A headlight on liquid biopsies: a challenging tool for breast cancer management

Breast cancer is the most frequent carcinoma and second most common cause of cancer-related mortality in postmenopausal women. The acquisition of somatic mutations represents the main mechanism through which cancer cells overcome physiological cellular signaling pathways (e.g., PI3K/Akt/mTOR, PTEN, TP53). To date, diagnosis and metastasis monitoring is mainly carried out through tissue biopsy and/or re-biopsy, a very invasive procedure limited only to certain locations and not always feasible in clinical practice. In order to improve disease monitoring over time and to avoid painful procedure such as tissue biopsy, liquid biopsy may represent a new precious tool. Indeed, it represents a basin of “new generation” biomarkers that are spread into the bloodstream from both primary and metastatic sites. Moreover, elevated concentrations of circulating tumor DNA (ctDNA) as well as circulating tumor cells (CTCs) have been found in blood plasma of patients with various tumor types. Nowadays, several new approaches have been introduced for the detection and characterization of CTCs and ctDNA, allowing a real-time monitoring of tumor evolution. This review is focused on the clinical relevance of liquid biopsy in breast cancer and will provide an update concerning CTCs and ctDNA utility as a tool for breast cancer patient monitoring during the course of disease

Molecular characterization of Giardia duodenalis cysts in the Oreto River (Sicily, Southern Italy)

<p><strong>Background</strong>: The presence of Giardia was investigated along the Oreto river between January 2008 and May 2009 with the aim of understanding the source of contamination by molecular typing of cysts.</p><p><strong>Methods</strong>: A total of 38 water samples (10 collected from the river mouth, 24 from the whole Oreto basin and 4 sewage samples from the Monreale treatment plant) were processed. In addition, 22 faecal samples of livestock living close to the Oreto area, were analyzed. The presence of Giardia was determined by immunofluorescence assay and their genetic characterization was achieved by a nested PCR assay targeting the triosephosphate isomerase gene. <strong></strong></p><p><strong>Results</strong>: All water samples from the river mouth were positive for Giardia, even if the concentration of cysts fluctuated considerably among sampling occasions. Our investigation showed that the Vadduneddu and Altofonte torrents, two influents of the river, were the principal sources of contamination. Moreover, the genotypes of Vadduneddu torrent were the same as those detected in human wastewater taken from the activated sludge plant of Monreale city. Assemblages A and B were found in water samples with a predominance of Assemblage A, subtype AII. Assemblage E was only found in a single calf isolate. <strong></strong></p><p><strong>Conclusions</strong>: The data show that the high cyst counts regularly detected in the Oreto river are due to contamination with wastewater of human origin. This finding is relevant for public health, particularly because river water is used for agricultural purposes.</p&gt

Efficacy of ruxolitinib retreatment in a patient with high-risk myelofibrosis using the international prognostic scoring system

Primary myelofibrosis (PMF) is a myeloproliferative neoplasm in which clonal proliferation of hematopoietic stem cells and bone marrow fibrosis coexist.1 Patients may eventually die due to leukemic progression, which occurs in up to 20% of cases, or because of cardiovascular comorbidities or cytopenia, which causes susceptibility to infections and bleeding

COMPARISON OF CLINICAL AND LABORATORY DATA, INCLUDING JAK-2 46/1 HAPLOTYPE, BETWEEN PATIENTS WITH IDIOPATHIC ERYTHROCYTOSIS AND POLYCYTHEMIA VERA.

Background Idiopathic erythrocytosis (IE) is a relatively rare finding characterized by an increased red blood cell mass without an identifiable cause. Diagnosis of IE is based on the exclusion of primary and secondary erythrocytosis including JAK2-wild-type polycythemia Vera (PV). Aims In the current study, we report clinical features and laboratory data able to discriminate IE from PV, at diagnosis Methods We have here analyzed clinical and laboratory parameters, including Jak-2 46/1 haplotype, from patients with a confirmed diagnosis of IE and PV, followed from January 2010 to December 2016. Data were statistically analyzed, nominal variables were compared with X2 test and continuous variables with the Mann-Whitney test. Results Overall, 40 patients with IE and 93 patients with PV were included in the current analysis (Table 1). Splenomegaly and itch were reported only in one patient with IE. History of thrombosis and cardiovascular events was positive in one case with IE. Jak-2 (V617F) and exon 12 mutations were negative in all patients with IE, while Jak-2 46/1 haplotype was found at heterozygous state in 18 patients and at homozygous state in 2 patients with IE. Conclusion In the current study, we highlight peculiar clinical and laboratory findings of IE, in comparison with Polycythemia Vera. As shown by available studies, Hb and HCT level do not easily discriminate between the two categories of patients while gene panels may be useful to improve diagnostic accuracy of IE. We have here first observed the presence of Jak-2 46/1 haplotype in approximately half patients with IE, even in absence of JAk-2 mutations; the homozygous status was statistically different among PV and IE patients. The role of such association deserves further specific studies

CARDIOVASCULAR RISK IN ESSENTIAL THROMBOCYTHEMIA AND POLYCYTHEMIA VERA: THROMBOTIC RISK AND SURVIVAL

Thromboembolic and bleeding events pose a severe risk for patients with Polycythemia Vera (PV) and Essential Thrombocythemia (ET). Many factors can contribute to promoting the thrombotic event due to the interaction between platelets, leukocytes, and endothelium alterations. Moreover, a significant role can be played by cardiovascular risk factors (CV.R) such as cigarette smoking habits, hypertension, diabetes, obesity and dyslipidemia. In this study, we evaluated the impact that CV.R plays on thrombotic risk and survival in patients with PV and ET

Splenomegaly Impacts Prognosis in Essential Thrombocythemia and Polycythemia Vera: A Single Center Study

Splenomegaly is one of the major clinical manifestations of primary myelofibrosis and is common also in other chronic Philadelphia-negative myeloproliferative neoplasms, causing symptoms and signs and affecting quality of life of patients diagnosed with these diseases. We aimed to study the impact that such alteration has on thrombotic risk and on the survival of patients with essential thrombocythemia and patients with Polycythemia Vera (PV). We studied the relationship between splenomegaly (and its grade), thrombosis and survival in 238 patients with et and 165 patients with PV followed at our center between January 1997 and May 2019

EFFECT OF miR-21, miR-182 AND let-7i ON TSP-1 EXPRESSION IN COLON CANCER CELL LINE

Background: MicroRNAs (miRNAs) are small non-coding RNAs that regulate the expression of different genes, including genes involved in cancer progression, angiogenesis and metastasis. Thrombospondin-1 (TSP-1) has been shown to contrast angiogenesis in vivo. TSP-1 expression levels are inversely correlated with tumor vascularity and metastasis in colon cancer. Bio-informatic statistical analysis indicated that TSP-1 is hypothetical target of miR-21, miR-182, overexpressed in CRC, and let-7i which expression is down-regulated in this tumor. In this work we investigated whether TSP-1 expression could be regulated by miR-21, mir-182 and let-7i in HT29 colon cancer cell line. Methods: To investigated whether miR-21, mir-182 and let-7i directly modulates TSP-1 expression, we transfected HT29 cell line with pre-mir21, pre-mir182 and pre-let7i by using siPortNeo FX tranfection agent and after 48h we evaluated TSP-1 mRNA, using Quantitative Real Time-PCR, and intracellular and secreted protein level performed by Western blotting and ELISA. To confirm the modulation of TSP-1 by miRNAs we transfected HT29 cell line with anti-mir to target the mature form of miR-21, miR182 and let-7i. Results: Using Real-Time PCR we did not find any variation of TSP-1 mRNA expression levels after transfection with pre-mir21 in HT29 cell line, but we observed a down-regulation of cytosolic and secreted protein by Western blot and ELISA. In cells transfected with pre-mir182 we did not observe any down-regulation both TSP-1 mRNA and cytosolic and secreted protein. Finally, we did not find any variation of TSP-1 level in cells transfected with let- 7i. Results were confirmed by transfection with anti-mir21, anti- mir182 and anti-let7i and, using the same method, we evaluated TSP-1 expression. Conclusions: Data suggest that mir-182 induces degradation of TSP-1 mRNA in HT29 cell line, whereas mir-21 affects probably by blockage of TSP-1 translation. Let-7i does not seem involved in regulation of TSP-1 expression in HT29 cells. Understanding the molecular mechanism by which miRNAs regulate TSP-1 expression could be used to restore TSP-1 expression to contrast angiogenic events in colon cancer

Management of Ponatinib in Patients with Chronic Myeloid Leukemia with Cardiovascular Risk Factors

Cardiovascular (CV) adverse events are considered common complications of ponatinib treatment. Recently, it has been demonstrated that ponatinib dose reductions in definite settings can obtain optimal responses and lower ponatinib-related CV events. In this study, we describe the management of 5 patients with chronic myeloid leukemia treated with ponatinib, from second to fourth line of tyrosine kinase inhibitor therapy, carrying high pre-ponatinib CV risk, who obtained optimal molecular response and developed no CV adverse event during follow-up. Among these 5 patients, 2 had diagnosis of ischemic heart disease and underwent percutaneous angioplasty, 2 had type 2 diabetes and arterial hypertension, and 1 had only arterial hypertension. Median follow-up for ponatinib therapy is 1,039 days (34.6 months). Median dosage administered is 30 mg a day. SCORE charts were used to estimate risk of CV death in 10 years and Charlson Comorbidity Index was applied to estimate age-adjusted risk of death related to comorbidities. Strict cardiologic follow-up (complete evaluation every 3 to 6 months) and maximum effort in the control of CV modifiable risk factors are strongly recommended in the management of ponatinib treatment in patients at high risk for CV events and may allow the use of ponatinib in patients belonging to CV risk category

The Essential Thrombocythemia, Thrombotic Risk Stratification, and Cardiovascular Risk Factors

Essential thrombocythemia is a rare hematological malignancy with good overall survival, but moderate to high risk of developing arterial or venous thrombosis lifelong. Different thrombotic risk scores for patients with essential thrombocythemia have been proposed, but only one of them (the IPSET-t scoring system) takes into account the classical cardiovascular risk factors as one of the scoring items. Currently, in clinical practice, the presence of cardiovascular risk factors in patients with diagnosis of ET rarely determines the decision to initiate cytoreductive therapies. In our study, we compared different risk models to estimate the thrombotic risk of 233 ET patients and the role of specific driver mutations and evaluated the impact that conventional cardiovascular risk factors (hypertension, cigarette smoking, diabetes, obesity, and dyslipidaemia) have on thrombotic risk in patients with ET. Perspective studies conducted on a polycentric large cohort of patients should be conducted to estimate the impact of cardiovascular risk factors in determining thrombosis in ET patients, evaluating the opportunity of initiating a cytoreductive therapy in patients with cardiovascular risk factors, even if classified into low to moderate risk groups according to other scoring systems