Background Domestication appears to modify the morphology and physiology of the central nervous system. Owing to the limited availability of proteomic and cytoarchitectural data for comparisons between wild and domesticated species, we conducted a comparative analysis of the cytoarchitecture and protein profile of the primary motor cortex (M1), the key region controlling motor activity, in pigs and wild boars to assess the effects of domestication. Methods M1 samples used in this study were obtained from brains previously analyzed in an earlier investigation. From six pig and wild boar brains fixed in paraformaldehyde, the right and left M1 regions were isolated. Paraffin sections, 5 µm thick, were prepared for histological and immunohistochemical analyses, and 10 µm sections were used for proteomic analysis. In M1, cortical thickness, cell density, and the density of parvalbumin-positive neurons were quantified, while proteomic analysis was performed to characterize the M1 protein profile. Results Our results revealed a lower density of parvalbumin-expressing interneurons compared with wild boars. Moreover, proteomic analyses showed an overexpression in wild boars of proteins involved in oxidative stress protection and synaptic plasticity. Conclusions These findings indicate that domestication may have influenced the cytoarchitecture of the pig M1. The reduced number of parvalbumin-expressing interneurons may reflect a modification in neuronal network properties in pigs compared with wild boars. In contrast, the proteomic profile of wild boars revealed an enrichment of proteins associated with oxidative stress protection and synaptic plasticity, supporting structural distinctions in the M1. Collectively, these results suggest that the wild boar may exhibit more finely tuned regulation of motor control, supported by enhanced mechanisms to sustain neuronal activity and viability.

This study involved 45 finishing Limousin young bulls (initial BW 350 ± 15 kg; aged 250 ± 20 days) reared under commercial conditions and randomly assigned for 250 days to three dietary treatments: a control diet without olive cake (CTR), and diets including partially destoned dried olive cake at 10% (OC10) or 15% (OC15) of dry matter. The effects of the dietary inclusion of partially destoned dried olive cake (DOC) on the physicochemical properties, intramuscular fatty acid composition, nutritional lipid indices, and antioxidant profile of the Longissimus dorsi muscle were evaluated. Meat pH was measured at 45 min, 24 h, and 7 days of aging. Instrumental color, proximate composition, fatty acid profile, lipid nutritional indices, total phenolic content (TPC), individual polyphenols, and antioxidant capacity were determined. Data were analyzed by ANOVA using PROC GLIMMIX in SAS, with orthogonal contrasts to evaluate linear and quadratic responses to olive cake inclusion. Treatments did not affect post mortem pH, and color differences observed at 24 h were not present after 7 days of aging. Crude protein content was lower in OC-fed groups, while total lipid content tended to increase; sodium chloride concentration was higher in OC15 meat. Olive cake supplementation reduced hypercholesterolemic saturated fatty acids and increased stearic, oleic, and polyunsaturated fatty acids, including α-linolenic acid, EPA, and DPA, improving PUFA/SFA and UFA/SFA ratios and reducing atherogenic and thrombogenic indices. Despite a higher peroxidability index, OC groups showed greater muscle TPC and antioxidant capacity, the presence of detectable hydroxytyrosol and tyrosol suggests a possible contribution of dietary olive phenolics or their metabolites, although the exact mechanisms underlying their appearance in muscle tissue remain to be fully elucidated. Overall, partially destoned DOC can be included up to 15% of the finishing diet to improve beef nutritional and functional quality within sustainable circular systems.

Aging represents a decline in function over time. Polyphenols are dietary antioxidants that could exert beneficial effects on aging. This study investigated the effects of dietary supplementation with two extracts from Thymbra spicata L. aerial parts on the lifespan, body-weight control, and antioxidant responses of Drosophila melanogaster cohorts distinguished for age and sex. The aqueous extract extended the lifespan of young female flies, and this pro-longevity potential was associated with a reduction in the body-weight gain. Furthermore, the aqueous extract promoted the antioxidant defense when young females were exposed to Paraquat by reducing oxidative stress and mortality. This study provides the first insight into the beneficial effects of T. spicata supplementation in an animal model, suggesting that the healthy effect could mainly depend on strengthening the antioxidant defenses which reflect on the lifespan. Further investigations will better clarify the mechanisms and potential of T. spicata as nutraceutics in anti-aging strategies.

Development and aging significantly impact the cellular levels of Coenzyme Q (CoQ), which is associated with both pathological and physiological conditions. Aim of this study was to describe the CoQ status throughout the lifetime of Drosophila melanogaster, a well-established model in aging studies. CoQ(9) and CoQ distribution was analysed across different body segments and various life stages in both male and female flies. The results indicate that CoQ(9) is the predominant isoform in every phase of flies' life cycle, with the highest concentrations observed in the thorax. We noted distinct trends in CoQ distribution during aging, which varied according to sex and body segments (head, thorax, and abdomen). Supplementation with two concentrations of CoQ(9) and CoQ(10) (15 mu M and 75 mu M) for 2 weeks induced a segment- and sex-specific CoQ uptake. Although 75 mu M CoQ(10) was more effective in modulating the CoQ status, lifelong treatment with this concentration did not affect the longevity of the flies.

Dietary fiber has been shown to have multiple health benefits, including a positive effect on longevity and the gut microbiota. In the present study, Drosophila melanogaster has been chosen as an in vivo model organism to study the health effects of dietary fiber supplementation (DFS). DFS extended the mean half-life of male and female flies, but the absolute lifespan only increased in females. To reveal the underlying mechanisms, we examined the effect of DFS on gut microbiota diversity and abundance, local gut immunity, and the brain proteome. A significant difference in the gut microbial community was observed between groups with and without fiber supplementation, which reduced the gut pathogenic bacterial load. We also observed an upregulated expression of dual oxidase and a modulated expression of Attacin and Diptericin genes in the gut of older flies, possibly delaying the gut dysbiosis connected to the age-related gut immune dysfunction. Brain proteome analysis showed that DFS led to the modulation of metabolic processes connected to mitochondrial biogenesis, the RhoV-GTPase cycle, organelle biogenesis and maintenance, membrane trafficking and vesicle-mediated transport, possibly orchestrated through a gut-brain axis interaction. Taken together, our study shows that DFS can prolong the half-life and lifespan of flies, possibly by promoting a healthier gut environment and delaying the physiological dysbiosis that characterizes the ageing process. However, the RhoV-GTPase cycle at the brain level may deserve more attention in future studies.

Inonotus obliquus (Chaga) is a medicinal mushroom with several pharmacological properties that is used as a tea in traditional Chinese medicine. In this study, Chaga water extract was digested in vitro to mimic the natural processing and absorption of its biocomponents when it is consumed as functional beverage, and its anticancer activities were evaluated in breast cancer (BC) cell lines, representing HER2-positive and triple-negative subtypes. After chemical characterization by liquid chromatography/mass spectrometry (HR-QTOF) analysis, the effect of Chaga biocomponents on cell viability and cell cycle progression was assessed by MTT assay, FACS analysis, and Western blot. Dihydrofolate reductase (DHFR) activity was measured by an enzymatic assay. Four highly bioactive triterpenoids (inotodiol, trametenolic acid, 3-hydroxy-lanosta-8,24-dien-21-al, and betulin) were identified as the main components, able to decrease BC cell viability and block the cell cycle in G0/G1 by inducing the downregulation of cyclin D1, CDK4, cyclin E, and phosphorylated retinoblastoma protein. DHFR was identified as their crucial target. Moreover, bioactive Chaga components exerted a synergistic action with cisplatin and with trastuzumab in SK-BR-3 cells by inhibiting both HER2 and HER1 activation and displayed an immunomodulatory effect. Thus, Inonotus obliquus represents a source of triterpenoids that are effective against aggressive BC subtypes and display properties of targeted drugs.

The present study, employing a comparative proteomic approach, analyzes the protein profile of pig claustrum (CLA), putamen (PU), and insula (IN). Pig brain is an interesting model whose key translational features are its similarities with cortical and subcortical structures of human brain. A greater difference in protein spot expression was observed in CLA vs PU as compared to CLA vs IN. The deregulated proteins identified in CLA resulted to be deeply implicated in neurodegenerative (i.e., sirtuin 2, protein disulfide-isomerase 3, transketolase) and psychiatric (i.e., copine 3 and myelin basic protein) disorders in humans. Metascape analysis of differentially expressed proteins in CLA vs PU comparison suggested activation of the α-synuclein pathway and L1 recycling pathway corroborating the involvement of these anatomical structures in neurodegenerative diseases. The expression of calcium/calmodulin-dependent protein kinase and dihydropyrimidinase like 2, which are linked to these pathways, was validated using western blot analysis. Moreover, the protein data set of CLA vs PU comparison was analyzed by Ingenuity Pathways Analysis to obtain a prediction of most significant canonical pathways, upstream regulators, human diseases, and biological functions. Interestingly, inhibition of presenilin 1 (PSEN1) upstream regulator and activation of endocannabinoid neuronal synapse pathway were observed. In conclusion, this is the first study presenting an extensive proteomic analysis of pig CLA in comparison with adjacent areas, IN and PUT. These results reinforce the common origin of CLA and IN and suggest an interesting involvement of CLA in endocannabinoid circuitry, neurodegenerative, and psychiatric disorders in humans.

IntroductionThe single nucleotide polymorphism (SNP) rs4644 at codon 64 of galectin-3 (gal-3, gene name: LGALS3), specifying the variant proline (P64) to histidine (H64), is known to affect the protein’s functions and has been associated with the risk of several types of cancer, including differentiated thyroid carcinoma (DTC).Materials and methodsTo deepen our understanding of the biological effects of this SNP, we analyzed the proteome of two isogenic cell lines (NC-P64 vs. NA-H64) derived from the immortalized non-malignant thyrocyte cell line Nthy-Ori, generated through the CRISPR-Cas9 technique to differ by rs4644 genotype. We compared the proteome of these cells to detect differentially expressed proteins and studied their proteome in relation to their transcriptome.ResultsFirstly, we found, consistently with previous studies, that gal-3-H64 could be detected as a monomer, homodimer, and heterodimer composed of one cleaved and one uncleaved monomer, whereas gal-3-P64 could be found only as a monomer or uncleaved homodimer. Moreover, results indicate that rs4644 influences the expression of several proteins, predominantly upregulated in NA-H64 cells. Overall, the differential protein expression could be attributed to the altered mRNA expression, suggesting that rs4644 shapes the function of gal-3 as a transcriptional co-regulator. However, this SNP also appeared to affect post-transcriptional regulatory mechanisms for proteins whose expression was oppositely regulated compared to mRNA expression. It is conceivable that the rs4644-dependent activities of gal-3 could be ascribed to the different modalities of self-dimerization.ConclusionOur study provided further evidence that rs4644 could affect the gal-3 functions through several routes, which could be at the base of differential susceptibility to diseases, as reported in case-control association studies.

This study provides the first comprehensive evaluation of the bioactive potential of acorn flour extracts (Quercus robur, Fagaceae) prepared at different temperatures (20, 60, 80 and 100 °C), focusing on polyphenolic content, antioxidant properties and enzyme inhibitory activities. Through HPLC-ESI-MS/MS analysis, 36 bioactive compounds were identified, with the extract at 60 °C showing the highest concentrations of key polyphenols, notably gallic acid (210,008.9 mg·kg−1) and ellagic acid (45,469.6 mg·kg−1). This extract also exhibited a high antioxidant activity and significant inhibition of glucosidase and acetylcholinesterase, suggesting potential benefits for diabetes management and neuroprotection. The results indicate that extraction temperature affects bioactivity, with the 60 °C extract standing out as a promising candidate for nutraceutical, pharmaceutical, and cosmeceutical applications due to its rich polyphenol profile and potent biological properties.

Intense exercise can cause inflammation and oxidative stress due to the production of reactive oxygen species. These pathophysiological processes are interdependent, and each one can induce the other, creating a vicious circle. A placebo-controlled blind study was carried out in show jumping horses (n. 16) to evaluate the effects of a commercial dietary supplement (Dolhorse® N.B.F. Lanes srl, Milan, Italy) containing Verbascum thapsus leaf powder (1.42%), Curcuma longa (14.280 mg/kg), and Boswellia serrata (Roxb ex Colebr) (14.280 mg/kg) extracts. Before and after 10 days of dietary supplementation, blood samples were collected to evaluate the protein levels, antioxidants, and inflammatory responses by proteomic analysis or real-time Reverse Transcriptase-Polymerase Chain Reaction (real-time RT-PCR). A total of 36 protein spots, connected to 29 proteins, were modulated by dietary supplementation, whereas real-time RT-PCR revealed a significant downregulation of proinflammatory cytokines (interleukin 1α (p < 0.05) and interleukin-6 (0.005), toll-like receptor 4 (p < 0.05), and IKBKB (p < 0.05) in supplemented sport horses. Immunoglobulin chains, gelsolin, plasminogen, vitamin D binding protein, apolipoprotein AIV, and filamin B were overexpressed, whereas haptoglobin, α-2-HS-glycoprotein, α2-macroglobulin, afamin, amine oxidase, 60S acidic ribosomal protein, and complement fragments 3, 4, and 7 were reduced. No effect was observed on the antioxidant defense systems. The present results suggest this phytotherapy may reinforce the innate immune responses, thus representing a valid adjuvant to alleviate inflammation, which is a pathophysiological process in sport horses.

Olive pomace (OP) represents one of the main by-products of olive oil production, which still contains high quantities of health-promoting bioactive compounds. In the present study, three batches of sun-dried OP were characterized for their profile in phenolic compounds (by HPLC-DAD) and in vitro antioxidant properties (ABTS, FRAP and DPPH assays) before (methanolic extracts) and after (aqueous extracts) their simulated in vitro digestion and dialysis. Phenolic profiles, and, accordingly, the antioxidant activities, showed significant differences among the three OP batches, and most compounds showed good bioaccessibility after simulated digestion. Based on these preliminary screenings, the best OP aqueous extract (OP-W) was further characterized for its peptide composition and subdivided into seven fractions (OP-F). The most promising OP-F (characterized for its metabolome) and OP-W samples were then assessed for their potential anti-inflammatory properties in ex vivo human peripheral mononuclear cells (PBMCs) triggered or not with lipopolysaccharide (LPS). The levels of 16 pro-and anti-inflammatory cytokines were measured in PBMC culture media by multiplex ELISA assay, whereas the gene expressions of interleukin-6 (IL-6), IL-10 and TNF-α were measured by real time RT-qPCR. Interestingly, OP-W and PO-F samples had a similar effect in reducing the expressions of IL-6 and TNF-α, but only OP-W was able to reduce the release of these inflammatory mediators, suggesting that the anti-inflammatory activity of OP-W is different from that of OP-F.

Nutrition has relevant consequences for human health and increasing pieces of evidence indicate that medicinal mushrooms have several beneficial effects. One of the main issues in Western countries is represented by the challenges of aging and age-related diseases, such as neurodegenerative disorders. Among these, Parkinson’s disease (PD) affects 10 million people worldwide and is associated with α-synuclein misfolding, also found in other pathologies collectively called synucleinopathies. Here, we show that aqueous extracts of two edible mushrooms, Grifola frondosa and Hericium erinaceus, represent a valuable source of β-glucans and exert anti-aging effects in yeast. Their beneficial effects are mediated through the inhibition of the Ras/PKA pathway, with increased expression of heat shock proteins, along with a consistent increase of both mean and maximal lifespans. These fungal extracts also reduce the toxicity of α-synuclein heterologously expressed in yeast cells, resulting in reduced ROS levels, lower α-synuclein membrane localization, and protein aggregation. The neuroprotective activity of G. frondosa extract was also confirmed in a PD model of Drosophila melanogaster. Taken together, our data suggest the use of G. frondosa and H. erinaceus as functional food to prevent aging and age-related disorders, further supporting the neuro-healthy properties of these medicinal mushroom extracts.

This study investigated the effects of dietary linseed and physiological state on blood concentrations of some metabolic hormones and indicators, in transition dairy ewes. From 21 d before lambing to 60 d post-partum, ewes were provided with one of three isoenergetic and iso-nitrogenous pelleted concentrates, without (CTR, n = 21), or with 100 (EL-10, n = 22) or 200 g/kg (EL-20, n = 22) of extruded linseed. Animals were fed alfalfa hay ad libitum and had access to mixed pasture. Plasma thyroid hormones and insulin concentrations were not affected by diet and significantly changed by time. The last week of pregnancy T3 (2.27 +/- 0.15 ng/mL) and T4 (102.63 +/- 2.85 ng/mL) were lower than at 4 weeks before lambing (T3: 3.27 +/- 0.27 ng/mL; T4: 125.89 +/- 2.63 ng/mL). After lambing, T3 rose to be highest at 4 and 8 weeks of lactation (3.41 +/- 0.22). Mean insulin peaked 2 weeks after lambing (0.31 +/- 0.02 ng/mL) vs. late pregnancy (0.22 +/- 0.01 ng/mL) and progressing lactation (0.18 +/- 0.01 ng/mL). Plasma Leptin concentra-tion (2.43 +/- 0.03 ng/mL) was not affected by diet nor time. All the blood metabolites and enzymes investigated showed significant time x treatment interaction. Differences of several haematological parameters were found in EL vs. CTR (cholesterol, triglycerides, total protein, bilirubin, ALP, ALT), however, in most cases the values were fairly within the reference physiological ranges. Metabolic hormones are confirmed to be mainly linked to the different physiological states, energy intake and variations of energy balance, without clear effects by different sources of energy and quality of dietary lipids.

Twenty-two pluriparous single-foetus-bearing Angora goats, housed 3 weeks pre-partum in mid-December, and in transition from pregnancy to lactation were used to test the efficacy of postpartum supplementation (Group S: 11 goats) or not (Group C: 11 goats) with 300 g/head/day of whole field bean seeds (WFBS) to a diet based on mixed hay ad libitum and natural pasture.The goats were maintained with the suckling of their kids for 90 days.Significant decreases in liveweight (LW) and body condition score (BCS), were documented post-parturition in both groups and were then maintained throughout.These suggest a mobilisation in body tissue that was not mitigated by WFBS supplementation.However, the faster growth of the suckled kids and greater elongation of mohair fibre recorded (Group S > Group C), suggested improvement in partition towards production indices of lactation and hair fibre deposition, respectively.Greater concentrations of total thyroid hormones, T3 and T4 in supplemented does were recorded.Overall mean (pre-prandial) plasma insulin concentrations, not different between treatments, decreased (p < 0.5) from pre-partum concentration, and showed a pattern in time, similar to that of LW and BCS.Insulin appeared not to contribute to the suggested lactational and hair growth effects and possibly contributed to transition by a homeorhetic-type reduction in concentration post-partum.Mean plasma insulin-like growth factor-1 (IGF-1) concentrations trended lower due to supplementation from 3 weeks after kidding.It is concluded that WFBS supplementation was efficacious in improving production indices of the transition Angora goats.The feeding of different quantities of WFBS should be investigated. HIGHLIGHTSSupplementation of transition Angora goats with field bean did not affect the loss of body weight or body condition score occurring after kidding.Supplementation with field bean induced higher concentrations of blood thyroid hormones, without affecting post-partum reductions in insulin.Supplemented dams had a greater mohair elongation rate and their kids grew more, by likely preferential partition to hair growth and lactation, rather than other body tissues.

Aging is a multifactorial phenomenon characterized by degenerative processes closely connected to oxidative damage and chronic inflammation. Recently, many studies have shown that natural bioactive compounds are useful in delaying the aging process. In this work, we studied the effects of an in vivo supplementation of the stilbenoid pterostilbene on lifespan extension in Drosophila melanogaster. We found that the average lifespan of flies of both sexes was increased by pterostilbene supplementation with a higher effect in females. The expression of longevity related genes (Sir2, Foxo, and Notch) was increased in both sexes but with different patterns. Pterostilbene counteracted oxidative stress induced by ethanol and paraquat and up-regulated the antioxidant enzymes Ho e Trxr-1 in male but not in female flies. On the other hand, pterostilbene decreased the inflammatory mediators dome and egr only in female flies. Proteomic analysis revealed that pterostilbene modulates 113 proteins in male flies and only 9 in females. Only one of these proteins was modulated by pterostilbene in both sexes: vacuolar H[+] ATPase 68 kDa subunit 2 (Vha68-2) that was strongly down-regulated. These findings suggest a potential role of pterostilbene in increasing lifespan both in male and female flies by mechanisms that seem to be different in the two sexes, highlighting the need to conduct nutraceutical supplementation studies on males and females separately in order to give more reliable results.

Neurodegenerative diseases are driven by several mechanisms such as inflammation, abnormal protein aggregation, excitotoxicity, mitochondrial dysfunction and oxidative stress. So far, no therapeutic strategies are available for neurodegenerative diseases and in recent years the research is focusing on bioactive molecules present in food. In particular, extra-virgin olive oil (EVOO) phenols have been associated to neuroprotection. In this study, we investigated the potential antioxidant and neuroprotective activity of two different EVOO extracts obtained from Quercetano cultivar trees grown in two different areas (plain and hill) of the Tuscany region (Italy). The different geographical origin of the orchards influenced phenol composition. Plain extract presented a higher content of phenyl ethyl alcohols, cinnammic acids, oleacein, oleocanthal and flavones; meanwhile, hill extract was richer in lignans. Hill extract was more effective in protecting differentiated SH-SY5Y cells from peroxide stress thanks to a marked upregulation of the antioxidant enzymes heme oxygenase 1, NADPH quinone oxidoreductase 1, thioredoxin Reductase 1 and glutathione reductase. Proteomic analysis revealed that hill extract plays a role in the regulation of proteins involved in neuronal plasticity and activation of neurotrophic factors such as BDNF. In conclusion, these data demonstrate that EVOOs can have important neuroprotective activities, but these effects are strictly related to their specific phenol composition.

Objectives: To determine the draft genome sequence and analyse the genetic features of a Staphylococcus pseudintermedius clinical isolate according to the main typing schemes available, with a special focus on antibiotic resistance. Methods: The strain was isolated from a case of otitis externa in a dog. Its identity and pattern of antibiotic susceptibility were determined using an automated system. The genome was sequenced using an Illumina platform. MLST, SCCmec typing, resistome, and mobile genetic elements were derived by comparative analysis using available specific databases. Results: S. pseudintermedius CAM1 isolate has a chromosome size of 2 652 610 bp. It showed a wide pattern of phenotypic resistance, comprising beta-lactams, macrolides and lincosamides, aminoglycosides, fluoroquinolones, tetracyclines, and trimethoprim-sulfamethoxazole. The genetic determinants of the underlying mechanisms were all found by in silico analysis of the genome. The mecA gene for methicillin resistance was harboured by the Vc type of the SCCmec. MLST of the strain was st551. Conclusion: By comparison with the MLST database of S. pseudintermedius and data from published molecular epidemiology studies, CAM1 is the first st551 strain recorded in Italy and, in the context of an already extremely wide antibiotic resistance pattern, it harbours also the tetK gene, the prevalence of which is rare in MDR S. pseudintermedius.

Spent coffee grounds (SCGs), waste products of coffee beverage production, are rich in organic compounds such as phenols. Different studies have demonstrated phenol beneficial effects in counteracting neurodegenerative diseases. These diseases are associated with oxidative stress and neuroinflammation, which initiates the degeneration of neurons by overactivating microglia. Unfortunately, to date, there are no pharmacological therapies to treat these pathologies. The aim of this study was to evaluate the phenolic content of 4 different SCG extracts and their ability to counteract oxidative stress and neuroinflammation. Caffeine and 5-O-caffeoylquinic acid were the most abundant compounds in all extracts, followed by 3-O-caffeoylquinic acid and 3,5-O-dicaffeoylquinic acid. The four extracts demonstrated a different ability to counteract oxidative stress and neuroinflammation in vitro. In particular, the methanol extract was the most effective in protecting neuron-like SH-SY5Y cells against H2O2-induced oxidative stress by upregulating endogenous antioxidant enzymes such as thioredoxin reductase, heme oxygenase 1, NADPH quinone oxidoreductase, and glutathione reductase. The water extract was the most effective in counteracting lipopolysaccharide-induced neuroinflammation in microglial BV-2 cells by strongly reducing the expression of proinflammatory mediators through the modulation of the TLR4/NF-κB pathway. On these bases, SCG extracts could represent valuable nutraceutical sources for the treatment of neurodegeneration.

Allium subhirsutum, known as hairy garlic, is a bulbous plant widespread in the Mediterranean area and locally used as a food and spice. In the present study, the chemical profile of the ethanolic extracts from bulbs (BE) and aerial parts (APE) were analyzed by HPLC-ESI-MSn, and antioxidant properties were evaluated by DPPH, ABTS and TEAC assays. The traditional use in the diet, and the well documented biological activity of Allium species suggest a potential as a new nutraceutical. For this reason, the potential usefulness of this food can be considered in the treatment and prevention of degenerative Alzheimer disease. For this reason, acetylcholinesterase inhibitory property was investigated. Furthermore, due to the observed presence of sulfur-containing and phenolic constituents, the cytotoxicity on tumor cells line was investigated. Results revealed significant AChE inhibitory activity for BE and APE. Both extracts exhibited also moderate antioxidant properties in the in vitro assays. Finally, limited cytotoxic activity was observed towards Human colon carcinoma and adenocarcinoma cell line, with differences between the individual parts tested. HPLC-ESI-MSn analysis showed that hairy garlic is a good source of sulphur compounds, flavonoids and phenylpropanoids derivatives, thus being a valid alternative to the common garlic (A. sativum). This work opens new opportunities for the application of A. subhirsutum as a health-promoting food.