Mulberry extracts (Morus spp., Morasceae), composition, activity and application

 Upotrebom različitih ekstragenasa izvršena je ekstrakcija ploda, lista i korena belog (Morus alba L.) i crnog duda (Morus nigra L. ). Ekstrakcija etanolom je izvršena pri predhodno određenim optimalnim uslovima ekstrakcije (70% koncentracija etanola, temperatura 60°C i odnos rastvarač:droga 20 ml/g). Ispitan je hemijski sastav ekstrakata različitih delova duda. Ispitivanja hemijskog sastava etanolnih ekstrakata obuhvatala su određivanje sadržaja ukupnih fenola, ukupnih flavonoida, ukupnih i monomernih antocijana, kvalitativni i kvantitativni sastav polifenolnih komponenata, kao i sadržaj mikro-, makro- i toksičnih elemenata. Najvedi sadržaj ukupnih fenolnih jedinjenja određen je u ekstraktu korena M. nigra (186 mg EHK/g), a najvedi sadržaj flavonoidnih jedinjenja u ekstraktu lista M. nigra (67 mg ER/g). U ekstraktima lista duda po svom sadržaju u odnosu na ukupan sadržaj polifenolnih jedinjenja ističe se kafena kiselina (od 53,34 do 62,11%), dok se u ekstraktima korena izdvaja hlorogenska kiselina (udeo vedi od 74%). U pogledu mineralnog sastava ekstrakata relativno veliki sadržaj Ca i Mg je karakterističan za obe vrste ekstrakata (vedi od 1.400 μg/g). Mikroelementi u svim ispitanim uzorcima duda po opadajudem nivou, su Fe, B, Zn, Cu i Mn. U ekstraktima dobijenim superkritičnim CO2 određen je sadržaj masnih kiselina i ukupnih karotenoida. U ispitivanim ekstraktima dominante su nezasidene masne kiseline, linolna i linolenska kiselina, a značajan je udeo i zasidene masne kiseline (palmitinske kiseline). Ispitivanje delovanja ekstrakata obuhvatalo je evaluaciju antioksidativne aktivnosti pradenjem sposobnosti neutralizacije slobodnih radikala, redukcionog potencijala i inhibicije lipidne peroksidacije. Po svim primenjenim metodama najvedu aktivnost je pokazao ekstrakt lista crnog duda. U završnoj fazi rada ispitana je in vitro citotoksična aktivnost ekstrakata duda, njihovim delovanjem na rast tri delijske linije: Hep2, RD i L2OB. Ekstrakti lista M. nigra su se pokazali kao najpotentniji inhibitori rasta na sve tri delijske linije (IC50 B> Zn> Cu> Mn. In the extracts obtained by supercritical CO2 fatty acids and total carotenoids were determined. In these extracts dominant unsaturated fatty acids were linoleic and linolenic acid. Also, significant proportion of saturated fatty acid (palmitic acid) has been detected. Testing of extracts activity included the evaluation of antioxidant activity applying the methods based on: neutralize free radicals, reducing power potential and the inhibition of lipid peroxidation. All applied methods showed the highest activity of black mulberry leaf extract. In the final phase of work in vitro cytotoxic activity of mulberry extracts was examined. Their effect on growth of three cell lines: Hep2, RD, and L2OB were analysed. Extracts of M. nigra proved to be the most potent inhibitors of growth in all three cell lines (IC50<30 mg/g). On the basis of all conducted tests it has been proven that M. nigra extracts are products with the best characteristics

Isolation and GC Analysis of Fatty Acids: Study Case of Stinging Nettle Leaves

Urtica dioica L. is perennial plant with remarkable medical properties and has been widely used in folk medicine and as a food. Fatty acids presented in its leaves were extracted applying three different techniques: classic, automated Soxhlet, and supercritical fluid extraction (SFE). SFE was performed at three different pressures (100, 200, and 300 bar) and two different temperatures (40 and 60°C). Obtained extract was analyzed using GC-FID analytical technique in order to obtain fatty acid profile samples. The highest yield was obtained in the case of automated Soxhlet extraction (218.907 mg/g), while the lowest was observed in the case of classic extraction (15.031 mg/g). Soxhlet approach provided the highest yield of saturated fatty acids (173.348 mg/g), while supercritical fluid extraction gave better results in the case of unsaturated fatty acids (60.062 mg/g). Deeper analysis of SFE extracts revealed relationship between extraction parameters (temperature and pressure) and yields of fatty acids where lower pressure ensured higher yield of unsaturated while higher pressure gave higher yield of saturated acids. On the other hand, increasing in temperature at isobaric conditions negatively influenced the yield of fatty acids except in the case of 300 bar where yield was higher at 60°C

Association of functional variants of phase i and ii genes with chronic obstructive pulmonary disease in a Serbian population

Background: Chronic obstructive pulmonary disease (COPD) is a complex disorder characterized by increased oxidative stress. Functional genetic variants of phase I and II genes are implicated in oxidants antioxidants imbalance and may be involved in COPD development. In this study, we aimed to investigate the role of cytochrome P450 (CYP), glutathione S-transferase (GST) and microsomal epoxide hydrolase (mEH) functional variants in the pathogenesis of COPD in a Serbian population. Methods: The genotypes of 122 COPD patients and 100 controls with normal lung function were determined for CYP1A1 *1A/*2A, CYP2E1 *1A/*5B, GSTM1 null, GSTT1 null GSTP1 11e105Val, mEH Tyr113His and mEH His139Arg gene variants. Results: Results obtained showed that GSTM1 null variant was significantly more represented in COPD patients than in controls (61.5% vs. 47.0%; OR=1.80; p=0.042). Also, a significant difference was observed for combinations of GSTM1 null and GSTP1 105Val/(Val) (38.5% vs. 24.0%; OR=1.98; p=0.029), as well as for CYP1A1 *1A/*2A, GSTM1 null and mEH 113His/(His) genotypes (7.4% vs. 1.0%; OR=7.88; p=0.025). Conclusions: These are the first data concerning the analysis of the variants of phase I and II genes in the pathogenesis of COPD in a Serbian population. Results obtained in this study open up the possibility for thorough analyses of the role of genetic factors in COPD on larger cohorts. Also, they implicate the importance of previously described genetic associations with COPD in our population, as well as reveal a new one, not reported so far

Association of functional variants of phase I and II genes with chronic obstructive pulmonary disease in a Serbian population

Uvod: Hronična opstruktivna bolest pluća (HOBP) jeste složeno oboljenje koje karakteriše povišen oksidativni stres. Funkcionalne varijante gena faze I i II ksenobiotičkog metabolizma mogu uticati na ravnotežu oksidanti-antioksidanti i mogu dovesti do razvoja HOBP. Cilj ove studije je bio ispitivanje uloge funkcionalnih genskih varijanti u genima za citohrom P450 (CYP), glutation S-transferazu (GST) i mikrozomalnu epoksidnu hidrolazu (mEH) u patogenezi HOBP u srpskoj populaciji. Metode: U ovoj studiji analizirane su genske varijante CYP1A1 *1A/*2A, CYP2E1 *1A/*5B, GSTM1 null, GSTT1 null, GSTP1 Ile105Val, mEH Tyr113His i mEH His139Arg u grupi obolelih od HOBP koja je obuhvatala 122 ispitanika i kontrolnoj grupi koja je obuhvatala 100 ispitanika sa normalnom funkcijom pluća. Rezultati: Dobijeni rezultati su pokazali da je GSTM1 null varijanta statistički značajno povišena u grupi obolelih od HOBP u poređenju sa kontrolnom grupom (61,5% i 47,0%; or = 1,80; p = 0,042). Takođe, uočena je značajna razlika u zastupljenosti kombinacije genotipova GSTM1 null i GSTP1 105Val/(Val) (38,5% i 24,0%; or = 1,98; p = 0,029), kao i kombinacije CYP1A1 *1A/*2A, GSTM1 null i mEH 113H is/(H is) (7,4% i 1,0% ; or = 7 ,88; p = 0,025). Zaključak: Ovo su prvi podaci o ulozi genskih varijanti gena faze I i II u patogenezi HOBP u srpskoj populaciji. Rezultati dobijeni u ovoj studiji otvaraju mogućnost za detaljniju analizu uloge genetičkih faktora u HOBP na većim grupama ispitanika. Pored toga, podaci dobijeni u našoj studiji potvrđuju važnost genetičkih determinanti povezanih sa HOBP u prethodnim studijama, ali takođe otkrivaju nove genetičke faktore, koji nisu objavljeni do sada.Background: Chronic obstructive pulmonary disease (COPD) is a complex disorder characterized by increased oxidative stress. Functional genetic variants of phase I and II genes are implicated in oxidants-antioxidants imbalance and may be involved in COPD development. In this study, we aimed to investigate the role of cytochrome P450 (CYP), glutathione S-transferase (GST) and microsomal epoxide hydrolase (mEH) functional variants in the pathogenesis of COPD in a Serbian population. Methods: The genotypes of l2 2 COPD patients and 100 controls with normal lung function were determined for CYP1A1 *1A /*2A , CYP2E1 *1 A /*5B , GSTM1 null, GSTT1 null GSTP1 Ile105Val, mEH Tyr113His and mEH His139Arg gene variants. Results: Results obtained showed that GSTM1 null variant was significantly more represented in COPD patients than in controls (61.5% vs. 47.0%; or = 1.80; p = 0.042). Also, a significant difference was observed for combinations of GSTM1 null and GSTP1 105Val/(Val) (38.5% vs. 24.0%; or = 1.98; p = 0.029), as well as for CYP1A1 *1A/*2A, GSTM1 null and mEH 113His/(His) genotypes (7.4% vs. 1.0%; or = 7.88; p = 0.025). Conclusions: These are the first data concerning the analysis of the variants of phase I and II genes in the pathogenesis of COPD in a Serbian population. Results obtained in this study open up the possibility for thorough analyses of the role of genetic factors in COPD on larger cohorts. Also, they implicate the importance of previously described genetic associations with COPD in our population, as well as reveal a new one, not reported so far

Carcass Quality and Fatty Acids Profile of the Fatteners of Swallow-Belly Mangalitsa Breed Reared in Outdoor System

The objective of this study was to determine carcass quality of fatteners of Swallow-belly Mangalitsa breed reared in outdoor system. In m. longissimus dorsi (MLD) samples chemical composition, cholesterol content and fatty acids profile was determined. The study included 22 castrated males reared in the oak forest from the early spring to late autumn, with minimal corn harvest (approx. 0.5 kg per day). During the winter, the fatteners were kept in wooden stables and fed with cereals (corn and barley mixture in a ratio of 70:30, approx. 3 kg per day). The data was carried out by statistical package SAS 9.1.3 (SAS Inst. Inc., 2002-2003) and basic statistical parameters are showed (Mean±SD). At the end of fattening, the pigs were about one year old, and slaughtered at 92.0±16.7 kg live body weight. The average lean content in the carcasses was 35.9±2.8 %, while the content of intermuscular fat in MLD was 6.2±1.2 %. In the MLD samples average content of proteins, water, fat and ash were 20.7±0.5 %, 70.4±0.5 %, 7.7±0.5 % and 1.0±0.1 %, respectively. Average cholesterol content was 41.4±3.8 mg/100g. Shares of saturated fatty acids (ΣSFA) in MLD fat was 35.6±1.2 %, monounsaturated fatty acids (ΣMUFA) was 56.6±1.8 % and polyunsaturated fatty acids (ΣPUFA) was 6.9±1.2%. Within ΣSFA the most common was palmitic acid (C16:0, 24.7±0.6 %), wile within ΣMUFA it was oleic acid (C18:1c-9, 46.9±1.1 %) and within ΣPUFA it was linoleic acid (C18:2n-6, 5.6±1.0 %). ΣPUFA and ΣSFA ratio was 0.2±0.0, while omega-6 and omega-3 ratio was 25.1±5.6. Showed ratios are not desirable from the nutritional aspect of human diet

Polyaniline thin films in sensors for detection of toxic welding vapors

In this paper the possibilities of applying sensors based on filters with surface acoustic waves (SAW) for the detection of harmful products in welding processes are analyzed. This review discusses the sensing mechanism and configurations of the sensors. The principles of SAW sensors are considered with special emphasis on the application of conducting polyaniline nanocomposites as sensitive thin layers in sensors for the detection of CO, NO2, and COCl2. The literature suggests the use of In2O3 as a dopant for CO and NO2 detection sensors, and ethylenediamine and phenylenediamine in COCl2 sensors. The paper presents original results of modeling of sensors that are designed for detection of CO and NO2 for which there are experimental data in the literature. The results obtained based on this model are in excellent agreement with results from the literature, which shows the validity of modeling. Based on the developed modeling methods, calculations were done for sensors for the detection of these gases in which the structure was the surface of quartz. From these modeling results it can be concluded that the use of the quartz substrate provides better sensitivity. In addition, the quartz sensors are thermally stable.U radu su analizirane mogućnosti primene senzora u čijoj osnovi se nalaze filtri sa površinskim akustičkim talasom (PAT), za detekciju štetnih produkata u procesu zavarivanja. Objašnjeni su principi rada PAT senzora sa posebnim osvrtom na primenu elektroprovodnih polianilinskih nanokompozitnih materijala – polianilin/In2O3 i etilendiamin i fenilendiamin polianilinskih nanokompozita kao osetljivih tankih slojeva za detekciju CO, NO2 i COCl2. Prikazan je deo originalnih rezultata modelovanja PAT senzora koji su namenjeni detekciji pomenutih gasova. Rezultati dobijeni na bazi sopstvenog modela poređeni su sa rezultatima iz literature. Pokazano je da se ovim na činom modelovanja može uspešno odrediti koncentracija štetnih gasova u okruženju

Tumor necrosis factor alpha and alpha-1 antitrypsin gene variants in Serbian pediatric arterial ischemic stroke patients

Etiologija arterijskog ishemijskog moždanog udara (AIS) kod dece je veoma kompleksna i razlikuje se od one kod odraslih. Iako je redak, moždani udar kod dece predstavlja značajan uzrok mortaliteta i morbiditeta. Sve je više podataka o ulozi genetičkih faktora, uključujući tu i medijatore inflamacije, u nastanku i ishodu moždanog udara. U ovoj studiji, odabrali smo da ispitamo ulogu polimorfizma -308G/A u genu za faktor nekroze tumora-alfa (eng. Tumor Necrosis Factor α -TNFα), kao i mutacija S i Z u genu za alfa1-antitripsin (AAT)u etiologiji moždanog udara kod dece. Polimorfizam -308G/A u genu za TNFα dovodi do povećanja koncentracije ovog proteina u plazmi, što bi moglo da doprinese patologiji moždanog udara. Pokazano je i da povišena koncentracija AAT može da predstavlja rizik za nastanak moždanog udara kod dece. S obzirom da mutacije S i Z u genu za AAT dovode do smanjenja koncentracije ovog proteina u plazmi, one bi mogle da imaju protektivnu ulogu kada je u pitanju moždani udar.Genske varijante TNFα (-308G/A) i AAT (S i Z) su ispitivane u grupi od 26 dece sa AIS i 100 odraslih osoba PCR/RFLP metodom.Nije nađena statistički značajna razlika u učestalosti -308G/A TNFα polimorfizma između pacijenata i kontrola, tako da u našoj grupi pacijenata TNFα najverovatnije nije imao značajnu ulogu u razvoju bolesti. Ni kod jednog pacijenta nije pronađena nijedna od ispitivanih mutacija u genu za AAT, što je bilo u skladu sa potencijalnom protektivnom ulogom ovih varijanti.AIS je multifaktorijalno oboljenje, u čijoj patologiji veliki broj gena ima -ulogu, tako da je potrebna dalja analiza zajedničkog delovanja većeg broja genskih varijanti da bi se rasvetlila njihova uloga kao genetičkih faktora rizika i njihovog uticaja na razvoj i ishod moždanog udara.The etiology of arterial ischemic stroke (AIS) in children is complex, and different from that in adults. Although rare, stroke in children is an important cause of mortality and morbidity. There is increasing evidence that genetic factors, including inflammation mediators, have a role in occurrence and outcome of stroke. We have chosen to assess the role of polymorphism -308G/A in the promoter of tumor necrosis factor α (TNFα) gene and S and Z mutations in alpha 1-antitrypsin (AAT) gene in the etiology of stroke in children. TNFα polymorphism affects plasma levels of this proinflamatory cytokine, and this could contribute to stroke pathology. It has been shown that increased AAT concentration may present a risk for AIS in children. Since S and Z mutations in AAT gene reduce its levels in plasma they could have a protective role in pediatric stroke. In this study twenty six children with AIS and 100 unrelated individuals from Serbian general population were investigated by PCR/RFLP for these gene variations. No statistically significant difference was observed between patients and general population in distribution of genotypes for -308G/A TNFα polymorphism, so its contributory role in the etiology of stroke was not evident in our group of patients. None of the tested AAT gene mutations were found in patients, which is in concordance with the proposed protective role of deficient AAT variants. AIS is a multifactorial disease, with many genes having a modest role in its pathophysiology, so further analyses of their combined effect are needed to elucidate genetic risk factors in the etiology and outcome of stroke in pediatric patients

Targeting autophagy to modulate cell survival: a comparative analysis in cancer, normal and embryonic cells

Autophagy is linked to multiple cancer-related signaling pathways, and represents a defense mechanism for cancer cells under therapeutic stress. The crosstalk between apoptosis and autophagy is essential for both tumorigenesis and embryonic development. We studied the influence of autophagy on cell survival in pro-apoptotic conditions induced by anticancer drugs in three model systems: human cancer cells (NCI-H460, COR-L23 and U87), human normal cells (HaCaT and MRC-5) and zebrafish embryos (Danio rerio). Autophagy induction with AZD2014 and tamoxifen antagonized the pro-apoptotic effect of chemotherapeutics doxorubicin and cisplatin in cell lines, while autophagy inhibition by wortmannin and chloroquine synergized the action of both anticancer agents. This effect was further verified by assessing cleaved caspase-3 and PARP-1 levels. Autophagy inhibitors significantly increased both apoptotic markers when applied in combination with doxorubicin while autophagy inducers had the opposite effect. In a similar manner, autophagy induction in zebrafish embryos prevented cisplatin-induced apoptosis in the tail region while autophagy inhibition increased cell death in the tail and retina of cisplatin-treated animals. Autophagy modulation with direct inhibitors of the PI3kinase/Akt/mTOR pathway (AZD2014 and wortmannin) triggered the cellular response to anticancer drugs more effectively in NCI-H460 and zebrafish embryonic models compared to HaCaT suggesting that these modulators are selective towards rapidly proliferating cells. Therefore, evaluating the autophagic properties of chemotherapeutics could help determine more accurately the fate of different cell types under treatment. Our study underlines the importance of testing autophagic activity of potential anticancer agents in a comparative approach to develop more rational anticancer therapeutic strategies.Related to published version: [https://imagine.imgge.bg.ac.rs/handle/123456789/1004]This is the peer reviewed version of the paper: Divac Rankov, A., Ljujić, M., Petrić, M., Radojković, D., Pesić, M., & Dinić, J. (2017). Targeting autophagy to modulate cell survival: A comparative analysis in cancer, normal and embryonic cells. Histochemistry and Cell Biology, 148(5), 529–544. [https://doi.org/10.1007/s00418-017-1590-4