Colletotrichum fioriniae prouzrokovač antraknoze plodova kruške

Kruška (Pyrus communis L.) je jedna od najstarijih i najrasprostranjenijih voćarskih kultura. Plodovi kruške su izrazito bogati nutrijentima i niskog glikemijskog indeksa, pa se preporučuju u ishrani svih kategorija stanovništva. U periodu vegetacije i nakon berbe propadanje plodova kruške mogu izazvati brojni biljni patogeni, među kojima su i gljive roda Colletotricum. Tokom jeseni 2021. godine iz skladišta u okolini Smedereva sakupljeni su plodovi kruške (cv. Santa Maria) sa simptomima antraknoze. Na površini svih inficiranih plodova bile su prisutne tamne, kružne, udubljene nekrotične lezije sa koncentrično raspoređenim acervulima i narandžastom konidijalnom masom. Izolacija patogena je obavljena standarnim fitopatološkim metodama na podlozi od krompir-dekstroznog agara (KDA). Svi dobijeni izolati su nakon 7 dana na KDA i temperaturi od 25⁰C ispoljili uniformne morfološke osobine. Formirane kolonije su bile svetlosive do svetloružičaste, sa naličja tamnoružičaste do vinastocrvene boje. Prosečan porast kolonija je iznosio 64±2 mm. Konidije su bile hijalinske, fuziformne, neseptirane, glatkih zidova, sa oba zaoštrena kraja, veličine 12,8 - 16,6 × 3,5 – 4,4 µm (n=100); a apresorije pojedinačne, tamnobraon boje, okrugle do nepravilnog oblika, glatkih zidova, veličine 5,5 – 9,6 × 4,8 – 7,2 µm (n=20). Morfološke karakteristike izolata su odgovarale literaturnom opisu C. acutatum species complex. Molekularna identifikacija reprezentativnog izolata SZ-21-36 je obavljena na osnovu ITS regiona rDNA korišćenjem prajmera ITS1/ITS4; dela gena za beta tubulin (Ben A) primenom prajmera Bt2a/Bt2b; gliceraldehid 3 - fosfat dehidrogenaze (GAPDH) sa prajmerima GDF1/GDR1, i dela aktin gena (ACT) upotrebom prajmera ACT512F/ACT783R. Dobijeni PCR produkti su sekvencirani, a nukleotidne sekvence deponovane u GenBank bazi podataka. BLAST analizom je utvrđeno da su sekvence ON171625 (ITS) i ON186698 (ACT) 99% slične, a sekvence ON186696 (BenA) i ON186697 (GAPDH) identične sa sekvencama istih regiona izolata vrste C. fioriniae deponovanih u GenBank bazi. Multilokus filogenetska analiza zasnovana na navedenim molekularnim markerima je potvrdila rezultate BLAST-a, i svrstala izolat SZ-21-36 sa ostalim izolatima C. fioriniae iz GenBank baze podataka (100% bootstrap podrška). Provera patogenosti izolata obavljena je inokulacijom sa 50 μl suspenzije spora (1 × 106 konidija/ml) u prethodno napravljene povrede na zdravim plodovima kruške (cv. Santa Maria). Kontrolni plodovi su inokulisani sterilnom destilovanom vodom. Nakon 10 dana inkubacije u vlažnoj komori na 25°C na svim inokulisanim plodovima kruške razvili su se tipični simptomi antraknoze, dok su kontrolni plodovi bili bez simptoma. Kulture dobijene reizolacijom bile su istih morfoloških karakteristika čime je potvrđena patogenost testiranih izolata. U ovom istraživanju je konvencionalnim i molekularnim metodama prvi put u Srbiji identifikovana vrsta C. fioriniae - prouzrokovač antraknoze plodova kruške

Površinski epitel jajnika hipotireoidnih novorođenih i neonatalnih pacova - iz perspektive PCNA i kaspaze-3

Introduction. The ovarian surface epithelium (OSE) undergoes intensive regeneration and remodelling after each ovulation during the whole reproductive period. This process increases the risk of one of the most common ovarian tumors in women and the female dog. Considering the fact that maternal hypothyroidism highly impacts cell proliferation and cell death during folliculogenesis in the early neonatal period, we aimed to analyze its effect on OSE morphology and dynamics. Materials and Methods. The study was performed on newborn (24-h-old) and neonatal (4-day-old) female rats, a randomized trial between the control and hypothyroid groups, born under controlled circumstances and hypothyroid mothers, respectively. Their ovaries were analyzed histologically and processed to determine the OSE cell height as an average value of four measurement points. Also, the immunopositivity of the proliferating cell nuclear antigen (PCNA) and caspase-3 were assessed semiquantitatively. Results and Conclusions. No major structural differences of OSE were found between groups within the given ages except for a slight increment of OSE cell height and incompleteness of apical cell membrane with cytoplasmic projections in hypothyroid animals. PCNA immunopositivity of the OSE cells was higher in ovaries of hypothyroid animals of both ages in comparison to the controls. Moreover, only scarce OSE cells were caspase-3 positive in both groups and ages, with no difference in immunopositivity. Our study confirms the impact of hypothyroidism in the early postnatal period on morphology and proliferation rate of OSE cells, with no effect on caspase-3 dependent cell removal, which may serve as a premise for future investigation of potential carcinogenesis, in terms of prevention and treatment of ovarian cancer.Uvod. Tokom reproduktivnog perioda, nakon svake ovulacije, površinski epitel (PE) jajnika se intenzivno regeneriše i remodelira. Ovaj proces povećava rizik od nastajanja jednog od najčešćih tumora jajnika kod žena i kuja. Imajući u vidu činjenicu da hipotireoidizam majki u velikoj meri utiče na proliferaciju i ćelijsku smrt u toku folikulogeneze u ranom postnatalnom periodu, cilj rada je bio da analiziramo efekat hipotireoidizma majki na morfologiju i dinamiku PE jajnika potomaka. Materijal i metode. U eksperimentu su korišćeni tek rođeni (24 časa stari) i neonatalni (4 dana stari) mladunci koji potiču od kontrolnih i hipotireoidnih majki. Morfometrijskom analizom određivana je prosečna visina ćelija PE, izmerena na četiri mesta na svakom ispitivanom preseku jajnika. Takođe, imunopozitivnost proliferativnivnog ćelijskog nuklearnog antigena (PCNA) i kaspaze 3 je određivana semikvantitativno. Rezultati i zaključci. Veće promene u strukturi PE jajnika nisu zapažene između grupa, osim blagog povećanja visine ćelija i nepotpune apikalne membrane sa projekcijama citoplazme kod hipotireoidnih životinja. Imunopozitivnost PCNA je značajno povećana u jajnicima hipotireoidnih životinja obe starosne grupe u odnosu na kontrole, dok je imunopozitivnost na kaspazu 3 slabo izražena kod svih životinja. Naše istraživanje je pokazalo da hipotireoidizam u ranom postnatalnom periodu dovodi do promena u morfologiji i proliferaciji ćelija PE, bez efekta na uklanjanje ćelija delovanjem kaspaze 3. Ovaj model može da posluži u budućim istraživanjima potencijalne karcinogeneze u cilju prevencije i tretmana kancera jajnika

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

Significance of molecular diagnostics in human papilloma virus (HPV) determination

HPV infection is considered to be the most important etiologic factor in cervical cancer development. In this retrospective study, which included the period from 2000 to 2012, the results of two molecular techniques used in the detection of HPV infection among women of the South Bačka District were analyzed. By using the technique of in situ hybridization and the rPCR method, the proportion of high-risk HPV among women with normal cytology was determined to be 19.8% and 32.7%, respectively, and among women with abnormal cytology 43.1% and 61%, respectively. Among the analyzed women, HPV type 16 was the most prevalent, followed by HPV types 31, 51 and 18. Application of molecular HPV diagnosis is valuable because it increases the sensitivity of the screening test, so that the application of both tests to detect cervical cancer is a true prevention of malignancy

Identifikacija optimizovanom taqman probom qpcr metodom gena rezistencije na scrapie kod ovaca na teritoriji Republike Srbije

Scrapie is an infectious neurodegenerative disease affecting the central nervous system of sheep and goats that belongs to transmissible spongiform encephalopathies. The disease is caused by the accumulation of proteinase-resistant isoform of the prion protein. The sheep predisposition to scrapie is associated with polymorphisms of the PrP gene. Genetic susceptibility to scrapie is mainly related to codons 136, 154, and 171. ARR sheep are strongly scrapie resistant and VRQ genotype is the most susceptible. Many countries have scrapie eradication programs based on using rams with resistant genotype. The eradication program has not yet been implemented in the Republic of Serbia. To examine the genetic makeup of sheep in Serbia related to scrapie, we optimized TaqMan probes of real-time polymerase chain reaction (qPCR) technique for three codons. Blood samples from 100 sheep were analyzed by qPCR and the majority of the examined sheep were AA homozygous for the 136 codon. For codon 154 the most frequent genotype was RR and for codon 171 the most frequent genotype was QQ.Ovčija svrabež je infektivno neurodegenerativno oboljenje iz grupe transmisivnih encefalopatija koje uzrokuje promene u moždanom tkivu ovaca i koza. Oboljenje je uzrokovano akumulacijom proteinaza-rezistentne izoforme prion proteina. Osetljivost ovaca na ovo oboljenje uslovljena je polimorfi zmom PrP gena. Genetska otpornost na ovčiju svrabež prevashodno je uslovljena kodonima 136, 154 i 171. Najotpornije su ovce sa ARR genotipom, dok se ovce sa VRQ genotipom smatraju najosetljivijim. Mnoge zemlje su sprovele programe eradikacije ovčije svrabeži bazirane na korišćenju priplodnih ovnova koji imaju rezistentan genotip. Ovakav program još nije sproveden u Republici Srbiji. U cilju ispitivanja genetskog statusa ovaca u Srbiji u pogledu otpornosti na ovčiju svrabež, optimizovali smo TaqMan probu, kvantitativne lančane reakcije polimeraze (qPCR) tehnike za sva tri kodona. Ispitivanjem uzoroka krvi od 100 ovaca primenom RT-PCR ustanovljeno je da je većina ovaca AA homozigot za kodon 136. Za kodon 154 najčešći genotip je RR, a za kodon 171 genotip QQ

Reduction of the matrix effects in the gold and silver analysis using the ICP-MS technique

Inductively coupled plasma mass spectrometry (ICP-MS) has been an indispensable technique in the analysis of geological samples in the last few decades. This technique can determine a large number of trace elements, such as the precious metals and rare earth elements. However, the technique is limited by the spectral and non-spectral interferences, which can affect the results of analysis. The aim of this study was to reduce the non-spectral interferences by the method of sample dilution (1000 - 10000). A sample containing gold-bearing minerals was subjected to a digestion in aqua regia, and determination was performed on a NexION 1000 PerkinElmer ICP-MS instrument. The results showed that the differences in gold and silver concentrations in the standard and He modes were insignificant at a dilution of 10000 indicating that the matrix effect was reduced applying a higher dilution. The results of XRD analysis have showed that the sulphide minerals pyrite (FeS2 ) and covellite (CuS) were very abundant in the tested samples, so the digestion of these samples with aqua regia has yielded the solutions containing iron and copper as the most abundant components of the matrix

Laboratorijski profil sinovijalne tečnosti kod šest radnih konja različite starosti

Inflamatorni i degenerativni procesi na zglobovima konja se mogu postepeno razvijati, pri čemu klinički znaci patoloških promena često nisu vidljivi. Cilj ove studije je bio da se na osnovu rutinskog pregleda sinovijalne tečnosti (ST) poreklom iz metakarpo/ metatarzofalangealnih (MCP/MTP) zglobova radnih konja, koji su bili upućeni na klanje i njenih biohemijskih karakteristika utvrdi da li je postojala lokalna inflamacija. Studija je sprovedena na 6 konja starosti 3 do 12 godina (medijana 8), odnosno 24 MCP/MTP zgloba. Post mortem, iz svakog zgloba je uzorkovana ST. Kliničkim pregledom pre klanja, nisu uočene promene u izgledu i funkciji mišićno-skeletnog sistema. Urađen je rutinski pregled ST kojim je opisana boja, prozirnost, viskoznost i broj ćelija. Određena je koncentracija sledećih biohemijskih parametara: ukupnih proteina, albumina, aktivnost alkalne fosfataze (ALP), aspartat aminotransferaze (AST), kreatin kinaze (CK), kao i koncentracije haptoglobina (Hp) i ceruloplazmina (Cp). Kod svih uzoraka, ST je bila bistra, providna, viskozna i blede do bledo žute boje. Najmlađi konj, star tri godine, je u sva četiri pregledana zgloba imao ukupan broj ćelija 200-890/μL, što odgovara fiziološkim vrednostima. Dva konja su imala povećan broj ćelija na jednom, odnosno dva zgloba (1 400-2 560/μL), dok su tri konja imala promene na sva četiri zgloba (1 100-2 200/μL). Ispitivani biohemijski parametri se nisu razlikovali u uzorcima ST iz zglobova sa ukupnim brojem ćelija u okviru fizioloških vrednosti i onih kod kojih je broj ćelija bio povećan. Pozitivne, umerene korelacije su postojale između koncentracije proteina i aktivnosti ALP, koncentracije proteina i albumina, aktivnosti AST i CK. Ispitivanjem ST radnih konja, dokazano je da inflamatorne promene u MCP/MTP zglobovima nisu prisutne. Nalaz povećanog broja ćelija u ST bez inflamatornih promena zahteva dodatna ispitivanja

Chemometric approach to quality characterization of milk-based kombucha beverages

Mliječni kombuča napici su dobiveni fermentacijom mlijeka kombučom. Hijerarhijska klaster analiza (HCA) i analiza glavnih komponenti (PCA) primijenjene su u cilju utvrđivanja razlika između analiziranih uzoraka, kao i identifikacije sličnosti ili različitosti između njih, u prostoru eksperimentalno određenih vrijednosti varijabli. Linearna diskriminantna analiza (LDA) provedena je na neobrađenim podacima u cilju pronalaženja pravila za dodjeljivanje novog uzorka nepoznatog porijekla odgovarajućoj grupi uzoraka. Hijerarhijska klaster analiza pokazala je da je dominantan faktor razlikovanja analiziranih uzoraka kombuča napitaka sadržaj mliječne masti, a zatim slijede sadržaji ukupnih nezasićenih, jednostruko nezasićenih (MUFA) i višestruko nezasićenih (PUFA) masnih kiselina. Uzorci s 0,8 i 1,6 % mliječne masti pripadaju istom klasteru u prostoru analiziranih varijabli zbog sličnosti u njihovoj antioksidativnoj aktivnosti prema DPPH radikalu. Linearnom diskriminantnom analizom utvrđeno je da je najveća razlika u kvaliteti kada se u obzir uzmu vrijednosti pH i antioksidativna aktivnost prema DPPH radikalu između grupa proizvoda sa čajem primorskog vriska i koprivom, dok je najveća sličnost između grupa proizvoda s majčinom dušicom i paprenom metvicom.Milk-based kombucha beverages were obtained conducting kombucha lead fermentation of milk. In order to discriminate the analysed samples and to detect similarities or dissimilarities among them in the space of experimentally determined variables, hierarchical cluster analysis (HCA) and principal component analysis (PCA) were applied. Linear discriminant analysis (LDA) was conducted on the raw data set in order to find a rule for allocating a new sample of unknown origin to the correct group of samples. In the space of the variables analysed by HCA, the dominant discriminating factor for the studied samples of kombucha beverages is the milk fat (MF) content, followed by total unsaturated fatty acids content (TUFA), monounsaturated fatty acids content (MUFA) and polyunsaturated fatty acids content (PUFA). The samples with 0.8 and 1.6% milk fat belong to the same cluster in the space of the analysed variables due to similarities in their AADPPH. It was determined by LDA that there was the biggest difference in quality between the groups of products with winter savoury and stinging nettle, while the highest similarity is between groups of products with wild thyme and peppermint regarding their pH values and antioxidant activity expressed as AADPPH

Ovarian surface epithelium of hypothyroid newborn and neonatal rats: From PCNA and caspase-3 perspective

Introduction. The ovarian surface epithelium (OSE) undergoes intensive regeneration and remodeling after each ovulation during the whole reproductive period. This process increases the risk of one of the most common ovarian tumors in women and the female dog. Considering the fact that maternal hypothyroidism highly impacts cell proliferation and cell death during folliculogenesis in the early neonatal period, we aimed to analyze its effect on OSE morphology and dynamics. Materials and Methods. The study was performed on newborn (24-h-old) and neonatal (4-day-old) female rats, a randomized trial between the control and hypothyroid groups, born under controlled circumstances and hypothyroid mothers, respectively. Their ovaries were analyzed histologically and processed to determine the OSE cell height as an average value of four measurement points. Also, the immunopositivity of the proliferating cell nuclear antigen (PCNA) and caspase-3 were assessed semiquantitatively. Results and Conclusions. No major structural differences of OSE were found between groups within the given ages except for a slight increment of OSE cell height and incompleteness of apical cell membrane with cytoplasmic projections in hypothyroid animals. PCNA immunopositivity of the OSE cells was higher in ovaries of hypothyroid animals of both ages in comparison to the controls. Moreover, only scarce OSE cells were caspase-3 positive in both groups and ages, with no difference in immunopositivity. Our study confirms the impact of hypothyroidism in the early postnatal period onmorphology and proliferation rate of OSE cells, with no effect on caspase-3 dependent cell removal, which may serve as a premise for future investigation of potential carcinogenesis, in terms of prevention and treatment of ovarian cancer