Influence of pro- and anti -inflammatory factors on production of extracellular matrix proteases uPA and MMP9 in mouse macrophages

Макрофази су централне ћелије имунског система јер поседују јединствене особине: учествују у одржавању физиолошке хомеостазе ткива и организма, процени врсте могуће претње по организам и покретању адекватног имунског одговора ангажовањем одговарајућих компоненти имунског система, али и његовом завршетку када више није неопходан, активно учествују у обнављању оштећених ткива и тако омогућавају повратак хомеостазе. Остваривање овако разноврсних улога омогућује висока прилагодљивост ових ћелија спољашњим сигналима. Различити подтипови макрофага имају одлучујућу улогу у различитим фазама инфламације. Класични М1 макрофази, изложени дејству про-инфламацијских цитокина, као што је интерферон γ, и/или продуктима микроорганизама, као што је липополисахарид (LPS), имају главну улогу у индукцији инфламације и елиминацији патогена, док алтернативни М2 макрофази, у одговору на стимулацију анти-инфламацисјким/имунореглаторним цитокинима IL-4, IL-13, IL-10, TGF-β или глукокортикоидима, обустављају инфламацију и кључни су за обнављање ткива и зарастање рана. Током инфламације одвијају се значајне промене на нивоу протеина ванћелијског матрикса (ЕСМ, extracellular matrix), које су строго регулисане протеазама ЕСМ, чија се продукција остварује у дефинисаном просторно-временском контексту. Секрецијом матриксних протеаза (ММР) као што је ММР9, М1 макрофази учествују у деградацији ЕСМ и разградњи ткива, што олакшава улазак других инфламацијских ћелија у оштећено ткиво. Истовремено се покреће каскада коагулације током које се формира привремена структура ЕСМ од фибрина, која се касније у фази резолуције разграђује, а ЕСМ се обнавља. Урокиназа (uPA) је серинска протеаза која разградњом плазминогена у плазмин остварује главну улогу у процесу разградње фибринске мреже. Осим тога, uPA је важан регулатор ткивне инфламације, процеса зарастања рана и ткивне регенерације. Отуда је важно расветлити молекулски механизам регулације ММР9 и uPA током промене поларизације макрофага из М1 у М2. На основу ових података, претпостављено је да се у инфламацијским макрофазима ММР9 и uPA регулишу на различит начин...Macrophages are one of the major effector cells in inflammation. They are responsible for the recognition and processing of foreign materials, activation of the adaptive immune response, regulation of tissue repair/regeneration, and overall homeostasis maintenance. Multiple functions of macrophages are facilitated by their high plasticity in response to environmental or intrinsic signals. Two polarized macrophage modalities are critical in the regulation of different stages of tissue repair after injury. Classical M1 macrophages, exposed to pro-inflammatory cytokines, IFNs and microbial products, such as lipopolysaccharide (LPS), have major role in inducing inflammation and clearing of pathogens, whereas alternative M2 macrophages, in response to IL-4 and IL- 13, IL-10, TGF-β or glucocorticoids, resolve inflammation and are crucial in tissue remodeling and wound healing. In addition to cellular changes, reorganization/remodeling of critical extracellular matrix (ECM) proteins also occurs in injured tissues. During this ECM reorganization, several important ECM proteinases are triggered in a space-temporal fashion through tightly regulated mechanisms. Proinflammatory M1 macrophages are able to secrete matrix metalloproteinases, such as MMP9, which help in degradation of the ECM in affected tissue, thus facilitating the recruitment of inflammatory cells to the site of tissue injury. Moreover, an antifibrinolytic-coagulation cascade, that activates clotting and development of a provisional ECM, is also triggered, and later replaced by normal ECM during the resolution of inflammation. The urokinase-type plasminogen activator (uPA) is a serine protease that plays a major role in fibrinolytic processes, where it converts plasminogen to plasmin which further degrades coagulation components. Moreover, uPA is a key regulator of tissue inflammation and wound-healing processes and tissue regeneration, and is also secreted by macrophages. Considering macrophage’s importance in tissue repair, it is crucial to elucidate how MMP9 and uPA are involved in M1-M2 transition, as well as what are the underlying molecular mechanisms..

Mesenchymal stem cells isolated from human periodontal ligament

Mesenchymal stem cells (MSCs) were isolated from human periodontal ligament (hPDL-MSCs) and characterized by their morphology, clonogenic efficiency, proliferation and differentiation capabilities. hPDL-MSCs, derived from normal impacted third molars, possessed all of the properties of MSC, including clonogenic ability, high proliferation rate and multi-lineage (osteogenic, chondrogenic, adipogenic, myogenic) differentiation potential. Moreover, hPDL-MSCs expressed a typical MSC epitope profile, being positive for mesenchymal cell markers (CD44H, CD90, CD105, CD73, CD29, Stro-1, fibronectin, vimentin, alpha-SMA), and negative for hematopoietic stem cell markers (CD34, CD11b, CD45, Glycophorin-CD235a). Additionally, hPDL-MSCs, as primitive and highly multipotent cells, showed high expression of embryonic markers (Nanog, Sox2, SSEA4). The data obtained provided yet further proof that cells with mesenchymal properties can be obtained from periodontal ligament tissue. Although these cells should be further investigated to determine their clinical significance, hPDL-MSCs are believed to provide a renewable and promising cell source for new therapeutic strategies in the treatment of periodontal defects

Mesenchymal stem cell properties of dental pulp cells from deciduous teeth

In the present study we have isolated and identified mesenchymal stem cells (MSCs) from the exfoliated deciduous teeth dental pulp (DP-MSCs), as plastic-adherent, spindle-shaped cells with a high proliferative potential. Immunophenotype analyses revealed that DP-MSCs were positive for mesenchymal cell markers (CD90, CD44, CD105, STRO-1, vimentin and α-SMA), and negative for hematopoietic stem cell markers (CD11b, CD33, CD34, CD45, CD235a). DPMSCs were also capable of differentiating into adipogenic, chondrogenic, myogenic and osteogenic lineages, fulfilling the functional criterion for their characterization. These results demonstrate that DP-MSCs offer a valuable, readily accessible source to obtain and store adult stem cells for future use

Kultivacija matičnih i progenitorskih ćelija hematopoeze iz kostne srži hrčka

Hamster, a hibernating animal, is an important experimental model in research on the influence of hypothermia on different physiological processes. A simple procedure for cultivation and identification of hamster hematopoetic stem cells (HSC) and hematopoetic progenitor cells (HPC) is a premise for a successful investigation upon hypothermia effects on hematopoiesis. The aim of this work was to evaluate the utilization of commercially available methylcellulose media (MC) and recombinant mouse and human cytokines for hamster HSC and HPC assays, in order to enable further studies on these cells. Hamster bone marrow mononuclear cells (BMMNC) were plated in MC containing cytokines that support mouse or human HPC growth. Also, BMMNC were resuspended in cytokine supplemented liquid media and incubated for 5 weeks with a four day monitoring of viable cell number. We demonstrated that hamster hematopoietic progenitor cells committed for erythroid lineage and myeloid lineage successfully formed recognizable colonies in both mouse and human MC, while multipotent progenitor cells formed colonies only in mouse MC. We also defined conditions for the evaluation of hamster HSC activity in liquid cultures, based on continuous 5 weeks HSC proliferation. The obtained results verify the utilization of mouse specific MC for further research on hamster HPC biology during hypothermia.Fiziološka hibernacija u koju hrčci ulaze prilikom izlaganja niskim temperaturama, čini ove životinje zanimljivim eksperimentalnim modelom za ispitivanje hematopoeze u uslovima hipotermije. Preduslov za ovo ispitivanje je postojanje jednostavne metode za kultivaciju i identifikaciju hematopoetskih ćelija hrčka. Cilj ovog rada je bio da se ispita mogućnost kultivacije progenitorskih ćelija hematopoeze hrčka u kompletnoj metil celulozi dizajniranoj za kultivaciju mišijih i humanih hematopoetskih ćelija, kao i da se odrede optimalni uslovi za kultivaciju matičnih ćelija hematopoeze hrčka u tečnoj kulturi. Mononuklearne ćelije kostne srži hrčka su posađene u metil celulozu i u tečnu kulturu. Oba medijuma su sadržala kombinacije rekombinantnih mišijih i/ili humanih citokina. Kolonije progenitorskih ćelija opredeljenih za mijelopoezu i opredeljenih za eritropoezu su se formirale u metil celulozi dizajniranoj za kultivaciju mišijih i humanih hematopoetskih ćelija, dok su se primitivnije kolonije sastavljene od oba tipa ćelija (mijeloidna i eritrocitna loza) formirale samo u metil celulozi dizajniranoj za kultivaciju mišijih hematopoetskih ćelija. Osim toga, populacija matičnih ćelija hematopoeze hrčka je proliferisala u tečnim kulturama tokom 5 nedelja bez znakova opadanja proliferativnog potencijala. Ova istraživanja pokazuju da se primenjene metode mogu uspešno koristiti za ispitivanje hematopoeze kod hrčka

GEML: A Grammatical Evolution, Machine Learning Approach to Multi-class Classification

In this paper, we propose a hybrid approach to solving multi-class problems which combines evolutionary computation with elements of traditional machine learning. The method, Grammatical Evolution Machine Learning (GEML) adapts machine learning concepts from decision tree learning and clustering methods and integrates these into a Grammatical Evolution framework. We investigate the effectiveness of GEML on several supervised, semi-supervised and unsupervised multi-class problems and demonstrate its competitive performance when compared with several well known machine learning algorithms. The GEML framework evolves human readable solutions which provide an explanation of the logic behind its classification decisions, offering a significant advantage over existing paradigms for unsupervised and semi-supervised learning. In addition we also examine the possibility of improving the performance of the algorithm through the application of several ensemble techniques

Influence of pro- and anti -inflammatory factors on production of extracellular matrix proteases uPA and MMP9 in mouse macrophages

Макрофази су централне ћелије имунског система јер поседују јединствене особине: учествују у одржавању физиолошке хомеостазе ткива и организма, процени врсте могуће претње по организам и покретању адекватног имунског одговора ангажовањем одговарајућих компоненти имунског система, али и његовом завршетку када више није неопходан, активно учествују у обнављању оштећених ткива и тако омогућавају повратак хомеостазе. Остваривање овако разноврсних улога омогућује висока прилагодљивост ових ћелија спољашњим сигналима. Различити подтипови макрофага имају одлучујућу улогу у различитим фазама инфламације. Класични М1 макрофази, изложени дејству про-инфламацијских цитокина, као што је интерферон γ, и/или продуктима микроорганизама, као што је липополисахарид (LPS), имају главну улогу у индукцији инфламације и елиминацији патогена, док алтернативни М2 макрофази, у одговору на стимулацију анти-инфламацисјким/имунореглаторним цитокинима IL-4, IL-13, IL-10, TGF-β или глукокортикоидима, обустављају инфламацију и кључни су за обнављање ткива и зарастање рана. Током инфламације одвијају се значајне промене на нивоу протеина ванћелијског матрикса (ЕСМ, extracellular matrix), које су строго регулисане протеазама ЕСМ, чија се продукција остварује у дефинисаном просторно-временском контексту. Секрецијом матриксних протеаза (ММР) као што је ММР9, М1 макрофази учествују у деградацији ЕСМ и разградњи ткива, што олакшава улазак других инфламацијских ћелија у оштећено ткиво. Истовремено се покреће каскада коагулације током које се формира привремена структура ЕСМ од фибрина, која се касније у фази резолуције разграђује, а ЕСМ се обнавља. Урокиназа (uPA) је серинска протеаза која разградњом плазминогена у плазмин остварује главну улогу у процесу разградње фибринске мреже. Осим тога, uPA је важан регулатор ткивне инфламације, процеса зарастања рана и ткивне регенерације. Отуда је важно расветлити молекулски механизам регулације ММР9 и uPA током промене поларизације макрофага из М1 у М2. На основу ових података, претпостављено је да се у инфламацијским макрофазима ММР9 и uPA регулишу на различит начин...Macrophages are one of the major effector cells in inflammation. They are responsible for the recognition and processing of foreign materials, activation of the adaptive immune response, regulation of tissue repair/regeneration, and overall homeostasis maintenance. Multiple functions of macrophages are facilitated by their high plasticity in response to environmental or intrinsic signals. Two polarized macrophage modalities are critical in the regulation of different stages of tissue repair after injury. Classical M1 macrophages, exposed to pro-inflammatory cytokines, IFNs and microbial products, such as lipopolysaccharide (LPS), have major role in inducing inflammation and clearing of pathogens, whereas alternative M2 macrophages, in response to IL-4 and IL- 13, IL-10, TGF-β or glucocorticoids, resolve inflammation and are crucial in tissue remodeling and wound healing. In addition to cellular changes, reorganization/remodeling of critical extracellular matrix (ECM) proteins also occurs in injured tissues. During this ECM reorganization, several important ECM proteinases are triggered in a space-temporal fashion through tightly regulated mechanisms. Proinflammatory M1 macrophages are able to secrete matrix metalloproteinases, such as MMP9, which help in degradation of the ECM in affected tissue, thus facilitating the recruitment of inflammatory cells to the site of tissue injury. Moreover, an antifibrinolytic-coagulation cascade, that activates clotting and development of a provisional ECM, is also triggered, and later replaced by normal ECM during the resolution of inflammation. The urokinase-type plasminogen activator (uPA) is a serine protease that plays a major role in fibrinolytic processes, where it converts plasminogen to plasmin which further degrades coagulation components. Moreover, uPA is a key regulator of tissue inflammation and wound-healing processes and tissue regeneration, and is also secreted by macrophages. Considering macrophage’s importance in tissue repair, it is crucial to elucidate how MMP9 and uPA are involved in M1-M2 transition, as well as what are the underlying molecular mechanisms..

Attitudes of oncologists, family doctors, medical students and lawyers to euthanasia

The purpose of this survey was to define attitudes and opinions of two types of physicians, medical students and lawyers in the area of euthanasia and related issues and problems. A questionnaire was used as the source of data. There were four groups of test persons: oncologists, home care physicians (family doctors), third-year medical students and lawyers. The questionnaire included 22 questions, 4 of which concerned general characteristics of tested persons (including religious belief), while 18 referred to the problems of euthanasia. The total number of tested persons was 123. 55 men and 68 women with a median age of 38 +/- 11 years (+/-SD). There were 30 test persons in the group of oncologists, 31 in the group of family doctors, 31 in the group of third-year students, and 31 in the group of lawyers. Between 97% and 100% of individuals gave scored responses to most items. More than half of the individuals (57%) were against euthanasia, and 61% are against the legalization of euthanasia. The views of doctors and medical students were similar (2/3 against) and significantly different from the view of lawyers (2/3 for, P lt 0.01). The legalization of euthanasia is favored by 61% of lawyers, in contrast to 43%, 30% and 23% of oncologists, family doctors and medical students, respectively. Overall, 31% sais they would apply euthanasia if they were asked for it, and 36% that would if it had been legalized. Lawyers are twice as willing to perform euthanasia as students or physicians. The least ready to apply euthanasia are physicians working as oncologists (only 1 in 5). Compared with oncologists, one-third of home-care physicians would perform euthanasia anyway, whether legalized or not. Most of the test persons were of the opinion that euthanasia should be performed in the case of children born with a severe anomaly. None of the tested groups considered invalidity or being a burden to the family important reasons for the termination of somebody's life. Approximately 40% of responders believed that the decision for euthanasia should be made by the patient alone. Only lawyers were of the opinion that the misuse of euthanasia could be controlled. Our study shows that it is probably more important to determine factors associated with behavior pertaining to euthanasia in physicians working closely with suffering patients. Reducing suffering and launching a hospice movement and palliative care services might be the most appropriate way to deal with the problem of euthanasia