Annurca apple polyphenol extract selectively kills MDA-MB-231 cells through ROS generation, sustained JNK activation and cell growth and survival inhibition

Polyphenols represent the most studied class of nutraceuticals that can be therapeutics for a large spectrum of diseases, including cancer. In this study, we investigated for the first time the antitumor activities of polyphenol extract from Annurca apple (APE) in MDA-MB-231 triple negative breast cancer cells, and we explored the underlying mechanisms. APE selectively inhibited MDA-MB-231 cell viability and caused G2/M phase arrest associated with p27 and phospho-cdc25C upregulation and with p21 downregulation. APE promoted reactive oxygen species (ROS) generation in MDA-MB-231 cells while it acted as antioxidant in non-tumorigenic MCF10A cells. We demonstrated that ROS generation represented the primary step of APE antitumor activity as pretreatment with antioxidant N-acetylcysteine (NAC) prevented APE-induced G2/M phase arrest, apoptosis, and autophagy. APE downregulated Dusp-1 and induced a significant increase in JNK/c-Jun phosphorylation that were both prevented by NAC. Moreover, downregulation of JNK by its specific inhibitor SP600125 significantly diminished the anticancer activity of APE indicating that ROS generation and sustained JNK activation represented the main underlying mechanism of APE-induced cell death. APE also inhibited AKT activation and downregulated several oncoproteins, such as NF-kB, c-myc, and beta-catenin. In light of these results, APE may be an attractive candidate for drug development against triple negative breast cancer

Phase angle and Mediterranean diet in patients with acne: Two easy tools for assessing the clinical severity of disease

Acne is a chronic, inflammatory and debilitating skin disorder. Dietary factors and nutritional status are among the exacerbating factors of acne. Phase angle (PhA), a direct measure of Bioelectrical Impedance Analysis (BIA), represents an indicator of the chronic inflammatory state. The Mediterranean diet (MD) is a healthy dietary pattern that can exert anti-inflammatory effects in several inflammatory diseases. We aimed to investigate the difference in PhA and adherence to the MD and their associations with the severity of acne in a sample of naïve treatment patients with acne compared to control group

Distribution pattern of hepatitis C virus genotypes and correlation with viral load and risk factors in chronic positive patients.

Objective: Hepatitis C virus (HCV) has emerged as a leading cause of chronic hepatitis, liver cirrhosis and hepatocellular carcinoma worldwide. The purpose of this study was to describe the distribution pattern of HCV genotypes in chronic hepatitis patients in the Campania region of southern Italy and estimate their association with risk factors and viral load. Materials and Methods: 404 consecutive HCV ribonucleic acid-positive patients were included in the study. HCV genotyping was carried out by the HCV line probe assay test and viral load estimation by the TaqMan real-time PCR system. Results: The predominant genotype was 1 (63.6%), followed by genotype 2 (29.4%), 3 (6.2%) and 4 (0.8%). Subtype 1b was more frequent in females than in males. Conversely, genotype 3 was more frequent in males. No significant difference was observed in age distribution of HCV genotypes. Surgery and dental therapy were the most frequent risk factors for genotype 1 and intravenous drug abuse and tattooing for genotype 3. Patients with genotype 1 more frequently showed high HCV viral load when compared to those with genotypes 2 and 3. Conclusion: The present study revealed that HCV genotypes 1 and 2 accounted for over 95% of all HCV infections in the Campania region, and genotype 1 was more frequently associated with a higher viral load when compared to genotypes 2 and 3

Effects of Annurca Apple (Malus pumila cv Annura) Polyphenols on Breast Cancer Cells

Abstract: Background: Dietary micronutrients have been proposed as effective inhibitory agents for cancer initiation, progression, and incidence. Among them, polyphenols, present in different foods and beverages, have retained attention in recent years. Apples are among the most consumed fruits worldwide, and several studies suggest that apple polyphenols could play a role in the prevention of degenerative diseases. Aims and Objectives: The present study aimed at evaluating the effects of Annurca flesh polyphenols extract (AFPE) effects of proliferation on MCF-7 cells. Methods: The data indicated that apple polyphenolic compounds had a significant antiproliferative action on MCF-7 cells and 500μM EqC AFPE induced a cell cycle arrest at G2/M. AFPE was also capable of inducing morphological changes as evidenced by nuclear condensation. Results: The cellular, morphological, and molecular data unequivocally suggested that induction of cellular apoptosis was mainly responsible for the previously observed antiproliferation-induced AFPE on MCF-7 cells. Conclusion: Taken together, AFPE that acts at a low micromolar range against breast cancer cells may be considered as a promising candidate for anticancer therapy

Site-directed mutagenesis gives insights into substrate specificity of Sulfolobus solfataricus purine-specific nucleoside hydrolase

Nucleoside hydrolases (NHs) are a class of metalloproteins that catalyze the irreversible hydrolysis of the N-glycosidic bond of β-ribonucleosides forming ribose and purine or pyrimidine base. In the hyperthermophilic archaeon Sulfolobus solfataricus, two NHs have been purified and extensively characterized. Although these enzymes show different substrate specificity, one for purines, the other for pyrimidines, their architectures are similar to that of non-specific NHs previously identified in protozoa. The detailed inspection into the active site of the two homologous enzymes obtained by homology modeling allowed us to infer the role of specific residues in substrate specificity. We report here the site-directed mutagenesis of the Sulfolobus solfataricus purine-specific inosine-adenosine-guanosine nucleoside hydrolase (SsIAG-NH). The double (L221Y/N228V) mutant of SsIAG-NH was expressed in E. coli and purified, and its activity with different substrates was compared to that of the wild-type enzyme. The double substitution modifies the catalytic pattern of the wild-type enzyme affecting both its substrate specificity and catalytic efficiency. Kinetic data obtained for the double mutant are in good agreement with modeling predictions

Purine nucleoside phosphorylases from hyperthermophilic Archaea require a CXC motif forstability and folding

5'-Deoxy-5'-methylthioadenosine phosphorylase II from Sulfolobus solfataricus (SsMTAPII) and purine nucleoside phosphorylase from Pyrococcus furiosus (PfPNP) are hyperthermophilic purine nucleoside phosphorylases stabilized by intrasubunit disulfide bonds. In their C-terminus, both enzymes harbour a CXC motif analogous to the CXXC motif present at the active site of eukaryotic protein disulfide isomerase. By monitoring the refolding of SsMTAPII, PfPNP and their mutants lacking the C-terminal cysteine pair after guanidine-induced unfolding, we demonstrated that the CXC motif is required for the folding of these enzymes. Moreover, two synthesized CXC-containing peptides with the same amino acid sequences present in the C-terminal regions of SsMTAPII and PfPNP were found to act as in vitro catalysts of oxidative folding. These small peptides are involved in the folding of partially refolded SsMTAPII, PfPNP and their CXC-lacking mutants, with a concomitant recovery of their catalytic activity, thus indicating that the CXC motif is necessary to obtain complete reversibility from the unfolded state of the two proteins. The two CXC containing peptides are also able to reactivate scrambled RNaseA. The data obtained in the present study represent the first example of how the CXC motif improves both stability and folding in hyperthermophilic proteins with disulfide bonds

Biological activity of MelAnnurca flesh apple biophenols

Background. The apple is among the most consumed fruits in the world and several studies suggest that apple polyphenols could play a role in preventing degenerative diseases. Recent studies have shown that polyphenols are able a high antioxidant and/or anti-proliferative power, therefore their food intake could play a decisive role in the prevention of various pathologies, in particular those associated with the production of free radicals. The Annurca apple (MelAnnurca), a variety from southern Italy, is called the "queen of apples" thanks to its remarkable organoleptic qualities: taste, flavor and aroma. The Annurca apple is a constituent component of the Mediterranean diet and its potential health benefit could be attributed at a large amount of bioactive components; in fact, this apple is characterized by an extremely high content of polyphenols. Objective: The aim of this paper was to review the most recent literature regarding the health benefits of Annurca apples and their phytochemicals. In particular, this review highlighted the effects of the flesh of this fruit on different types of human cells. Methods: A literature research was performed using the keywords “Annurca”, “apple”, “flesh”, “fruit”, “polyphenols”, “nutrition”, “nutraceuticals”, individually or all together, in Scopus, Web of Science and PubMed. Results: The MelAnnurca apple has a higher content of bioactive compounds (polyphenols) than other apples, making it an ideal source of nutraceuticals. Both the cytotoxic activity and the antioxidant effect of the extracts of polyphenols obtained from its flesh have been highlighted. Conclusion These results give new insights for a future implementation of the production chain of Annurca apple in the area of Campania, Southern Italy. However, although evidence to support the health benefits of MelAnnurca apple polyphenols is rapidly accumulating, further human studies may be needed before the public is convinced and willing to incorporate the apple Annurca into their diet, accepting the idea that the integration of polyphenols has a beneficial effect on the health of the human body