Genetics of autoimmune diseases

Autoimunosne bolesti su vrlo kompleksne bolesti koje se javljaju kod sve većeg broja ljudi. Budući da nastaju kao posljedica genetičkih poremećaja, ali i utjecaja okolišnih faktora, javljaju se sa vrlo velikim spektrom simptoma što otežava njihovu karakterizaciju i pronalazak pravog uzroka zbog kojeg se bolest javlja. Zato se samo ugrubo dijele na organ-specifične i sustavne. U njihovom razvoju je, osim stanica s poremećenim funkcijama, vrlo važan i utjecaj komponenti komplementa i citokina i kemokina koji u velikoj mjeri utječu na simptome autoimunosnih bolesti. Hashimotov tiroiditis, mijastenija gravis i dijabetes mellitus tipa 1 su neki od primjera organ-specifičnih autoimunosnih bolesti, dok su sistemski eritematozni lupus, reumatoidni artritis i multipla skleroza predstavnici sustavnih autoimunosnih bolesti. Većina informacija o njihovoj genetičkoj podlozi je dobivena istraživanjima na razini genoma (engl. genome-wide association studies, GWAS), no takva istraživanja ne mogu odrediti sa potpunom sigurnošću koji genski lokusi su kritični za razvoj određene bolesti pa će u daljnjim istraživanjima biti ključno odrediti koji su to i koja je njihova funkcija.Autoimmune diseases are very complex diseases that keep affecting more and more people. Since they are the result of genetic disorders and enviromental factors, they appear with a large spectrum of symptoms which makes their characterization and discovering their real cause even more difficult. That is why they are only roughly divided into organ-specific and systemic. Besides cells having disrupted functions, influence of components of complement and cytokines and chemokines is very important in their development and affect symptoms of the diseases. Hashimoto’s thyroiditis, myasthenia gravis and type 1 diabetes mellitus are some of the organ-specific diseases, while systemic lupus erythematosus, rheumatoid arthritis and multiple sclerosis are representatives od systemic autoimmune diseases. Most of the information of their genetic background comes from the genome-wide association studies, however, such research cannot precisely predict which genetic locuses are critical for development of certain diseases, therefore, in further research it will be key to determine which locuses are critical and their functions

In vitro effects of ascorbic acid on viability and metabolism of patients’ osteosarcoma stem cells

Stagnation in novelties of osteosarcoma (OS) treatment indicates the need for new therapeutic methods. OS cancer stem cells (OS-CSC) are taught to have the ability to self-renew and develop mechanisms of anticancer drug resistance, and this is why it is difficult to eradicate them. Their metabolism has been recognized as a potential target of therapeutic action. Ascorbic acid (AA) is considered to act pro-oxidative against OS-CSC in vitro by oxidative effect and by inhibition of glycolysis. This study examined an in vitro impact of AA on OS-CSC metabolism isolated from patients\u27 biopsies, with the aim of better understanding of OS-CSC metabolism and the action of AA on OS-CSC. OS-CSC were isolated using a sphere culture system and identified as stem cells using Hoechst 33342 exclusion assay. Determination of the dominant type of metabolism of OS-CSC, parental OS cells, human mesenchymal stem cells (hMSC) and U2OS OS lineage before and after AA treatment was done by Seahorse XF (Agilent). Cytotoxicity of high-dose AA was confirmed by the MTT test and was proven for all the examined cell types as well as HEK293. Seahorse technology showed that OS-CSC can potentially use both glycolysis and oxidative phosphorylation (OXPHOS), and can turn to glycolysis and slow metabolic potential in unfavorable conditions such as incubation in AA

Macroporous silicon-wollastonite scaffold with Sr/Se/Zn/Mg-substituted hydroxyapatite/chitosan hydrogel

The scaffolds, which morphologically and physiologically mimic natural features of the bone, are of a high demand for regenerative medicine. To address this challenge, bioactive porous silicon/wollastonite (SC) scaffold has been developed for potential bone tissue engineering applications. Additive manufacturing through the selective laser melting approach has been exploited to fabricate computer-aided designed scaffolds with a pore size of 400 μm. To increase the biocompatibility and osteogenic properties of SC scaffolds, the hydrogel based on a mixture of four mono-substituted hydroxyapatites (sHAp) and biopolymer chitosan (CHT) has been incorporated into SC by impregnation and freeze-gelation method. The pore size of 400 μm of SC has provided enough space for the impregnation of polymer solution and composite (CHT/sHAp) suspension to form highly porous hydrogel within pores. By the combination of SC and CHT/sHAp, both cell attachment and homogeneous proliferation on SC scaffold as well as mechanical properties of CHT/sHAp hydrogel have been improved.</p

Synthesis and In Vitro Characterization of Ascorbyl Palmitate-Loaded Solid Lipid Nanoparticles

Antitumor applications of ascorbic acid (AA) and its oxidized form dehydroascorbic acid (DHA) can be quite challenging due to their instability and sensitivity to degradation in aqueous media. To overcome this obstacle, we have synthesized solid lipid nanoparticles loaded with ascorbyl palmitate (SLN-AP) with variations in proportions of the polymer Pluronic F-68. SLNs were synthesized using the hot homogenization method, characterized by measuring the particle size, polydispersity, zeta potential and visualized by TEM. To investigate the cellular uptake of the SLN, we have incorporated coumarin-6 into the same SLN formulation and followed their successful uptake for 48 h. We have tested the cytotoxicity of the SLN formulations and free ascorbate forms, AA and DHA, on HEK 293 and U2OS cell lines by MTT assay. The SLN-AP in both formulations have a cytotoxic effect at lower concentrations when compared to ascorbate applied the form of AA or DHA. Better selectivity for targeting tumor cell line was observed with 3% Pluronic F-68. The antioxidative effect of the SLN-AP was observed as early as 1 h after the treatment with a small dose of ascorbate applied (5 µM). SLN-AP formulation with 3% Pluronic F-68 needs to be further optimized as an ascorbate carrier due to its intrinsic cytotoxicity

Analysis of intestine microbiome of Alzheimer's disease rat model

Alzheimerova bolest (AB) je progresivna neurodegenerativna bolest i glavni uzrok kognitivnih poremećaja i poremećaja u ponašanju industrijaliziranog društva. Os mikrobiom-crijeva-mozak sastoji se od dvosmjernog komuniciranja između središnjeg i enteričkog živčanog sustava i povezuje emocionalne i kognitivne centre u mozgu s perifernom funkcijom crijeva. U ovom istraživanju je korišten štakorski model AB (n=10), koji je izazvan intraperitonealnim injiciranjem aluminijevog klorida (10 mg/kg štakora) i D-galaktoze (60 mg/kg štakora) tijekom 28 dana. Iz crijevnog sadržaja štakora analiziran je broj kolonija bakterija rodova Lactobacillus i Bifidobacter, enzimska aktivnost mikrobioma te je sekvencirana DNA. Podaci sekvenciranja su obrađeni metodom hijerarhijskog klasteriranja i metodom analize glavnih komponenti. Dobiveni rezultati pokazali su: a) smanjeni broj probiotičkih bakterija u modelu AB u odnosu na kontrolnu skupinu; b) povećanu enzimsku aktivnost β-galaktozidaze, β-glukozidaze i β-glukuronidaze u modelu AB u odnosu na kontrolnu skupinu c) da nije došlo do velike promjene u sastavu crijevnog mikrobioma, ali je došlo do značajnih promjena u 46 vrsta bakterija koje ukupno čine 11,61% crijevnog mikrobioma kontrolne skupine. Temeljem dobivenih rezultata može se zaključiti da su smanjeni udio Lactobacillus i Bifidobacter, povećana enzimska aktivnost β-galaktozidaze, β-glukozidaze i β-glukuronidaze i smanjenje biomase crijevnog mikrobioma povezani s upalnim procesom izazvanim kod štakorskog modela AB.Alzheimer's disease (AD) is progressive neurodegenerative disease and leading cause of cognitive and behaviour impairments of industrialized society. Microbiome-gut-brain axis is bidirectional communication between central and enteric nervous system, thus connecting emotional and cognitive centers in brain with peripheral gut function. In this research rat model of AD (n=10) was induced by intraperitoneal injection of aluminium chloride (10 mg/kg rat) and D-galactose (60 mg/kg rat) during 28 days. Dilutions were made from rat's colon content and streaked on selective plates for isolation of Lactobacillus and Bifidobacter, activity of bacterial enzymes was analyzed and DNA isolated for sequencing. Sequencing data was analyzed by hierarchical clustering and principal component analysis. Results showed: a) reduced number of probiotic bacteria in AD model compared to control group; b) increased activity of β-galactosidase, β-glucosidase i β-glucuronidase in AD model compared to control group c) there was no great changes in composition of intestine microbiome, but 46 bacterial species, which make 11,61% of intestine microbiome of control group, significantly changed. Based on these results it can be concluded that reduced number of Lactobacillus and Bifidobacter, increased enzyme activity of β-galactosidase, β-glucosidase i β-glucuronidase and reduced intestine biomass are connected with inflammation induced in AD rat model

Analysis of Charlotte Brontë's Jane Eyre

Ovaj diplomski rad jedna je od mogućih analiza i interpretacija romana Jane Eyre koji je napisan u devetnaestom stoljeću u Velikoj Britaniji. Djelo odražava razvoj glavne protagonistice koja teži neovisnosti i ravnopravnosti što ju čini progresivnom i atipičnom junakinjom viktorijanskih romana. Rad nastoji pokazati da je Brontë autorica koja je pisala o važnim temama koje se dotiču reprezentacije žene u viktorijanskom dobu i pitanja ženskih prava. Osim toga, naratološka analiza romana dokazala je da djelo možemo promatrati u svjetlu modernog pripovijedanja. Nadalje, literatura koja je korištena u radu podrazumijeva primarnu literaturu, odnosno roman Jane Eyre, i sekundarnu literaturu koja se odnosi na kritička djela domaće i anglofone pripadnosti. Djelo pokazuje težak položaj viktorijanske žene, ali također progovara i o činjenici da i žene mogu birati, kako u obrazovnom pogledu tako i u pogledu bračnog partnera. Ta činjenica potvrđuje da je Brontëin glavni lik aktivan i progresivan, ali treba napomenuti da ju karakteristike poput imena, izgleda, ponašanja i načina odijevanja čine „običnom“ protagonisticom s kojom se čitatelji mogu poistovjetiti. Između ostaloga, u romanu pronalazimo i personifikaciju vremena koja pruža uvid u Janeine osjećaje i njezin unutarnji svijet.This paper is one of the possible analyses and interpretations of the novel Jane Eyre, which was written in the nineteenth century in Great Britain. The paper reflects the development of the main protagonist who strives for independence and equality, which makes her a progressive and atypical heroine of Victorian novels. The paper tries to show that Brontë is an author who wrote about important topics that touch on the representation of women in the Victorian era, and the issue of women’s rights. In addition, the narratological analysis of the novel proved that we can observe this literary work in the light of modern storytelling. Furthermore, the bibliography used in the paper includes primary sources, namely the novel Jane Eyre, and secondary sources that refer to critical works of domestic and anglophone origin. This paper shows the difficult position of the Victorian woman, but also speaks about the fact that women can choose, both in terms of education and in terms of marriage partner. This fact confirms that Brontë's main character is active and progressive, but it should be noted that the characteristics such as name, appearance, behavior, and way of dressing make her an “ordinary” protagonist with whom readers can identify. Among other things, in the novel we also find the personification of time that provides an insight into Jane's feelings and her inner world

Ženska radna etika u romanima Male žene, Dječaci gospođe Jo i Staromodna djevojka Louise May Alcott

This paper provides an overview of three popular children’s nineteenth-century novels written by Louisa May Alcott. One of the ideas that the author advocates for are women’s rights and the idea of a more inclusive educational system. Focusing on the concept of “female work ethic”, I show how Alcott approached this topic in the framework of children’s literature. Certain examples from the novels provide a better insight into the core of this discussion, and each novel was influenced by the historical circumstances that took place, which is why there is an evident difference between the older and the younger generation of female characters in these novels. Besides the major topic of this paper, this analysis also touches upon other relatable notions of the 19th century, such as woman’s position in society, family life and relationships, etc. It is also worth mentioning influential messages that the author provides in her works, and to name just a few, Alcott for instance promotes the importance of family, labor in general, and inner values. Moreover, notions such as materialism, inclination towards money, fame and foreign fashions are condemned as undesirable traits in women. I also show how the author discusses the combination of older and modern values which her characters embody. Although the paper is dedicated to women’s labor, some minor parts of the discussion also tackle male characters whose role is instrumental for reinforcing certain arguments which concern female work ethic. I also discuss the introduction of new topics and concepts which Alcott included in her novels, and this refers to the new cultural concept called “tomboy” whose traits are presented in the characters of Jo and Nan, and it is also necessary to mention the idea of literary celebrity and the notion of privacy. Furthermore, these concepts enabled this paper to show the challenges and the complexities of women’s position in Victorian and postbellum America

Ženska radna etika u romanima Male žene, Dječaci gospođe Jo i Staromodna djevojka Louise May Alcott

This paper provides an overview of three popular children’s nineteenth-century novels written by Louisa May Alcott. One of the ideas that the author advocates for are women’s rights and the idea of a more inclusive educational system. Focusing on the concept of “female work ethic”, I show how Alcott approached this topic in the framework of children’s literature. Certain examples from the novels provide a better insight into the core of this discussion, and each novel was influenced by the historical circumstances that took place, which is why there is an evident difference between the older and the younger generation of female characters in these novels. Besides the major topic of this paper, this analysis also touches upon other relatable notions of the 19th century, such as woman’s position in society, family life and relationships, etc. It is also worth mentioning influential messages that the author provides in her works, and to name just a few, Alcott for instance promotes the importance of family, labor in general, and inner values. Moreover, notions such as materialism, inclination towards money, fame and foreign fashions are condemned as undesirable traits in women. I also show how the author discusses the combination of older and modern values which her characters embody. Although the paper is dedicated to women’s labor, some minor parts of the discussion also tackle male characters whose role is instrumental for reinforcing certain arguments which concern female work ethic. I also discuss the introduction of new topics and concepts which Alcott included in her novels, and this refers to the new cultural concept called “tomboy” whose traits are presented in the characters of Jo and Nan, and it is also necessary to mention the idea of literary celebrity and the notion of privacy. Furthermore, these concepts enabled this paper to show the challenges and the complexities of women’s position in Victorian and postbellum America

Analysis of intestine microbiome of Alzheimer's disease rat model

Alzheimerova bolest (AB) je progresivna neurodegenerativna bolest i glavni uzrok kognitivnih poremećaja i poremećaja u ponašanju industrijaliziranog društva. Os mikrobiom-crijeva-mozak sastoji se od dvosmjernog komuniciranja između središnjeg i enteričkog živčanog sustava i povezuje emocionalne i kognitivne centre u mozgu s perifernom funkcijom crijeva. U ovom istraživanju je korišten štakorski model AB (n=10), koji je izazvan intraperitonealnim injiciranjem aluminijevog klorida (10 mg/kg štakora) i D-galaktoze (60 mg/kg štakora) tijekom 28 dana. Iz crijevnog sadržaja štakora analiziran je broj kolonija bakterija rodova Lactobacillus i Bifidobacter, enzimska aktivnost mikrobioma te je sekvencirana DNA. Podaci sekvenciranja su obrađeni metodom hijerarhijskog klasteriranja i metodom analize glavnih komponenti. Dobiveni rezultati pokazali su: a) smanjeni broj probiotičkih bakterija u modelu AB u odnosu na kontrolnu skupinu; b) povećanu enzimsku aktivnost β-galaktozidaze, β-glukozidaze i β-glukuronidaze u modelu AB u odnosu na kontrolnu skupinu c) da nije došlo do velike promjene u sastavu crijevnog mikrobioma, ali je došlo do značajnih promjena u 46 vrsta bakterija koje ukupno čine 11,61% crijevnog mikrobioma kontrolne skupine. Temeljem dobivenih rezultata može se zaključiti da su smanjeni udio Lactobacillus i Bifidobacter, povećana enzimska aktivnost β-galaktozidaze, β-glukozidaze i β-glukuronidaze i smanjenje biomase crijevnog mikrobioma povezani s upalnim procesom izazvanim kod štakorskog modela AB.Alzheimer's disease (AD) is progressive neurodegenerative disease and leading cause of cognitive and behaviour impairments of industrialized society. Microbiome-gut-brain axis is bidirectional communication between central and enteric nervous system, thus connecting emotional and cognitive centers in brain with peripheral gut function. In this research rat model of AD (n=10) was induced by intraperitoneal injection of aluminium chloride (10 mg/kg rat) and D-galactose (60 mg/kg rat) during 28 days. Dilutions were made from rat's colon content and streaked on selective plates for isolation of Lactobacillus and Bifidobacter, activity of bacterial enzymes was analyzed and DNA isolated for sequencing. Sequencing data was analyzed by hierarchical clustering and principal component analysis. Results showed: a) reduced number of probiotic bacteria in AD model compared to control group; b) increased activity of β-galactosidase, β-glucosidase i β-glucuronidase in AD model compared to control group c) there was no great changes in composition of intestine microbiome, but 46 bacterial species, which make 11,61% of intestine microbiome of control group, significantly changed. Based on these results it can be concluded that reduced number of Lactobacillus and Bifidobacter, increased enzyme activity of β-galactosidase, β-glucosidase i β-glucuronidase and reduced intestine biomass are connected with inflammation induced in AD rat model

Analysis of intestine microbiome of Alzheimer's disease rat model

Alzheimerova bolest (AB) je progresivna neurodegenerativna bolest i glavni uzrok kognitivnih poremećaja i poremećaja u ponašanju industrijaliziranog društva. Os mikrobiom-crijeva-mozak sastoji se od dvosmjernog komuniciranja između središnjeg i enteričkog živčanog sustava i povezuje emocionalne i kognitivne centre u mozgu s perifernom funkcijom crijeva. U ovom istraživanju je korišten štakorski model AB (n=10), koji je izazvan intraperitonealnim injiciranjem aluminijevog klorida (10 mg/kg štakora) i D-galaktoze (60 mg/kg štakora) tijekom 28 dana. Iz crijevnog sadržaja štakora analiziran je broj kolonija bakterija rodova Lactobacillus i Bifidobacter, enzimska aktivnost mikrobioma te je sekvencirana DNA. Podaci sekvenciranja su obrađeni metodom hijerarhijskog klasteriranja i metodom analize glavnih komponenti. Dobiveni rezultati pokazali su: a) smanjeni broj probiotičkih bakterija u modelu AB u odnosu na kontrolnu skupinu; b) povećanu enzimsku aktivnost β-galaktozidaze, β-glukozidaze i β-glukuronidaze u modelu AB u odnosu na kontrolnu skupinu c) da nije došlo do velike promjene u sastavu crijevnog mikrobioma, ali je došlo do značajnih promjena u 46 vrsta bakterija koje ukupno čine 11,61% crijevnog mikrobioma kontrolne skupine. Temeljem dobivenih rezultata može se zaključiti da su smanjeni udio Lactobacillus i Bifidobacter, povećana enzimska aktivnost β-galaktozidaze, β-glukozidaze i β-glukuronidaze i smanjenje biomase crijevnog mikrobioma povezani s upalnim procesom izazvanim kod štakorskog modela AB.Alzheimer's disease (AD) is progressive neurodegenerative disease and leading cause of cognitive and behaviour impairments of industrialized society. Microbiome-gut-brain axis is bidirectional communication between central and enteric nervous system, thus connecting emotional and cognitive centers in brain with peripheral gut function. In this research rat model of AD (n=10) was induced by intraperitoneal injection of aluminium chloride (10 mg/kg rat) and D-galactose (60 mg/kg rat) during 28 days. Dilutions were made from rat's colon content and streaked on selective plates for isolation of Lactobacillus and Bifidobacter, activity of bacterial enzymes was analyzed and DNA isolated for sequencing. Sequencing data was analyzed by hierarchical clustering and principal component analysis. Results showed: a) reduced number of probiotic bacteria in AD model compared to control group; b) increased activity of β-galactosidase, β-glucosidase i β-glucuronidase in AD model compared to control group c) there was no great changes in composition of intestine microbiome, but 46 bacterial species, which make 11,61% of intestine microbiome of control group, significantly changed. Based on these results it can be concluded that reduced number of Lactobacillus and Bifidobacter, increased enzyme activity of β-galactosidase, β-glucosidase i β-glucuronidase and reduced intestine biomass are connected with inflammation induced in AD rat model