THE WAYS OF ADJUSTING HOSPITALS FOR CHILDREN TO THE NEEDS OF THE CHILD – THE EXAMPLE OF MEYER CHILDREN HOSPITAL IN FLORENCE, ITALY

Web aplikacija za vizualizaciju hijerarhijskih podataka je razvijana kako bi se olakšalo vizualizaciju podataka u kojima postoji relacija ili hijerarhija. Mogućnost različitih prostornih raspoređivanja čvorova daje dodatni uvid podatke. Tako Breadthfirstsearch layout omogućava pregled hijerarhije unutar usmjerenog grafa, te jednostavnu izolaciju jednog ili više korijena, ako postoje unutar grafa. Izolacijom predaka pojedinog čvora možemo dobiti sve izravne prethodnike nekog čvora te tako izolirati podgraf koji sadrži samo one čvorove i bridove koji su bitni u kontekstu odabranog čvora. Filtriranje po kategorijama čvorova pruža uvid u to kakve veze postoje unutar nekog podskupa podataka s danim obilježjem, to jest kategorijom. Postoji veliki skup raznovrsnih podataka koji se mogu prilagoditi i prikazati pomoću ovakve aplikacije, a to može biti kao što je prikazano u primjeru, skup znanja iz nekog udžbenika. Dodatno se mogu prikazati skupovi podataka poput prometne, telekomunikacijske, plinske, vodovodne ili električne mreže. Osim toga može se koristiti u razvoju softvarea za prikaz Entity-Relationship$^1$ modela ili kao dijagram da se vide međuovisnosti pojedinih biblioteka ili paketa koji su potrebni za razvoj aplikacije. Tu svakako spada i mogućnost kreiranja grafa povezanosti između web stranica pa bi to bilo izrazito zanimljivo napraviti za Wikipediu, koja ima mnoštvo poveznica između stranica koje bi predstavljale čvorove. Aplikacija se može koristiti i za kreiranje skupa podataka tako da se korištenjem sučelja kreiraju novi projekti, grafovi, informacije i veze među njima koje se potom automatski spremaju u strukturu koja se može vizualizirati, a isto tako i uređivati i nadograđivati. Prostor za širenje funkcionalnosti aplikacije postoji na svim razinama. Od optimizacije koda, do dodavanja novih funkcionalnosti koje se mogu koristiti za bolju vizualizaciju skupa podataka. Prostor leži i u povezivanju aplikacije s metodama strojnog učenja koje otvaraju jednu novu dimenziju. Primjer bi bio određeni skup pojmova, te klasični tekst bez ikakvih posebnih tagova. Tada bi se korištenjem tehnika vezanih za NLP$^2$ mogla napraviti obrada teksta te vizualizirati povezanost danih pojmova unutar teksta. Izazovi za ovakvu aplikaciju su prvenstveno u veličini podataka i njihovoj pripremi. Ostaju otvorena pitanja kako bi se aplikacija ponašala u slučaju vizualizacije grafova koji sadrže milijune čvorova te još više bridova. Skup podataka nad kojim bi se mogla provesti takva testiranja i proučavanja je Wikipedia, koja je javno dostupna za preuzimanje u obliku komprimirane xml datoteke. 1: https://en.wikipedia.org/wiki/Entity-relationship_model 2: https://en.wikipedia.org/wiki/Natural_language_processingWeb application for visualization of hierarchically structured data was developed in order to enable visualization of any data that is structured in relational or hierarchical order. By using different graph layouts user can get an extra insight in the data. Breadthfirstsearch layout enables one to see if there is a hierarchy inside the directed graph and to isolate all root nodes inside it, if there is any. By isolating ancestors of one node, application can show all direct predecessors and by doing so user can see the subgraph that contains only the nodes and edges important for the chosen node. Filtering data by categories will enable user to see if there are any connections inside a specific subset of data. There is a vast array of different data sets that can be adapted and visualized by using an application such as this, and it can be done in the same way as it was shown in example by adapting the knowledge set from schoolbook. But it can also visualize data from traffic, telco, gas, electrical or water grid, or in software development for visualization of Entity-Relationship model, or as a diagram for modules dependency in software. There is also a potential for visualization of web, showing web pages and links between them and it would be extremely interesting to test on Wikipedia, as it has millions of articles and many more millions of links. Application can also be used for creation of data through user interface by creating projects, graphs, nodes, edges and categories that will be automatically saved in relational or hierarchical structure that can be visualized, updated or upgraded. There are many other functionalities that could be implemented in different parts of application and it is spread from optimizing code to adding new options for visualization. Other direction for improvement is connecting the application with machine learning solutions. For example, one could use a given set of data without any connections, and then create connections by extracting them with NLP from text. This would enable user to create edges between existing nodes, and by doing so build a new graph structure. There are also many challenges for application such as this one and they are primarily in size od data set and in preparation of that data set for use in application. There are still some open questions, some of them are directly related to visualization of data sets with millions of nodes and edges. Those questions could be answered by processing Wikipedia, which can be downloaded and processed

CD44 and tumor-derived extracellular vesicles (TEVs) : possible gateway to cancer metastasis

Cancer metastasis, the final stage of tumor progression, is a complex process governed by the interplay of multiple types of cells and the tumor microenvironment. One of the aspects of this interplay involves the release of various factors by the tumor cells alone or by forcing other cells to do so. As a consequence of these actions, tumor cells are prepared in favorable conditions for their dissemination and spread to other sites/organs, which guarantees their escape from immunosurveillance and further progression. Tumor-derived extracellular vesicles (TEVs) represent a heterogeneous population of membrane-bound vesicles that are being actively released by different tumors. The array of proteins (i.e., receptors, cytokines, chemokines, etc.) and nucleic acids (i.e., mRNA, miR, etc.) that TEVs can transfer to other cells is often considered beneficial for the tumor’s survival and proliferation. One of the proteins that is associated with many different tumors as well as their TEVs is a cluster of differentiation 44 in its standard (CD44s) and variant (CD44v) form. This review covers the present information regarding the TEVs-mediated CD44s/CD44v transfer/interaction in the context of cancer metastasis. The content and the impact of the transferred cargo by this type of TEVs also are discussed with regards to tumor cell dissemination

A Temperature Sensitive Mutation in Cactin Causes a G1 Phase Arrest in Toxoplasma gondii

Thesis advisor: Marc Jan GubbelsThesis advisor: Thomas ChilesThe length of the tachyzoite cell cycle, in particular G1, is an important virulence factor in Toxoplasma gondii. Cdk and Cyclin activities ultimately control the cell cycle; however, the checkpoint control mechanisms diverge from higher eukaryotes and are poorly understood. In order to elucidate these mechanisms, temperature sensitive (ts) mutants were generated by chemical mutagenesis. One of these mutants, called FV-P6, dies within one cell cycle in the G1 phase upon transfer from the permissive (35°C) to the restrictive temperature (40°C). Cosmid complementation identified the gene responsible for this G1 arrest as a `Cactin' ortholog. A single point mutation in this gene that resulted in an amino acid substitution from Tyrosine to Histidine at position 661 in the highly conserved C-terminus was shown to underlay the temperature sensitive effect. Cactin is highly conserved across eukaryotes and plays a role in embryonic development of metazoa although its mechanism of action is poorly understood. In agreement with the predicted nuclear localization signal in the N-terminus, expression of a fluorescent reporter gene fusion resulted in nuclear localization. Genome-wide expression profiling analysis of mutant and wild type at the permissive and restrictive temperatures confirmed the G1 arrest and furthermore demonstrated up-regulation of bradyzoite and Toxoplasma cat life cycle stage genes, hinting at TgCactin's role as a repressor. Since DNA binding domains or enzymatic domains are absent in TgCactin, TgCactin must act in a complex. Native blue gel electrophoresis demonstrated that TgCactin is present in large complexes of 720 and 800 kDa. A yeast two-hybrid screen (YTH) identified 40 potential TgCactin-interacting proteins of which 10 were selected for further validation. Eight out of these ten candidates are involved in DNA/RNA processes pertaining to transcription and translation, respectively. One-on-one YTH interactions between mutated and N-terminal deletion mutants of TgCactin and the above 10 interactors were abolished except for a single RNA helicase. Studies in Toxoplasma of four of these interactors demonstrated that only the RNA helicase localized to the nucleus; however, co-immunoprecipitation experiments to demonstrate that this protein is present in a complex with TgCactin were inconclusive. Furthermore, TgCactin self interactions identified domains necessary for TgCactin-TgCactin binding. Taken together, these findings indicate that TgCactin likely functions as a repressor of gene expression, possibly through an epigenetic mechanism reminiscent of an RNA/DNA helicase- based system in plants.Thesis (PhD) — Boston College, 2010.Submitted to: Boston College. Graduate School of Arts and Sciences.Discipline: Biology

Clinical analysis of 52 obstetric hysterectomies

Objectives: Analysis of obstetric (peripartum and postpartum) hysterectomies with regard to their frequency, indications, complications, and risk factors. Material and methods: The study included 52 women operated between 1985–2012. Obstetric hysterectomies were performed in 39 (75%) and 13 (25%) women, respectively. The results were statistically analyzed as arithmetic mean and standard deviation (SD). Results: Peri- and postpartum hysterectomies accounted for 0.123% of all births (0.092% and 0.031%, respectively). Mean pa­tient age, length of pregnancy, and number of deliveries was 32.6 years [SD ± 6.2], 38.1 weeks [SD ± 7.0], and 3.2 [SD ± 2.4], respectively. In the study group, 92.31% of the women were multiparous, and 86.54% gave birth by cesarean section and had a history of CS. Placental pathology accounted for 44.4% of indications for hysterectomy. Blood transfusion was required in 94.2% of the cases, symptoms of hypovolemic shock were observed in 21.2%, and ICU admission was required in 15.4% of the patients. Relaparotomy was necessary in 4 (7.7%) cases. Intrauterine fetal death occurred in 4 (7.6%) cases and extremely poor neonatal status was observed in 4.1% of the newborns. Conclusions: Hemorrhage due to placental pathology was the most frequent indication for obstetric hysterectomy. Risk factors for obstetric hysterectomy included multiparity, history of CS, recent CS, and age > 35 years. Postpartum hysterec­tomy accounted for 25% of the obstetric surgeries

Tumour-derived microvesicles contain interleukin-8 and modulate production of chemokines by human monocytes

Background: Tumour-derived microvesicles (TMVs) may interact with cells of the immune system. Our previous observations indicated that TMVs modulate production of cytokines and reactive oxygen species (ROS) by monocytes. This study was designed to determine the role of TMVs in stimulation of chemokine production by human monocytes. Materials and Methods: Chemokines at the mRNA and protein level were detected by real-time PCR and by Western blot, respecively. Chemokine release and chemotaxis of blood leukocytes were analysed by flow cytometry. Matrigel assay was used to determine angiogenesis in a NOD-SCID mice model. Results: TMVs induced secretion of interleukin-8 (CXCL8), monocyte chemoattractant protein-1 (CCL2), macrophage inflammatory protein-1α (CCL3) and MIP-1β (CCL4), and regulated on activation normal T-cells expressed and secreted (CCL5) chemokines and accumulation of their mRNA in monocytes. Moreover, TMVs enhanced angiogenesis in NOD-SCID mice by delivering chemokines and via stimulation of monocytes. In addition, TMVs may be storage for chemokines thus inducing chemotaxis of blood leukocytes. Conclusion: These results further support the role of TMVs in modulation of monocyte biological activity

The role of CD44H molecule in the interactions between human monocytes and pancreatic adenocarcinoma-derived microvesicles

Introduction. CD44H is a transmembrane molecule important for cell-cell and cell-extracellular matrix interactions. In monocytes, CD44H is implicated in phagocytosis of particles coated by hyaluronan (HA). HA fragments were shown to induce chemokine secretion by monocytes. Tumour derived microvesicles (TMVs), which are small membrane fragments derived from tumour cells can carry fragments of HA. The aim of the study was to examine whether monocyte’s CD44H is involved in the engulfment of pancreatic adenocarcinoma-derived microvesicles and in the production of chemokines induced by TMVs. Materials and methods. TMVs engulfment and chemokines’ secretion stimulated with TMVs were determined in control human monocytes and cells incubated with anti-CD44H monoclonal antibody (mAb) by flow cytometry and ELISA, respectively. Phosphorylation of STAT3, transcription factor essential for chemokines’ production and CD44 signal transduction, was determined by Western blotting. Results. Blocking of CD44H by anti-CD44H mAb on monocytes decreased the engulfment of TMVs and thesecretion of CCL4 and CCL5, but had no effect on CCL2, CCL3 and CXCL8. STAT-3 phosphorylation inmonocytes incubated with TMVs after CD44 blocking was also reduced. Conclusion. The results suggest that tumour-derived microvesicles (TMVs) may carry bioactive cargo(s) which induces STAT3 dependent signalling pathway in human monocytes via CD44 molecules

Tumour-derived microvesicles (TMV) mimic the effect of tumour cells on monocyte subpopulations

Background: Monocytes/macrophages may be affected by tumour cells via cell-to-cell contact, soluble factors and by tumour-derived microvesicles (TMV). Previous observations indicate that TMV interact with monocytes and alter their immunophenotype and activity. This study was designed to determine interactions of TMV with subpopulations (CD14++CD16– and CD14+CD16++) of human monocytes. Methods: Engulfment of TMV by subsets of monocytes was analysed by flow cytometry. Moreover cytokine release and production of reactive oxygen intermediates (ROI) and reactive nitrogen intermediates (RNI) by CD14++CD16– and CD14+CD16++ cells after TMV stimulation was determined. Results: It was found that TMV are engulfed more efficiently by CD14++CD16– than CD14+CD16++ cells. TMV-activated CD14++CD16– cells produce more ROI and interleukin -10 (IL-10) than CD14++CD16+. CD14+CD16++ cells following TMV stimulation showed an increased release of tumour necrosis factor alpha, IL-12p40 and RNI. Conclusion: TMV significantly modulate biological activity of monocyte subsets with a pattern similar to tumour cells. Therefore, TMV mimic the activating effect of tumour cells on monocytes as assessed by release of cytokines, ROI and RNI