Creation of synthetic life in laboratory

Ljudi su oduvijek bili fascinirani životom i pokušali su ga stvoriti više nego jednom kroz povijest. Pojavom novog područja biologije, sintetske biologije, koje kombinira znanost i inženjerstvo; je to postalo moguće. Ovaj rad daje kratki pregled događaja koji su doveli znanstvenike iz J. Craig Venter Instita (JCVI) do objave rezultate u kojima opisuju uspješno konstruiranje prve samoreplicirajuće sintetske bakterijske stanice. Tim znanstvenika je sintetizirao modificirani kromosom iz genoma bakterije Mycoplasma mycoides veličine 1.08 milijuna parova baza. Sintetska stanica je nazvana Mycoplasma mycoides JCVI-syn1.0 i dokaz je da genomi mogu biti dizajnirani u računalu, kemijski stvoreni u laboratoriju te transplantirani u domaćinsku stanicu gdje će stvoriti novu samoreplicirajuću stanicu kontroliranu samo sintetičkim genomom.People have always been fascinated by life and have tried to create it more than once throughout the history. With the appearance of new area of biology, the synthetic biology, which combines science and engineering; that become possible. This work gives a short review of events that led to publishing results of scientists at the J. Craig Venter Institute (JCVI) describing the successful construction of the first self-replicating, synthetic bacterial cell. The team of scientists synthesized the 1.08 million base pair chromosome of a modified Mycoplasma mycoides genome. The synthetic cell is called Mycoplasma mycoides JCVIsyn1.0 and is the proof of principle that genomes can be designed in the computer, chemically made in the laboratory and transplanted into a recipient cell to produce a new self-replicating cell controlled only by the synthetic genome

Urinary tract infections in children - nursing intervention

Pojam uroinfekcija označava prodor mikroorganizama u tkiva mokraćnog sustava. U najvećoj mjeri su to bakterijske infekcije, tj. infekcije u kojima se nađe značajan broj bakterija u mokraći. Bakterija E. coli uzročnik je 80 % upala mokraćnog sustava, koje se ubrajaju u najčešće bakterijske infekcije u dječjoj dobi. Češće obolijevaju djevojčice jer je mokraćna cijev kratka, ravna i široka pa bakterije lakše prodiru u mokraćni mjehur. Učestalost infekcija mokraćnog sustava također ovisi o dobi i spolu djeteta. Najveća je u male djece. Simptomi kod novorođenčeta često ne upućuju na mokraćni sustav, već se primjećuje slabije napredovanje na tjelesnoj težini, teškoće u hranjenju, žutica, povraćanje, proljev, grčevi u trbuhu, a temperatura moţe biti povišena, normalna ili čak sniţena. Do dvije godine starosti simptomi uroinfekcije su opći i nespecifični: klonulost, bezvoljnost, odbijanje jela i pića, povraćanje i povišena tjelesna temperatura. Nadalje, simptomi se mogu očitovati kao abdominalna i suprapubična bol, učestalo mokrenje, dizurija. Rezultati istraţivanja koji su provedeni na osnovi dokumentacije na Odjelu pedijatrije u Požegi, govore da su djevojčice te koje u većini slučajeva obolijevaju od urinarnih infekcija. Postotak urinarnih infekcija na Odjelu pedijatrije u OŽB Požega čini 3,3 % od ukupno hospitalizirane djece. Od velike je vaţnosti utvrditi ranu dijagnozu i započeti liječenje, jer ponavljane i neliječene uroinfekcije mogu dovesti do trajnog oštećenja bubrega. Medicinska sestra prikuplja podatke o znakovima i simptomima bolesti ovisno o dobi djeteta, od djeteta i roditelja. Utvrđuje obrasce praţnjenja crijeva zato što konstipacija može povećati rizik za uroinfekcije kod određene djece. Fizički pregled uključuje mjerenje tjelesne temperature, tlaka, pregled abdomena, procjenu osjetljivosti lumbalne loţe te pregled vanjskih genitalija (abnormalnosti). Osim vađenja krvi, važno je uzeti i urin za kemijsku analizu i urinokulturu i to kod veće djece mokrenjem na zahtjev, a kod manje kateterizacijom mokraćnog mjehura ili suprapubičnom kateterizacijom. Dužnost sestre je umiriti roditelje i informirati ih o tome zašto se neke procedure moraju provesti kroz neko vrijeme. Potrebno je poticati roditelje da verbaliziraju svoju zabrinutost i nastojati stvoriti odnos povjerenja. Također je potrebno što manje narušavati djetetovu uobičajenu rutinu – npr. majka treba nastaviti dojiti dijete. Ako dijete nema snage za sisanje, potrebno je da se majka izdoji i da se dijete hrani na bočicu izdojenim majčinim mlijekom. Dopuštajući roditeljima da participiraju u njezi djeteta, osiguravamo im mogućnost kontrole u neizvjesnoj situaciji. Sestrinska skrb za djecu koja nisu hospitalizirana uključuje skrb za pravovremenu i točnu primjenu antibiotika, osiguravanje udobnosti, dobru hidraciju djeteta, pripremu djeteta za dijagnostičke procedure i nadziranje odgovora na primijenjene mjere i moguće komplikacije. Djeca i obitelj trebaju biti educirani kako prevenirati uroinfekciju i spriječiti moguće oštećenje bubrega. U konačnici evaluacija medicinske sestre glede provedenih intervencija i očekivanog ishoda mora odgovarati na sljedeća pitanja – je li dijete još uvijek učestalo mokri, ima li dizurične tegobe, je li urinokultura negativna, uzimaju li djeca dovoljnu količinu tekućine s obzirom na dob, je li mokrenje uredno, je li djetetu propisana kontinuirana antibiotska terapija te znaju li roditelji i starija djeca opisati simptome i mjere koje će poduzeti ako se infekcija ponovi. Uloga medicinske sestre u zbrinjavanju djece s uroinfekcijom vrlo je važna jer sestra svojim radom doprinosi ranom utvrđivanju bolesti, učestvuje u liječenju i edukaciji te prevenciji, što je vrlo važno jer je poznato da ponavljane i neliječene infekcije mokraćnog sustava djece mogu dovesti do trajnog oštećenja bubrega i bubrežne funkcije

Credit Rating As a Factor of Stability in the Global Capital Market

Credit rating has an outstanding importance on the capital market. Opinions and assessments of rating agencies help us to improve growth, stability and efficiency of international and domestic markets, which now include over 80 trillion dollars of rated bonds and other securities with the fixed income. The contribution of the credit agencies to the market stability and efficiency is reflected in their ability to provide accurate, clear and reliable assessments of the solvency of participants on the financial markets. An adequate and proper risk assessment of securities contributes to stability. In order to achieve a given goal and to satisfy its purpose, the assessments should be based on a fundamental understanding of the key components of the credit risk. Also, in order to ensure a reliable framework for making investment decisions, the rating agencies are obliged to offer and to provide a wide range of securities, which are based on a global comparability of rating symbols and on the support given by the credit rating assignment committee and by the other relevant decision making bodies. Markets for structured products could not have developed without the quality assurance provided by CRAs. When analyzing a securitization program CRAs examine legal and structural protections provided to investors. Since the globalization is an inevitable phenomenon in today’s world the importance of the credit rating becomes more noticeable. On the other hand, the rating agencies have an obligation to reanalyze their decision making models in order to contribute to the reliability of the evaluation

Expression and biochemical and functional characterization of R-MC 17 antigen in rat

Razvoj T limfocita u timusu je rezultat niza dvosmernih komunikacija između timocita i ćelija mikrosredine, posredovanih pretežno adhezivnim molekulima. U Institutu za medicinska istaživanja VMA je napravljen panel monoklonskih antitela (mAt) R-MC serije, koja prepoznaju timusne epitelne ćelije (TEC). Cilj ovog rada je bio biohemijska, imunohistohemijska i funkcionalna karakterizacija antigena koga detektuje R-MC 17 mAt. Ovo mAt detektuje antigen od 205 kDa u timusu pacova. Na osnovu poređenja biohemijskih karakteristika anti-DEC 205 mAt (HD83) i unakrsne imunoprecipitacije antigena iz lizata timusa pacova pomoću ova dva mAt zaključeno je da R-MC 17 mAt detektuje pacovski DEC-205 molekul. R-MC 17 antigen je jako ispoljen na kortikalnim TEC, medularnim timusnim dendritskim ćelijama (TDC), DC u parakorteksu i interfolikularnim zonama limfnog čvora i DC u periarteriolarnom limfnom omotaču slezine. Slaba imunoreaktivnost je detektovana na limfocitima u folikulima limfnog čvora i slezine i makrofagama u crvenoj pulpi slezine. Takođe, ekspresija DEC-205 molekula je detektovana na oko 40% ćelija TEC linije sa karakteristikama ćelija „dadilja“. R-MC 17 mAt podstiče vezivanje i emperipoezu timocita od strane ove epitelne linije. Istovremeno, ekspresija R-MC 17 antigena je podjednaka na subpopulacijama TDC pacova (CD11b+ i CD11b-), i ne menja se pod uticajem GM-CSF-a. Oba anti-DEC 205 mAt (R-MC 17 mAt i HD83 mAt) stimulišu apoptozu timocita u kokulturi sa TDC i ovaj proces je povezan sa povećanim vezivanjem timocita za TDC. R-MC 17 mAt inhibira proliferaciju singenih timocita i alogenih T limfocita pacova u kokulturi sa TDC u prisustvu anti-TCR (R7.3) mAt. Intratimusna aplikacija R-MC 17 mAt dovodi do smanjenja celularnosti timusa pacova što je povezano sa indukcijom apoptoze timocita. Fenotipska analiza subpopulacija timocita nakon intratimusne aplikacije R-MC 17 mAt pokazuje smanjenje ukupnog broja CD4+CD8+ i CD4+CD8-, a povećanje ukupnog broja CD4-CD8+ timocita, dok je ukupan broj zrelih CD5+TCRjako+ timocita nepromenjen. Sveukupno, ovi rezultati prvi put opisuju potencijalnu funkciju R-MC 17 antigena u timusu.Opisani mehanizmi mogu biti povezani sa procesima intratimusne diferencijacije timocita i prezentacije vi autoantigena u timusu od strane kortikalnih TEC i medularnih TDC, a što u krajnjem ishodu ima uticaj na procese selekcije (delecije autoreaktivnih klonova timocita).Intrathymic development of T lymphocyte is mainly governed by an array of bidirectional interactions between thymocytes and thymic microenvironment, which are mostly provided by the adhesive molecules. The monoclonal antibody (mAb) panel of R-MC series, designed against thymic epithelial cells (TEC), was developed at the Institute of medical research (MMA). The aim of this doctoral thesis was focused on biochemical, immunohistochemical and functional characterization of the R-MC 17 antigen, detected by R-MC 17 mAb. R-MC 17 mAb recognizes the 205 kDa molecule expressed in the rat thymus. Based on similarity in biochemical features with an anti- DEC-205 mAb (HD83), and cross-immunoprecipitation of antigen from the thymic lysate with these two mAbs, R-MC 17 was considered to be the antibody specific for the rat DEC-205 molecule. Cortical TEC, medullary thymic dendritic cells (TDC), lymph node dendritic cells (DC) in paracortex and interfollicular zones, and spleen paraarteriolar-lymphoid sheaths DC, are highly positive with R-MC 17. Lymphocytes in lymph node and spleen follicles and red pulp macrophages in spleen, show weak R-MC 17 reactivity. About 40% of TEC-line cells with nursing characteristics express R-MC 17 antigen as well. R-MC 17 stimulates thymocyte binding and emperipoesis displayed by this cell line. Concomitantly, R-MC 17 antigen is equally expressed on two rat TDC subpopulations (CD11b+ and CD11b-), and cultivation with GM-CSF has no effect. RMC 17, as well as HD83, stimulates thymocyte apoptosis in co-culture with TDC, and this process is associated with the increased binding of thymocytes to TDC. R-MC 17 mAb inhibits proliferation of syngeneic thymocytes and allogeneic T lymphocytes in anti-TCR (R7.3)-stimulated cocultures with TDC. Intrathymic injection of R-MC 17 mAb decreases thymic cellularity, which was related to the induction of thymocyte apoptosis. Phenotypic analysis of thymocyte populations after intrathymic application of R-MC 17 mAb, indicates decrease of the absolute number of CD4+CD8+ and CD4+CD8- , and increase of CD4-CD8+ thymocyte subpopulations, while the absolute number of mature CD5+TCRhigh thymocytes stays unchanged. Taken together, these results show, for the first time, a potential function of R-MC 17 antigen in rat thymus. Mechanisms viii described, are potentially involved in the processes of intrathymic differentiation and the presentation of autoantigen in the thymus by the cortical TEC and medullary TDC, having final implications in the processes of thymic selection (deletion of autoreactive thymocytes)

HEALTH KNOWLEDGE AND OPINIONS OF PARENTS OF CHILDREN WITH BRONCHITIS

Uvod: Bronhitis je upala sluznice dušnika i bronha, a uzrokuju je virusi i rjeđe bakterije. Navedenu bolest mogu dobiti sve dobne strukture od dojenčadi do osoba starije životne dobi. Bronhitis može biti akutni i kronični. Zdravstveno znanje roditelja izravno utječe na sposobnost preuzimanja kontrole nad vlastitim zdravljem i zdravljem svoje djece. Ciljevi: Ovim istraživanjem se željelo ispitati zdravstvena znanja i stavove roditelja djece oboljele od bronhitisa. Ispitano je njihovo znanje o bronhitisu, simptomima bolesti, uzrocima bolesti, periodu po-javljivanja, učestalosti oblijevanja njihovog djeteta od bronhitisa, znanje o korištenju pomagala za ublažavanje simptoma bolesti, te dijagnostičkim postupcima i načinu liječenja bronhitisa. U studiji su uspoređene i navike ponašanja roditelja djece oboljele od bronhitisa u svakodnevnom životu vezane uz pušenje i redovito prozračivanje prostora u kojem boravi njihovo dijete. Metode: U istraživanju je sudjelovalo 115 roditelja koji su doveli djecu sa simptomima bronhitisa u pedijatrijsku ambulantu Opće županijske bolnice Požega. Istraživanje je provedeno uz pomoć anonim-nog anketnog upitnika kreiranog za ovo istraživanje, a upitnik je sadržavao 35 pitanja, od kojih se 4 pitanja odnose na demografske karakteristike ispitanika (spol, dob, mjesto stanovanja i stručnu spremu), dok se ostatak pitanja odnosi na znanje i stavove ispitanika o bronhitisu te o navikama ponašanja u svakodnevno životu. Rezultati: Pokazalo se kako je zdravstveno znanje roditelja djece oboljele od bronhitisa najviše pov-ezano sa stupnjem obrazovanja, dobi roditelja i mjestom stanovanja, a najmanje sa spolom roditelja. Roditelji su pokazali dobro znanje o bolesti, simptomima, uzrocima bolesti, te dijagnostičkim postupcima i načinu liječenja, dok su značajno manje znanje pokazali o pušenju i njegovom štetnom utjecaju na zdravlje njihovog djeteta. Zaključak: Zdravstveno znanje i stavovi roditelja imaju veliki utjecaj na umanjivanje rizika od nastanka bronhitisa i na učestalost njegovog pojavljivanja.Objective: The aim of this study is to examine knowledge and attitudes about health of parents whose children suffer from bronchitis. Their knowledge of bronchitis, symptoms of illness, causes of illness, period of occurrence, frequency of their child\u27s bronchitis, knowledge of the use of aids to alleviate the symptoms of the disease, as well as diagnostic procedures and the methods of treating bronchitis were examined. The study also compares the behavioral habits of parents whose children suffer from bron-chitis in their everyday life which are connected to smoking and regular ventilation of their child\u27s liv-ing space. Methods: 115 parents who brought their children with bronchitis symptoms to the pediatric depart-ment of the General County Hospital in Požega participated in the research. The survey was conducted by an anonymous questionnaire composed by the author, and the questionnaire contained 35 questions, four of which are related to the demographic characteristics of the respondents (sex, age, place of resi-dence and vocation), while the remaining questions are related to knowledge and attitudes of respond-ents about bronchitis and the behavioral habits in everyday life. Results: The results of the research have shown that the knowledge of health of parents whose chil-dren suffer from bronchitis is most related to the degree of education, the age of the parents and the place of residence, and least to the parents\u27 sex. Parents have shown good knowledge of the disease, symptoms, causes of the disease, and diagnostic procedures and treatment, while significantly less knowledge has been shown about smoking and its adverse impact on the health of their child. Conclusion: Parents\u27 knowledge and attitudes about health have a major influence on diminishing the risk of bronchitis and the frequency of its occurrence

Prognostic Value of Venoarterial Carbon Dioxide Gradient in Patients with Severe Sepsis and Septic Shock

Aim To investigate the changes in the venoarterial carbondioxide gradient (V-a Pco2) and its prognostic value for survival of patients with severe sepsis and septic shock. Methods The study was conducted in General Hospital Holy Spirit from January 2004 to December 2007 and included 71 conveniently sampled adult patients (25 women and 46 men), who fulfilled the severe sepsis and septic shock criteria and were followed for a median of 8 days (interquartile range, 12 days). The patients were divided in two groups depending on whether or not they had been mechanically ventilated. Both groups of patients underwent interventions with an aim to achieve hemodynamic stability. Mechanical ventilation was applied in respiratory failure. Venoarterial carbon dioxide gradient was calculated from the difference between the partial pressure of arterial CO2 and the partial pressure of mixed venous CO2, which was measured with a pulmonary arterial Swan-Ganz catheter. The data were analyzed using Kaplan-Meier survival analysis, along with a calculation of the hazard ratios. Results There was a significant difference between nonventilated and ventilated patients, with almost 4-fold greater hazard ratio for lethal outcome in ventilated patients (3.85; 95% confidence interval, 1.64-9.03). Furthermore, the pattern of changes of many other variables was also different in these two groups (carbon dioxide-related variables, variables related to acid-base status, mean arterial pressure, systemic vascular resistance, lactate, body mass index, Acute Physiology and Chronic Health Evaluation II, Simplified Acute Physiology II Score, and Sepsis-related Organ Failure Assessment score). Pco2 values (with a cut-off of 0.8 kPa) were a significant predictor of lethal outcome in non-ventilated patients (P = 0.015) but not in ventilated ones (P = 0.270). Conclusion V-a Pco2 was a significant predictor of fatal outcome only in the non-ventilated group of patients. Ventilated patients are more likely to be admitted with a less favorable clinical status, and other variables seem to have a more important role in their outcome

Efekti aspirina na apoptozu neutrofilnih granulocita

Neutrophils are a part of the immune system, and they are involved in host defence against microorganisms. Neutrophil granulocytes have the shortest lifespan among leukocytes, which can be modulated by cytokines and pharmacological agents. The effect of aspirin on apoptosis of inflammatory granulocytes has not been studied in detail yet, and therefore was the chosen subject of this study. Inflammatory granulocytes have been isolated from polyvinyl sponges implanted under the skin of Albino Oxford (AO) rats. Inflammatory cells that were isolated 20 hours later were more than 95% neutrophil granulocytes. The cells were cultivated 24 h with different concentrations of aspirin ranging from 1 µM to 10 mM. After the cultivation period, apoptosis of neutrophils was assessed by morphological criteria, as well as by flow cytometry (after staining the cells with propidium iodide). We found that at concentrations from 0,1 mM to 2,5 mM aspirin inhibited apoptosis of granulocytes, but at 10 mM aspirin induced apoptosis of these cells.Neutrofilni granulociti su deo imunog sistema, uključenog u odbranu od mikroorganizama. Oni imaju najkraći životni vek među leukocitima, koji se može modulisati citokinima i farmakološkim agensima, a do sada nije ispitivan efekat aspirina na apoptozu inflamatornih granulocita. Zbog toga je u ovoj studiji ispitivan efekat aspirina na apoptozu inflamatornih neutrofilnih granulocita pacova. Inflamatorni granulociti su izolovani iz polivinilskih sunđera, potkožno implantiranih, pacovima Albino Oxford (AO) soja. Inflamatorne ćelije, izolovane 20 sati kasnije, najvećim delom (više od 95 %) predstavljaju neutrofilne granulocite. Ove ćelije su kultivisane 24 sata sa aspirinom u koncentracijama od 1 µM do 10 mM. Posle ovog perioda supernatanti su sakupljani i korišćeni za merenje koncentracije NO. Ćelije su bojene propidijum jodidom i apoptoza je analizirana na protočnom citofluorimetru, kao i pomoću morfoloških kriterijuma. Ustanovljeno je da u koncentracijama od 0,1 do 2,5 mM aspirin inhibira, a samo u visokim koncentracijama (10 mM) indukuje apoptozu ovih ćelija. Aspirinom indukovana apoptoza je bila u pozitivnoj korelaciji sa smanjenom produkcijom NO

Assessment of the Biological Effects of Pellia endiviifolia and its Constituents in Vitro

Liverworts are characterized by a high content of bioactive compounds reported to show antimicrobial, anticancer, and antioxidant properties. In this study, the biological effects of the methanol extract of the liverwort Pellia endiviifolia and its constituents, bis-bibenzyls perrottetin E, 10′-hydroxyperrottetin E, and 10,10′-dihydroxyperrottetin E, were investigated using human peripheral blood cells as a model system. The assessment of the investigated compounds comprised testing their genotoxicity, apoptotic potential, and redox modulating activities. The genotoxicity testing indicated that medium (25 µM) and high concentrations (100 µM) of the investigated compounds displayed genotoxic and antiproliferative effects in human lymphocytes as revealed by significant, concentration-dependent enhancement of the micronuclei incidence and decrease in the cytokinesis-block proliferation index compared to the control (P .05). The observed genotoxic, antiproliferative, and proapoptotic effects of the investigated compounds make them suitable for further comprehensive studies related to their possible applications as anticancer agents

The importance of tests applied to evaluate the effectiveness of antiplatelet therapy in patients with recurrent coronary stent thrombosis

Background. Stent thrombosis is potentially lethal complication with huge economic burden. The role of insufficient response to antiplatelet therapy is still unclear reason for its occurrence. Case report. We presented 54-year-old man with recurrent stent thrombosis on the 4th, 9th and 12th day after the primary percutaneous coronary intervention in spite of double antiaggregation therapy (aspirin+ clopidogrel). All possible procedural causes were excluded and reimplantation of intracoronary stent was insufficient to resolve the problem, so four platelet tests were performed: flow cytometry, Platelet Function Analyzer-100 test, aggregometry, and determination of gene polymorphism for P2Y12 receptor (directly involved in the mechanism of thienopyridine), and GPIIbIIIa receptor (final receptor in aggregation). The patient was the carrier of the major haplotype H1H1 for P2Y12 receptor and minor A1A2 for GPIIbIIIa receptor. The results of all the performed tests showed insufficient antiplatelet effect of aspirin and sufficient response to thienopyridin (not to clopidogrel, but to ticlopidine). Conclusion. Performance of platelet function tests is necessary in the case of major adverse cardiac events especially stent thrombosis, after implantation of intracoronary stent

Support for students with disabilities in higher education in the Republic of Croatia

Glavni je cilj ovog rada dobiti uvid u oblike i kvalitetu podrške namijenjenu studentima s invaliditetom u visokoškolskom obrazovanju u Republici Hrvatskoj. Uvid je dobiven prilikom analiziranja postojeće dokumentacije i relevantnih rezultata istraživanja. Prilikom analiziranja osam sveučilišta u Republici Hrvatskoj (Split, Zadar, Rijeka, Dubrovnik, Pula, Zagreb, Osijek i Slavonski Brod), dobiven je uvid o tome kako je pozicionirana podrška studentima s invaliditetom u sveučilišnim dokumentima i u pravnim dokumentima Republike Hrvatske te djelovanje i ustrojstvo Ureda za studente s invaliditetom.The main goal of this paper is to gain insight into the forms and quality of support intended for students with disabilities in higher education in the Republic of Croatia. The insight was obtained during the analysis of existing documentation and relevant research results. During the analysis of eight universities in the Republic of Croatia (Split, Zadar, Rijeka, Dubrovnik, Pula, Zagreb, Osijek and Slavonski Brod), an insight was gained into how support for students with disabilities is positioned in university documents and in legal documents of the Republic of Croatia, as well as the operation and organization. Office for students with disabilities