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Determination of sibutramine content in food supplements by HPLC
Get PDFU današnje vrijeme znatno su rašireni dodaci prehrani sa svrhom pomaganja mršavljenju. Djelovanje takvih suplemenata se temelji na laţnom osjećaju sitosti. Djelatna tvar u lijekovima za mršavljenje je sibutramin koji djeluje na mehanizam ponovne resorpcije serotonina i norepinefrina. Sibutramin je prema strukturi sliĉan amfetaminima i stoga spada u strogo kontrolirane djelatne tvari. Unazad 15-tak godina pojavio se veliki broj sluĉajeva srĉanih udara i moţdane kapi osoba koje su uzimale lijekove ĉija je djelatna tvar sibutramin. Nakon provedenih istraţivanja Europska agencija za lijekove (EMA) je obustavila distribuciju lijeka po Europi. Ubrzo nakon povlaĉenja lijekova s trţišta sibutramin se pojavio u raznim dodacima prehrani poput zelene kave, ĉajeva za mršavljenje i sl. koji su deklarirani kao prirodni proizvodi. Krivotvorenje lijekova i dodataka prehrani je sve veći globalni problem koji moţe rezultirati narušavanjem sigurnosti i zdravlja pacijenata.
Navedeno istraţivanje provedeno je identifikacijom i odreĊivanjem sadrţaja sibutramina u dodacima prehrani s hrvatskog trţišta tehnikom tekućinske kromatografije. Poznato je da uzimanje 10 mg sibutramina dnevno dovodi do znaĉajnih nuspojava poput povećanog krvnog tlaka, povećanog broja otkucaja srca, srĉanog i moţdanog udara.
Ispitano je 8 uzoraka dodataka prehrani za pomoć pri mršavljenju prikupljenih na hrvatskom trţištu. Niti jedan od ispitanih uzoraka nije pokazao pozitivne rezultate na sibutramin.Nowadays, dietary supplements whose purpose is to reduce body weight are considerably widespread. The action of such supplements is based on a false sense of satiety. The active drug for weight loss is sibutramine that acts on the mechanism of re-absorption of serotonin and norepinephrine. Sibutramine, according to a structure similar to amphetamines, therefore belongs to strictly controlled substances. In the last 15 years, a large number of cases of heart attacks and brain drops occurred in people who consumed drugs whose active substance is sibutramine. After the research has been carried out, the European Medicines Agency has suspended the distribution of drugs across Europe. Soon after the withdrawal of the drugs from the market, substantial concentrations of sibutramine were found in various dietary supplements such as green coffee, weight loss teas, etc., declared as natural products. Counterfeiting of medicines and food supplements is an increasing global problem that can result in disruption of patient safety and health.
This research will be conducted by identification and content determination of sibutramine in dietary supplements from the Croatian market by liquid chromatography technique(HPLC). It is well known that consuming 10 mg of sibutramine daily leads to significant side effects such as increased blood pressure, increased heart rate, heart attack and stroke.
Eight samples of dietary supplements collected on the Croatian market were tested and none of the respondents gave positive results on sibutramine
Spinal cord stem cells and molecules that control them
No full textOzljede leđne moždine predstavljaju veliki medicinski problem jer trenutno za njih nema efikasne terapije. Prilikom ozljede dolazi do oštećenja motoneurona i prekida u prijenosu živčanih impulsa. Terapeutski potencijal u liječenju ozljeda leđne moždine imaju neuralne matične stanice leđne moždine, kojima posebno obiluje lumbarna regija. Kao najbolji modeli za istraživanje ozljeda leđne moždine pokazali su se glodavci i posebno niži kralježnjaci koji imaju mogućnost potpune regeneracije leđne moždine nakon ozljede. Ependimalne stanice koje okružuju središnji kanal leđne moždine sastoje se od nekoliko podtipova stanica, a pojedini podtipovi posjeduju svojstva matičnih stanica. U istraživanju su analizirane stanice ependimalne i subependimalne regije leđne moždine, njihovi specifični markeri te njihova uloga u fiziološkim i patofiziološkim stanjima. Na primjer, detaljno je opisana izuzetna važnost ovih stanica u stvaranju glijalnog ožiljka nakon ozljede leđne moždine. Također, u radu su opisana i razmotrena istraživanja koja su dokazala da se aktivnost ependimalnih stanica nakon ozljede mijenja ovisno o starosti organizma. Nadalje, opisana su i druga brojna istraživanja koja su usmjerena prema genetičkom reprogramiranju ovih stanica kako bi se povećala njihova proliferacija i diferencijacija u živčane stanice s ciljem pronalaska terapija ne samo za ozljede leđne moždine, već i za druge bolesti poput multiple skleroze ili amiotrofične lateralne skleroze.Spinal cord injury represent a major medical problem because there is currently no effective therapy. In the event of injury there is a loss of motoneuron function and interruption in the transmission of nerve impulses. Therapeutic potential in the treatment of spinal cord injuries have neural stem cells of the spinal cord, particularly abundant in the lumbar region. Rodents and lower vertebrates are the best models for spinal cord injury research because they have the possibility of complete regeneration of the spinal cord after injury. The ependymal cells surrounding the central spinal cord are made up of several cell subtypes, including cells that possess stem cell properties. In this paper, the cells of the ependymal and subependimal spinal cord regions, their specific markers, and their role in physiological and pathophysiological conditions are described in detail. For example, the importance of these cells in the formation of the glial scar after spinal cord injury. Also, the paper describes and analyzes studies that have shown that the activity of ependymal cells after injury changes depending on the age of the organism and other numerous studies aimed the genetic reprogramming of these cells in order to increase their proliferation and differentiation in nerve cells in order to find therapy not only for spinal cord injuries, but also for other diseases such as multiple sclerosis or amyotrophic lateral sclerosis
The role of tumor microenvironment in development of chemoresistance
Get PDFMnoga istraživanja potvrdila su kompleksnost tumora kojeg ne čine samo tumorske stanice, nego i mikrookoliš koji se sastoji od stromalnih stanica (fibroblasta, imunih stanica, endotelnih stanica, mezenhimalnih matičnih stanica, adipocita) i izvanstaničnog matriksa. Putem raznih faktora i enzima koje luče, kao što su IL-6, TGF- β, MMPs, HIF-1 potiču proliferaciju tumora, angiogenezu i kemorezistenciju. CAFs su najmnogobrojnije stanice u tumorskom mikrookolišu te bi se njih trebalo posebno uzeti u obzir pri istraživanju novih terapija. Mikrookoliš utječe na agresivnost i metastaziranje tumora, a kroz interakcije sa stanicama raka putem različitih signalnih puteva omogućava uvjete za proliferaciju tumora i zaštitu stanica raka od kemoterapeutika.Many reports revealed complexity of tumor which is composed not only of cancer cells, but also of microenvironment which contains stromal cells (fibroblasts, immune cells, endothelial cells, mesenchymal stem cells, adypocytes), extracellular matrix. By release of different soluble factors and enzymes, such as IL-6, TGF- β, MMPs, HIF-1 they promote tumor proliferation, angiogenesis and chemoresistance. CAFs are the most numerous cells in microenvironment so there should be special attention payed to them when creating new cancer therapies. Tumor microenvironment can affect how aggressive and metastatic tumor can get and by interacting with cancer cells through different signaling pathways can promote tumor proliferation and protection of tumor cells from chemotherapeutics
Analiza broja jezgrica u primarnim mišjim megakariocitima
No full textJezgra kao središte pohrane genetičke informacije sadrži svoje pododjeljke među kojima je jezgrica najistraživanija i najistaknutija. Jezgrica nije obavijena membranom već se formira oko rDNA regija na akrocentričnim kromosomima. U skladu s time je i njezina funkcija – dorada i transkripcija rRNA pomoću RNA polimeraze I što posljedično dovodi do slaganja ribosomskih podjedinica. Obzirom na funkciju jezgrica se može podijeliti u tri morfološke regije, a to su fibrilarni centar (FC), guste fibrilarne komponente (DFC) i granularne komponente (GC). No, osim njezine primarne funkcije pronađene su joj i dodatne uloge u drugim fiziološkim procesima.
U ovom radu analizirane su jezgrice u primarnim megakariocitima. Točnije gledan je odnos između površine megakariocita, broja lobula jezgri i broja jezgrica. Rezultati su pokazali da sa sazrijevanjem megakariocita raste broj jezgrica, dok u krajnje zrelim, velikim stanicama jezgrice nisu prisutne. Obzirom da su megakariociti rijetke stanice koštane srži i visokospecijalizirani prekursori trombocita, dodatnim informacijama o jezgri i jezgrici u megakariocitima može biti korisno u izučavanju raznih bolesti koje zahvaćaju megakariocite, ali posljedično i trombocite.The nucleus as the main storage of genetic information is comprised of compartments, among which the nucleolus is the best-studied and the most prominent one. The nucleolus is not surrounded with a membrane but is formed around rDNA regions on acrocentric chromosomes. Consequently, its function - the upgrading and transcription of rRNAs by RNA polymerase I - results in the formation of ribosomal subunits. Given the function, the nucleolus can be divided into three morphological regions, namely fibrillar center (FC), dense fibrillar component (DFC) and granular components (GC). Apart from its primary function in ribosome biogenesis, it has additional roles in other physiological processes.
In this work we analysed the nucleoli of the primary megakaryocytes. More specifically, the relationship between the megakaryocyte surface, the number of lobulus nuclei, and the number of nucleoli was evaluated. Results show that with maturation of megakaryocytes the number of nucleoli increases, while in the most mature cells nucleoli are not visible. Since megakaryocytes are rare bone marrow cells and represent highly specialized platelet precursors, additional information on nuclei and nucleolus in megakaryocytes may be useful in further studies of various diseases affecting megakaryocytes and therefore platelets
In Vitro Photodynamic Activity of N-Methylated and N-Oxidised Tripyridyl Porphyrins with Long Alkyl Chains and Their Inhibitory Activity in Sphingolipid Metabolism
No full textA series of N-methylated and N-oxidised tripyridyl porphyrins were synthesised, characterised, and their PDT activity was studied with six cell lines. All the tested porphyrins with a long alkyl chain, except one, were more efficient for PDT than an N- methylated hydrophilic porphyrin and N-oxidised porphyrin without the long alkyl chain. Generally, N-methylated tripyridyl porphyrins were more active than those N-oxidised, but IC50 values for phototoxicity of two N-oxides, named TOPyP3-C17H33O and TOPyP3-C17H35, were still in the nanomolar concentration range for most of the tested cell lines. However, TOPyP3- C17H35 did not show phototoxicity on human foreskin fibroblast cells. Two methylated amphiphilic porphyrins, named TMPyP3-C17H33 and TMPyP4-C17H35, showed significant dark toxicity, whereas none of the oxidopyridyl porphyrins were toxic without light activation. The selected photosensitisers were shown to be apoptosis inducers, and had inhibitory effects on the clonogenic growth of HCT116 and HeLa cells. All three N-methylated amphiphilic porphyrins significantly reduced the migratory potential of HCT116 cells. Porphyrins TMPyP3- C17H35 and TOPyP3-C17H35 reduced the activity of acid ceramidase, whereas TOPyP3-C17H33O had a significant inhibitory effect on sphingosine kinase 1 activity in HeLa cells. Compounds with this dual activity were shown to be the most promising photosensitisers, with potential to treat invasive cancers
Antibody biotinylation and its application in flow cytometry
No full textProtutijela nalaze široku primjenu u medicini i biotehnološkim istraživanjima zbog sposobnosti specifičnog vezanja na različite molekule (antigene). Mogu se koristiti u detekciji specifičnih antigena za dijagnozu bolesti, u terapiji imunodeficijencija, raka ili zaraznih bolesti te u raznim biomedicinskim istraživačkim i dijagnostičkim tehnikama: Western blot, imunohistokemija, protočna citometrija, itd. U kliničkoj se praksi sve više radi na stvaranju monoklonskih protutijela koja su mnogostruko učinkovitija i specifičnija od standardnih poliklonskih antiseruma. Kako bi se olakšala njihova detekcija u protočnoj citometriji, protutijela se obilježavaju fluorokromima ili biotinom. U ovom radu biotinom je obilježeno protutijelo anti-m04.17 koje prepoznaje m04 protein mišjeg citomegalovirusa (MCMV) te je ispitana kvaliteta i primjenjivost ovako obilježenog protutijela za detekciju MCMV-om inficiranih stanica u protočnoj citometriji. Dobiveni rezultati pokazali su da je biotinilacija uspješno provedena, da nije uzrokovala uništenje paratopa antitijela te da je protutijelo anti-m04 specifično za m04 protein MCMV-a.Antibodies have a wide use in medicine and biotechnological research due to their ability to bind various molecules (antigens) with high specificity. They can be used for detecting specific antigens for diagnosis of diseases, treatment of some immune deficiencies, cancer, infectious diseases and in different biomedical techniques: Western blotting, immunohistochemistry, flow citometry, etc. Monoclonal antibodies are of special interest for clinical practice since they are more efficient and have higher specificity compared to standard polyclonal antiserums. Antibodies can be labeled with fluorochromes or biotin to make it easier for detection in flow citometry. In this work antibody anti-m04.17 that recognizes m04 protein of mouse cytomegalovirus (MCMV) was labeled with biotin. After labeling, the quality and applicability of such labeled antibody for detection of cells infected with MCMC in flow citometry was tested. The results have shown that biotinylation was succesfull and did not cause the destruction of antibody's paratope and that the antibody anti-m04 is specific for m04 protein of MCMV
The influence of methylamine and infectious doses on the results of infection of Dictyostelium discoideum and Acanthamoeba castellanii with Francisella novicida
Get PDFFrancisella tularensis is a gram-negative facultative intracellular bacterial pathogen that in humans and animals causes the fulminating zoonotic disease tularemia. Free-living amoeba, such as A. castellanii and D. discoideum, serve as environmental hosts of various intracellular pathogens. Epidemiology of F. tularensis is commonly associated with water-borne transmission in some parts of the world, including mosquitoes and amoebae as the host reservoir of bacteria in aquatic environment resources. In vitro studies demonstrated intracellular replication of F. tularensis within D. discoideum and A. castellanii cells. It has been established that crucial step in Francisella infection is entering the target cells by receptor mediated phagocytosis. This process has been shown to be meaningful in macrophages, arthropod-derived cells and amoeba cells. We used a water-borne amoeba A. castellanii and D. discoideum as surrogate macrophages as amoeba model systems to study influence of metabolic inhibitor methylamine on results of intracellular and extracellular life of F. novicida. For the first time we showed that methylamine has inhibitory effect on F. novicida entry and replication in D. discoideum and A. castellanii. The influence of different levels of infection was also examined and showed a dose-dependent effect on intracellular and extracellular infection. Moreover, methylamine was analyzed to observe whether in concentration of 100 mM induce cytopathogenicity in amoebae mentioned before. We conclude that F. novicida cannot entry and replicates in D. discoideum and A. castellanii after treated with 100 mM methylamine and that this concentration of inhibitor is not toxic to amoeba itself and can be used to inhibit receptor-mediated phagocytosis during Francisella infection.Francisella tularensis je gram-negativni fakultativno unutarstanični bakterijski pathogen koji uzrokuje tularemiju u ljudi i životinja. Amebe kao što su A. castellanii i D. discoideum služe kao stanice domaćina za različite unutarstanične patogene u prirodi. U određenim dijelovima svijeta epidemiologija F. tularensis je usko povezana s prijenosom bolesti putem vode, što uključuje komarce i amebe kao glavni izvor bakterije u vodenom okolišu. In vitro studije su pokazale unutarstaničnu replikaciju F. novicida u stanicama D. discoideum i A. castellanii. Utvrđeno je da ključan korak u infekciji je ulazak bakterije u ciljanu stanicu mehanizmom receptorom posredovane fagocizote. Pokazalo se da je taj proces od velike značajnosti u makrofazima, stanicama člankonožaca i amebama. Amebe A. castellanii i D. discoideum, kao surogat makrofaga, koristili smo kao modele za istraživanje utjecaja metaboličkog inhibitora metilamina na razmnožavanje F. novicida. Prvi smo pokazali da metilamin ima inhibitorni učinak na ulazak i replikaciju F. novicida u stanice D. discoideum i A. castellanii. Također, istraživan je utjecaj infektivne doze bakterija na unutarsničnu i izvastaničnu replikaciju Francisella, kao i utjecaj metilamina u koncentraciji 100 mM na citotoksičnost u ameba. Naši rezultati pokazuju da F. novicida ne ulazi i replicira se u stanicama D. discoideum i A. castellanii koje su prethodno tretirane sa 100 mM metialminom. Navedena koncentracija nije toksična za same stanice te se može korisiti za sprečavanje receptorom posredovane fagocitoze tijekom Francisella infekcije
Optimisation of rat faecal microbiota isolation method for proteomic analysis purposes
No full textProteomska istraživanja mikrobiote fecesa postaju sve značajnija zbog rastućih spoznaja o povezanosti različitih bolesti i disbalansa mikrobiote crijeva. Radi jednostavnosti, ali i potrebe kontrole prehrane te sastava mikrobiote, u istraživanjima se najčešće koriste modelni organizmi. Štakor u usporedbi s laboratorijskim mišem, ima sličnija farmakokinetička i farmakodinamička svojstva ljudima, te se stoga koristi kao pogodan modelni organizam u istraživanjima crijevne mikrobiote. Crijevna mikrobiota spada u najkompleksnije zajednice mikroorganizama koju čine bakterije, kvasci, virusi i ostali mikrobiote. U proteomskim istraživanjima crijevne mikrobiote kompleksnosti uzorka pridodaju dijelovi neprobavljene hrane i proteini domaćina koji u visoko osjetljivim proteomskim analizama, poput one koja se provodi spektrometrijom masa, prekrivaju prisutnost proteina izoliranih iz mikrobiote. Iz tog razloga važno je dalje razvijati i optimizirati već postojeće metode za izolaciju crijevne mikrobiote. Metode izolacije koje su se pokazale djelotvornima u dosadašnjim istraživanjima uključuju diferencijalna centrifugiranja i centrifugiranja u gradijentu. U ovom radu je provedena optimizacija postojećih metoda za izolaciju proteina mikrobiote iz fecesa, a početni uzorak je bio duboko zamrznuti feces štakora. U prvom se koraku optimizirao protokol za homogenizaciju fecesa, a zatim kod diferencijalnih metoda centrifugiranja i centrifugiranja u gradijentu mijenjani su parametri broja, jačine i vrste centrifugiranja, a kod centrifugiranja u gradijentu, mijenjana je koncentracija sredstva za stvaranje gradijenta, Histodenza. Kontrola efikasnosti izolacije vršena je bojanjem po Gramu i metilenskim modrilom uz promatranje mikroskopskih preparata svjetlosnim mikroskopom. Također je provedeno kemijsko i mehaničko liziranje stanica mikrobiote s ciljem izolacije proteina. Proteini su zatim kvantificirani i razdvojeni uz pomoć jednodimenzionalne (1D) gel elektroforeze s ciljem provjere uspješnosti njihove izolacije. Metoda centrifugiranja u gradijentu pokazala se kao najefikasnija metoda za izolaciju mikrobiote iz duboko zamrznutog fecesa pri čemu su bili zadovoljeni parametri razvnovrsnosti izolirane mikrobiote, a mikroskopski vidljivo onečišćenje, osobito ono uzrokovano neprobavljenim dijelovima hrane, bilo je minimalno.Proteome studies of feces microbiome are becoming more relevant due to the increasing number of evidences linking various diseases and disbalance of gut microbiota. Because of the possibility to control food intake and microbiome flora, as well as simplicity, model organisms are often used in microbiome studies. In comparison to laboratory mice, rats have pharmacokinetic and pharmacodynamic properties more similar to the human and therefore are used as a model organism in studies of gut microbiota. Intestinal microbiota is a complex community consisting of bacteria, yeasts, viruses and other. Undigested food components and host proteins obscure presence of microbial proteins in sensitive proteome analyses, such as mass spectrometry. For that reason, it is important to optimize existing methods for isolation of gut microbiota. Differential centrifugation and density gradient centrifugation have shown to be effective methods of isolation in previous studies. In this study, we have conducted optimization of existing methods used in isolation of fecal microbiota. Microbiota has been isolated from frozen rat feces. The first step was to optimize a protocol for feces homogenization, followed by optimization of centrifugation parameters. Parameters that were optimized included centrifugal force and the number of centrifugation steps, as well as previously mentioned centrifugation types. Histodenz, which is used as a density gradient media for density gradient centrifugation, was also tested in different concentrations. Efficacy of isolation was tested using Gram and methylene blue staining, detected with a light microscope. Chemical and mechanical disruption of the microbiota was performed to isolate proteins, followed by isolation and quantification using one-dimensional (1D) gel electrophoresis. Gradient centrifugation has shown to be the most efficient method in isolation of microbiota from frozen feces due to the preservation of microbiota composition and success rate of isolation, whereas contamination caused by undigested food fragments was minimal
FUNCTION AND INTRACELLULAR LOCALIZATION OF HERPES SIMPLEX VIRUS 1 ICP34.5 PROTEIN
No full textHerpes simpleks virus tipa 1 jedan je od najraširenijih virusa kod ljudi. Uzrokuje trajnu infekciju, a razlikujemo litičku i latentnu fazu infekcije. Latentnu fazu karakterizira mirovanje virusa u neuronima, nakon čega kao posljedica stresa ili smanjenog imunog statusa domaćina, virus može preći u litičku fazu replikacije u epitelnim stanicama.
Virus HSV-1 je razvio mnoge mehanizme pomoću kojih suprimira obrambene odgovore domaćina te omogućava efikasnu virusnu replikaciju. Jedan od vrlo važnih proteina uključenih u modulaciju staničnog odgovora je ICP34.5 koji je i centralni protein ovog istraživanja. ICP34.5 je multifunkcionalni protein i važan faktor neurovirulencije u ljudi. Ulazi u interakciju s mnogim staničnim proteinima kao što su NOP53, PP1, PKR koji predstavljaju važne faktore u obrani stanice od unutarstaničnih mikroorganizama kao što je HSV-1.
Svrha ovog istraživanja je eksperimentalno utvrđivanje lokalizacije ovog proteina te povezivanje s već poznatim funkcijama. Plazmid pcDNA 3xFLAG 34.5 korišten je za transfekciju stanica. pcDNA 3xFLAG 34.5 je transfeciran u humane epitelne stanice retine (RPE-1). Lokalizacija proteina je određena pomoću fluorescentne mikroskopije. Dobiveni rezultati pokazuju da je ICP34,5 ponajviše eksprimiran unutar jezgre stanica što može govoriti o važnosti njegovih funkcija u jezgri, dok je u manjoj mjeri eksprimiran i u citoplazmi stanica. Osim toga, utvrdili smo da količina plazmida kojim su transfecirane ciljne stanice može imati znatan utjecaj na interpretaciju lokalizacije eksprimiranog proteina u stanicama.Herpes simplex virus type 1 is one of the most widespread human viruses. The HSV-1 infection causes persistent infection which can be divided into two phases: a productive and latent phase. Latent phase is characterized as non-productive phase placed in neurons. Viral reactivation often occurs as a result of stress or immunocompromised state of the host. During viral reactivation, HSV-1 goes to a productive phase which is placed in epithelial cells and is characterized by viral replication.
HSV-1 has developed a range of mechanisms by which it modulates host defense and provides efficient viral replication. One of the crucial proteins included in modulating host defense is ICP34.5 which is also the central protein of this study. ICP34.5 is a multifunctional protein and important neurovirulence factor. It interacts with many cellular proteins such as NOP53, PP1, PKR, etc. and ensures efficient viral replication.
The purpose of this study is to determine experimentally the localization of ICP34.5 and link to its functions. The plasmid pcDNA 3xFLAG 34.5 with viral gene γ34.5 was used to express the protein of interest, and it contains three sequences of a FLAG tag, HSV-1 γ34.5 gene, and promoter from cytomegalovirus. pcDNA 3xFLAG 34.5 was transfected into human retinal epithelial cells (RPE-1). The localization of the ICP34.5 protein was analyzed using fluorescent microscopy. The results show that ICP34,5 is mostly expressed in the nucleus of transfected cells, which indicates its functions in the nucleus. It was also mildly expressed in the cell cytoplasm. In addition, we have observed that the amount of transfected plasmid shows a significant influence on the interpretation of the protein localization
The role of M116 gene product in tropism of murine cytomegalovirus
No full textHumani citomegalovirus (HCMV) široko je rasprostranjeni virus u ljudskoj populaciji koji, nakon primarne, uspostavlja latentnu infekciju i ostaje prisutan u organizmu domaćina tijekom cijelog života. Iako u imunološki kompetentnih osoba ne izaziva bolest, HCMV je važan i opasan oportunistički patogen u osoba s oslabjelim ili nezrelim imunološkim sustavom kao što su novorođenčad ili osobe na imunosupresivnoj terapiji. HCMV inficira samo ljude te se stoga u pokusima sa eksperimentalnim životinjama koriste drugi, srodni animalni citomegalovirusi. Od njih, najčešće korišten je mišji citomegalovirus (MCMV). Nedavno provedena transkriptomska analiza MCMV-a ukazala je na to da su najjače eksprimirane regije genoma MCMV-a vrlo slabo okarakterizirane. Jedna od njih je regija M116. Nedavno je također pokazano da gen HCMV-a UL116 kodira za glikoprotein koji se nalazi na površini viriona i stupa u interakciju sa proteinom gH koji je važan za prianjanje i ulazak virusa u stanicu te stoga može utjecati i na virusni tropizam. Stoga sam u sklopu ovog rada proveo istraživanje utječe li delecija gena M116 na tropizam MCMV-a in vitro. Korištenjem mutanta MCMV-a deletirane genske regije M116 i njegovog divljeg tipa provjerio sam je li mutant atenuiran u odnosu na divlji tip i na kojim vrstama stanica. Fenotipizirao sam stanice na kojima je virus atenuiran i provjerio postoji li razlika u ekspresiji mišjih upalnih citokina.Human cytomegalovirus (HCMV) is a widely-spread virus in the human population which, after the primary, establishes latent infection and remains present in the host organism throughout its life. Although in immunologically competent individuals does not cause the disease, HCMV is an important and dangerous opportunistic pathogen in people with a low or immature immune system, such as infants or people on immunosuppressive therapy. HCMV infects only humans and, therefore, other related animal cytomegaloviruses are used in studies with experimental animals. Of these, murine cytomegalovirus (MCMV) is most commonly used. Recently conducted transcriptional MCMV analysis indicated that the most strongly expressed regions of the MCMV genome are characterized very poorly. One of them is the M116 region. Recently, it has also been shown that the HCMV gene UL116 encodes the glycoprotein located on the virion surface and interacts with the gH protein that is important for the adherence and entry of the virus into the cell and hence can also affect viral tropism. Therefore, as part of this paper, research has been carried out to investigate the deletion of M116 gene on MCMV tropism in vitro. Using the MCMV mutant with deleted M116 gene region and its wild type I checked whether the mutant was attenuated in relation to the wild type and on which types of cells. I phenotypized the cells at which the virus was attenuated and checked for any difference in the expression of mouse inflammatory cytokines