Cell-Free DNA Methylation: The New Frontiers of Pancreatic Cancer Biomarkers' Discovery

Pancreatic ductal adenocarcinoma (PDAC) is among the most lethal cancer types world-wide. Its high mortality is related to the difficulty in the diagnosis, which often occurs when the disease is already advanced. As of today, no early diagnostic tests are available, while only a limited number of prognostic tests have reached clinical practice. The main reason is the lack of reliable biomarkers that are able to capture the early development or the progression of the disease. Hence, the discovery of biomarkers for early diagnosis or prognosis of PDAC remains, de facto, an unmet need. An increasing number of studies has shown that cell-free DNA (cfDNA) methylation analysis represents a promising non-invasive approach for the discovery of biomarkers with diagnostic or prognostic potential. In particular, cfDNA methylation could be utilized for the identification of disease-specific signatures in pre-neoplastic lesions or chronic pancreatitis (CP), representing a sensitive and non-invasive method of early diagnosis of PDAC. In this review, we will discuss the advantages and pitfalls of cfDNA methylation studies. Further, we will present the current advances in the discovery of pancreatic cancer biomarkers with early diagnostic or prognostic potential, focusing on pancreas-specific (e.g., CUX2 or REG1A) or abnormal (e.g., ADAMTS1 or BNC1) cfDNA methylation signatures in high risk pre-neoplastic conditions and PDAC

Neutron Star Radius-to-mass Ratio from Partial Accretion Disc Occultation as Measured through Fe Kα\alpha Line Profiles

We present a new method to measure the radius-to-mass ratio (R/M) of weakly magnetic, disc-accreting neutron stars by exploiting the occultation of parts of the inner disc by the star itself. This occultation imprints characteristic features on the X-ray line profile that are unique and are expected to be present in low mass X-ray binary systems seen under inclinations higher than ~65 degrees. We analyse a NuSTAR observation of a good candidate system, 4U 1636-53, and find that X-ray spectra from current instrumentation are unlikely to single out the occultation features owing to insufficient signal-to-noise. Based on an extensive set of simulations we show that large-area X-ray detectors of the future generation could measure R/M to ~2{\div}3% precision over a range of inclinations. Such is the precision in radius determination required to derive tight constraints on the equation of state of ultradense matter and it represents the goal that other methods too aim to achieve in the future.Comment: 17 pages, 8 figures; this is a pre-print edition of an article that has been accepted for publication in the Astrophysical Journa

Optimizing the supercritical fluid extraction process of bioactive compounds from processed tomato skin by-products

Abstract A supercritical fluid extraction (SC-CO2) was used to extract high-quality oil from tomato skin by-products. The effects of pressure and extraction time on oil yield was investigated in the study. Lycopene and β-carotene content as well as p-coumaric acid, ferulic acid, caffeic acid, chlorogenic acid, vanillic acid, epicatechin, naringenin, catechin, quercetin and luteolin were estimated. The highest oil yield of 79.00% was obtained after 80 min with a pressure of 550 bar. The resulting oleoresin in carotenoids with lycopene and β-carotene content respectively of 0.86 and 1.5 mg/100 g, this oleoresin was found to be the richest. Naringenin was the most abundant flavonoid identified with a maximum content in oleoresin extracted at 550 bar (84.04 mg/kg DW) followed by caffeic acid (26.60 mg/kg DW). A moderate radical scavenging potential was further observed. Overall, results highlight that pressure is a key parameter for the extraction bioactive oleoresin from tomato skin by-products

Role of IL-6/STAT3 Axis in Resistance to Cisplatin in Gastric Cancers

Gastric cancer, the second most common cause of death worldwide, is characterized by poor prognosis and low responsiveness to chemotherapy. Indeed, multidrug resistance, based mainly on cellular and molecular factors, remains one of the most limiting factors of the current approach to gastric cancer (GC) therapy. We employed a comprehensive gene expression analysis through data mining of publicly available databases to assess the role of the signal transducer and activator of transcription 3 (STAT3) in gastric cancer drug efficiency. It has been proposed that gastric cancer cells are less sensitive to these drugs because they develop resistance to these agents through activating alternative signalling pathways responsible for overcoming pharmacological inhibition. Our study evaluated the hypothesis that activating STAT3 signalling in response to cisplatin reduces the reaction to the drug. Consistent with this hypothesis, inhibition of interleukin 6 (IL-6)/STAT3 in combination therapy with cisplatin prevented both STAT3 activation and more lethality than induction by a single agent. The data suggest that the IL-6/STAT3 axis block associated with cisplatin treatment may represent a strategy to overcome resistance

Pancreatic Progenitor Commitment Is Marked by an Increase in Ink4a/Arf Expression

The identification of the molecular mechanisms controlling early cell fate decisions in mammals is of paramount importance as the ability to determine specific lineage differentiation represents a significant opportunity for new therapies. Pancreatic Progenitor Cells (PPCs) constitute a regenerative reserve essential for the maintenance and regeneration of the pancreas. Besides, PPCs represent an excellent model for understanding pathological pancreatic cellular remodeling. Given the lack of valid markers of early endoderm, the identification of new ones is of fundamental importance. Both products of the Ink4a/Arf locus, in addition to being critical cell-cycle regulators, appear to be involved in several disease pathologies. Moreover, the locus’ expression is epigenetically regulated in ES reprogramming processes, thus constituting the ideal candidates to modulate PPCs homeostasis. In this study, starting from mouse embryonic stem cells (mESCs), we analyzed the early stages of pancreatic commitment. By inducing mESCs commitment to the pancreatic lineage, we observed that both products of the Cdkn2a locus, Ink4a and Arf, mark a naïve pancreatic cellular state that resembled PPC-like specification. Treatment with epi-drugs suggests a role for chromatin remodeling in the CDKN2a (Cycline Dependent Kinase Inhibitor 2A) locus regulation in line with previous observations in other cellular systems. Our data considerably improve the comprehension of pancreatic cellular ontogeny, which could be critical for implementing pluripotent stem cells programming and reprogramming toward pancreatic lineage commitment

Ferulago nodosa subsp. geniculata (Guss.) Troia & Raimondo from Sicily: isolation of essential oil and evaluation of its bioactivity

Ferulago nodosa (L.) Boiss. (Apiaceae) is a species occurring in the Balkan-Tyrrhenian area. The object of the present study is Sicilian F. nodosa subsp. geniculata (Guss.) Troia & Raimondo, classified as an endemic F. nodosa subspecies. Aerial parts of this plant species were subjected to hydrodistillation to obtain an essential oil. A total of 93 compounds were identified with 2,3,6- trimethyl benzaldehyde (19.0%), spathulenol (9.0%), (E)-caryophyllene (5.4%), and caryophyllene oxide (5.4%) as the main components. The biological activities of F. nodosa essential oil were also investigated. This oil showed an interesting antioxidant potential in a 2,2′-Azino-bis(3- ethylbenzothiazoline-6-sulfonic acid) (ABTS) test (IC50 of 14.05 μg/mL). Additionally, hypoglycemic and antilipidemic effects were evaluated. Lipase enzyme was inhibited with an IC50 value of 41.99 μg/mL. Obtained data demonstrated that F. nodosa could be considered a promising source of bioactive compounds useful for the treatment and management of obesity

Influence of Organic and Conventional Agricultural Practices on Chemical Profile, In Vitro Antioxidant and Anti-Obesity Properties of Zingiber officinale Roscoe

Zingiber officinale Roscoe (Zingiberaceae), common known as ginger, is extensively used as spice worldwide in cooking and to prepare beverage. Moreover, it was used to treat a wide range of diseases including metabolic syndrome (MetS). Mets is a group of risk factors, including insulin resistance and consequently impaired glucose tolerance, dyslipidaemia, obesity, and hypertension. It is estimated that that MetS affects 25% of the population [1]. The efficacy of natural products especially derived from vegetables and spice largely consumed worldwide is a topic of great interest not only to cure but also to prevent the onset of the disease. In this study the influence of organic (OR) and conventional (CONV) agricultural practices on chemical profile and nutraceutical properties of Zingiber officinale Roscoe spice was evaluated. A multi-target approach was used to test the antioxidant activity by using DPPH, ABTS, β-carotene bleaching, and FRAP assays. The anti-obesity effect was investigated through inhibition of lipase and carbohydrate hydrolyzing enzymes a-amylase and a-glucosidase [2]. Ginger bioactive compounds were extracted by ultrasound assisted maceration process with ethanol. OR Ginger (Z5) showed the highest TPC and TFC with values of 39.27 and 15.38 ug/g DW. This sample resulted the most active in all applied antioxidant test with particular reference to ABTS test where Z5 showed a stronger activity with IC50 value of 0.81 ug/mL in comparison to the positive control ascorbic acid (1.70 mg/mL). RACI and GAS statistical approach confirmed the Z5 highest antioxidant potency. Moreover, Z5 exhibited a promising lipase inhibitory activity with IC50 value quite similar to the positive control orlistat (IC50 values of 34.48 vs 37.42 ug/mL). Collectively, our results demonstrated the impact of agricultural practices on ginger health properties. However, further in vivo studies will be needed to confirm the potential in humans and prove the safety of the products. [1] Rochlani, Y.; Pothineni, N.V.; Kovelamudi, S.; Mehta, J.L. Metabolic syndrome: Pathophysiology, management, and modulation by natural compounds. Ther. Adv. Cardiovasc. Dis. 2017, 11, 215–225. [2] Leporini, M.; Loizzo, M.R.; Sicari, V.; Pellicano, T.M.; Reitano, A.; Dugay, A.; Deguin, B.; Tundis, R. Citrus × Clementina Hort. Juice Enriched with Its By-Products (Peels and Leaves): Chemical composition, in vitro bioactivity, and impact of processing. Antioxidants 2020, 9, 298

Effect of Feijoa Sellowiana Acetonic Extract on Proliferation Inhibition and Apoptosis Induction in Human Gastric Cancer Cells

Gastric cancer (GC) still represents a relevant health problem in the world for both incidence and mortality rates. Many studies underlined that natural products consumption could reduce GC risk, indicating flavonoids as responsible for the beneficial eects through the modulation of several biological processes, such as the inhibition of cancer antioxidant defense and induction of apoptosis. Since Feijoa sellowiana fruit is known to contain high amounts of flavonoids, among which is flavone, we evaluated the antiproliferative and proapoptotic eects of F. sellowiana acetonic extract on GC cell lines through MTS and Annexin-V FITC assays. Among three GC cell lines tested, SNU-1 results being sensitive to both the F. sellowiana acetonic extract and synthetic flavone, which was used as the reference treatment. Moreover, we evaluated their antioxidant eects, assessing the activity of the antioxidant enzymes supeoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) in polymorphonuclear cells. We found a significant increase of their activity after exposure to both F. sellowiana acetonic extract and flavone, supporting the idea that a diet that includes flavone-rich fruits could be of benefit for health. In addition to this antioxidant eect on normal cells, this study indicates, for the first time, an anticancer eect of F. sellowiana acetonic extract in GC cells

CHM/REP1 Transcript Expression and Loss of Visual Function in Patients Affected by Choroideremia

PURPOSE. To evaluate the disease progression in patients with clinical and genetic diagnoses of choroideremia during a long-term follow-up and to investigate the relationship between pathogenic variants in the CHM/REP1 gene and disease phenotypes. METHODS. We performed a retrospective longitudinal study on 51 affected men by reviewing medical charts at baseline and follow-up visits to extract the following ocular findings: best-corrected visual acuity, Goldmann visual field, optical coherence tomography, microperimetry. Data obtained from the analysis of DNA and mRNA were reevaluated for genetic classification of patients. RESULTS. The longitudinal analysis showed a significant (P < 0.001) worsening of best-corrected visual acuity with a mean rate of 0.011 logMar per year before 50 years and 0.025 logMar per year after 50 years. Similarly, V4e Goldmann visual field area significantly (P ≤ 0.01) decreased at a mean rate of 2.7% per year before 40 years and 5.7% after 40 years. Moreover, we observed a significant (P < 0.05) decrease of macular sensitivity with a mean rate of 5.0% per year and a decrease of mean macular thickness with a mean rate of 0.8% per year. We classified our patients into two groups according to the expression of the CHM/ REP1 gene transcript and observed that mutations leading to mRNA absence are associated with an earlier best-corrected visual acuity and Goldmann visual field loss. CONCLUSIONS. Our analysis of morphological and functional parameters in choroideremia patients showed a slow disease progression, particularly in the first decades of life. Overall, reevaluation of clinical and molecular data suggests exploring the genotype–phenotype relationship based on CHM/REP1 transcript expression. PURPOSE. To evaluate the disease progression in patients with clinical and genetic diagnoses of choroideremia during a long-term follow-up and to investigate the relationship between pathogenic variants in the CHM/REP1 gene and disease phenotypes. METHODS. We performed a retrospective longitudinal study on 51 affected men by reviewing medical charts at baseline and follow-up visits to extract the following ocular findings: best-corrected visual acuity, Goldmann visual field, optical coherence tomography, microperimetry. Data obtained from the analysis of DNA and mRNA were reevaluated for genetic classification of patients. RESULTS. The longitudinal analysis showed a significant (P < 0.001) worsening of best-corrected visual acuity with a mean rate of 0.011 logMar per year before 50 years and 0.025 logMar per year after 50 years. Similarly, V4e Goldmann visual field area significantly (P ≤ 0.01) decreased at a mean rate of 2.7% per year before 40 years and 5.7% after 40 years. Moreover, we observed a significant (P < 0.05) decrease of macular sensitivity with a mean rate of 5.0% per year and a decrease of mean macular thickness with a mean rate of 0.8% per year. We classified our patients into two groups according to the expression of the CHM/ REP1 gene transcript and observed that mutations leading to mRNA absence are associated with an earlier best-corrected visual acuity and Goldmann visual field loss. CONCLUSIONS. Our analysis of morphological and functional parameters in choroideremia patients showed a slow disease progression, particularly in the first decades of life. Overall, reevaluation of clinical and molecular data suggests exploring the genotype–phenotype relationship based on CHM/REP1 transcript expression