CD271 downregulation promotes melanoma progression and invasion in 3-dimensional models and in zebrafish

CD271 is a neurotrophin receptor variably expressed in melanoma. While contradictory data are reported on its role as a marker of tumor initiating cells, little is known on its function in tumor progression. CD271 expression was higher in spheroids derived from freshly isolated cells of primary melanomas and in primary WM115 and WM793-B cell lines, while it decreased during progression to advanced stages in cells isolated from metastatic melanomas and in metastatic WM266-4 and 1205Lu cell lines. Moreover, CD271 was scarcely detected in the highly invasive spheroids (SKMEL28 and 1205Lu). CD271, originally expressed in the epidermis of skin reconstructs, disappeared when melanoma started to invade the dermis. SKMEL8 CD271(-) cells showed greater proliferation and invasiveness in vitro, and were associated with a higher number of metastases in zebrafish, as compared to CD271(+) cells. CD271 silencing in WM115 induced a more aggressive phenotype in vitro and in vivo. On the contrary, CD271 overexpression in SKMEL28 cells reduced invasion in vitro, and CD271 overexpressing 1205Lu cells was associated with a lower percentage of metastases in zebrafish. A reduced cell-cell adhesion was also observed in absence of CD271. Taken together, these results indicate that CD271 loss is critical for melanoma progression and metastasis

Comparative elemental analysis of dairy milk and plant-based milk alternatives

Together with essential elements, toxic elements can also be found in food. In this study, we analysed the content of 41 elements in milk from mammals (cow, goat, and donkey) and plant-based milk alternatives (from soy, rice, oat, spelt, almond, coconut, hazelnut, walnut, cashew, hemp, and quinoa) using inductively coupled plasma mass spectrometry and cold vapour generation atomic fluorescence (for Hg). The analytical methods were validated using both milk certified reference materials and recovery experiments for different milk samples, obtaining satisfactory results in all cases. Only cow and goat milks were important sources of all major mineral elements like Ca, K, Mg, Na and P, and some minor elements like Se and Zn, while soy milk contained significant amounts of Cu and Fe, coconut milk contained Cr and Se, and hemp milk contained Mo. The level of toxic trace elements, including As, Cd, Hg, and Pb was very low in all analysed samples and did not pose any threat to consumers. The study is of significance for consumers of plant-based beverages from nutritional and food safety point of view. © 2020 Elsevier Lt

Do DLX3 and CD271 Protect Human Keratinocytes from Squamous Tumor Development?

Well-regulated epidermal homeostasis depends on the function of different classes of factors, such as transcription regulators and receptors. Alterations in this homeostatic balance may lead to the development of cutaneous squamous tumorigenesis. The homeobox transcription factor DLX3 is determinant for a p53-dependent regulation of epidermal differentiation and modulates skin carcinogenesis. The maintenance of skin homeostasis also involves the action of neurotrophins (NTs) and their receptors, Trk and CD271. While Trk receptor overexpression is a hallmark of cancer, there are conflicting data on CD271 expression and function in cutaneous SCC (cSCC). Previous studies have reported NT receptors expression in head and neck SSC (HNSCC). We show that CD271 is expressed at low levels in primary cSCC cells and the number of CD271+ cells correlates with cell cohesion in SCC spheroids. In normal epidermis, CD271 is expressed in proliferative progenitor cells and DLX3 in terminally differentiated keratinocytes. Brain-derived neurotrophic factor (BDNF) and neurotrophin 3 (NT3) increase DLX3 expression. In the absence of a functional BDNF receptor TrkB in keratinocytes, we hypothesize that the BDNF-dependent DLX3 response could be mediated via CD271. Altogether, our results support a putative CD271-DLX3 connection in keratinocytes, which might be crucial to preventing squamous skin cancer

Qualitative and quantitative determination of water in airborne particulate matter

Abstract. This paper describes the optimization and validation of a new simple method for the quantitative determination of water in atmospheric particulate matter (PM). The analyses are performed by using a coulometric Karl-Fisher system equipped with a controlled heating device; different water contributions are separated by the application of an optimized thermal ramp (three heating steps: 50–120 °C, 120–180 °C, 180–250 °C). The analytical performance of the method was verified by using standard materials containing 5.55% and 1% by weight of water. The recovery was greater than 95%; the detection limit was about 20 μg. The method was then applied to NIST Reference Materials (NIST1649a, urban particulate matter) and to real PM10 samples collected in different geographical areas. In all cases the repeatability was satisfactory (10–15%). When analyzing the Reference Material, the separation of four different types of water was obtained. In real PM10 samples the amount of water and its thermal profile differed as a function of the chemical composition of the dust. Mass percentages of 3–4% of water were obtained in most samples, but values up to about 15% were reached in areas where the chemical composition of PM is dominated by secondary inorganic ions and organic matter. High percentages of water were also observed in areas where PM is characterized by the presence of desert dust. A possible identification of the quality of water released from the samples was tried by applying the method to some hygroscopic compounds that are likely contained in PM (pure SiO2, Al2O3, ammonium salts, carbohydrates and dicarboxylic acids) and by comparing the results with those obtained from field samples

Investigating Cutaneous Squamous Cell Carcinoma in vitro and in vivo: Novel 3D Tools and Animal Models

Cutaneous Squamous Cell Carcinoma (cSCC) represents the second most common type of skin cancer, which incidence is continuously increasing worldwide. Given its high frequency, cSCC represents a major public health problem. Therefore, to provide the best patients' care, it is necessary having a detailed understanding of the molecular processes underlying cSCC development, progression, and invasion. Extensive efforts have been made in developing new models allowing to study the molecular pathogenesis of solid tumors, including cSCC tumors. Traditionally, in vitro studies were performed with cells grown in a two-dimensional context, which, however, does not represent the complexity of tumor in vivo. In the recent years, new in vitro models have been developed aiming to mimic the three-dimensionality (3D) of the tumor, allowing the evaluation of tumor cell-cell and tumor-microenvironment interaction in an in vivo-like setting. These models include spheroids, organotypic cultures, skin reconstructs and organoids. Although 3D models demonstrate high potential to enhance the overall knowledge in cancer research, they lack systemic components which may be solved only by using animal models. Zebrafish is emerging as an alternative xenotransplant model in cancer research, offering a high-throughput approach for drug screening and real-time in vivo imaging to study cell invasion. Moreover, several categories of mouse models were developed for pre-clinical purpose, including xeno- and syngeneic transplantation models, autochthonous models of chemically or UV-induced skin squamous carcinogenesis, and genetically engineered mouse models (GEMMs) of cSCC. These models have been instrumental in examining the molecular mechanisms of cSCC and drug response in an in vivo setting. The present review proposes an overview of in vitro, particularly 3D, and in vivo models and their application in cutaneous SCC research

On-line separation and determination of trivalent and hexavalent chromium with a new liquid membrane annular contactor coupled to inductively coupled plasma optical emission spectrometry

We describe a new on-line sensitive and selective procedure for the determination of trivalent and hexavalent chromium in liquid samples by a tailor-made contactor (TMC), specifically a liquid membrane annular TMC, coupled with inductively coupled plasma with optical detection. The TMC was designed and developed to integrate the extraction and stripping phases of the analyte in one module to minimize the membrane solvent’s consumption and maximize the speed of transport through the liquid membrane. Moreover, the particular geometry studied, which consists of two coaxial hollow fibers, allows the TMC to be used for both separating and preconcentrating purposes. Both (−)-N-dodecyl-N-methylephedrinium bromide (30 mM) in dichloroethane and HNO3 (0.75 M) were used as the liquid membrane and receiving solution, re-spectively. The proposed method’s performance was evaluated in terms of the hexavalent chro-mium extraction efficiency and the coefficient of variation percentages; these were higher than 85% and less than 5%, respectively. In addition, the proposed procedure was applied to two real sam-ples: a tap water sample and an eluate from solid urban waste. In both cases, the analytical per-formances were good and comparable to those obtained using synthetic standard solutions

Phenolic content and antioxidant activity of einkorn and emmer sprouts and wheatgrass obtained under different radiation wavelengths

Abstract Sprouted seeds represent intriguing ready-to-eat micro-scale vegetables for the healthy food market, since they are tasty and rich in bioactive compounds. However, sprouts have been recently proposed as a source for the extraction and purification of several phytochemicals to be used in food supplementation or pharmaceutics. Recently, there has been an industrialization of sprout production, carried out indoor, often with use of artificial light, which have implications on biomass yield and composition, and on energetic and economic costs. This work investigates the effects of different radiation wavelengths from light emitting diodes (LED) on free and bound phenolics and antioxidant activity of sprouts and wheatgrass of einkorn (Triticum monococcum L. ssp. monococcum) and emmer ([(Triticum turgidum L. spp. dicoccum, (Schrank ex Schubler) Thell.)]). After 3 days of grain incubation in the dark, three light treatments were applied, labelled as BLUE (447 and 470 nm), RED (627 and 655 nm), and SUN (447, 470, 505, 530, 590, 627, 655 nm), for a same total photon flux density (PFD) of 200 μmol m−2 s−1. Sprouts were harvested at 5 days after sowing (DAS) and wheatgrass at 9 DAS. The effect of light was generally not significant for sprouts, much greater and species-specific for wheatgrass: BLUE in einkorn and RED in emmer generally increased free and total content of polyphenol (PC), tannins (TC), flavonoid (FC) and phenolic acids (PAs). The antioxidant activity was increased by BLUE in einkorn and decreased by RED in both species. BLUE and RED resulted energy saving compared to SUN

Apparent diffusion coefficient values in borderline breast lesions upgraded and not upgraded at definitive histopathological examination after surgical excision

Purpose: The study aims were to evaluate if the apparent diffusion coefficient (ADC) value could distinguish between breast lesions classified as B3 at core needle biopsy (CNB) that show or do not show atypia or malignancy at definitive histopathological examination (DHE) after surgical excision. Material and methods: From January 2013 to December 2017, 141 patients with a B3 breast lesion underwent magnetic resonance imaging and were included in the study. The ADC value was assessed drawing a ROI outlining the entire lesion, evaluating the mean ($ADC_{mean}$) and minimum ADC values ($ADC_{min}$). Results: Both $ADC_{mean}$ and ADCmin values showed a statistically significant difference between B3 lesions without and with malignancy or, for B3a lesions, atypia at DHE. They both showed a statistically significant difference also between B3a lesions without or with atypia or malignancy at DHE, but only $ADC_{min}$ (not $ADC_{mean}$) showed statistically significant difference between B3b lesions without or with malignancy at DHE. Conclusions: The ADC value could help distinguish between B3a lesions without or with atypia/malignancy at DHE after surgical excision and between B3b lesions without or with malignancy at DHE. Therefore, it could be used to help guide the diagnostic-therapeutic pathway of these lesions, particularly of B3a lesion

A previously unreported function of beta1B integrin isoform in caspase-8-dependent integrin-mediated keratinocyte death

Integrins regulate adhesive cell-matrix interactions and mediate survival signals. On the other hand, unligated or free cytoplasmic fragments of integrins induce apoptosis in many cell types (integrin-mediated death). We have previously shown that b1 integrins expression protects keratinocyte stem cells from anoikis, while the role of the b1B integrin isoform has never been clarified. Here we report that suspended keratinocytes undergo apoptosis via the activation of caspase-8, independently of Fas/Fas Ligand system. Indeed, anti-b1 integrin neutralizing antibodies induced apoptosis in short-hairpin-RNA-Fas-Associated-Death-Domain treated cells. Moreover, before and during suspension, caspase-8 directly associated with b1 integrin, that in turn internalized and progressively degraded, shedding the cytoplasmic domain. b1B was expressed only in the cytoplasm in a perinuclear fashion and remained unaltered during suspension. At 24 hrs, as b1A located close to the nucleus, b1B co-localized with b1A and co-immunoprecipitated with caspase-8. Caspase-8 was activated earlier in b1B integrin transfected keratinocytes, and these cells underwent a higher rate of apoptosis than mock cells. By contrast, caspase-8 was not activated in siRNA b1B transfected cells. These results indicate that when b1A is unligated, b1B is responsible for “integrin-mediated death” in human keratinocytes