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

DISTRIBUTION AND ECOLOGY OF LOWLAND PINE MARTEN (MARTES MARTES L. 1758)

The pine marten (Martes martes) has been long considered a prototypical forest-specialist, but recent studies carried out in western Europe have pointed out that the species is more generalist in terms of habitat preferences than previously reported. In the western plain of the River Po residual woods mainly consist of small patches or riparian woods merged in intensively cultivated areas; nonetheless in this apparently unsuitable area the number of roadkill pine martens has increased exponentially since the beginning of the century. The expansion of the pine marten in agricultural lowlands provided an invaluable opportunity for investigating its ecological requirements and the environmental factors that shape this species\u2019 range. To assess the actual level of pine marten penetration in lowland areas of north-western Italy, a non-invasive, faecal mDNA-based genetic method was applied at landscape scale, demonstrating that the pine marten has colonised the overall study area north of the River Po, which probably is acting as a barrier. Pine marten distribution suggests that expansion has mainly followed a north-to-south direction, following the course of major rivers as natural corridors of expansion. The unexpected high frequency of haplotypes of the central-northern European phylogroup supported the hypothesis of Alpine or trans-Alpine populations as sources of pine marten expansion. Based on presence-absence data, Species Distribution Models were applied to identify the main factors driving the colonisation by the pine marten of the western River Po plain and predict its potential south- and eastwards expansion. The distribution of residual wood patches and availability of suitable riparian corridors played a major role in shaping the potential expansion area of the pine marten in agricultural lowlands. This was high only for the western part of the River Po plain and, secondarily, for the pine wood patches bordering the Adriatic coast. The hypothesis that lowland woods may represent marginal habitats for the pine marten was not supported by the data collected about both its density and macronutrient balance. In the valley of the River Ticino, microsatellite analysis of faecal DNA allowed to point out one of the highest densities ever recorded in continental Europe. The high genetic relatedness of identified individuals suggested that the agricultural matrix may act as a barrier to the dispersal of juveniles. Genotyping allowed also to test for the effectiveness of a camera-trapping-based method (Random Encounter Model) for assessing pine marten density. The REM underestimated population size by ca. 40%. Dietary macronutrient ratios of pine marten differed from the target, as assessed by the meta-analysis of available diet data for Europe, to an almost negligible extent, confirming that plasticity in feeding behaviour plays a major role in pine marten expansion in agricultural areas. Genetic surveys confirmed that pine marten expansion is coinciding with a reduction in stone marten range or abundance. When syntopic, stone marten exclusion from woodland by the pine marten is a common output, although the mechanisms underlying the higher competitive ability of the latter are still unclear. By camera-trapping, we tested the hypothesis that spatial and temporal segregation may favour the coexistence of the pine marten with the other mesocarnivores to a wider extent than that of the stone marten. Results from two study areas suggested that pine marten higher competitive ability with respect to the stone marten could be the indirect consequence of its greater plasticity in activity patterns. As different guild compositions result into different interspecific relations among coexisting species, our findings need to be further tested across different communities and study areas

Association between benzodiazepine prescriptions and potential risk factors of adverse drug reactions among elderly and very elderly: Findings from Friuli Venezia Giulia region, Italy

4noBackground: benzodiazepines (BDZs) highly increase the risk of adverse drug reactions (ADRs), especially in the elderly with certain medical conditions. Methods: point-prevalence study in December 2017; 2,456 patients ≥65 years were included from 14 regional public and private hospitals, 24 long-term chronic care facilities (LTCCFs) and 20 nursing homes (NHs). Data regarded BDZ prescriptions and comorbidities, co-prescriptions, or concurrent diseases, which could increase the risk of ADRs in BDZ users. Univariate and multivariate logistic regression analyses were used to assess associations between ADRs in BDZ users vs. non-users, as well as within users. Results: 671 patients were prescribed BDZ, whose half were in NHs. Being prescribed 10 or more drugs was twice more common among BDZ users compared to non-users. BDZs were prescribed for long-term to 80% of patients in NHs, while proportions were halved in hospitals and LTCCFs. In the analyses within BDZ users, patients in NHs and LTCCFs were at higher risk of dementia and concurrent use of opioid analgesic and antipsychotics compared to hospitalized patients. Conclusions: the use of BDZs in older patients is common in FVG. A great extent of comorbidities and concurrent medications at higher risk of ADRs was found. Analyses in different healthcare settings can allow to drive evidence-based interventions in order to discourage the use of BDZs and monitor the raise of ADRs.nonenoneCastelpietra G.; Balestrieri M.; Brusaferro S.; Arnoldo L.Castelpietra, G.; Balestrieri, M.; Brusaferro, S.; Arnoldo, L

Combining electrostatic, hindrance and diffusive effects for predicting particle transport and separation efficiency in deterministic lateral displacement microfluidic devices

Microfluidic separators based on Deterministic Lateral Displacement (DLD) constitute a promising technique for the label-free detection and separation of mesoscopic objects of biological interest, ranging from cells to exosomes. Owing to the simultaneous presence of different forces contributing to particle motion, a feasible theoretical approach for interpreting and anticipating the performance of DLD devices is yet to be developed. By combining the results of a recent study on electrostatic effects in DLD devices with an advection-diffusion model previously developed by our group, we here propose a fully predictive approach (i.e., ideally devoid of adjustable parameters) that includes the main physically relevant effects governing particle transport on the one hand, and that is amenable to numerical treatment at affordable computational expenses on the other. The approach proposed, based on ensemble statistics of stochastic particle trajectories, is validated by comparing/contrasting model predictions to available experimental data encompassing different particle dimensions. The comparison suggests that at low/moderate values of the flowrate the approach can yield an accurate prediction of the separation performance, thus making it a promising tool for designing device geometries and operating conditions in nanoscale applications of the DLD technique

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

Influence of days after calving and thermal stress on the efficacy of a progesterone-based treatment in acyclic italian mediterranean buffalo

The aim of this study was to evaluate the efficacy of a progesterone-based treatment on anoestrus in buffaloes. Primiparous acyclic buffaloes (n = 276), were divided into three classes according to their days in milk (DIM): from 50 to 90 (Class I; n = 86), from 91 to 150 (Class II; n = 102) and from 150 to 200 (Class III; n = 88). Animals were synchronized using P4 vaginal implants, followed by timed artificial insemination (TAI). They were then allowed to enter into a larger group of buffaloes for natural mating 15 days after AI was performed, and pregnancy status was monitored from then on at 15-day intervals. Finally, the temperature–humidity index (THI) was calculated. Statistical analysis was performed by ANOVA by means and both multiple and linear regression. The total pregnancy rate (PR) was 87.7%, with no differences among DIM classes (88.0, 92.4, and 80.0% in Classes I, II, and III, respectively). However, the PR at TAI tended to be higher (p = 0.07) in buffaloes in Class II. The follicle (FL) area in Class II buffaloes was larger (p < 0.01) than that of the other classes. No influence of the THI on the total PR was recorded. The pregnancy outcome at TAI was affected by the FL area (odds ratio = 2.237; p < 0.05) and body condition score (BCS) (odds ratio = 1.256; p < 0.05). In conclusion, treatment with vaginal P4 optimizes pregnancy rates in anoestrus buffaloes, particularly when the animals are in mid-lactation and show an optimal BCS. Furthermore, the THI does not seem to affect the efficiency of the progesterone treatment

Strategies to reduce embryonic mortality in buffalo cows

The aim of the present study was to examine whether treatment with a GnRH agonist, hCG or P4 on Day 25 after AI increased P4 concentrations and reduced the incidence of embryonic mortality (EM) in pregnant buffaloes mated in mid-winter in a Mediterranean environment. The trial was carried out in two farms characterized, in previous years, by low (LEM Group), 153 buffaloes (DIM=150±7 days), and high (HEM Group), 284 buffaloes (DIM=163±5 days), incidence of embryo mortality. Animals were synchronized by Ovsynch-TAI Program and artificially inseminated. On day 25, pregnant buffaloes were randomly assigned to four groups: Control (no treatment), GnRH agonist (buserelin acetate, 12.6 μg), hCG (1500 IU) and P4 (341 mg of P4 i.m. every 4 days for three times). Progesterone (pg/ml) was determined in milk whey on Days 10, 20 and 25 after AI in all buffaloes and in Days 30 and 45 only in buffaloes pregnant on day 25 and assigned to four groups of treatment. Pregnancy diagnosis was undertaken on Day 45 by ultrasound. All treatments increased P4 milk whey and reduced embryonic mortality in buffalo cows bred in the farm characterized by high EM