Inhibition of proliferation, migration and invasion of human non-small cell lung cancer cell line A549 by phlomisoside F from Phlomis younghusbandii Mukerjee

Purpose: To determine the effect of phlomisoside F (PMF) on the proliferation, migration and invasion of human non-small cell lung cancer cell line A549 and explore the possible mechanisms. Methods: The anti-proliferative effect of PMF on A549 cells was determined by CCK-8. Subsequently, migration and invasion were evaluated by Transwell and Transwell with matrigel assays, respectively. Furthermore, cell cycle and apoptosis were assessed by flow cytometry, while the mechanisms of action were determined by Western blotting. Results: PMF exhibited significant anti-proliferative effect on A549 cells in concentration-dependent and time-dependent manners, with half maximal inhibitory concentration (IC50) of 54.51 μM. Treatment with PMF (10, 20 and 40 μM) for 48 h resulted in significantly decreased migration and invasion in A549 cells. In addition, PMF at concentrations of 25, 50 and 75 μM induced cell cycle arrest in G0/G1phase and enhanced cell apoptosis in A549 cells. Furthermore, caspase-3, caspase-9 and Bax protein expressions were up-regulated while Bacl-2 and COX-2 protein expressions were significantly downregulated at 10, 20 and 40 μM concentrations of PMF. Conclusion: PMF suppresses A549 cell growth, migration and invasion. The mechanism may be related to the induction of mitochondria-mediated apoptosis pathway via regulation of caspase-3, caspase-9, Bcl-2 and Bax expressions, and inhibition of PGE2 synthesis by reducing COX-2 expression.

Withaferin A promotes proliferation and migration of brain endothelial cells

Purpose: To investigate the effect of withaferin A (WFA) on the proliferation and migration of brain endothelial cells. Methods: BALB-5023 mouse microvascular cells were treated with a range of withaferin A (WFA) concentrations from 10 to 100 ng/mL. Dojindo’s CCK-8 cell proliferation kit was used for the analysis of cell proliferation. Transwell cell culture inserts were used to determine the migration potential of WFAtreated endothelial cells. Absorbance was measured at 450 nm on an enzyme-linked immunosorbent (ELISA) reader. Results: The results revealed a significant increase in the proliferation and migration of endothelial cells following treatment with a low concentration (30 ng/mL) of WFA compared with the higher concentration (> 10 ng/mL). The effect was further enhanced when WFA was used in combination with soluble Fas ligand (sFasL). Autocrine signaling of vascular endothelial growth factor (VEGF) by endothelial cells was significantly increased following treatment with WFA or in combination with sFasL. WFA increased the expression of Fas on endothelial cells, suggesting the involvement of sFasL in the proliferation and migration of brain endothelial cells. Conclusion: Thus, WFA promotes the proliferation and migration of endothelial cells through increase in the expression of Fas and secretion of VEGF.

Prototype and proof of concept for a display system in retail stores

Selling devices on retail stores comes with the big challenge of grabbing the customer’s attention. Nowadays people have a lot of offers at their disposal and new marketing techniques must emerge to differentiate the products. When it comes to smartphones and tablets, those devices can make the difference by themselves, if we use their computing power and capabilities to create something unique and interactive. With that in mind, three prototypes were developed during an internship: a face recognition based Customer Detection, a face tracking solution with an Avatar and interactive cross-app Guides. All three revealed to have potential to be differentiating solutions in a retail store, not only raising the chance of a customer taking notice of the device but also of interacting with them to learn more about their features. The results were meant to be only proof of concepts and therefore were not tested in the real world.

Conception of a YBCO Superconducting ZFC-Magnetic Bearing Virtual Prototype

This paper presents the conception of an original superconducting Frictionless Zero Field Cooling bearing virtual prototype. In previous work also shown in this conference, a viability study of a Zero Field Cooling-superconducting bearing concept was conducted. It showed that the virtual prototype is feasible. Moreover, the simulation studies showed that a Zero Field Cooling superconducting track provides not only effective lateral stability but also higher levitation forces than the commonly used Field Cooling tracks. In this paper the new Zero Field Cooling -bearing virtual prototype is modeled in 3D. The virtual prototype was designed having in mind: i) a future implementation in high density polyurethane, for low temperature robustness; ii) future manufacturing in a three axes CNC milling machine and; iii) future implementation of some parts using an additive manufacturing technique.

Biochemical characterization of systemic bacteria in bananas, sensitivity to antibiotics and plant phytotoxicity during shoot proliferation.

2016

Prognostic significance of pulmonary multifocal neuroendocrine proliferation with typical carcinoid

Background. Clinical significance of multifocal pulmonary neuroendocrine proliferations (MNEP), including tumorlets and pulmonary neuroendocrine cell hyperplasia, in association with Typical Carcinoid (TC), is still debated. Methods. A large retrospective series of TC with long-term follow-up data prospectively collected from two institutions was evaluated. Recurrence or new TC development was followed-up. Patients with TC alone and MNEP+TC were compared. Results. 234 TC patients undergone surgery were included: 41 MNEP+TC (17.5%) and 193 TC alone (82.5%). In the MNEP+TC group older age (p<0.001), peripheral tumors (p=0.0032), smaller tumor size (p=0.011) and lymph-nodal spread (p=0.02) were observed in comparison with TC group. Relapses occurred in 8 patients (19.5%) in the MNEP+TC group and in 7 (3.6%) of the TC group. The 10-years progression-free survival were 96.1% in TC and 83.8% in MNEP+TC (p<0.001). After matching, in 36 pairs of patients a significantly higher 5-years progression-free survival was calculated for TC group (p<0.01). Furthermore the odds of belonging to MNEP+TC group was higher with work-related exposure to inhalant agents (p=0.008), asthma/bronchitis (p=0.002), emphysema, fibrosis and inflammatory status (p=0.032), micronodules on the chest CT scan and respiratory insufficiency (p=0.036). Conclusions. The identification of MNEP requires careful pathological examination and postoperative follow-up. MNEP seems to be an adverse prognostic factor in patients with synchronous TC. Therefore, suspicion of MNEP during the pre-operative assessment should not be underestimated, enabling changes in the surgical strategy.

Altered transcriptional and epigenetic regulation affects brain cells proliferation and differentiation in the ultra-rare genetic demyelinating disease AGC1 deficiency: a study on in vitro models

AGC1 deficiency is a rare demyelinating disease caused by mutations in the SLC25A12 gene, which encodes for the mitochondrial glutamate-aspartate carrier 1 (AGC1/Alarar), highly expressed in the central nervous system. In neurons, impairment in AGC1 activity leads to reduction in N-acetyl-aspartate, the main lipid precursor for myelin synthesis (Profilo et al., 2017); in oligodendrocytes progenitors cells, AGC1 down regulation has been related to early arrest proliferation and premature differentiation (Petralla et al., 2019). Additionally, in vivo AGC1 deficiency models i.e., heterozygous mice for AGC1 knock-out and neurospheres from their subventricular zone, respectively, showed a global decrease in cells proliferation and a switch in neural stem cells (NSCs) commitment, with specific reduction in OPCs number and increase in neural and astrocytic pools (Petralla et al., 2019). Therefore, the present study aims to investigate the transcriptional and epigenetic regulation underlying the alterations observed in OPCs and NSCs biological mechanisms, in either AGC1 deficiency models of Oli-neu cells (murine immortalized oligodendrocytes precursors cells), partially silenced by a shRNA for SLC25A12 gene, and SVZ-derived neurospheres from AGC1+/- mice. Western blot and immunofluorescence analysis revealed significant variations in the expression of transcription factors involved in brain cells’ proliferation and differentiation, in association with altered histone post-translational modifications, as well as histone acetylases (HATs) and deacetylases (HDACs) activity/expression, suggesting an improper transcriptional and epigenetic regulation affecting both AGC1 deficiency in vitro models. Furthermore, given the large role of acetylation in controlling in specific time-windows OPC maturation (Hernandez and Casaccia; 2015), pharmacological HATs/HDACs inhibitions were performed, confirming the involvement of chromatin remodelling enzymes in the altered proliferation and early differentiation observed in the AGC1 deficiency models of siAGC1 Oli-neu cells and AGC1+/- mice-derived neurospheres.

Characterization of a prototype for cone beam breast computed tomography

The project aims to experiment the Cone Beam Breast Computed Tomography technique using a standard digital mammography system. The work is focused on the definition of a protocol of quality measurements for the pre-clinical evaluation of the machine. The paper is developed in two parts. The first is specifically concerned with the methods used to define the image quality and dosimetry aspects specific for digital mammography devices. A complete characterization of the system has been performed according to the applicable IEC standards to assure the performances of the equipment and define the quality levels. Due to the lack of a quality control protocol dedicated to CBBCT mammography scanner, a new equivalent test procedure has been proposed. The second part of the paper is focused on the evaluation, through quantitative and visual analyzes, of the CBCT exam feasibility in the hardware and software conditions currently proposed by IMS Giotto. The prototype was in fact developed differing from the technical choices of competing companies and developed for a different intended use. The main difference with respect to the existing breast CT scanners is the possibility of performing on the same system the CBBCT scanning but also all the mammographic techniques. In this thesis, we aim to assess whether, in the current setup, considering a dosimetric range very close to that used in the clinic, the tests produce results that can be considered acceptable or at least indicative of the feasibility of the entire project from a commercial point of view. For this purpose, the final reconstruction images, obtained by two previously developed software, are analyzed.

Study of the performance of the prototype EIC dRICH readout based on SiPM with thermoelectric cooling

L'Electron-Ion Collider è un futuro acceleratore di particelle che approfondirà la nostra conoscenza riguardo l'interazione forte tramite la collisione di elettroni con nuclei e protoni. Uno dei progetti attualmente considerati per la costruzione del rivelatore, il dual-radiator RICH, prevede l'impiego di due radiatori Cherenkov, sui quali verranno montati dei fotorivelatori per rilevare l'emissione della luce Cherenkov e risalire alla massa delle particelle. L'opzione di base per questi rivelatori sono i sensori al silicio SiPM. Questo lavoro di tesi si basa sullo studio delle prestazioni di un prototipo per l'acquisizione dei dati rilevati dai SiPM che sfrutta l'effetto termoelettrico per raffreddare la zona in cui sono situati i sensori. L'analisi dei dati acquisiti ha portato alla conclusione che le prestazioni del prototipo sono confrontabili con quelle misurate all'interno di una camera climatica quando si trovano alla stessa temperatura.

Model-Based Contrastive Explanations for XAIP: Towards a General Model and Prototype

Planning is an important sub-field of artificial intelligence (AI) focusing on letting intelligent agents deliberate on the most adequate course of action to attain their goals. Thanks to the recent boost in the number of critical domains and systems which exploit planning for their internal procedures, there is an increasing need for planning systems to become more transparent and trustworthy. Along this line, planning systems are now required to produce not only plans but also explanations about those plans, or the way they were attained. To address this issue, a new research area is emerging in the AI panorama: eXplainable AI (XAI), within which explainable planning (XAIP) is a pivotal sub-field. As a recent domain, XAIP is far from mature. No consensus has been reached in the literature about what explanations are, how they should be computed, and what they should explain in the first place. Furthermore, existing contributions are mostly theoretical, and software implementations are rarely more than preliminary. To overcome such issues, in this thesis we design an explainable planning framework bridging the gap between theoretical contributions from literature and software implementations. More precisely, taking inspiration from the state of the art, we develop a formal model for XAIP, and the software tool enabling its practical exploitation. Accordingly, the contribution of this thesis is four-folded. First, we review the state of the art of XAIP, supplying an outline of its most significant contributions from the literature. We then generalise the aforementioned contributions into a unified model for XAIP, aimed at supporting model-based contrastive explanations. Next, we design and implement an algorithm-agnostic library for XAIP based on our model. Finally, we validate our library from a technological perspective, via an extensive testing suite. Furthermore, we assess its performance and usability through a set of benchmarks and end-to-end examples.

RED-Bridge: A Multiprotocol Tool Prototype for IoT Sensors

The IoT is growing more and more each year and is becoming so ubiquitous that it includes heterogeneous devices with different hardware and software constraints leading to an highly fragmented ecosystem. Devices are using different protocols with different paradigms and they are not compatible with each other; some devices use request-response protocols like HTTP or CoAP while others use publish-subscribe protocols like MQTT. Integration in IoT is still an open research topic. When handling and testing IoT sensors there are some common task that people may be interested in: reading and visualizing the current value of the sensor; doing some aggregations on a set of values in order to compute statistical features; saving the history of the data to a time-series database; forecasting the future values to react in advance to a future condition; bridging the protocol of the sensor in order to integrate the device with other tools. In this work we will show the working implementation of a low-code and flow-based tool prototype which supports the common operations mentioned above, based on Node-RED and Python. Since this system is just a prototype, it has some issues and limitations that will be discussed in this work.

Pipkiiα Is Widely Expressed In Hematopoietic-derived Cells And May Play A Role In The Expression Of Alpha- And Gamma-globins In K562 Cells.

Characterized for the first time in erythrocytes, phosphatidylinositol phosphate kinases (PIP kinases) belong to a family of enzymes that generate various lipid messengers and participate in several cellular processes, including gene expression regulation. Recently, the PIPKIIα gene was found to be differentially expressed in reticulocytes from two siblings with hemoglobin H disease, suggesting a possible relationship between PIPKIIα and the production of globins. Here, we investigated PIPKIIα gene and protein expression and protein localization in hematopoietic-derived cells during their differentiation, and the effects of PIPKIIα silencing on K562 cells. PIPKIIα silencing resulted in an increase in α and γ globins and a decrease in the proliferation of K562 cells without affecting cell cycle progression and apoptosis. In conclusion, using a cell line model, we showed that PIPKIIα is widely expressed in hematopoietic-derived cells, is localized in their cytoplasm and nucleus, and is upregulated during erythroid differentiation. We also showed that PIPKIIα silencing can induce α and γ globin expression and decrease cell proliferation in K562 cells.

Obesity And Inflammation And The Effect On The Hematopoietic System.

Bone marrow is organized in specialized microenvironments known as 'marrow niches'. These are important for the maintenance of stem cells and their hematopoietic progenitors whose homeostasis also depends on other cell types present in the tissue. Extrinsic factors, such as infection and inflammatory states, may affect this system by causing cytokine dysregulation (imbalance in cytokine production) and changes in cell proliferation and self-renewal rates, and may also induce changes in the metabolism and cell cycle. Known to relate to chronic inflammation, obesity is responsible for systemic changes that are best studied in the cardiovascular system. Little is known regarding the changes in the hematopoietic system induced by the inflammatory state carried by obesity or the cell and molecular mechanisms involved. The understanding of the biological behavior of hematopoietic stem cells under obesity-induced chronic inflammation could help elucidate the pathophysiological mechanisms involved in other inflammatory processes, such as neoplastic diseases and bone marrow failure syndromes.

Antiangiogenic And Finasteride Therapies: Responses Of The Prostate Microenvironment In Elderly Mice.

The aim of this study was to evaluate the structural and molecular effects of antiangiogenic therapies and finasteride on the ventral prostate of senile mice. 90 male FVB mice were divided into: Young (18 weeks old) and senile (52 weeks old) groups; finasteride group: finasteride (20mg/kg); SU5416 group: SU5416 (6 mg/kg); TNP-470 group: TNP-470 (15 mg/kg,) and SU5416+TNP-470 group: similar to the SU5416 and TNP-470 groups. After 21 days, prostate ventral lobes were collected for morphological, immunohistochemical and Western blotting analyses. The results demonstrated atrophy, occasional proliferative lesions and inflammatory cells in the prostate during senescence, which were interrupted and/or blocked by treatment with antiangiogenic drugs and finasteride. Decreased AR and endostatin reactivities, and an increase for ER-α, ER-β and VEGF, were seen in the senile group. Decreased VEGF and ER-α reactivities and increased ER-β reactivity were verified in the finasteride, SU5416 groups and especially in SU5416+TNP-470 group. The TNP-470 group showed reduced AR and ER-β protein levels. The senescence favored the occurrence of structural and/or molecular alterations suggesting the onset of malignant lesions, due to the imbalance in the signaling between the epithelium and stroma. The SU5416+TNP-470 treatment was more effective in maintaining the structural, hormonal and angiogenic factor balance in the prostate during senescence, highlighting the signaling of antiproliferation via ER-β.

An Efficient Protocol For The Generation Of Monocyte Derived Dendritic Cells Using Serum-free Media For Clinical Applications In Post Remission Aml Patients.

Protocols for the generation of dendritic cells (DCs) using serum as a supplementation of culture media leads to reactions due to animal proteins and disease transmissions. Several types of serum-free media (SFM), based on good manufacture practices (GMP), have recently been used and seem to be a viable option. The aim of this study was to evaluate the results of the differentiation, maturation, and function of DCs from Acute Myeloid Leukemia patients (AML), generated in SFM and medium supplemented with autologous serum (AS). DCs were analyzed by phenotype characteristics, viability, and functionality. The results showed the possibility of generating viable DCs in all the conditions tested. In patients, the X-VIVO 15 medium was more efficient than the other media tested in the generation of DCs producing IL-12p70 (p=0.05). Moreover, the presence of AS led to a significant increase of IL-10 by DCs as compared with CellGro (p=0.05) and X-Vivo15 (p=0.05) media, both in patients and donors. We concluded that SFM was efficient in the production of DCs for immunotherapy in AML patients. However, the use of AS appears to interfere with the functional capacity of the generated DCs.