Teaching Non-Euclidean Geometries in High-School: an experimental study.

In these years it is becoming quite clear that the development of a young student into an adult citizen requires a solid scientific background. Facing the challenges of a quickly changing world where political decisions are not only concerned with economics or ethics, but also with climate sciences, medicine, etc., requires a good education. Citizens are required to exert logical thinking and know the methods of science in order to adapt, to understand and to develop as persons. At the core of all these required skills sits mathematics, with its formal methods to develop knowledge. Learning the axiomatic method is fundamental to understand how hard sciences work, and helps in consolidating logical thinking, which will be useful for the entire life of a student. In my experience as a secondary school teacher, I have tried to understand how students perceive mathematics and what difficulties they encounter. One observation I often made was that the axiomatic study of geometry was a problematic topic for students, even for those with an interest for mathematics. For this reason, I decided to focus my PhD work on the teaching and learning of geometry, focusing explicitly on its axiomatic foundations, in order to concentrate on those aspects that foster the development of a logical thinking, of the ability of proving a thesis, etc., which are necessary, as said, for the growth of a modern citizen. Axiomatic geometry exposes the students to plural worlds, where the choice of a few base axioms heavily influences the properties of the objects that can be observed. The students, who are used to an intuitive study of geometry in the Euclidean plane, can benefit from the discovery of non-Euclidean geometries in several regards. First, they are shown that different non-Euclidean geometries exist, then they discover how these geometries can be developed by slightly modifying the axioms. Finally, they can be taught how these are useful to model real problems. The importance of teaching non-Euclidean geometries in high school has been debated for decades and several experiences have been conducted with students in the past. However, all these works were often of qualitative nature, the experimental protocols were poorly documented, and the statistical data was missing. The main objective of this thesis is to investigate, by means of quantitative experimental protocols, the viability and effects of teaching an introductory course on non-Euclidean geometries to high-school students. The experimental nature of this study required the classroom work to be concise and limited to a short number of seminars and workshops. Several experiences are described, involving several high school classes and a total of 154 students and 57 teachers. These have been used to evaluate and refine the teaching tools and topics that are covered in this thesis and are reported in detail for use in future experiences. Statistical methods and evaluation questionnaires are discussed to assess the effectiveness of the approach, which will prove necessary in larger-scale experiments. The outline of the thesis follows. In Chapter 1, a more elaborate motivation and introduction to the topics of the thesis are given. Chapter 2 reports a brief history of the development of Euclidean and non-Euclidean geometries and then discusses whether the birth and the development of non-Euclidean geometries constitute a revolution in mathematics. Chapter 3 discusses the teaching of geometry in secondary schools, with a specific interest in the Italian education system. Chapter 4 gives an informative introduction to the main aspects of the study and open questions; summarise and critiques the studies that have been conducted on the teaching of non-Euclidean geometries; and states the research questions that are investigated in the present thesis: - RQ1:What features of a short introductory course in non-Euclidean geometries are effective in engaging high-school students? - RQ2: To what extent do students gain a new perspective on the concept of axiomatic system? - RQ3: How well do students learn the taught concepts of non-Euclidean geometries? - RQ4: To what extent do students’ critical thinking and proof skills improve over the duration of the course? - RQ5: Do students’ beliefs about mathematics change over the duration of the course? Chapter 5 discusses all the details about the experimental phase. Specifically, this chapter describes and justifies the research methods and justifies the choice of adopting an essentially positivist paradigm using quantitative methods; discusses a preliminary experimentation conducted to investigate a suitable methodology, and – in more detail – a second experimentation. In addition, this chapter contains two sections dealing, respectively, with topics related to the experimentations: an experience with high-school teachers; and the description of the necessary adaptations for the distance learning imposed by Covid-19 restrictions. While Covid-19 restrictions impaired the possibility of a large-scale experiment, it allowed me to observe some peculiarities of the distance learning paradigm that must be accounted for when conducting geometry seminars online. Chapter 6 discusses the results of the data analysis, and provide an interpretation of the data shown and many conjectures for future developments. Chapter 7 concludes the doctoral dissertation

Axitinib induces DNA damage response leading to senescence, mitotic catastrophe, and increased NK cell recognition in human renal carcinoma cells.

Tyrosine kinase inhibitors (TKIs) including axitinib have been introduced in the treatment of renal cell carcinoma (RCC) because of their anti-angiogenic properties. However, no evidence are presently available on a direct cytotoxic anti-tumor activity of axitinib in RCC.Herein we reported by western blot analysis that axitinib treatment induces a DNA damage response (DDR) initially characterized by γ-H2AX phosphorylation and Chk1 kinase activation and at later time points by p21 overexpression in A-498 and Caki-2 RCC cells although with a different potency. Analysis by immunocytochemistry for the presence of 8-oxo-7,8-dihydro-2'-deoxyguanosine in cellular DNA and flow cytometry using the redox-sensitive fluorescent dye DCFDA, demonstrated that DDR response is accompanied by the presence of oxidative DNA damage and reactive oxygen species (ROS) generation. This response leads to G2/M cell cycle arrest and induces a senescent-like phenotype accompanied by enlargement of cells and increased senescence-associated β-galactosidase activity, which are abrogated by N-acetyl cysteine (NAC) pre-treatment. In addition, axitinib-treated cells undergo to cell death through mitotic catastrophe characterized by micronucleation and abnormal microtubule assembly as assessed by fluorescence microscopy.On the other hand, axitinib, through the DDR induction, is also able to increase the surface NKG2D ligand expression. Accordingly, drug treatment promotes NK cell recognition and degranulation in A-498 RCC cells in a ROS-dependent manner.Collectively, our results indicate that both cytotoxic and immunomodulatory effects on RCC cells can contribute to axitinib anti-tumor activity

Effects of moderate and high rates of biochar and compost on grapevine growth in a greenhouse experiment

Biochar is used as soil amendment and enhancer of plant growth, but the mechanisms involved in grapevine are not understood. In this study, the short-term effects of amendments were evaluated in a trial combining three substrates (biochar, compost, peat-based media) with three doses(30, 70, 100%) along a time sequence on 1-year-old bare root cuttings of grapevine. Amendments were analyzed for elemental composition. Soil pH, electrical conductivity (EC), chlorophyll (CHL), flavonoids (FL), anthocyans (ANT) and nitrogen balance index (NBI) were measured.Biochar differed from other amendments for stable C structures, where nutrients and lignin residues were high in compost. Biochar increased soil pH, whereas biochar plus compost mixture augmented EC. The amended plants had detrimental effects on root, true and lateral leaves. Nevertheless, at the lowest rate biochar increased the primary shoot and total scion to root biomass ratio. Among biochemicals, ANT and NBI were mostly affected by biochar, while compost gave only slight increments. Thus, although biochar rate was not adequate for the shedding in open field our results suggest that biochar might be useful in nursery when used at low dosages

Adsorption and degradation of three pesticides in a vineyard soil and in an organic biomix

none5noA soil and an organic biomix (soil/vine branch/garden compost 20/40/40) were used in this lab experiment to evaluate adsorption and degradation parameters for three pesticides (chlorpyrifos, metalaxyl and cymoxanil) used in a vineyard. Adsorption in the biomix material was higher than in the soil for the three pesticides and chlorpyrifos was the most adsorbed pesticide. The role of the organic carbon is essential for enhancing the adsortion of the three pesticides, especially for the most apolar chlorpyrifos. Degradation was generally faster in the biomix material than in the soil although the process was slower in the case of chlorpyrifos if compared with the other two chemicals, due to a more toxic eect of this pesticide on soil microflora and a larger adsorption of this pesticide on the organic biomix that reduces its availability for dissipation. Amendment with cheap and available organic wastes or a grass-covered management of soil in the vineyard could reduce the impact of pesticides in the vineyard ecosystem and contribute to the sustainable management of chemicals in the environment.openCostantino Vischetti, Elga Monaci, Cristiano Casucci, Arianna De Bernardi, Alessandra CardinaliVischetti, Costantino; Monaci, Elga; Casucci, Cristiano; DE BERNARDI, Arianna; Cardinali, Alessandr

Vegetated ditches for the mitigation of pesticides runoff in the po valley

In intensive agricultural systems runoff is one of the major potential diffuse pollution path- ways for pesticides and poses a risk to surface water. Ditches are common in the Po Valley and can potentially provide runoff mitigation for the protection of watercourses. The effec- tiveness depends on ditch characteristics, so there is an urgent need for site-specific field trials. The use of a fugacity model (multimedia model) can allows recognition of the mitiga- tion main processes. A field experiment was conducted in order to evaluate the mitigation capacity of a typical vegetated ditch, and results were compared with predictions by a fugacity model. To evaluate herbicide mitigation after an extreme runoff, the ditch was flooded with water containing mesotrione, S-metolachlor and terbuthylazine. Two other sub- sequent floods with uncontaminated water were applied 27 and 82 days later to evaluate herbicides release. Results show that the ditch can immediately reduce runoff concentration of herbicides by at least 50% even in extreme flooding conditions. The half-distances were about 250 m. As a general rule, a runoff of 1 mm from 5 ha is mitigated by 99% in 100 m of vegetated ditch. Herbicides retention in the vegetated ditch was reversible, and the second flood mobilized 0.03-0.2% of the previous one, with a concentration below the drinking water limit of 0.1 \u3bcg L-1. No herbicide was detected in the third flood, because the residual amount in the ditch was too low. Fugacity model results show that specific physical-chemi- cal parameters may be used and a specific soil-sediment-plant compartment included for modelling herbicides behaviour in a vegetated ditch, and confirm that accumulation is low or negligible for herbicides with a half-life of 40 days or less. Shallow vegetated ditches can thus be included in a general agri-environment scheme for the mitigation of pesticides runoff together with wetlands and linear buffer strips. These structures are present in the land- scape, and their environmental role can be exploited by proper management

High-throughput analysis of the RNA-induced silencing complex in myotonic dystrophy type 1 patients identifies the dysregulation of miR-29c and its target ASB2

Myotonic dystrophy type 1 (DM1) is a multi-systemic disorder caused by abnormally expanded stretches of CTG DNA triplets in the DMPK gene, leading to mutated-transcript RNA-toxicity. MicroRNAs (miRNAs) are short non-coding RNAs that, after maturation, are loaded onto the RISC effector complex that destabilizes target mRNAs and represses their translation. In DM1 muscle biopsies not only the expression, but also the intracellular localization of specific miRNAs is disrupted, leading to the dysregulation of the relevant mRNA targets. To investigate the functional alterations of the miRNA/target interactions in DM1, we analyzed by RNA-sequencing the RISC-associated RNAs in skeletal muscle biopsies derived from DM1 patients and matched controls. The mRNAs found deregulated in DM1 biopsies were involved in pathways and functions relevant for the disease, such as energetic metabolism, calcium signaling, muscle contraction and p53-dependent apoptosis. Bioinformatic analysis of the miRNA/mRNA interactions based on the RISC enrichment profiles, identified 24 miRNA/mRNA correlations. Following validation in 21 independent samples, we focused on the couple miR-29c/ASB2 because of the role of miR-29c in fibrosis (a feature of late-stage DM1 patients) and of ASB2 in the regulation of muscle mass. Luciferase reporter assay confirmed the direct interaction between miR-29c and ASB2. Moreover, decreased miR-29c and increased ASB2 levels were verified also in immortalized myogenic cells and primary fibroblasts, derived from biopsies of DM1 patients and controls. CRISPR/Cas9-mediated deletion of CTG expansions rescued normal miR-29c and ASB2 levels, indicating a direct link between the mutant repeats and the miRNA/target expression. In conclusion, functionally relevant miRNA/mRNA interactions were identified in skeletal muscles of DM1 patients, highlighting the dysfunction of miR-29c and ASB2

Axitinib induces DNA damage response leading to senescence, mitotic catastrophe, and increased NK cell recognition in human renal carcinoma cells

Tyrosine kinase inhibitors (TKIs) including axitinib have been introduced in the treatment of renal cell carcinoma (RCC) because of their anti-angiogenic properties. However, no evidence are presently available on a direct cytotoxic anti-tumor activity of axitinib in RCC.Herein we reported by western blot analysis that axitinib treatment induces a DNA damage response (DDR) initially characterized by γ-H2AX phosphorylation and Chk1 kinase activation and at later time points by p21 overexpression in A-498 and Caki-2 RCC cells although with a different potency. Analysis by immunocytochemistry for the presence of 8-oxo-7,8-dihydro-2'-deoxyguanosine in cellular DNA and flow cytometry using the redox-sensitive fluorescent dye DCFDA, demonstrated that DDR response is accompanied by the presence of oxidative DNA damage and reactive oxygen species (ROS) generation. This response leads to G2/M cell cycle arrest and induces a senescent-like phenotype accompanied by enlargement of cells and increased senescence-associated β-galactosidase activity, which are abrogated by N-acetyl cysteine (NAC) pre-treatment. In addition, axitinib-treated cells undergo to cell death through mitotic catastrophe characterized by micronucleation and abnormal microtubule assembly as assessed by fluorescence microscopy.On the other hand, axitinib, through the DDR induction, is also able to increase the surface NKG2D ligand expression. Accordingly, drug treatment promotes NK cell recognition and degranulation in A-498 RCC cells in a ROS-dependent manner.Collectively, our results indicate that both cytotoxic and immunomodulatory effects on RCC cells can contribute to axitinib anti-tumor activity

A simplified genomic profiling approach predicts outcome in metastatic colorectal cancer

The response of metastatic colorectal cancer (mCRC) to the first-line conventional combination therapy is highly variable, reflecting the elevated heterogeneity of the disease. The genetic alterations underlying this heterogeneity have been thoroughly characterized through omic approaches requiring elevated efforts and costs. In order to translate the knowledge of CRC molecular heterogeneity into a practical clinical approach, we utilized a simplified Next Generation Sequencing (NGS) based platform to screen a cohort of 77 patients treated with first-line conventional therapy. Samples were sequenced using a panel of hotspots and targeted regions of 22 genes commonly involved in CRC. This revealed 51 patients carrying actionable gene mutations, 22 of which carried druggable alterations. These mutations were frequently associated with additional genetic alterations. To take into account this molecular complexity and assisted by an unbiased bioinformatic analysis, we defined three subgroups of patients carrying distinct molecular patterns. We demonstrated these three molecular subgroups are associated with a different response to first-line conventional combination therapies. The best outcome was achieved in patients exclusively carrying mutations on TP53 and/or RAS genes. By contrast, in patients carrying mutations in any of the other genes, alone or associated with mutations of TP53/RAS, the expected response is much worse compared to patients with exclusive TP53/RAS mutations. Additionally, our data indicate that the standard approach has limited efficacy in patients without any mutations in the genes included in the panel. In conclusion, we identified a reliable and easy-to-use approach for a simplified molecular-based stratification of mCRC patients that predicts the efficacy of the first-line conventional combination therapy

MRE11 inhibition highlights a replication stress-dependent vulnerability of MYCN-driven tumors

MRE11 is a component of the MRE11/RAD50/NBS1 (MRN) complex, whose activity is essential to control faithful DNA replication and to prevent accumulation of deleterious DNA double-strand breaks. In humans, hypomorphic mutations in these genes lead to DNA damage response (DDR)-defective and cancer-prone syndromes. Moreover, MRN complex dysfunction dramatically affects the nervous system, where MRE11 is required to restrain MYCN-dependent replication stress, during the rapid expansion of progenitor cells. MYCN activation, often due to genetic amplification, represents the driving oncogenic event for a number of human tumors, conferring bad prognosis and predicting very poor responses even to the most aggressive therapeutic protocols. This is prototypically exemplified by neuroblastoma, where MYCN amplification occurs in about 25% of the cases. Intriguingly, MRE11 is highly expressed and predicts bad prognosis in MYCN-amplified neuroblastoma. Due to the lack of direct means to target MYCN, we explored the possibility to trigger intolerable levels of replication stress-dependent DNA damage, by inhibiting MRE11 in MYCN-amplified preclinical models. Indeed, either MRE11 knockdown or its pharmacological inhibitor mirin induce accumulation of replication stress and DNA damage biomarkers in MYCN-amplified cells. The consequent DDR recruits p53 and promotes a p53-dependent cell death, as indicated by p53 loss- and gain-of-function experiments. Encapsulation of mirin in nanoparticles allowed its use on MYCN-amplified neuroblastoma xenografts in vivo, which resulted in a sharp impairment of tumor growth, associated with DDR activation, p53 accumulation, and cell death. Therefore, we propose that MRE11 inhibition might be an effective strategy to treat MYCN-amplified and p53 wild-type neuroblastoma, and suggest that targeting replication stress with appropriate tools should be further exploited to tackle MYCN-driven tumors

Sorafenib induces cathepsin B-mediated apoptosis of bladder cancer cells by regulating the Akt/PTEN pathway. The Akt inhibitor, perifosine, enhances the sorafenib-induced cytotoxicity against bladder cancer cells

Sorafenib, a tyrosine kinase inhibitor, has been demonstrated to exert anti-tumor effects. However, the molecular mechanisms underlying its effects on bladder cancer remain unknown. Here, we evaluated the mechanisms responsible for the sorafenib-induced anti-tumor effects on 5637 and T24 bladder cancer cells. We demonstrated that sorafenib reduces cell viability, stimulates lysosome permeabilization and induces apoptosis of bladder cancer cells. These effects are dependent by the activation of cathepsin B released from lysosomes. The sorafenib-increased cathepsin B activity induced the proteolysis of Bid into tBid that stimulates the intrinsic pathway of apoptosis characterized by mitochondrial membrane depolarization, oxygen radical generation and cytochrome c release. Moreover, we found that cathepsin B enzymatic activity, induced by sorafenib, is dependent on its dephosphorylation via PTEN activation and Akt inactivation. Pretreatment with orthovanadate rescued bladder cancer cells from apoptosis. In addition, the Akt inhibitor perifosine increased the sensitivity of bladder cancer cells to sorafenib-induced cytotoxicity. Overall, our results show that apoptotic cell death induced by sorafenib in bladder cancer cells is dependent on cathepsin B activity and involved PTEN and Akt signaling pathways. The Akt inhibitor perifosine increased the cytotoxic effects of sorafenib in bladder cancer cells