The hypothesis to use microRNAs (miRNAs) circulating in the blood as cancer biomarkers was formulated some years ago based on promising initial results. After some exciting discoveries, however, it became evident that the accurate quantification of cell-free miRNAs was more challenging than expected. Difficulties were linked to the strong impact that many, if not all, pre- and post- analytical variables have on the final results. In this study, we used currently available high-throughput technologies to identify miRNAs present in plasma and serum of patients with breast, colorectal, lung, thyroid and melanoma tumors, and healthy controls. Then, we assessed the absolute level of nine different miRNAs (miR-320a, miR-21-5p, miR-378a-3p, miR-181a-5p, miR-3156-5p, miR-2110, miR-125a-5p, miR-425-5p, miR-766-3p) in 207 samples from healthy controls and cancer patients using droplet digital PCR (ddPCR) technology. We identified miRNAs specifically modulated in one or more cancer types, according to tissue source. The significant reduction of miR-181a-5p levels in breast cancer patients serum was further validated using two independent cohorts, one from Italy (n = 70) and one from US (n = 90), with AUC 0.66 and 0.73 respectively. This study finally powers the use of cell-free miRNAs as cancer biomarkers and propose miR-181a-5p as a diagnostic breast cancer biomarker.

Industrial Hemp, Cannabis sativa L., is characterized by low content of THC (<0.2%). An edible oil with excellent nutritional proprieties is obtained from cold pressing of hempseed. Since Hempseed oil is not reported in a Regulation yet, in order to ensure quality parameters, it is necessary to optimize standard methods (taken from the Regulation for EVOO). In this work, the standard method of quality parameters (free fatty acidity, peroxide number, and anisidine number) were miniaturized and optimized for Hempseed oil matrix. The miniaturized methods result in being sustainable, in environmental and economical perspectives, by using a smaller amount of chemicals (e.g., reagents, solvents), also reducing waste production and the sample needed in relation to the high cost of the Hempseed oil (60–70 €/L). The standard methods of miniaturization, carried out by using the Central Composite Design, allow for great saving of sample (5.35 g vs. 29 g) and reagents (up to 50%).

Leptomeningeal carcinomatosis (LC), previously known as carcinomatous meningitis, is a devastating neurologic complication of cancer. LC is associated with major neurologic disability and high mortality and occurs in 3% to 34% of all cancer patients. 1,2 An increasing incidence of brain and leptomeningeal metastases has been reported 3 during the history of patients with human epidermal growth factor receptor 2 (HER2) -overexpressing breast cancer. This clinical condition is the result of the neurotropism of HER2-overexpressing breast cancer cells combined with the high antitumoral activity of systemic administrations of trastuzumab without preventive effect in the brain and leptomeninges since trastuzumab does not effectively reach the cerebrospinal fluid (CSF) when given intravenously. 4 Intrathecal (IT) administration of trastuzumab has been tested in some patients with HER2-positive breast cancer and with LC occurring after a systemic treatment. [5][6][7][8] Recently, tumor specimens from 3,807 patients with gastric cancer were centrally tested for HER2 status. The rate for HER2-positive gastric cancer was 22%, which is similar to the rate for HER2-positive breast cancer 9 and, for the first time, positive results of a phase III trial of the efficacy of trastuzumab were reported for gastric cancer. 9,10 We report here the case of a patient with HER2-overexpressing gastric cancer who developed LC, and we demonstrated the HER2-positive status of malignant cells in the CSF. To the best of our knowledge, this is the first case report of LC in HER2-positive gastric cancer.A 77-year-old woman who was previously well complained of abdominal discomfort, anorexia, fatigue, and weight loss for the past 6 months. She was admitted to the Oncology-Hematology Department of the Hospital of Piacenza in Piacenza, Italy, on August 3, 2009, after she developed a disseminated intravascular coagulation (DIC). General physical examination revealed pallor, diffuse cutaneous hemorrhage with ecchymoses and petechiae, no adenopathy, and no hepatosplenomegaly. Laboratory tests revealed anemia with a hemoglobin level of 7 g/dL and a reduction in the platelet count to 5,000/mL. Other laboratory tests (prothrombin time, activated partial thromboplastin time, fibrinogen, fibrin split products, and D-dimer) were consistent with the diagnosis of DIC.Abdominal ultrasound revealed the presence of small lymphadenopathies in the perigastric region; computed tomography (CT) scan of the chest was unremarkable, although a CT scan of the abdomen confirmed the findings of the ultrasound test: perigastric lymphadenopathies 2 cm in diameter. Gastroscopy with biopsy revealed gastric adenocarcinoma. A bone marrow biopsy was performed that demonstrated metastasis from gastric cancer. The immunohistochemistry

The recurrent genetic anomalies used to classify prostate cancer (PC) into distinct molecular subtypes have limited relevance for clinical practice. In consideration of WHO 2016 histological classification, which includes the introduction of Gleason Score 4 for patients with cribriform component and the definition of intraductal carcinoma as a new entity, a retrospective pilot study was conducted to investigate, by histological review, if there were any variations of Gleason Score and the incidence of intraductal carcinoma and cribriform pattern, intended as “phenotypic” markers of potentially lethal PC, among metastatic castration-sensitive PC (mCSPC) and metastatic castration-resistant PC (mCRPC) samples. Potentially predictive factors were also assessed. Among 125 cases, a variation in the Gleason Score was reported in 26% of cases. A cribriform (36%) or intraductal (2%) pattern was reported in a higher percentage. Of them, a primary Gleason pattern 4 was reported in 80% of cases. All patients with intraductal carcinoma present a BRCA2 mutation, also found in 80% of cases with a cribriform pattern. This pilot study documented some hypothesis-generating data, as the evaluation of de novo mCSPC and mCRPC as phenotypic/biologic model to be translated in clinical practice. A cribriform pattern/intraductal carcinoma might be a marker of potentially lethal PC. The high incidence of TP53 and BRCA2 mutations in de novo mCSPC may also have a therapeutic implication.

The hypothesis to use microRNAs (miRNAs) circulating in blood as cancer biomarkers was formulated some years ago based on promising initial results. After some exiting discoveries, however, it became evident that the accurate quantification of cell-free miRNAs (i.e. that retrieved in serum or plasma) was more challenging than expected. Difficulties were linked to the strong impact that many, if not all, pre- and post- analytical variables (i.e. tissue preparation, storage condition, extraction method, quantification technique, normalization approach) have on the final result. In this study, we used currently available high-throughput technologies to identify miRNAs present in plasma and serum of patients with breast, colorectal, lung and melanoma tumors, and healthy controls. Then, we used droplet digital PCR (ddPCR) technology to get an accurate absolute quantification of specific cell-free miRNAs. We assessed the level of 8 different miRNAs (miR-320a, miR-21-5p, miR-378a-3p, miR-181a-5p, miR-3156-5p, miR-2110, miR-125a-5p, miR-425-5p, miR-766-3p) in 180 samples from healthy controls and cancer patients. We identified miRNAs specifically modulated in one or more cancer types. Plasma and serum from the same patient provided different results in terms of absolute miRNA amount and modulation. The significant reduction of miR-181a-5p levels in serum of breast cancer patients was further validated using two independent cohorts, one from Italy (n = 70) and one from US (n = 90). This study finally powers the use miR-181a-5p as a breast cancer biomarker. Citation Format: Manuela Ferracin, Laura Lupini, Irene Salamon, Elena Saccenti, Barabara Zagatti, Alessandra Mangolini, Maria Vittoria Zanzi, Paolo Carcoforo, Andrea Rocchi, Giorgio Cavallesco, Antonio Frassoldati, Alan B. Hollingsworth, Massimo Negrini. How to fish a good micro-marker out from a worthless lake: The case of cell-free miR-181a-5p and breast cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3964. doi:10.1158/1538-7445.AM2015-3964