Analiza antiapoptotskog proteina bcl-2 u skvamocelularnom karcinomu usne regije

Aim: The aim of this study was to analyze the presence of the anti-apoptotic protein bcl-2 in oral squamous cell carcinoma and determine its potential role in the development and progression of this type of tumor. Materials and methods: The expression of bcl-2 was determined in 28 paraffin blocks of oral squamous cell carcinoma using the immunohistochemical method. The percentage of the immuno-reactive cells in positively stained tumor regions was determined using the microscopic analysis and Ozaria software. Results: Positive immunohistochemical test was observed in 19 out of 28 samples (68%) as follows: in 11 samples there was a low (+), in four a moderate (++) and in the last four a high percentage (+++) of stained cells. In the group of patients at the low stage of the disease (T2), 50% of tumor samples showed bcl-2 protein expression whereas in the higher stages (T3 and T4) of positively stained samples, this percentage was 67%. There was a trend of an increasing number of cells with positive bcl-2 staining in the tumors of higher clinical stages but not the level of bcl-2 protein expression. Conclusion: Both parameters, the presence of bcl-2 staining and the percentage of cells with bcl-2 immunoexpression, may act as additional prognostic parameters that indicate an increased proliferative tumor potential.Cilj ove studije bio je analiza prisustva antiapoptotskog proteina bcl-2 u skvamocelularnom karcinomu usne regije i procena njegove eventualne uloge u razvoju i progresiji ove vrste tumora. Materijal i metode: Na uzorku od 28 parafinskih blokova skvamocelularnog karcinoma usne regije, imunohistohemijskom metodom ispitan je ekspresioni status bcl-2 proteina. Mikroskopskom analizom i primenom softvera- Ozaria određen je procenat imunoreaktivnih ćelija u pozitivno obojenim tumorskim regijama. Rezultat: Pozitivnu imunohistohemijsku obojenost pokazalo je 19 od 28 uzoraka (68%) i to: 11 je bilo sa niskim (+), 4 sa srednjim (++) i 4 sa visokim procentom (+++) obojenih ćelija. U grupi pacijenata niskog stadijuma (T2) 50 % uzoraka tumora je pokazivalo ekspresiju bcl-2 proteina dok je u višim stadijumima (T3 i T4) pozitivnih uzoraka bilo 67%. Postojao je trend porasta broja ćelija sa pozitivnom bcl-2 obojenošću kod tumora u višim kliničkim stadijumima, ali ne i povećan nivo ekspresije bcl-2 proteina. Zaključak: Oba parametra, prisustvo bcl-2 obojenosti i procenat ćelija sa bcl-2 imunoekspresijom, mogu predstavljati dopunske prognostičke parametre koji ukazuju na povećan proliferativni potencijal tumora

Primjene vrijednosti retencijskih sila CEKA ankera

Retention o f a partial denture is the resistance of retaining elements to forces which endeavour to dislodge it from its base. Basic retention is obtained by means o f clasps, attachments, diaphragms, telescopic crowns, magnets etc. The aim of the present study was to determine the dynamics o f reduced retention forces in CEKA attachments (OL 694). An instrument, especially constructed for the purpose, was used to measure retention forces, and the test was carried out with and without the addition o f saliva. The results of the study, which are presented graphically and in tables, and described by exponential law, show that at the commencement o f the test the value of retention force in the attachment, without the addition o f saliva, amounted to approximately 10.2 N, and after 1200 cycles o f insertion and removal, 3.4 N. With the addition o f saliva the retention force o f the attachment amounted to 9.92 N at the commencement o f the test and after 1200 cycles o f insertion and removal, 5.12 N. A fundamental factor which has a cardinal effect on the extent of the retention force is wear and fatigue o f the material. Although in this study no significant differences in the reduction o f retention fo rces were found than had been anticipated, it is recommended that special attention should be paid to the choice of retention technique in partial dentures.Retencija djelomične proteze jest otpor retencijskih elemenata silama koje je nastoje pomaknuti s ležišta. Osnovna retencija dobiva se s pomoću kvačica, ankera, prečki, teleskop krunica, magneta i dr. Ovim radom nastojala se utvrditi dinamika smanjenja retetncijske sile kod CEKA ankera (OL 694), Ispitivalo se je s pomoću aparature za mjerenje retencijske sile, koja je konstruirana u tu svrhu. Ispitivanje je rađeno s nazočnošću sline i bez nje. Rezultati istraživanja - koji su prikazanai grafički i tablicama, a opisani eksponencijslnim zakonom - pokazuju da je vrijednost retencijske sile kod etečmena bez nazočnosti sline na početku ispitivanja iznosila prosječno 10.2 N. Nakon 1200 ciklusa stavljanja i vađenja, izmjerena je sila 3,4 N. Promjena retencijske sile etečmena s nazočnošću sline iznosila je od početnih 9,92 N do 5,12 N nakon 1200 ciklusa stavljanja i vađenja. Čimbenik koji bitno utječe na veličinu retencijske sile jest trošenje i zamor materijala. Iako u ovom istraživanju nisu vidljive velike razlike u opadanju retencijskih sila od pretpostavljenih, može se preporučiti posebna pozornost izboru načina retencije djelomične proteze

STR loci D19S216, D20S502 and D20S842 analysis in the Serbian population using dentin DNA

Dentin provides a protective enclosure for genomic and mitochondrial DNA. In the present study, DNA was obtained from pulverized or ground teeth. The quality of the DNA extracted from the teeth of 70 unrelated individuals was tested in the context of assessing the allelic and genotypic frequencies of autosomal loci D19S216, D20S502 and D20S842, and calculating a number of parameters of population genetics and forensic interest. This study illustrates that teeth can be a convenient tissue to extract DNA from large numbers of individuals for population genetic studies as well as for forensic case work

Multiple Sclerosis and Cancers in Croatia – A Possible Protective Role of the »Mediterranean Diet«

Multiple sclerosis (MS) is an autoimmune disease triggered by a combination of genetic and environmental risk factors which are however individually insufficient to provoke the disease. Previous investigations studied the coexistence of cancer in MS patients, and only a few relations between the geographic distribution of MS and that of cancer. The aim of this research was to find an environmental link between the aetiology of MS and cancers in Croatia. Incidence and prevalence of MS in Croatia were compared with the incidence of the most frequent cancer sites: stomach cancer, cancer of the colon and the rectum, pancreatic cancer, lung cancer, cancer of the kidneys and brain cancer. Data for MS were collected from seven population-based epidemiologic studies which used Poser’ s diagnostic criteria and reported the number of cases and the magnitude of the studied population. Data for cancers were drawn from the Croatian National Cancer Registry. The analysis was done for single municipalities, grouped in their belonging regions or counties, and separately for the continental and the coastal area. For each rate a 95% confidence interval was calculated. The differences between rates were tested with the chi-square test as well. In addition, MS incidence or prevalence were correlated with the corresponding cancer incidence data. Pearson’s correlation coefficients were used to measure the correlation between both diseases. Calculations were done with the statistical package Statistica V 7.1. and the Smith’s Statistical Package freeware In the continental area of Croatia the mean annual incidence (per 100,000 inhabitants) of MS was nearly two folds higher than in the coastal area: 2.1 vs. 1.3 (p=0.0029). The difference was lower when expressed by prevalence: 46.5 vs. 36.7 (p=0.0601). Among the malignant neoplasms, in the continental area significantly higher incidence rates were found for stomach (32.9 vs. 20.8; p=3.14E-14) and lung cancer (55.8 vs. 46.4; p=1.21E-05), whilst colon cancer alone (20.4 vs. 15.7; p=9.44E-05) or colorectal cancer (38.3 vs. 31.6; p=8.18E-05) had a significantly higher incidence in the coastal area. The geographic distribution of MS expressed by incidence was significantly correlated with pancreatic (r=0,62024, df=23, p=0.00094) and lung cancer (r=0,46380, df=23, p=0.01953). This research adds further malignant neoplasms, possibly exposure-related, to the list of diseases with geographic distribution like MS. The similarity of MS distribution with the named malignancies is unlikely to be incidental. MS in Gorski Kotar and Slavonia seems to be associated with a diet rich in meat and fat. A diet rich in fat and meat and poor in vegetables is a risk factor for stomach, colorectum, pancreatic as well as lung cancers. Some authors have documented a possible protective role of the »Mediterranean diet« for the named cancers. Olive oil is the main source of fat in the »Mediterranean diet«. Oleocanthal, a phenolic compound of the extra-virgin olive oil was found to inhibit the cyclooxigenase enzymes which are involved in demyelination and tumorigenesis. We hypothesize that the »Mediterranean diet«, olive oil and particularly oleocanthal, to have a protective role in MS too

Mutacioni status gena p53 u oralnim skvamocelularnim karcinomima

Introduction. p53 gene is the most common tumor suppressor gene involved in pathogenesis oral squamous cell carcinoma (OSCC). Protein product of p53 gene contributes to cell cycle control and apoptosis. p53 gene mutations may lead to uncontrolled cell growth. The aim of this study was to determine the incidence of mutation in DNA-binding domain of p53 gene. Materials and Methods. In the 60 specimens, the presence of point mutation in exons 5, 6, 7 and 8 was detected using PCR-SSCP method. To confirm the presence of p53 mutation found by SSCP method, five samples were analyzed by sequencing of exon 5. Results. Point mutation affecting exons 5, 6, 7 and 8 were found in 60% of analyzed samples. A higher incidence of mutation was detected in exon 7 and 8 (60%), than in exon 5 and 6. Sequencing of exon 5, confirmed the presence of mutations revealed by SSCP method. Study of associations showed an increase of p53 mutations in poor differentiated and carcinoma of higher clinical stages. Conclusion. p53 gene is one of major factor in control of cell cycle and has important role in pathogenesis of oral squamous cell carcinoma.Uvod. TP53 je ključni tumor-supresorski gen uključen u patogenezu oralnih skvamocelularnih karcinoma (OSCK). Proteinski proizvod gena p53 zadužen je za kontrolu ćelijskog ciklusa i apoptozu, a mutacije u TP53 mogu dovesti do nekontrolisane proliferacije ćelija. Cilj ovog rada je bio da se utvrdi zastupljenost tačkastih mutacija u regionu gena p53 koji se vezuje za DNK, odnosno proceni uloga ovoga gena u patogenezi OSCK. Materijal i metode rada. U 60 uzoraka OSCK su korišćenjem metode lančane reakcije polimeraze i polimorfizma jednolančanih fragmenata DNK (PCR-SSCP) ispitane tačkaste mutacije u egzonima 5, 6, 7 i 8 gena p53. Pet slučajno odabranih uzoraka u kojima je otkrivena mutacija naknadno je podvrgnuto sekvenciranju radi potvrde validnosti metode PCR-SSCP. Rezultati. Tačkaste mutacije u nekom od analiziranih egzona gena p53 utvrđene su u 60% uzoraka OSCK. Veća učestalost mutacija zabeležena je u egzonima 7 i 8. Sekvenciranje je potvrdilo mutacije otkrivene metodom SSCP. Studija asocijacije pokazuje povećanje broja mutacija gena p53 kod slabo diferenciranih i karcinoma viših kliničkih stadijuma. Zaključak. Gen p53, jedan od glavnih kontrolora ćelijskog ciklusa, ima značajnu ulogu i u patogenezi karcinoma oralne duplje

Analysis of loss of heterozygosity of the tumor suppressor genes p53 and BRCA1 in ovarial carcinomas

Background/aim: Among the genes involved in ovarian carcinogenesis, there has been increased interest in tumor-suppressor genes p53 and BRCA1. Both of the genes make control of cell cycle, DNA repair and apoptosis. The p53 is a "genome guardian" inactivated in more than 50% of human cancers, while BRCA1 mutations are found mostly in breast and ovarian cancer. The aim of this investigation was to establish the frequency of loss of heterozygosity (LOH) in the regions of the genes p53 and BRCA1 in ovarian carcinomas, and to analyze the association of LOH with the disease stage and prognosis. Methods. We analyzed 20 patients with a confirmed diagnosis of epithelilal ovarian carcinoma. DNA for molecular-genetic analysis was extracted from the tumor tissue and blood as normal tissue of each person. Microsatellite markers of the regions of genes p53 and BRCA1 were amplified by PCR method. The determination of allelic status of microsatellites and detection of LOH was performed after PAA gel electroforesis. Results. Both of the analyzed microsatellite markers were informative in 13/20 (65%) cases. In the region of gene p53, LOH was established in 4/13 (30.7%) tumors. One of them had histological gradus G1, one had gradus G2, and two of them had gradus G3, while all were with the International Federation of Gynecology and Obstetrics (FIGO) IIIc stage. In the region of gene BRCA1, LOH was detected in 5/13 (38.5%) tumors. Four of them had histological gradus G2, and one had gradus G3, while by the (FIGO) classification one was with stage Ib, one was with stage IIIb, while the three were with stage IIIc. LOH in both of the analyzed regions was detected in one tumor (7.7%), with histological gradus G3 and the FIGO IIIc stage. Conclusion. The frequency of LOH in epthelial ovarian carcinomas was 30.7% and 38.5% for p53 and BRCA1 gene regions, respectively. Most of tumors with LOH had histological gradus G2 or G3, and the clinical FIGO stage IIIc, suggesting the association of this occurrence with a later phase of the disease

Ispitivanje mutagenog i genotoksičnog potencijala derivata tert-butilhinona SOS/umuC testom i alkalnim komet testom na MRC-5 i HepG2 ćelijskim linijama

Poslednjih godina, istraživanja u terapiji kancera usmerena su na korišćenje prirodnih proizvoda, naročito poreklom iz marinskih organizama. Hidrohinon avarol i hinon avaron, izolovani iz Dysidea avara pokazuju dobru antikancerogenu aktivnost, ali njihova proizvodnja nije dovoljna za potencijalnu kliničku primenu. Kao potencijalna zamena koristi se model baziran na tert-butilhinonu (TBQ) koji je sličan po strukturi avarolu i avaronu. Međutim, interakcije TBQ sa molekulom DNK mogu dovesti do neželjenih efekata, zbog čega je važno ispitati genotoksičnu aktivnost. U ovom radu ispitivan je genotoksični potencijal TBQ i njegovih alkilamino i aralkilamino derivata u različitim test sistemima. Interakcija sa molekulom DNK je prvo ispitivana u acelularnom modelu na plazmidnoj DNK. Prokariotski test sistem je bio SOS/umuC test na bakteriji Salmonella typhimurium TA1535/pSK1002, a eukariotski komet test na ćelijskim linijama HepG2 (humani karcinom jetre) i MRC-5 (zdravi fibroblasti pluća fetusa). U acelularnom modelu, rezultati su pokazali da TBQ i njegovi derivati ne interaguju direktno sa plazmidom pUC19. U SOS/umuC testu jedino TBQ pokazuje slab genotoksični potencijal i to pri najvećim testiranim koncentracijama. Genotoksični potencijal uočen je primenom komet testa, uglavnom pri najvećim koncentracijama testiranih jedinjenja, ali je on znatno niži u odnosu na benzo(a)piren i etopozid koji su u eksperimentu korišćeni kao pozitivne kontrole

Синтеза и биолошка активност алкилтио и арилтио деривата терц-бутилхинона

Biological activity of 2-tert-butyl-1,4-benzoquinone (TBQ) and its derivatives, 2-tert-butyl-5-(2-propylthio)-1,4-benzoquinone, 2-tert-butyl-5- -(propylthio)-1,4-benzoquinone, 2-tert-butyl-5,6-(ethylenedithio)-1,4-benzoquinone, 2-tert-butyl-5-(phenylthio)-1,4-benzoquinone and 2-tert-butyl-6-(phenylthio)- 1,4-benzoquinone, were tested for their antioxidant, antibacterial, toxic, cytotoxic and genotoxic potential. Using the DPPH test, all derivatives showed good antioxidant activity, better than ascorbic acid, and the 2-tert- -butyl-5-(propylthio)-1,4-benzoquinone derivative showed the strongest effect. Better antibacterial potential was observed against Gram-positive bacteria in the broth microdilution method in which the 2-tert-butyl-5-(phenylthio)-1,4- -benzoquinone derivative showed the strongest activity (MIC = 15.6 μM). The results of toxicity tests, using the Brine shrimp test, indicated that the derivatives lose their toxic potential compared to TBQ, except for 2-tert-butyl-6- -(phenylthio)-1,4-benzoquinone, which showed a 3 times stronger effect. Cytotoxicity was assessed by the MTT assay in 24 and 72 h treatments in MRC-5, HS 294T and A549 cell lines in threefold decreasing gradient (11, 33 and 100 μM). Modifications potentiate the cytotoxic effect, and the strongest effect was observed with the 2-tert-butyl-5,6-(ethylendithio)-1,4-benzoquinone derivative. In addition, the genotoxic potential was examined in the MRC-5 cell line using the comet assay. All tested derivatives of TBQ showed a genotoxic effect at all applied subtoxic concentrations. In general, the chemical modifications of TBQ enhanced its biological activity.Испитана је биолошка активност 2-терц-бутил-1,4-бензохинона (TBQ) и његових деривата: 2-терц-бутил-5-(изопропилтио)-1,4-бензохинона, 2-терц-бутил-5-(пропилтио)-1,4-бензохинона, 2-терц-бутил-5,6-(етиленедитио)-1,4-бензохинона, 2-терц-бутил- -5-(фенилтио)-1,4-бензохинона и 2-терц-бутил-6-(фенилтио)-1,4-бензохинона укључујући њихов антиоксидативни, антибактеријски, токсични, цитотоксични и генотоксични потенцијал. Применом DPPH теста, сви деривати су показали добру антиоксидативну активност, бољу од аскорбинске киселине, а најјаче дејство показао је дериват 2-терц-бутил-5-(пропилтио)-1,4-бензохинон. Бољи антимикробни потенцијал је примећен против Грам-позитивних бактерија методом микродилуције у бујону, где је дериват 2-терц-бутил-5-(фенилтио)-1,4-бензохинон показао најјачу активност (MIC = 15,6 μМ). Резултати испитивања токсичности, применом теста на Artemia salina, показују да деривати губе токсични потенцијал у односу на TBQ, осим 2-терц-бутил-6-(фенилтио)- -1,4-бензохинона, који је показао 3 пута јачи ефекат. Цитотоксичност је испитана МТТ тестом у третманима од 24 и 72 h на ћелијским линијама MRC-5, HS 294T и A549 у троструко опадајућем градијенту (11, 33 и 100 μМ). Модификације појачавају цитотоксични ефекат, а најјачи ефекат је примећен код деривата 2-терц-бутил-5,6-(етилендитио)-1,4-бензохинона. Поред тога, генотоксични потенцијал је испитан на ћелијској линији MRC-5 комет тестом. Сви испитивани деривати су показали генотоксични ефекат при свим примењеним субтоксичним концентрацијама. Генерално, хемијске модификације побољшавају биолошку активност 2-терц-бутил-1,4-бензохинона

Antimicrobial activity of the essential oil of wild-growing Micromeria thymifolia (Scop.) Fritsch

The genus Micromeria Benth. (Lamiaceae, Nepetoideae) includes about 130 species, often aromatc. The essential oil and extracts of some Micromeria species have significant antioxidant, antibacterial and antifungal activities. Micromeria thymifolia is endemic species of the Balkan peninsula. It has been traditionally used in the Mediterranean area as condiment and medicinal plant. The aim of this study was to investigate antimicrobial properties of essential oil of wild Micromeria thymifolia against four Gram negative bacteria (Escherichia coli SY252, Pseudomonas aeruginosa ATCC27853, Salmonella enterica ATCC13076 and human patogen Burkholderia cepacia ATCC25416), four Gram positive bacteria (Enterococcus fecalis ATCC29212, Staphylococcus aureus ATCC25923, Bacillus subtilis ATCC6633, Listeria innocua ATCC33090) and two fungi strains (Candida albicans ATCC10231 and Saccharomyces cerevisiae ATCC9763). The MICs of M. thymifolia essential oil against tested bacteria and fungi was assessed using microtitre plate-based antimicrobial assay. MHB was used as growth media for bacteria, with exception of L.innocua when BHI was used, YPD was used for fungi. The results of our investigation showed that the essential oil of wild-growing M. thymifolia possess significant antimicrobial activity against all tested strains except the P. aeruginosa

INVESTMENTS IN SERBIAN AGRICULTURE

The paper shows the relation between gross domestic product (GDP) and investments in the Republic of Serbia. The observed relation was analyzed at the economy level, as well as in agricultural particular. The observation period is from 2005 to 2020. The function of investments is overviewed through the capital ratio. In this way, it was established to what extent part of the newly created value is returned to the production process, both at the level of the entire economy and at the level of agricultural activity separately. The low participation of investments in the gross domestic product is highlighted, which indicates an unfavorable relationship towards economic and agricultural activity for the observed time period