LICHENS AS INDICATORS OF AIR QUALITY IN BALNEOLOGICAL CENTER PROLOM BANJA (SOUTHERN SERBIA)

This paper deals with using lichens as a bioindicators of the air quality and it was conducted on the territory of balneological center Prolom Banja (Southern Serbia). The exploration was conducted in the year 2019. The analysis of the sample from 15 investigated points indicates the presence of 72 lichen taxa, which shows that this area is rich in lichen species. For each investigated point, the index values of atmospheric purity (IAP) and index of human impact (IHI) were calculated. The IAP values varied in range from 40 to 56, while IHI values ranged between 8 and 24. Therefore, the map showing the air quality of the investigated area was made. There is a presence of “normal lichen zone” on the map which indicates that the air quality in this area is quite good. There are no significant air pollution sources in this area, so the level of pollution is considered low or very low. In the investigated area there are not stations for the monitoring of physico-chemical parameters of air quality. The investigation of air quality on the territory of Prolom Banja has not been done until now.

New Values of Teucrium species: in Vitro Study of Cytotoxic Activities of Secondary Metabolites

The cytotoxicity of seven Teucrium species, a long time ago used as a food spices, for beverages and teas preparing, as well as therapeutics for digestive and respiratory diseases, were examined against human cervix adenocarcinoma HeLa, human melanoma Fem-x, human chronic myelogenous leukemia K562 and human breast adenocarcinoma MDA-MB-361 cells. MTT assay was used for determination of target cell survival. The most prominent cytotoxic effect was observed against K562 cells, especially by T. scordioides, T. montanum and T. botrys. All Teucrium extracts showed good cytotoxic activity on HeLa cells, but very low cytotoxic effect on MDA-MB-361 cells. In addition, the cytotoxic activities of T. scordioides and T. montanum extract were tested on healthy resting and phytohaemagglutinin-stimulated peripheral blood mononuclear cells (PHA-stimulated PBMC). T. scordioides and T. montanum extracts at concentration of 200 µg/ml reduced the resting PBMC and PHA-stimulated PBMC survival up to 10% and 20%, while the reduction of K562 cell survival at the same concentration of extracts was 94% and 97%, respectively. These results point to selectivity in their antitumor actions. Teucrium species can be regarded as promising candidates for natural plant sources of effective biological compounds as a supplements in the food industry, as well as for therapeutic use

ارزیابی کیفیت آزمون های چهار گزینه ای دروس تئوری عمومي گروه اندودانتیکس دانشکده دندانپزشکی قزوین در طی سال های تحصیلی 1388 تا 1396

Svrha je ovoga rada bila ispitati citotoksični, proapoptotski, antimigratorni i pro-antioksidacijski učinak metanolnih, acetonskih i etil-acetatnih ekstrakata dvaju vrsta lišaja, i to: Pseudevernia furfuracea i Platismatia glauca na tumorske stanične linije debelog crijeva (HCT-116 i SW-480). Citotoksični efekti na tumorskim staničnim linijama debelog crijeva uspoređeni su s onima na staničnoj liniji normalnih humanih fibroblasta (MRC-5). Citotoksičnost je ispitana MTT testom, stanična migracija Transwell esejom, dok je apoptoza praćena fluorescentnom metodom pomoću akridin narančastog bojila i etidijevog bromida. Prooksidacijski/antioksidacijski učinak praćen je spektrofotometrijskim mjerenjem koncentracije redoks parametara. Ispitani ekstrakti imali su značajan citotoksični učinak na tumorske stanične linije, bez zapaženog utjecaja na normalnu staničnu liniju. Najjača citotoksičnost postignuta je nakon 72 h obrade staničnih linija ekstraktima vrste P. furfuracea (IC50= (21,2±1.3) µg/mL pri obradi HCT-116 stanica etil-acetatnim ekstraktom i IC50=(51.3±0.8) µg/mL pri obradi stanica SW-480 acetonskim ekstraktom). Ekstrakti P. furfuracea imali su značajan proapoptotski i prooksidacijski učinak, dok je metanolni ekstrakt te vrste imao najjači antimigratorni učinak na ispitane tumorske stanične linije. Svi ispitani ekstrakti vrste P. glauca imali su znatan citotoksični učinak na stanice HCT-116 nakon 72 h tretmana (IC50<40 μg/mL), dok su metanolni i acetonski ekstrakti imali citotoksični učinak na stanice SW-480 nakon 24 h, s izraženom proapoptotskom/nekrotskom aktivnošću. Svi navedeni efekti posljedica su induciranog oksidacijskog stresa u tretiranim stanicama. Može se zaključiti da ekstrakti lišaja bitno mijenjaju vijabilnost i migratorni potencijal ispitanih tumorskih staničnih linija karcinoma debelog crijeva. Dobiveni rezultati pokazuju da su stanice HCT-116 osetljivije na ispitane ekstrakte, pri čemu ekstrakti vrste P. furfuracea pokazuju bolji proapototski i antimigratorni učinak. Ispitane vrste lišaja mogu se smatrati izvorima antikancerogenih aktivnih tvari.The aim of this study is to investigate cytotoxic, proapoptotic, antimigratory and pro-antioxidant effects of methanol, acetone and ethyl acetate extracts of lichens Pseudevernia furfuracea and Platismatia glauca on colorectal cancer (HCT-116 and SW-480) cell lines. We compared the cytotoxic effects on colorectal cancer cells with the effects obtained from normal human fibroblast (MRC-5) cell line. Tetrazolium (MTT) test evaluated the cytotoxic effects, Transwell assay evaluated cell migration, acridine orange/ethidium bromide (AO/EB) fluorescent method followed the apoptosis, while prooxidant/antioxidant effects were determined spectrophotometrically through concentration of redox parameters. The tested extracts showed considerable cytotoxic effect on cancer cells with no observable cytotoxic effect on normal cells. Ethyl acetate and acetone extract of P. furfuracea induced the highest cytotoxicity (IC50=(21.2±1.3) μg/mL on HCT-116, and IC50=(51.3±0.8) μg/mL on SW-480 cells, respectively, after 72 h), with noteworthy apoptotic and prooxidant effects, and antimigratory potential of methanol extract. P. glauca extracts induced cytotoxic effects on HCT-116 cells after 72 h (IC50<40 μg/mL), while only methanol and acetone extracts had cytotoxic effects on SW-480 cells after 24 h, with proapoptotic/necrotic activity, as a consequence of induced oxidative stress. In conclusion, lichen extracts changed to a great extent cell viability and migratory potential of colorectal cancer cell lines. HCT-116 cells were more sensitive to treatments, P. furfuracea had better proapoptotic and antimigratory effects, and both investigated lichen species might be a source of substances with anticancer activity

Activity of a Catalase Enzyme in Plants from the Burned Areas of the Vidlič Mountain Beech Forest

In 2007, a catastrophic fire on the Vidlic Mountain in south-east Serbia occurred. It burned down nearly 1000 ha of forest. Study of biochemical and physiological parameters in plants which inhabit post fire areas and their comparison with control is of essential importance in estimating the impact of fire on plant characteristics and potential applications. After the fire pioneer and indigenous plants from habitats affected by fire have a characteristic metabolism. In this comparative study, the activity of enzyme catalase (ЕC 1.11.1.6, H2 O2 : H2 O2 oxidoreductase) was determined in plant species from a habitat affected by fire and the same plant species from the forest which had not been affected by fire as a control. Assessment of enzyme activity was carried out on the root, leaves and flowers of plant species Geranium macrorrhizum, Doronicum columnae, Aegopodium podagraria, Fagus moesiaca, Tussilago farfara, Glechoma hirsuta, Chelidonium majus and Primula veris. The first group of plant samples used for determination was obtained from the habitat that was affected by fire two years ago. Catalase activity was measured using the gasometric method and the values obtained for this activity were expressed as ml of O2 . The presented results show a significant increase in catalase activity in individuals from habitats affected by fire in relation to the control group. Increased catalase activity is a consequence of oxidative stress caused by chemical changes in soil that were generated by fire.Publishe

Platismatia glauca and Pseudevernia furfuracea lichens as sources of antioxidant, antimicrobial and antibiofilm agents

The antioxidative, antimicrobial and antibiofilm potentials of acetone, ethyl acetate and methanol extracts of lichen species Platismatia glauca and Pseudevernia furfuracea were evaluated. The phytochemical analysis by GC, GC/MS and NMR revealed caperatic acid, atraric acid, atranorin and chloroatranorin as the predominant compounds in Platismatia glauca. Atraric acid, olivetoric acid, atranorin and chloroatranorin were the major constituents in Pseudevernia furfuracea. The strong antioxidant capacities of the Platismatia glauca and Pseudevernia furfuracea extracts were assessed by their total phenolic and flavonoid contents and DPPH scavenging activities. The methanol extracts of both species exhibited the strongest antioxidant activities with the highest IC50 value for Pseudevernia furfuracea (95.33 μg/mL). The lichen extracts demonstrated important antibacterial activities against 11 bacterial strains with detectable MIC values from 0.08 mg/mL to 2.5 mg/mL for Platismatia glauca and from 0.005 mg/mL to 2.5 mg/mL for Pseudevernia furfuracea. While the antibacterial activities of Pseudevernia furfuracea were solvent–independent, the acetone and ethyl acetate extracts of Platismatia glauca showed higher antibacterial activities compared to its methanol extract. The methanol extracts of both species demonstrated significant antifungal activities against 9 fungal strains with detectable MIC values from 0.04 mg/mL to 2.5 mg/mL. The best antifungal activities were determined against Candida species in Pseudevernia furfuracea extracts with remarkable MIC values which were lower than the MIC values of the positive contol fluconazole. The acetone and ethyl acetate extracts of Platismatia glauca showed better antibiofilm activities on Staphylococcus aureus and Proteus mirabilis with BIC value at 0.63 mg/mL then its methanol extract. On the other hand, the methanol extract of Pseudevernia furfuracea was more potent with BIC value at 1.25 mg/mL on Staphylococcus aureus and 0.63 mg/mL on Proteus mirabilis compared to other types of extracts. Our study indicates a possible use of lichens Platismatia glauca and Pseudevernia furfuracea as natural antioxidants and preservatives in food, pharmaceutical and cosmetic industry

A Comparison of MGMT Testing by MSP and qMSP in Paired Snap-Frozen and Formalin-Fixed Paraffin-Embedded Gliomas

Comparative analysis of the conventional methylation-specific PCR (MSP) vs. the quantitative MSP (qMSP) assessment of the O6-methylguanine-DNA methyltransferase (MGMT) promoter methylation status in 34 snap-frozen (SF) glioma samples was performed. The accuracy of the semi-quantitative MSP was compared with the corresponding qMSP semi-quantitative values using two semi-quantitative cut-off values (0—unmethylated and 1—weakly methylated) to discriminate methylated from unmethylated samples. In the case of the cut-off value 0, MSP test showed 80.0% sensitivity and 78.9% specificity compared to the reference qMSP analysis. However, when using the cut-off value 1, the diagnostic accuracy of the MSP test was significantly higher (85.7% sensitivity, 85.2% specificity). Fleiss’ Kappa statistical analyses indicated moderate agreement (Fleiss’ Kappa Coefficient = 0.509; 70.59% agreement) between MSP and qMSP semi-quantitative measurements of MGMT promoter methylation in glioma patients, justifying the conventional MSP use in diagnostics and confirming its high reliability. Further, we aimed to compare the validity of SF and formalin-fixed paraffin-embedded (FFPE) glioma samples for MGMT testing. Statistical analyses indicated moderate overall agreement of FFPE glioma samples and SF MSP semi-quantitative measurements (Fleiss’ Kappa Coefficient = 0.516/0.509; 70.0% agreement) and emphasized their low reliability in the assessment of highly methylated MGMT promoter samples

Antioxidant, Antimicrobial and Antiproliferative Activities of Five Lichen Species

The antioxidative, antimicrobial and antiproliferative potentials of the methanol extracts of the lichen species Parmelia sulcata, Flavoparmelia caperata, Evernia prunastri, Hypogymnia physodes and Cladonia foliacea were evaluated. The total phenolic content of the tested extracts varied from 78.12 to 141.59 mg of gallic acid equivalent (GA)/g of extract and the total flavonoid content from 20.14 to 44.43 mg of rutin equivalent (Ru)/g of extract. The antioxidant capacities of the lichen extracts were determined by 2,2-diphenyl-1-picrylhydrazyl (DPPH) radicals scavenging. Hypogymnia physodes with the highest phenolic content showed the strongest DPPH radical scavenging effect. Further, the antimicrobial potential of the lichen extracts was determined by a microdilution method on 29 microorganisms, including 15 strains of bacteria, 10 species of filamentous fungi and 4 yeast species. A high antimicrobial activity of all the tested extracts was observed with more potent inhibitory effects on the growth of Gram (+) bacteria. The highest antimicrobial activity among lichens was demonstrated by Hypogymnia physodes and Cladonia foliacea. Finally, the antiproliferative activity of the lichen extracts was explored on the colon cancer adenocarcinoma cell line HCT-116 by MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) viability assay and acridine orange/ethidium bromide staining. The methanol extracts of Hypogymnia physodes and Cladonia foliacea showed a better cytotoxic activity than the other extracts. All lichen species showed the ability to induce apoptosis of HCT-116 cells

Lichens as biological indicators of air quality in the urban area of Kuršumlija (Southern Serbia)

In this work, we provided a first indication of Kuršumlija (town of Southern Serbia) air quality using epiphytic lichens known as bioindicators. The investigations were carried out from April to October 2014. The analysis of thalli samples (from 15 investigated points in Kuršumlija), indicates the presence of 42 lichens taxa from 23 genera. An assessment of air quality was made by using the Index of Atmospheric Purity (IAP) and Index of Human Impact (IHI). IHI values ranged between 15 and 36. IAP values ranged between 10 and 28. According to the IAP values, it has been found that there are three different air pollution zones: “normal”, “struggle” and “lichen desert”. The latest characterize the majority of the urban area. The aim of this study is to formulate an indication of air quality using lichens as bioindicators on the basis of lichens diversity and frequency. This is of great importance because there are no continuous physical and chemical measurements of air quality in this area

New Values of <i>Teucrium</i> species: <i>in Vitro</i> Study of Cytotoxic Activities of Secondary Metabolites

The cytotoxicity of seven Teucrium species, a long time ago used as a food spices, for beverages and teas preparing, as well as therapeutics for digestive and respiratory diseases, were examined against human cervix adenocarcinoma HeLa, human melanoma Fem-x, human chronic myelogenous leukemia K562 and human breast adenocarcinoma MDA-MB-361 cells. MTT assay was used for determination of target cell survival. The most prominent cytotoxic effect was observed against K562 cells, especially by T. scordioides, T. montanum and T. botrys. All Teucrium extracts showed good cytotoxic activity on HeLa cells, but very low cytotoxic effect on MDA-MB-361 cells. In addition, the cytotoxic activities of T. scordioides and T. montanum extract were tested on healthy resting and phytohaemagglutinin-stimulated peripheral blood mononuclear cells (PHA-stimulated PBMC). T. scordioidesand T. montanum extracts at concentration of 200 µg/ml reduced the resting PBMC and PHA-stimulated PBMC survival up to 10% and 20%, while the reduction of K562 cell survival at the same concentration of extracts was 94% and 97%, respectively. These results point to selectivity in their antitumor actions. Teucrium species can be regarded as promising candidates for natural plant sources of effective biological compounds as a supplements in the food industry, as well as for therapeutic use.</p

Epiphytic lichen Flavoparmelia caperata as a sentinel for trace metal pollution

Widely spread lichen specie Flavoparmelia caperata is used in a biomonitoring study for atmospheric trace metal pollution in natural ecosystems in Southeastern Serbia. The concentration and distribution pattern of 21 metals in lichens were determined by inductively coupled plasma atomic emission spectrometry. The difference observed between metal deposition in peripheral and central parts of lichen thalli reflected air quality changes in the last and previous years. These findings were confirmed with principal component analysis. Our study demonstrated the accumulation of Ba, K, Mg, Na, Tl and Zn in peripheral parts of thalli, while As, B, Cd, Cr, Cu, Fe, Ga, In, Li, Ni, Pb and Se were concentrated in central parts of thalli