Primarni bilijarni holangitis miševa izazvan bakterijom Novosphingobium aromaticivorans: uloga galektina-3 u aktivaciji inflamazoma

Značajno slabije izražen primarni bilijarni holangitis izazvan infekcijom bakterijom Novosphingobium aromaticivorans, sa manjim oštećenjem bilijarnih kanalića i vrlo slabo izraženom fibrozom, detektovan je u grupama Lgals3−/− i miševa tretiranih inhibitorom galektina-3, GM-CT-01. U jetrama wild type miševa inficiranih bakterijom Novosphingobium aromaticivorans detektovana je veća zastupljenost inflamacijskih makrofaga, NK, NKT i T ćelija u infiltratima. U infiltratima jetre Lgals3-/- i miševa tretiranih inhibitorom galektina-3, GM-CT-01, je nakon infekcije bakterijom Novosphingobium aromaticivorans, u infiltratima jetre zabeležena manja ekspresija inflamazoma NLRP3 i manja produkcija IL-1β u poređenju sa grupom wild type miševa. In vitro stimulacija wild type peritonealnih makrofaga bakterijom Novosphingobium aromaticivorans za posledicu ima veću ekspresiju NLRP3, aktivnost kaspaze-1 i produkciju IL-1β u poređenju sa stimulisanim peritonealnim makrofagima Lgals3−/− miševa. GM-CT-01 takođe redukuje aktivaciju DC i ekspresiju citokina IL-4, IL-1β i p40, zajedničke subjedinice IL-12 i IL-23 u DC in vitro stimulisanim bakterijom Novosphingobium aromaticivorans. Naši rezultati ukazuju na značaj galetkina-3 u stimulaciji inflamacije u RVS izazvanom infekcijom bakterijom Novosphingobium aromaticivorans, a koji se ogleda u aktivaciji DC i inflamazoma NLRP3 i posledičnoj produkciji IL-1β, i ukazuju da galektin-3 može da bude meta delovanja potencijalnih novih lekova. Galektin-3 je verovatno uključen u inflamacijski odgovor na komensalne bakterije creva što može da bude incijalni okidač za razvoj primarnog bilijarnog holangitisa.Marked attenuation of primary biliary cholangitis (PBC), manifested by the absence of bile duct damage and fibrosis, is detected in Lgals3−/− and Galectin-3 inhibitor, GM-CT-01, treated mice. Liver infiltrates of Novosphingobium aromaticivorans infected wild type mice had higher incidence of pro-inflammatory macrophages, NK, NKT, and T cells. Lgals3 deletion and treatment with Galectin-3 inhibitor, GM-CT-01, reduced liver damage and fibrosis, inflammatory mononuclear cell infiltrate, expression of NLRP3 inflammasome in the liver infiltrates and interleukin-1β (IL-1β) production in the livers of Novosphingobium aromaticivorans infected mice. In vitro stimulation of wild type peritoneal macrophages with Novosphingobium aromaticivorans caused increased NLRP3 expression, caspase-1 activity and IL-1β production, compared with Lgals3−/− cells. GM-CT-01 reduced activation of dendritic cells and expression of IL-4, IL-1β, and p40, common subunit of IL-12 and IL-23 in dendritic cells in vitro stimulated with Novosphingobium aromaticivorans. Our data highlight the importance of Galectin-3 in promotion of inflammation in Novosphingobium aromaticivorans induced PBC by enhancing the activation of dendritic cells and NLRP3 inflammasome leading to enhanced production of IL-1β, and indicate Galectin-3 as possible therapeutical target in autoimmune cholangitis. Galectin-3 appears involved in inflammatory response to gut commensals leading to PBC

The significance of the expression of galectin-3 in the pathogenesis of primary biliary cholangitis in mice

Izvod: Uticaj galektina-3 na patogenezu primarnog bilijarnog holangitisa do sada nije ispitivan, a rezultati ove studije jasno pokazuju da Gal-3 može da ima potpuno suprotne efekte na tok bolesti u dva razlišita modela pa izgleda sasvim dokumentovano da uticaj galektina-3 na razvoj i tok bolesti zavisi od dominantnog mehanizma indukcije bolesti. U modelu primarnog bilijarnog holangitisa indukovanog imunizacijom C57BL/6 miševa ksenobiotikom u adjuvansu Gal-3 ima protektivni efekat, ubalažavajći bolest verovatno tako što štiti holangiocite od apoptoze čime smanjuje dostupnost autoantigena i tako ograničava autoimunski proces. Suprotno, u modelu ove bolesti izazvane bakterijom Novosphingobium aromaticivorans odsustvo galektina-3 gotovo da čini miševe rezistentnim na razvoj RVS najverovatnije zbog neadekvatne aktivacije dendritskih ćelija bakterijom u odsustvu galektina-3, usled čega izostaje i aktivacija svih ostalih ćelija (NKT i NK ćelija, T limfocita) koje učestvuju u razvoju oštećenja bilijarnih kanala.Abstract: The impact of galectin-3 in the pathogenesis of primary biliary cholangitis has not been studied, and the results of this study clearly show that Gal-3 can have completely opposite effects on the course of the disease in two different induction models looks quite documented that the effect of galectin-3 in the development and course of the disease depends on the prevalent mechanism for the induction of the disease. In the model of primary of biliary cholangitis induced by immunizing C57BL/6 mice with xenobiotics in adjuvant, Gal-3 has a protective effect, mitigating disease probably by protecting from apoptosis holangiocite thus reducing the availability of autoantigens, thus limiting the autoimmune process. Contrary, in the model of this disease caused by bacteria Novosphingobium aromaticivorans absence of galectin-3 almost makes the mice resistant to the development of PBC most likely due to inadequate activation of dendritic cells by the bacteria in the absence of galectin-3, and subsequent reduced activation of other cells (NKT and NK cells, T lymphocytes) that participate in the development of biliary duct damage

Tumori ovarijuma u adolescentkinja i mladih žena

Phenomenon of heterogeneous ovary tumors is probably the result of activity of the whole series of various factors, which play significant role in genesis, development and classification of tumors, In the age of puberty and adolescence, as a transitional period from childhood to adulthood, it is not rarely seen that due to the influence of hormone regulation, social environment and psychophysical stability, the break of correlation of the mentioned factors can imperil reproductive health of young people. That is the moment when the illnesses of reproductive system become noteworthy for the first time. Thus the immense role of juvenile gynecology is to preserve reproductive health of adolescent girls by preventive health education. The most frequent illnesses of ovary are in tight connection with ovary tumor changes, which could have influence on the normal function of the ovary.Pojava raznovrsnih tumora ovarijalnog porekla, verovatno je posledica delovanja čitavog niza različitih faktora, koji imaju značajnu ulogu u pojavi, razvoju i klasifikaciji tumora. U pubertetu i adolescenciji, kao prelaznom periodu iz detinjstva u zrelo doba, pod uticajem hormonske regulacije, socijalne sredine i psihofizičke ravnoteže, najčešće može doći do prekida korelacije pomenutih faktora, a što može da ugrozi reproduktivno zdravije mladih. Bolesti reproduktivnog sistema tada prvi put postaju značajne. Zato je značaj i uloga juvenilne ginekologije da preventivno zdravstveno-vaspitnim radom i pravovremenim lečenjem sačuva reproduktivno zdravlje adolescentkinja, jer najčešća oboljenja ovarijuma su u tesnoj vezi sa tumoroznim promenama, koji mogu da utiču na normalnu funkciju ovarijuma

Analysis of cervical resistance during continuous controllable balloon dilatation: controlled clinical and experimental study

Background: Hydraulic dilatation is a novel method of cervical dilatation that is based on continuous controllable dilatation (CCBD) by the pumping of fluid into the balloon extension of the system. The main advantage of this procedure is that it allows control of and insight into the process of cervical dilatation. Methods: For the purposes of our research, we created a new and upgraded system for CCBD which consists of a programmed hydrostatic pump connected to a balloon extension. With regard to our aim to precisely measure and determine the location of the cervical resistance, we placed two pressure-measuring films, one on the top and one on the bottom of the balloon extension. This study included 42 patients in whom cervical resistance was measured before suction curettage. Results: Cervical dilatation and measurement of cervical resistance were successful in all patients. The analysis of the pressure-measuring films showed that the points of highest resistance were located in the zone of the internal cervical os and that these values were much higher than those in the zone of the external cervical os (0.402 versus 0.264 MPa at the upper pressure-sensitive film; 0.387 versus 0.243 MPa at the lower pressure-sensitive film). This study also showed that an increase in cervical resistance in the zone of the internal cervical os was followed by an increase in cervical resistance in the zone of the external cervical os. Conclusions: During CCBD, the internal cervical os is the centre of cervical resistance, and the values do not decline with the number of miscarriages or the number of previous births

O,O'-diethyl-(S,S)-ethylenediamine-N,N'-di-2-(3-cyclohexyl)propanoate dihydrochloride enhances influx of effective NK and NKT cells in murine breast cancer

Background/Aim. O,O'-diethyl-(S,S)-ethylenediamineN,N'-di-2-(3-cyclohexyl)propanoate dihydrochloride (DE-EDCP) has been found to possess promising cytotoxic activity against various tumor cell lines. Also, DE-EDCP reduces tumor progression by several mechanisms such as triggering tumor cell death and inhibition of cell proliferation. The aim of present study was to further evaluate antitumor activity of DE-EDCP by investigating effects on migratory potential of tumor cells and anti-tumor immune response. Methods. Migratory potential of DE-EDCP was evaluated by scratch wound assay. Female BALB/c mice were inoculated with 4T1 breast cancer cells and treatment with DE-EDCP started five days following orthotopic tumor implantation. The frequency and phenotype of tumor-infiltrating natural killer (NK) and natural killer T (NKT) cells were analyzed by flow cytometry. Results. DE-EDCP inhibited migratory potential of highly metastatic 4T1 cells. DE-EDCP facilitated accumulation of CD3+CD49+ NKT cells and CD3-CD49+ NK cells in tumor microenvironment. DE-EDCP treatment led to significant decrement of tumor infiltrating anergic NKT cells expressing cytotoxic T-lymphocyte-associated protein 4 (CTLA-4), killer cell lectin like receptor G1 (KLRG-1) and programmed cell death protein-1 (PD-1). Mice given DE-EDCP had significantly increased percentages of tumoricidal fas ligand (FasL) positive NK cells. Conclusion. DE-EDCP inhibits murine breast cancer progression through direct effects on tumor cells and by facilitating anti-tumor immunity. DE-EDCP enhances accumulation, promotes tumoricidal phenotype and maintenances responsiveness of NK and NKT cells in 4T1 murine breast cancer

Can miRNA expression patterns predict radiotoxicity in patients with glioblastoma?

Background: Standard treatment of the glioblastoma patients is implemented with Stupp’s protocol since 2005, but 2-years survival of the patients are still at low percentage. Even with implementation of modern radiotherapy techniques, acute radiotoxicity is still observed in these patients. Micro RNAs (miRNAs) expression patterns are introduced in recent years as factors that might predict survival in patients with glioblastoma, as well as cancer treatment related side effects. Material and methods: The data used in this review were conducted from research papers in PUBMED/MEDLINE databases with a special focus on role of miRNAs in cancer and with emphasis on brain tumors correlated to acute radiotoxicity. Results: Inflammation due to oxidative stress via free radicals is one of the mechanisms of radiation injury of irradiated cells and may correlate with toxicity. MiRNAs expression in prostate cancer patients may predict acute radiotoxicity, while miRNAs expression levels that could predict radiation toxicity in glioblastoma are still under investigation. However, it has been shown that miRNA levels like miR-16, miR 21, miR-19a and miR-22 are increased after radiation in glioma, while levels of miR-107, miR-181a are decreased. Conclusion: Recent studies have shown that miRNAs may have role as potent indicator of radiotoxicity in cancer patients. To date, there is no available data about acute brain radiotoxicity and its correlation with expression levels of mRNAs in patients with GB. There is a emerge need for further investigation on role of radioresponsive miRNAs in glioblastoma patients with acute radiotoxicity.5th Congress of the Serbian Association for Cancer Research with International Participation SDIR-5, Virtual event, December 3, Belgrade, 2021

Auto imunska bolest štitaste žlezde - patogeneza Graves-ove bolesti i Hashimoto tireoiditisa

It is generally accepted that autoimmune thyroid disorders, Graves disease and Hashimoto thyroiditis, differ in pathogenesis and clinical implications. In Graves disease the basic pathogenetic mechanism is B lymphocyte activation which produce auto antibodies specific for TSH receptor (TSHR) which binding to thyrocyte membrane causes their long-termed stimulation which gives as a result the occurrence of hyperthyrosis. On the other hand in Hashimoto thyroiditis lymphocyte accumulation occurs and they cause gradual thyrocyte damage and hypothyrosis development. However, it was found that the reisa certain genetic predisposition for both autoimmune thyroid diseases and that they can appear among several members of the same family. Besides in the serum of the patients with Graves disease and Hashimoto thyroiditis the presence of autoantibodi esspecific for dominant thyroid autoantigenes: TSHR, thyroperoxidase (TPO) and thyroglobulin (Tg) can be found as indicators of the auto immune process in thyroid gland. For these reasons both autoimmune diseases of thyroid gland sometimes are marked with the common name: autoimmune thyroid disease. As cell and molecule mechanisms included in initiation of the autoimmune process in thyroid gland that define the type and natural course of disease have not completely been explained, in this review the data from the literature considering pathogenesis of autoimmune diseases of thyroid gland have been shown. After introductory considerations, dominant autoantigenes and autoantibodies (as indicators of autoimmune process in thyroid gland) are shown in details, as well as mechanisms included in effector phase of autoimmune process which cau se the thyroid cell damage. The role of disturbance in regulation of the apoptosis process is especially analyzed as they could effect the development of autoimmune diseases of thyroid gland.Prihvaćeno je shvatanje da se autoimunske bolesti štitaste žlezde Graves-ova bolest i Hashimoto tireoiditis razlikuju po patogenezi i kliničkim posledicama. U Graves-ovoj bolesti je osnovni patogenetski mehanizam aktivacija Blimfocita koji produkuju auto antitela specifična za TSH receptor (TSHR), čije vezivanje za membranu tireocita uzrokuje njihovu dugotrajnu stimulaciju, sa posledičnim nastankom hipertireoze. S druge strane, u Hashimoto tireoiditi su nastaje akumulacija limfocita koji prouzrokuju postepeno oštećenje tireocita i nastanak hipotireoze. Međutim utvrđeno je da za obe autoimunske tireoidne bolesti postoji određena genetska predispozicija i da se one mogu javiti kod više članova u istoj porodici. Osim toga, u serumu obolelih od Graves-ove bolesti i Hashimoto tireoiditi sa se može pokazati prisustvo autoantitela specifičnih za dominantne tireoidne autoantigene (receptor za TSH, tireoperoksidazu i tireoglobulin) koji predstavljaju pokazatelje autoimunskog procesa u štitastoj žlezdi. Iz tih razloga se obe autoimunske bolesti štitaste žlezde nekad označavaju zajedničkim nazivom: autoimunska bolest štitaste žlezde. Budući da ćelijski i molekulski mehanizmi koji su uključeni u inicijaciju autoimunskog procesa u štita stoj žlezdi, i opredeljuju vrstu i prirodni tok bolesti, nisu potpuno rasvetljeni, u ovom radu su prikazani podaci iz literature koji se odnose na patogenezu autoimunskih bolesti štitaste žlezde. Nakon uvodnih razmatranja, detaljno su prikazani dominantni autoantigeni i autoantitela (kao pokazatelji autoimunskog procesa u štitastoj žlezdi), kao i mehanizmi uključeni u efektorsku fazu autoimunskog procesa koji uzrokuju oštećenje tireocita. Posebno je analizirana uloga poremećaja u regulaciji procesa apoptoze u nastanku autoimunskih bolesti štitaste žlezde

The content of estragole in essential oil and infusion of basil herb, Ocimum basilicum L.

Herba bosiljka se tradicionalno koristi kao karminativ, spazmolitik, blag sedativ i laktagog. Etarsko ulje herbe bosiljka sadrži fenilpropanoid estragol, koji je kancerogen i genotoksičan, a čija količina zavisi od hemotipa bosiljka. Količina estragola koja se unosi putem biljnih lekovitih proizvoda, prema preporuci Evropske agencije za lekove (EMA), ograničena je na 0,5 mg dnevno tokom 14 dana. Cilj rada bio je ispitivanje hemijskog sastava etarskih ulja, sadržaja estragola u etarskim uljima i infuzima herbe bosiljka, dostupne u Srbiji. Ispitivani su komercijalni uzorci monokomponentnih čajeva i uzorci herbe bosiljka gajenog u domaćinstvima u različitim delovima Srbije. Infuzi su pripremljeni prelivanjem uzoraka ključalom vodom (2 g/150 ml) i ceđenjem nakon 15 min. Izolovanje etarskog ulja iz herbe, kao i izolovanje estragola iz infuza izvršeno je destilacijom vodenom parom u aparaturi po Klevendžeru. Analiza etarskih ulja i destilata infuza izvršena je GC‐FID‐MS metodom. Sadržaj estragola u etarskim uljima i infuzima određen je metodom eksternog standarda. Sastav svih ispitivanih etarskih ulja odgovara evropskom hemotipu bosiljka, kojeg karakteriše visok sadržaj linalola ili linalola i estragola. Sadržaj estragola u ispitivanim etarskim uljima iznosio je 2,1‐565,4 mg/ml, a u infuzima 0,5‐11,3 μg/ml. Primenom ispitivanih infuza prema preporuci Deutsche Arzneimittel‐Codex (DAC) (2 g/150 ml, tri puta dnevno), dnevni unos estragola iznosio bi 0,2‐5,1 mg. Primenom infuza pojedinih uzoraka herbe bosiljka postoji mogućnost prekoračenjа dozvoljenоg dnevnog unosa estragola. Dobijeni rezultati ukazuju na značaj utvrđivanja sadržaja estragola u herbi bosiljka koja se koristi u obliku biljnih lekovitih proizvoda.Basil herb is traditionally used as carminative, spasmolytic, mild sedative and lactagogue. Basil essential oil contains phenylpropanoide estragole, which is carcinogenic and genotoxic, and its amount depends on Basil chemotype. The amount of estragole administered through herbal medicinal products, according to the recommendation of the European Medicines Agency (EMA), is limited to 0.5 mg daily during 14 days. The aim of this work was examination of chemical composition of essential oils, as well as determination of estragole content in essential oils and infusions of Basil herb available in Serbia. We have analyzed commercial samples of monocomponent Basil herbal teas and samples of Basil herb cultivated in different parts of Serbia. Infusions were prepared by steeping samples in boiling water (2 g/150 ml) for 15 min and subsequent filtering. Isolation of the herb essential oil, as well as isolation of estragole from infusions was carried out by hydrodistillation using a Clevenger‐type apparatus. Analyses of essential oils and infusion distillates were performed using GC‐FID‐MS. The content of estragole in essential oils and infusions was determined using external standard method. Chemical composition of all investigated essential oils corresponds to European chemotype of Basil that is characterized by high content of linalool or linalool and estragole. The content of estragole in essential oils ranged from 2.1 to 565.4 mg/ml, whereas infusions contained 0.5‐11.3 μg/ml of this compound. If the examined infusions are administered according to the recommendation of Deutsche Arzneimittel‐ Codex (DAC) (2 g/150 ml, three times daily), the daily intake of estragole would be 0.2‐ 5.1 mg. By administration of infusions of some Basil herb samples, there is a possibility of exceeding the permitted daily intake of estragole. The obtained results indicate the importance of determining the content of estragole in Basil herb used in the form of herbal medicinal products.VII Kongres farmaceuta Srbije sa međunarodnim učešćem: Zajedno stvaramo budućnost farmacije, Beograd, Srbija, 10-14. oktobar 2018

Latent Murine Cytomegalovirus Infection Contributes to EAE Pathogenesis

ABSTRACT Viral infection has been identified as the most likely environmental trigger of multiple sclerosis (MS). There are conflicting data regarding the role of cytomegalovirus (CMV) in MS pathogenesis. We utilised experimental autoimmune encephalomyelitis (EAE)-resistant BALB/c mice and murine cytomegalovirus (MCMV), the murine homolog of CMV, to examine the mechanism by which viral infection enhances autoimmune neuroinflammation. Mice subjected to latent neonatal MCMV infection developed the typical characteristics of EAE. Similar to MS, the MCMV-infected EAE-induced mice developed infiltrates in the central nervous system (CNS) composed of similar percentages of CD4+ and CD8+ T cells. The influx of both Th 1 and Th 17 cells into the CNS of MCMV- infected EAE-induced mice was observed. Interestingly, the development of autoimmune neuroinflammation after latent MCMV infection was accompanied by a significant influx of Tc17 cells (CD8+IL-17+ and CD8+RoRγt+) but not Tc1, cells. Our results suggest that latent MCMV infection affects the development of inflammatory lymphocytes that exhibit encephalitogenic potential, thereby mediating increased CNS pathology following EAE induction, and that CMV represents a possible environmental factor in the pathogenesis of MS and other autoimmune disease

Synthesis, characterization, biomolecular interactions, molecular docking, and in vitro and in vivo anticancer activities of novel ruthenium(III) Schiff base complexes

In order to discover new anticancer drugs, novel ruthenium(III) complexes [Ru(L)Cl(H2O)], where L is tetradentate Schiff base bis(acetylacetone)ethylendiimine (acacen, 1), bis(benzoylacetone)ethylendiimine (bzacen, 2), (acetylacetone)(benzoylaceton)ethylendiimine (acacbzacen, 3), bis(acetylacetone)propylendiimine (acacpn, 4), bis(benzoylacetone)propylendiimine (bzacpn, 5) or (acetylacetone)(benzoylaceton)propylendiimine (acacbzacpn, 6), were synthesized. The complexes 1 – 6 were characterized by elemental analysis, molar conductometry, and by various spectroscopic techniques, such as UV–Vis, IR, EPR, and ESI-MS. Based on in vitro DNA/BSA experiments, complexes 2 (bzacen) and 5 (bzacpn) with two aromatic rings showed the highest DNA/BSA-activity, suggesting that the presence of the aromatic ring on the tetradentate Schiff base ligand contributes to increased activity. Moreover, these two compounds showed the highest cytotoxic effects toward human, A549 and murine LLC1 lung cancer cells. These complexes altered the ratio of anti- and pro-apoptotic molecules and induced apoptosis of A549 cells. Further, complexes 2 and 5 reduced the percentage of Mcl1 and Bcl2 expressing LLC1 cells, induced their apoptotic death and exerted an antiproliferative effect against LLC1. Finally, complex 5 reduced the volume of mouse primary heterotopic Lewis lung cancer, while complex 2 reduced the incidence and mean number of metastases per lung. Additionally, molecular docking with DNA revealed that the reduced number of aromatic rings or their absence causes lower intercalative properties of the complexes in order: 2 > 5 > 6 > 3 > 4 > 1. It was observed that conventional hydrogen bonds and hydrophobic interactions contribute to the stabilization of the structures of complex-DNA. A molecular docking study with BSA revealed a predominance of 1 – 6 in binding affinity to the active site III, a third D-shaped hydrophobic pocket within subdomain IB