Perfluorooctanoic Acid (PFOA) Induces Redox Status Disruption in Swine Granulosa Cells

Perfluoroctanoid acid (PFOA) is employed in the production and processing of several plastic materials, mainly during the production of waterproof fabrics or non-stick cookware. PFOA is identified as a "Substance of Very High Concern", as it is classified as a "PBT" (Persistent, Bioac-cumulative and Toxic substance) because of its persistence in the environment and its potential accumulation in organisms. Thus, safe levels of exposure cannot be established and PFOA emis-sions should be minimized. PFOA has been recently linked to several health concerns in humans. In particular, a disruptive effect on redox status homeostasis has been documented, with a po-tential impairment of normal reproductive function which requires an adequate oxidative balance. Therefore, the aim of the present study was to evaluate the effects of PFOA (2, 20 and 200 ng/mL) on ovarian granulosa cells, a model of reproductive cells. The obtained results revealed that PFOA stimulated cell viability (P <0.05). Regarding the effects on free radical production, both O2-, NO and H2O2 were significantly inhibited (P <0.05) while the non-enzymatic antioxidant power was not significantly modified. Collectively, present results deserve attention since free radical mol-ecules play a crucial role in ovarian follicle development leading to a successful ovulatio

The effects of Silymarin on ovarian activity and productivity of laying hens

In a previous work we evaluated the effects of Silymarin, a natural antioxidant and hepatoprotective polyphenolic compound, on laying hens performances and health status. The aim of the present work was to further confirm the previous results and to investigate the effects of Silymarin on ovarian endocrine activity and productivity of laying hens as well as on egg traits. Eighteen laying hens were randomly allotted into 3 groups and observed for 20 weeks: control hens were fed control diet, while treated groups received supplementations of 200 (S200) and 400 (S400) ppm of Silymarin. S200 group showed improved eggs laying rate (+2.91%), feed conversion rate (-4.52%) and a significant (P<0.05) increase of dry matter content (+0.54%), total lipids (+0.72%) and total sterols (0.02%) of the eggs. Any significant difference was shown for Silymarin at the highest dose (400 mg/kg of feed). At the end of the trial the hens were sacrificed and hierarchical follicles were removed and cultured for 48h. Media were assayed for progesterone (P4) and estradiol-17 beta (E2). Average E2 production in- creased (P<0.05) from F5 (follicles with initial development) to F3 (follicles with intermediate develop- ment), then decreased (P<0.05) from F3 to F1 (pre-ovulatory follicles). Basal P4 production augmented (P<0.05) throughout all follicle growth. Both Silymarin treatment inhibited (P<0.05) steroidogenesis. Silymarin may act as an endocrine-modulating chemical affecting hen performances

The Phytoestrogen Quercetin Impairs Steroidogenesis and Angiogenesis in Swine Granulosa Cells In Vitro

Experimental evidence documents that nutritional phytoestrogens may interact with reproductive functions but the exact mechanism of action is still controversial. Since quercetin is one of the main flavonoids in livestock nutrition, we evaluated its possible effects on cultured swine granulosa cell proliferation, steroidogenesis, and redox status. Moreover, since angiogenesis is essential for follicle development, the effect of the flavonoid on Vascular Endothelial Growth Factor output by granulosa cells was also taken into account. Our data evidence that quercetin does not affect granulosa cell growth while it inhibits progesterone production and modifies estradiol 17β production in a dose-related manner. Additionally, the flavonoid interferes with the angiogenic process by inhibiting VEGF production as well as by altering redox status. Since steroidogenesis and angiogenesis are strictly involved in follicular development, these findings appear particularly relevant, pointing out a possible negative influence of quercetin on ovarian physiology. Therefore, the possible reproductive impact of the flavonoid should be carefully considered in animal nutrition

Evaluation of triclosan effects on cultured swine luteal cells

Triclosan is a chlorinated phenolic, used in many personal and home care products for its powerful antimicrobial effect. Several studies have shown triclosan toxicity and the American Food and Drug Administration (FDA) in 2016 has limited its use. It has been recently included in endocrine-disrupting chemicals (EDCs), a list of chemicals known for their ability to interfere with hormonal signaling with particular critical effects on reproduction both in animals and humans. In order to deepen the knowledge in this specific field, the present study was undertaken to explore the effect of different concentrations of triclosan (1, 10, and 50 µM) on cultured luteal cells, isolated from swine ovaries, evaluating effects on growth Bromodeoxyuridine (BrDU) incorporation and Adenosine TriPhosphate (ATP) production, steroidogenesis (progesterone secretion) and redox status (superoxide and nitric oxide production, enzymatic and non-enzymatic scavenging activity). A biphasic effect was exerted by triclosan on P4 production. In fact, the highest concentration inhibited, while the others stimulated P4 production (p &lt; 0.05). Triclosan significantly inhibited cell proliferation, metabolic activity, and enzymatic scavenger activity (p &lt; 0.05). On the contrary, nitric oxide production was significantly increased by triclosan (p &lt; 0.01), while superoxide anion generation and non-enzymatic scavenging activity were unaffected

Antiangiogenic resveratrol analogues by mild m-CPBA aromatic hydroxylation of 3,5-dimethoxystilbenes.

A mild treatment of the resveratrol analogue 3,5,4′-trimethoxystilbene 2 with m-CPBA afforded two hydroxylated methoxystilbenes 5 and 6 by direct aromatic hydroxylation. A similar protocol was applied to other stilbenes bearing a 3,5-dimethoxy moiety, namely tetramethoxystilbenes 7 and 10 to obtain respectively the hydroxylated analogues 8, 9 and 11, 12. The substrate 2 and the new compounds 5, 8 and 11 were evaluated as anti-angiogenic agents and proved significantly active in the range 1 – 100 μM

In Vitro Evaluation of Cytotoxicity and Proliferative Effects of Lyophilized Porcine Liver Tissue on HepG2 Hepatoma Cells and Adipose-Tissue-Derived Mesenchymal Stromal Cells

In recent years, nutritional supplements from different sources have been widely considered to support medical treatments in patients affected by chronic hepatopathies. Their potential therapeutic benefit has been recognized, but some evidence of safety issues has been reported. Recently it has been hypothesized that the liver could produce various of bioactive factors to maintain organ homeostasis and promote tissue healing. Thus, liver-specific preparations containing bioactive factors could provide a suitable substrate for in vitro study of liver tissue maintenance/healing, as a prospective regenerative medicine approach. Furthermore, they could represent a dietary supplement or nutraceutical for adjuvant therapies when correctly prepared and formulated. This work aims to provide data about the safety and biological activity of a freeze-dried porcine liver preparation. The lyophilized powder obtained from the whole organ has been tested in term of in vitro cell cytotoxicity (MTT assay) and proliferation assays (bromo-deoxyuridine incorporation and direct cell count) in two different cell types: human hepatoma HepG2 cell line and adipose-tissue-derived canine mesenchymal stromal cells (At-MSCs). At concentration levels between 100 to 500 g/mL, the lyophilized liver powder stimulated mitochondrial metabolism as assessed by MTT assay (p 0.001 for HepG2 and for At-MSCs) and induced an increase in bromo-deoxyuridine incorporation in both cell types (p 0.01 for HepG2 and p < 0.001 for At-MSCs). In addition, direct cell count demonstrated a higher proliferative activity in treated At-MSCs (p < 0.001). Although preliminary, these data suggest that the whole-liver powder is noncytotoxic in vitro and may represent a stimulus to cell metabolism and proliferation. Further studies are needed to detect the bioactive components of the supplement and characterize in deeper detail the cellular pathways that they can modulate

Evaluation of the oxidative status of periparturient mares supplemented with high amount of α-tocopherol

Aim of the study was to investigate the oxidative status during peripartum period in mares fed high amount of dietary α-tocopherol. α-Tocopherol, ferric reducing ability of plasma (FRAP) and reactive oxygen metabolites (d-ROMs) levels were measured in blood samples from 17 Thoroughbred mares at three intervals: (1) 20 days before the expected foaling date; (2) 12 h following parturition; (3) 7 days post-partum. The levels of α-tocopherol, d-ROMs and FRAP were retrospectively analysed in relation to the number of insemination services (Ins) after foaling performed per conception. The parameters α-tocopherol and d-ROMs evidenced minimal fluctuations during peripartum period while FRAP levels showed a linear decrease. The α-tocopherol did not show significant variations and was numerically higher in mares >10 years old while FRAP levels were significantly higher in older mares in the post-partum. Mares receiving three natural insemination services showed higher levels of FRAP in the pre-partum period (p = .009) and lower values of α-tocopherol (p = .015) in the postpartum compared to mares receiving one service. No d-ROMs level differences appeared among service classes. Oxidative stress is not present in periparturient mares fed 2.750 IU of supplementary α-tocopherol. Differences in redox metabolism are detectable between young and old mares. Plasma antioxidant potential is higher in older mares particularly in the post-partum period.Highlights Oxidative stress is not present in periparturient mares fed 2.750 IU of supplementary α-tocopherol. Antioxidant potential of plasma is higher in mares older than 10 years compared to younger mares. The increased plasma antioxidant potential seems related to an increase in services per conception

Effects of orexin B on swine granulosa and endothelial cells

In addition to the well-known central modulatory role of orexins, we recently demonstrated a peripheral involvement in swine granulosa cells for orexin A and in adipose tissue for orexin B (OXB). The aim of present research was to verify immunolocalization of OXB and its potential role in modulating the main features of swine granulosa cells. In particular, we explored the effects on granulosa cell proliferation (through the incorporation of bromodeoxyuridine), cell metabolic activity (as indirect evaluation by the assessment of ATP), steroidogenic activity (by immunoenzymatic examination) and redox status (evaluating the production of superoxide anion by means of the WST test, production of nitric oxide through the use of the Griess test and the non-enzymatic reducing power by FRAP test). Our data point out that OXB does not modify granulosa cell growth, steroidogenesis and superoxide anion generation. On the contrary, the peptide stimulates (p &lt; 0.05) nitric oxide output and non-enzymatic reducing power. Since new vessel growth is crucial for ovarian follicle development, a further aim of this study was to explore the expression of prepro-orexin and the effects of OXB on swine aortic endothelial cells. We found that the peptide is ineffective in modulating cell growth, while it inhibits redox status parameters. In addition, we demonstrated a stimulatory effect on angiogenesis evaluated in fibrin gel angiogenesis assay. Taken together, OXB appears to be potentially involved in the modulation of redox status in granulosa and endothelial cells and we could argue an involvement of the peptide in the follicular angiogenic event

Redox Status in Canine Leishmaniasis

World Health Organization defined Leishmaniasis as ‘one of the priority attention diseases’. Aiming to clarify some aspects of its pathogenetic mechanisms, our study has been focused on the assessment of redox status in dogs, the main reservoir for Leishmania infantum. Forty-five dogs from an endemic area in southern Italy were divided into four different groups (from mild disease with negative to low positive antibody levels to very severe disease with medium to high positive antibody levels) accordingly to the LeishVet group guidelines. Their plasma and/or sera were tested for Reactive Oxygen Species (ROS), namely superoxide anion (O2-), Reactive Nitrogen Species (RNS), such as nitric oxide (NO) and hydroperoxides (ROOH), as well as the activity of the detoxifying enzyme superoxide dismutase (SOD), and the total non-enzymatic antioxidant capacity, as determined by ferric reducing-antioxidant power (FRAP) assay, O2- generation was significantly (p &lt; 0.05) reduced in leishmaniasis affected dogs independently of the clinical stage, while NO production was stimulated (p &lt; 0.05) only in II and III stage patients. No difference could be found for the levels of hydroperoxides and SOD activity between healthy and pathological subjects. FRAP values, were lower in affected dogs but only in stage II. Taken together, although we demonstrate that several redox status parameters are altered in the plasma of dog affected by Leishmaniasis, the oxidative stress changes that are observed in this disease, are possibly mainly due to the cellular blood components i.e. neutrophils, responsible of the elimination of the parasite. Further studies are required to assess the clinical values of the collected data

Effects of dog-assisted therapies on cognitive mnemonic capabilities in people affected by Alzheimer's disease

Alzheimer’s disease (AD) is the most common cause of dementia in humans and, currently, a valid treatment is lacking. Our goal is to demonstrate the importance and benefits of the relationship with companion animals (considered as co-therapists), intended as a means of facilitating social relations and promoting evident wellbeing in AD patients. The study involved 30 randomly chosen patients with Alzheimer’s disease (group T) and three dogs. The group participated in a total of 24 animal-assisted interventions (AAIs) sessions over a span of 12 weeks, using the Mini-Mental State Examination (MMSE), Wellness and Cognitive Ability Questionnaire (Brief Assessment Cognition or BAC), and Alzheimer’s Disease Assessment Scale (ADAS) as assessment tests. A second group (group C), consisting of 10 people with AD, was enrolled as control group and underwent the same assessment tests but did not benefit from the presence of the dogs. Tests were carried out at time T0 (before starting sessions), T1 (end of sessions), and T2 (two months after last session). People belonging to group T achieved an overall improvement in their perceived state of wellbeing, even on a cognitive and mnemonic plane. However, two months after the end of the sessions, the test results in people suffering from AD decreased towards the baseline (T0). The study shows how such progress can be achieved through activities based on the relationship with an animal, as long as the animal is a steady presence in the life of the patient receiving the intervention. Dogs involved in other dog-assisted therapies have been found suitable also for assisting patients with AD