single treatments in HTLV-1 infected cells of different origin. Importantly, levels of RCD were considerably higher in infected cells in comparison with the uninfected ones. Inhibition of NF-κB activation following the combined treatment was confirmed by analysis of both gel-shift and functional activity of the NF-κB complex proteins, p65/p52. Moreover, a transcriptional analysis revealed that the addition of Bay 11-7085 to AZT treatment in HTLV-1-infected cells modified their transcriptional profile, by inducing the upregulation of some pro-apoptotic genes together with the downregulation of some anti-apoptotic genes. Our data suggest that addition of adequate concentrations of IκBα phosphorylation inhibitor to therapeutic regimens including AZT could be a promising strategy in ATL

The otter population of the River Ticino (N Italy) 20 years after its reintroduction

On the River Ticino (Piedmont and Lombardy regions, N Italy), the Eurasian otter Lutra lutra became extinct in the 1980s and was reintroduced in 1997. Since then, the status of the reintroduced population has been assessed only occasionally, in 2008 and 2010. Between 2016 and 2018, we conducted an extensive survey for otter signs along the whole Italian stretch of the River Ticino, following the \u2018Standard Method\u2019 for otter surveys. In 2016\u20132017, we found 101 spraints (mean marking intensity: 0.40 spraints/100 m), spread over a 97-km long stretch of the river. In 2018 only five spraints were collected, the two furthermost marking sites being 32 km apart. Genotyping of nDNA extracted from 21 faecal samples enabled the identification of six different individuals. The surveys led to drawing a reliable picture of otter distribution and population size, with evidence of otter occurrence on a longer than previously recorded stretch of the river. The results of the 2018 survey suggest that stochastic factors may still threaten the survival of reintroduced otters and would suggest a reinforcement of the population is required to increase its genetic diversity