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The protista Acanthamoeba is a free-living amoeba existing in various environments. A number of species among protista are recognized as human pathogens, potentially causing Acanthamoeba keratitis (AK), granulomatous amoebic encephalitis (GAE), and chronic granulomatous lesions. In this study, 10 rhizosphere samples were collected from maize and alfalfa plants in experimental station at Institute of Genetics, Microbiology and Biotechnology, Szent István University. We detected Acanthamoeba based on the quantitative real-time PCR assay and sequence analysis of the 18S rRNA gene. All studied molecular biological methods are suitable for the detection of Acanthamoeba infection in humans. The quantitative real-time PCR-based methods are more sensitive, simple, and easy to perform; moreover, these are opening avenue to detect the effect of number of parasites on human disease. Acanthamoeba species were detected in five (5/10; 50%) samples. All Acanthamoeba strains belonged to T4 genotype, the main AK-related genotype worldwide. Our result confirmed Acanthamoeba strains in rhizosphere that should be considered as a potential health risk associated with human activities in the environment.

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Absztrakt

A petefészek felszíni hámtumorainak mintegy 15–20%-a tartozik a borderline (vagy atípusosan proliferáló, ill. alacsony malignitási potenciálú) csoportba, és összességében 5–7% a mucinosus borderline tumorok előfordulási gyakorisága, a második leggyakoribb típus a serosus borderline tumorok után. A borderline tumorok komoly diagnosztikus és terápiás nehézséget jelentenek a patológus ill. a klinikus számára egyaránt. E daganatok mind szövettani megjelenésükben, mind prognózisukat tekintve köztes helyet foglalnak el a jóindulatú cystadenomák és a carcinomák között. Gyakran fertilis korban lévő nők betegsége indolens lefolyással. A prognózis jó, de a hagyományos kemoterápiával szemben rezisztensek. A legfőbb nehézséget az intraepithelialis carcinoma, a mikroinvázió és az expanzív invázió diagnosztizálása, a primer borderline tumorok colorectalis adenocarcinomák petefészekáttétjétől való elkülönítése okozza. Az Intézetünkben 2000 és 2008 között mucinosus borderline tumornak diagnosztizált 11 esetet újra átnéztük. Közülük 8 eset intestinalis típusú, a maradék 3 eset cervicalis (Müller-cső differenciációjú) típusú volt. 5 esetben lehetett intraepithelialis carcinomát és 5 esetben mikroinváziót találni. Munkánkban e diagnosztikus nehézségeket foglaljuk össze saját tapasztalataink és az irodalmi adatok alapján, néhány szóban kitérve a peritonealis és ovarialis pseudomyxomára.

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A petefészek-daganatok mintegy 15–20%-a tartozik a borderline csoportba (más néven atípusosan proliferáló tumor vagy alacsony malignitási potenciálú carcinoma). E tumorok diagnosztikája és kezelése sok problémát okoz a patológusok és klinikusok számára egyaránt. A borderline tumorok gyakran reproduktív korban lévő, fiatal nőkben alakulnak ki, jó prognózisúak, a betegség lefolyása indolens, de a hagyományos kemoterápiával szemben rezisztensek. A borderline tumorok különböző szöveti típusai közül a leggyakrabban a serosus borderline tumorok fordulnak elő, a nőgyógyászati patológiában a legtöbb diagnosztikai és differenciáldiagnosztikai problémát okozva. Intézetünkben 2000 és 2008 között 30 esetben diagnosztizáltunk serosus borderline tumort. 13 esetben típusos borderline tumor volt, 7 esetben mikropapilláris borderline tumort láttunk, 2 esetben mikroinvázióval és a maradék 8 esetben a borderline tumor jól differenciált serosus carcinomával kombinálódott. A 22 borderline tumoros esetből 17 esetben I-es stádiumú, a petefészekre, vagy petefészkekre lokalizálódó daganatról volt szó, a maradék 5 esetben nem-invazív implantátumok voltak a hasüregben. A jól differenciált serosus carcinomák mellett nem-invazív implantátumokat (3 eset) ill. invazív implantátumokat, metasztázisokat találtunk. A petefészek serosus borderline tumorainak fő diagnosztikai problémája a mikropapilláris mintázat értékelése, a mikroinvázió meghatározása és észlelése, a pseudo-borderline mintázatú jól differenciált serosus carcinomától való elkülönítés mellett az extraovariális manifesztációk, implantátumok dignitásának meghatározása. Munkánkban e főbb diagnosztikus problémákat tárgyaljuk saját tapasztalataink ill. a szakirodalom adatai alapján.

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Acanthamoeba species are free-living amebae that can be found in almost every range of environments. Within this genus, numerous species are recognized as human pathogens, potentially causing Acanthamoeba keratitis (AK). AK is a corneal disease that is predominantly associated with contact lens use, the epidemiology of which is related to the specific genotype of Acanthamoeba. This study reports seven (7/16; 43.75%) positive cases. Detection of Acanthamoeba in corneal scrapings is based on cultivation and polymerase chain reaction (PCR) combined with the molecular taxonomic identification method. By PCR, seven samples were positive; cultivation was successful for five samples, probably because of the low quantity of samples. Genotype identification was carried out with a real-time fluorescence resonance energy transfer PCR assay based on sequence analysis of the 18S rRNA gene, and sensitivity and specificity were evaluated in comparison with traditional parasitological techniques. All seven detected Acanthamoeba strains belonged to the T4 genotype, the main AK-related genotype worldwide. These results confirmed the importance of a complete diagnostic protocol, including a PCR assay, for the clinical diagnosis of AK from human samples. Genotyping allowed the identification of all isolates in the T4 group, thus demonstrating the prevalence of this genotype in Hungary.

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Simple real-time PCR assay with one set of primer and probe for rapid, sensitive qualitative and quantitative detection of Entamoeba histolytica has been used. Consensus sequences were used to amplify a species-specific region of the 16S rRNA gene, and fluorescence resonance energy transfer hybridization probes were used for detection in a LightCycler platform (Roche). The anchor probe sequence was designed to be a perfect match for the 16S rRNA gene of Entamoeba species, while the acceptor probe sequence was designed for Entamoeba histolytica, which allowed differentiation. The performed characteristics of the real-time PCR assay were compared with ELISA antigen and microscopical detection from 77 samples of individuals with suspected clinical diagnosis of imported E. histolytica infection. Stool and liver abscess pus samples were examined with analytical sensitivity of 5 parasites per PCR reaction. The melting curve means Tms (standard deviation) in clinical isolates were 54°C. The real-time assay was 100% sensitive and specific for differentiation of Entamoeba histolytica, compared with conventional ELISA or microscopy. This real-time PCR assay with melting curve analysis is rapid, and specific for the detection and differentiation of Entamoeba histolytica. The suitability for routine use of this assay in clinical diagnostic laboratories is discussed.

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Acta Microbiologica et Immunologica Hungarica
Authors: Erika Orosz, Nóra Szentmáry, Huba J. Kiss, Ágnes Farkas, István Kucsera, and Zoltán Zsolt Nagy

Acanthamoeba has a worldwide distribution in the environment and it is capable of causing a painful sight-threatening disease of the cornea designated as Acanthamoeba keratitis (AK). Nowadays, the cases of AK have surged all over the world along with its disease burden due to increasing use of contact lenses used not only for optical correction but also for cosmetic purposes. In our present work, epithelial abrasion of a 27-year-old female soft contact lens wearer with keratitis was examined. Genotype identification was carried out with a real-time fluorescence resonance energy transfer polymerase chain reaction (PCR) assay based on sequence analysis of the 18S rRNA gene. Genotyping allowed the identification of a T8 group isolate. The analysis confirmed the importance of a complete diagnostic protocol, including a PCR assay, for the clinical diagnosis of AK from human samples. Acanthamoeba T8 should be considered as potential causative organism in keratitis in human.

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Acta Microbiologica et Immunologica Hungarica
Authors: Erika Orosz, Ágnes Farkas, László Ködöböcz, Péter Becságh, József Danka, István Kucsera, and György Füleky

Acanthamoeba species are free-living amoebae that can be found in almost every range of environments. Within this genus, a number of species are recognized as human pathogens, potentially causing Acanthamoeba keratitis, granulomatous amoebic encephalitis, and chronic granulomatous lesions. Soil and water samples were taken from experimental station at Julianna Major of Plant Protection Institute of Centre for Agricultural Research, Hungarian Academy of Sciences (CAR HAS). We detected living Acanthamoeba spp. based on culture-confirmed detection combined with the molecular taxonomic identification method. Living Acanthamoeba spp. were detected in thirteen (65%) samples. The presence of Acanthamoeba spp. in the samples depends significantly on the rhizosphere plants. The most frequently identified living Acanthamoeba genotype was T4 followed by T11, T2/T6 and T17. Genotypes T4 and T11 of Acanthamoeba, are responsible for Acanthamoeba keratitis as well as granulomatous amoebic encephalitis, and should therefore be considered as a potential health risk associated with human activities in the environment.

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Acta Microbiologica et Immunologica Hungarica
Authors: Erika Orosz, Dorottya Kriskó, Lei Shi, Gábor L. Sándor, Huba J. Kiss, Berthold Seitz, Zoltán Zsolt Nagy, and Nóra Szentmáry

Genus Acanthamoeba is an opportunistic protozoan that is widely distributed in the environment. Within this genus, numerous species are recognized as human pathogens, potentially causing Acanthamoeba keratitis (AK). AK is a corneal disease, associated predominantly with contact lens (CL) wear; its epidemiology is related to the specific Acanthamoeba genotypes. This study reports seven CL wearer, Acanthamoeba PCR-positive patients with AK, diagnosed between January 2015 and 2018. Patients had the diagnosis of AK 1.36 months after first symptoms. Genotyping allowed the identification of six isolates of the T4 and one of the T8 genotypes. At first presentation, pseudendritiformic epithelopathy/dirty epithelium (four eyes, 57.1%), multifocal stromal infiltrates (five eyes, 71.4%), ring infiltrate (three eyes, 42.8%), and perineuritis (one eye, 14.3%) were observed. AK was healed without later recurrence in two eyes (28.5%) using triple-topical therapy, in three eyes (42.8%) following additional penetrating keratoplasty. In one patient (14.3%), AK recurred following successful application of triple-therapy and was treated successfully with repeated triple-topical therapy and in one patient (14.3%), no follow-up data were available after diagnosis. We could not observe correlation of genotype and clinical course or the necessity of corneal transplantation in our case series.

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Extraabdominalis desmoid tumorral megjelenő Gardner-syndromás beteg kezelésével szerzett hosszú távú tapasztalataink és irodalmi áttekintés

Long-term experience with therapy of a Gardner's syndrome female, first presenting with extra-abdominal desmoid tumor, and review of the literature

Magyar Sebészet
Authors: Zoltán Mátrai, János Papp, Csaba Polgár, Erika Hitre, István Köves, Edit Oláh, Judit Andi, Andrea Kiss, István Vámosi Nagy, László Tóth, and Zsolt Orosz

Absztrakt

A Gardner-syndroma a familiaris adenomatosus polyposis klinikai altípusa, autosomalis dominánsan öröklődő betegség, amelyet a gastrointestinalis traktus polyposisa és extraintestinalis elváltozások jellemeznek, mint multiplex osteomák, valamint bőr- és lágyrésztumorok. A Gardner-syndromához társuló desmoid tumorok terápiás kihívást jelenthetnek. A szerzők egy 17 éves nőbeteg esetét mutatják be, akinél a Gardner-syndroma a lumbalis régió agresszív desmoid tumorával jelent meg. A beteget a 80 hónapos utánkövetés során sebészileg, non-steroid gyulladáscsökkentőkkel, tamoxifennel és sugárterápiával kezeltük. Következtetésként elmondhatjuk, hogy familiaris adenomatosus polyposisban vagy Gardner-syndromában a desmoid tumorok megjelenése megelőzheti a betegség gastrointestinalis manifesztációját. Az ilyen betegeknél genetikai vizsgálatot és colonoscopiát kell végezni, megelőzendő a későbbi colorectalis rosszindulatú daganat kialakulását. A desmoid tumor multidisciplinaris kezelést igényel.

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Acta Microbiologica et Immunologica Hungarica
Authors: Lei Shi, Tanja Stachon, Lorenz Latta, Mohamed Ibrahem Elhawy, Gubesh Gunaratnam, Erika Orosz, Albrecht F. Kiderlen, Berthold Seitz, Markus Bischoff, and Nóra Szentmáry

We aimed to compare LDH release assay, trypan blue and fluorescent stainings, and non-nutrient Escherichia coli plate assay in determining treatment efficacy of antiamoebic agents against Acanthamoeba castellanii trophozoites/cysts, in vitro. 1BU trophozoites/cysts were challenged with 0.02% polyhexamethylene biguanid (PHMB), 0.1% propamidine isethionate (PD), and 0.0065% miltefosine (MF). Efficacies of the drugs were determined by LDH release and trypan blue assays, by Hoechst 33343, calcein-AM, and ethidium homodimer-1 fluorescent dyes, and by a non-nutrient agar E. coli plate assay. All three antiamoebic agents induced a significant LDH release from trophozoites, compared to controls (p < 0.0001). Fluorescent-dye staining in untreated 1BU trophozoites/cysts was negligible, but using antiamoebic agents, there was 59.3%–100% trypan blue, 100% Hoechst 33342, 0%–75.3% calcein-AM, and 100% ethidium homodimer-1 positivity. On E. coli plates, in controls and MF-treated 1BU trophozoites/cysts, new trophozoites appeared within 24 h, encystment occurred after 5 weeks. In PHMB- and PD-treated 1BU throphozoites/cysts, irregularly shaped, smaller trophozoites appeared after 72 h, which failed to form new cysts within 5 weeks. None of the enzymatic- and dye-based viability assays tested here generated survival rates for trophozoites/cysts that were comparable with those yielded with the non-nutrient agar E. coli plate assay, suggesting that the culture-based assay is the best method to study the treatment efficacy of drugs against Acanthamoeba.

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