Phenotypic and genotypic characteristics of Acinetobacter baumannii strains isolated from secondary and tertiary care hospitals in Serbia

Uvod: Poslednjih godina dolazi do dramatičnog porasta rezistencije Acinetobacter baumannii na karbapeneme i klonskog širenja karbapenem rezistentnih A. baumannii (CRAB) sojeva širom sveta. Metod: U ovoj studiji analizirani su klinički uzorci pacijenata hospitalizovanih širom Srbije tokom 2018. godine. Kriterijum za uključivanje sojeva u studiju bila je izolacija nerepetitivnih bakterija Acb kompleksa iz kliničkih uzoraka inficiranih pacijenata. A. baumannii je identifikovan detekcijom blaOXA-51 gena. Testiranje osetljivosti na antibiotike izvršeno je disk difuzionom i bujon mikrodilucionom metodom. CRAB izolati su testirani na prisustvo stečenih karbapenemaza (OXA-24, OXA-23, OXA-58, OXA-143, IMP, VIM, GIM, SPM, SIM, NDM, GES i KPC) PCR metodom i Sangerovim sekvenciranjem. Klonska povezanost kliničkih izolata ispitana je PFGE, MLST i WGS metodom. Rezultati: Od testiranih 280 bakterija Acb kompleksa njih 237 identifikovano je kao A. baumannii. Prevalencija CRAB izolata iznosila je 93,7%. Iako su izolati pokazali visoku osetljivost na kolistin (95,7%) i tigeciklin (75,1%), deset sojeva je bilo panrezistentno. Detektovani su sledeći geni koji kodiraju stečene karbapenemaze: blaOXA-24 (44,2%), blaOXA-23 (34,5%) i blaNDM-1(3,2%). PFGE je definisao šest različitih klastera. MLST analizom CRAB izolatima su dodeljena tri tipa sekvence: ST2, ST492 i ST636. WGS je ukazao na visok stepen genetičke srodnosti testiranih sojeva koji se grupišu u četri glavna klastera. Zaključak: Ova studija svedoči o visokoj prevalenciji rezistencije A. baumannii na karbapeneme. Ona je rezultat prisustva blaOXA-72, blaOXA-23 i blaNDM-1 gena među CRAB izolatima i njihove klonske propagacije u bolnicama širom Srbije.Background: The worldwide emergence and clonal spread of carbapenem-resistant Acinetobacter baumannii (CRAB) is of great concern. Methods: Non-redundant clinical samples obtained from hospitalized patients throughout Serbia were included in the multicenter study conducted in 2018. Acb complexes recovered from clinical samples obtained from inpatients with confirmed bacterial infections were further evaluated for the presence of A. baumannii. Identification to the species level was done by the detection of the blaOXA-51 gene. Susceptibility testing was done by disk diffusion and broth microdilution method. CRAB isolates were tested for the presence of acquired carbapenemases (OXA-24, OXA-23, OXA-58, OXA-143, IMP, VIM, GIM, SPM, SIM, NDM, GES and KPC) by PCR and Sanger sequencing. Clonal relatedness was assessed by PFGE, MLST and WGS. Results: Overall, A. baumannii was identified in 237 out of 280 Acb complex (84.6%). CRAB prevalence was found to be 93.7%. Although susceptibility was high for colistin (95.7%) and tigecycline (75.1%), ten isolates were classified as pandrug-resistant. The following carbapenemases-encoding genes were found: blaOXA-24 (44.2%), blaOXA-23 (34.5%) and blaNDM-1(3.2%). PFGE analysis revealed six different clusters. MLST analysis identified three STs: ST2, ST492 and ST636. WGS suggested high genetic relatedness among the tested isolates belonging to the four clusters. Conclusion: This study revealed extremely high proportions of carbapenem resistance among A. baumannii clinical isolates due to the emergence of blaOXA-72, blaOXA-23, and blaNDM-1 genes among CRAB isolates in Serbia and their clonal propagation

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

Thermal Diffusivity of Sintered 12CaOx7Al2O3

Poster presented at the 14th International Conference on Photoacoustic and Photothermal Phenomena, Cairo, Egypt, January 6-9, 200

Thermal diffusivity of sintered 12CaO×7AI2O3

Poster presented at the 14th International Conference on Photoacoustic and Photothermal Phenomena, Cairo, Egypt, January 6-9, 2007Paper: [http://dx.doi.org/10.1140/epjst/e2008-00423-7

The first nationwide multicenter study ofAcinetobacter baumanniirecovered in Serbia: emergence of OXA-72, OXA-23 and NDM-1-producing isolates

Background The worldwide emergence and clonal spread of carbapenem-resistantAcinetobacter baumannii(CRAB) is of great concern. The aim of this nationwide study was to investigate the prevalence of CRAB isolates in Serbia and to characterize underlying resistance mechanisms and their genetic relatedness. Methods Non-redundant clinical samples obtained from hospitalized patients throughout Serbia were included in the prospective, observational, multicenter study conducted from January to June 2018. Samples were initially screened for the presence ofAcinetobacter baumannii-calcoaceticus(Acb) complex using conventional bacteriological techniques. Acb complexes recovered from clinical samples obtained from inpatients with confirmed bacterial infections were further evaluated for the presence ofA. baumannii. Identification to the species level was done by the detection of thebla(OXA-51)gene andrpoBgene sequence analysis. Susceptibility testing was done by disk diffusion and broth microdilution method. CRAB isolates were tested for the presence of acquired carbapenemases(bla(OXA-24-like),bla(OXA-23-like,)bla(OXA-58-like),bla(OXA-143-like),bla(IMP),bla(VIM),bla(GIM),bla(SPM),bla(SIM),bla(NDM)) by PCR. Clonal relatedness was assessed by pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). Results Acb complex was isolated in 280 out of 2401 clinical samples (11.6%). Overall,A. baumanniiwas identified in 237 out of 280 Acb complex (84.6%). CRAB prevalence was found to be 93.7% (237/222). The MIC50/MIC(90)for imipenem and meropenem were 8/ gt 32 mu g/mL and 16/ gt 32 mu g/mL, respectively. Although susceptibility was high for colistin (95.7%;n = 227) and tigecycline (75.1%;n = 178), ten isolates (4.3%) were classified as pandrug-resistant. The following carbapenemases-encoding genes were found: 98 (44.2%)bla(OXA-24-like), 76 (34.5%)bla(OXA-23-like), and 7 (3.2%)bla(NDM-1). PFGE analysis revealed six different clusters. MLST analysis identified three STs: ST2 (n = 13), ST492 (n = 14), and ST636 (n = 10). Obtained results evaluated that circulating CRAB clones in Serbia were as follows:bla(OXA66)/bla(OXA23)/ST2 (32.4%),bla(OXA66)/bla(OXA23)/bla(OXA72)/ST2 (2.7%),bla(OXA66)/bla(OXA72)/ST492 (37.8%), andbla(OXA66)/bla(OXA72)/ST636 (27.1%). Conclusion This study revealed extremely high proportions of carbapenem resistance amongA. baumanniiclinical isolates due to the emergence ofbla(OXA-72),bla(OXA-23), andbla(NDM-1)genes among CRAB isolates in Serbia and their clonal propagation

Far infrared properties of iron doped single crystal PbTe

Far infrared reflectivity spectra of single crystal PbTe samples doped with 0.2 at% Fe were measured in the temperature range between 10 K and 300 K. These spectra show very sharp and narrow dips below 120 cm-1. The experimental data were numerically analyzed using a fitting procedure based on the plasmon-phonon interaction model. Two local modes were observed. The first one was noted between 130 cm-1 and 170 cm-1 depending on the temperature and the second more frequency stable one was noted at about 230 cm-1

Alteracije onkogena c-myc i c-erbB-2 u malignim tumorima jajnika

Introduction. According to clinical and epidemiological studies, ovarian cancer ranks fifth in cancer deaths among women. The causes of ovarian cancer remain largely unknown but various factors may increase the risk of developing it, such as age, family history of cancer, childbearing status etc. This cancer results from a succession of genetic alterations involving oncogenes and tumour suppressor genes, which have a critical role in normal cell growth regulation. Mutations and/or overexpression of three oncogenes, c-erbB-2, c-Myc and K-ras, and of the tumour suppressor gene p53, have been frequently observed in a sporadic ovarian cancer. Objective. The aim of the present study was to analyze c-Myc and c-erbB-2 oncogene alterations, specifically amplification, as one of main mechanisms of their activation in ovarian cancers and to establish a possible association with the pathogenic process. Methods. DNA was isolated from 15 samples of malignant and 5 benign ovarian tumours, using proteinase K digestion, followed by phenol-chloroform isoamyl extraction and ethanol precipitation. C-Myc and c-erbB-2 amplification were detected by differential PCR. The level of gene copy increase was measured using the Scion image software. Results. The amplification of both c-Myc and c-erbB-2 was detected in 26.7% of ovarian epithelial carcinoma specimens. Only one tumour specimen concomitantly showed increased gene copy number for both studied genes. Interestingly, besides amplification, gene deletion was also detected (26.7% for c-erbB-2). Most of the ovarian carcinomas with alterations in c-Myc and c-erbB-2 belonged to advanced FIGO stages. Conclusion. The amplification of c-Myc and c-erbB-2 oncogenes in ovarian epithelial carcinomas is most probably a late event in the pathogenesis conferring these tumours a more aggressive biological behaviour. Similarly, gene deletions point to genomic instability in epithelial carcinomas in higher clinical stages as the result of clonal evolution and selection.Uvod. Kliničke i epidemiološke studije su pokazale da je kancer jajnika peti po redu uzročnik smrti žena. Iako postoje mnogi predisponirajući faktori, kao što su starosna dob, porodična istorija kancera, sterilnost, broj rođene dece i dr., tačni uzroci nastanka tumora jajnika još nisu poznati. Zna se, međutim, da je kancer jajnika rezultat akumulacije različitih genskih alteracija, od kojih su najznačajnije mutacije, odnosno povišena ekspresija određenih onkogena, kao što su c-myc, c-erbB-2 i K-ras, i tumor-supresorskih gena, od kojih je najvažniji p53. Cilj rada. Cilj istraživanja je bio da se ispitaju alteracije c-myc i c-erbB-2 (pre svega njihove amplifikacije), najčešćeg mehanizma aktivacije ovih onkogena, u epitelnim karcinomima jajnika, i utvrde njihove potencijalne uloge u patogenezi ovog tipa neoplazmi u našoj populaciji. Metode rada. DNK je izolovana iz 15 uzoraka malignih tumora i pet uzoraka benignih tumora jajnika. Amplifikacije onkogena c-myc i c-erbB-2 ustanovljavane su metodom diferencijalne reakcije lančanog umnožavanja DNK (engl. differential polymerase chain reaction - dPCR). Nivo amplifikacije određen je nakon denzitometrijskog merenja traka na gelu primenom programa Scion image. Rezultati. Amplifikacija i gena c-myc i c-erbB-2 otkrivena je u četiri uzorka (26,7%) epitelnog karcinoma jajnika. Jedan od ispitivanih uzoraka je imao simultanu amplifikaciju oba gena. Većina uzoraka bila je visokog stadijuma prema kriterijumima Međunarodne federacije za ginekologiju i akušerstvo (International Federation of Gynecology and Obstetrics - FIGO). Zanimljivo je da je pored amplifikacije nezavisno ustanovljena i delecija gena c-erbB-2 u 26,7% karcinoma. Svi karcinomi sa delecijama su takođe pripadali visokim kliničkim stadijumima. Zaključak. Amplifikovani onkogeni c-myc i c-erbB-2 su odlika kasnih stadijuma epitelnih maligniteta i verovatno jedan od razloga njihovog agresivnog biološkog ponašanja. Slično tome, i delecija gena erb obeležava uznapredovale stadijume bolesti i ukazuje na genomsku nestabilnost koja se javlja u epitelnim karcinomima kao rezultat evolucije i selekcije različitih tumorskih klonova

Clinical, Dermatoscopical and Laboratory Essentials of Fish Tank Granuloma

In immunocompetent individuals Mycobacterium marinum (M. marinum) infection usually presents with localized skin lesions, better known as “swimming pool granuloma” or “fish tank granuloma” (FTG). When establishing the diagnosis of FTG, doctors encounter several problems in the clinical practice: granulomas are detected in less than two thirds of biopsies and acid-fast bacilli are identified only in a minority of cases. Majority of disseminated and aggressive FTG forms are unrecognized at the beginning, and occur in immunosuppressed patients, including organ transplant patients. Functional impairment or even amputation of the extremity, visceral involvement and lethal outcome have been reported. Although more than one thousand cases of FTG have been reported worldwide, dermatoscopy of FTG, as a diagnostic aid, has not been reported yet. Presenting the case of FTG of recent onset where, guided by dermatoscopy, microorganisms were isolated and identified from the biopsy material, we summarize the essentials of clinical and laboratory diagnostics of M. marinum infection

Characterization of KG-line of wheat by morphological characteristics

U ovom istraživanju je sprovedena karakterizacija perspektivnih genotipova ozime pšenice na osnovu deskriptora morfoloških osobina definisanih od strane Međunarodnog udruženja za zaštitu novih sorti biljaka (UPOV). Karakterizacija KG-linija pšenice je izvršena za 7 kvalitativnih osobina. Poljski ogledi su izvedeni u dvogodišnjem periodu (2013/2014. i 2014/2015.) na tri lokacije: Institut za krmno bilje u Kruševcu, Agroinstitut u Somboru, Centar za strna žita i razvoj sela u Kragujevcu. Ekspresija analiziranih svojstava ispitana je vizuelnim opažanjem i ocenjena odgovarajućim brojem prema UPOV deskriptoru za pšenicu. Većinu KG-genotipova odlikuje piramidalan klas bele boje sa prisutnim zupcima. Izuzetak čini genotip KG-162/7 koji ima paralelan klas, kao i genotip KG-191/5-13 sa vretenastim oblikom klasa. Takođe, za razliku od svih ostalih genotipova, genotip KG-60-3/3 karakteriše klas sa prisutnim osjem. Svojstvo donja plevica – širina ramena se kretala u rasponu od uskog, do veoma širokog, dok su svi genotipovi imali ravan oblik ramena donje plevice. Analizirani genotipovi pšenice pripadaju ozimom tipu.In this research, the characterization of promising winter wheat genotypes was carried out based on the descriptors of morphological traits defined by the International Association for the Protection of New Plant Varieties (UPOV). Characterization of KG-lines of wheat was carried out for 7 qualitative traits. Experimental trial was carried out in a two-year period (2013/2014 and 2014/2015) at three locations: Institute for Forage Crops in Kruševac, Agroinstitut in Sombor and Centre for Small Grains and Rural Development in Kragujevac. The expression of the analyzed properties was examined by visual observation and evaluated with a corresponding number according to the UPOV descriptor for wheat. Most KG-genotypes are characterized by a white pyramidal ear with scurs present. The exception is the genotype KG-162/7, which has a parallel spike, as well as the genotype KG-191/5-13, which has a spindle-shaped spike. Also, unlike all other genotypes, genotype KG-60-3/3 is characterized by a spike with a present awns. Lower glume characteristic – shoulder width ranged from narrow to very wide, while all genotypes had a flat lower glume shoulder shape. The analyzed wheat genotypes belong to the winter type

Kvalitet rasada zelene salate: uticaj belog i plavog svetlosnog tretmana

In the Balkans, the production of globally cultivated green leaf lettuce (Lactuca sativa L.) can be slow due to reduced quality and intensity of solar insolation. Our research aimed to investigate the effects of artificial illumination and the duration of light exposure on the growth and quality of two lettuce genotypes seedlings: Genesis and Jukebox. The research was conducted at the Institute for Vegetable Crops Smederevska Palanka, Serbia. Lettuce seeds were sown in 9 styrofoam containers per genotype, filled with commercial substrate. In the phase of three leaves, plants were placed in plant growth chambers under multispectral white (W) LEDs and monochromatic blue (B) LEDs (three containers per LED and genotype), while three containers were kept in the greenhouse and served as control. The plants were cultured in plant growth chambers for 25 days under a 9h/15h and 10h/14h (light/dark) photoperiod, to simulate outdoor conditions. Morphological growth parameters (number of leaves and plant weight) were measured on the 11th and 25th days. Chlorophyll content was measured on the 4th, 11th, 18th and 25th days. Plants grown under W and B LEDs had a significantly higher number of leaves, plant weight, and chlorophyll content than plants grown in a greenhouse. Using energy-efficient white and blue LED lights in lettuce seedling production during 18-25 days positively impacts seedling quality, and with this seedling production, the plants can achieve high yields and quality.Proizvodnja zelene salate (Lactuca sativa L.) na Balkanu može biti spora usled smanjenog kvaliteta i intenziteta sunčeve insolacije. Cilj ovog istraživanja bio je da se ispita uticaj veštačkog osvetljenja i trajanja izlaganja svetlosnim tretmanima na rast i kvalitet sadnica dva genotipa zelene salate: Genesis i Jukebox. Istraživanje je sprovedeno u Institutu za povrtarstvo Smederevska Palanka, Srbija. Zelena salata je zasejana u 9 kontejnera po genotipu, ispunjenih komercijalnim supstratom. U fazi tri lista, biljke su prebačene u komore za rast biljaka i gajene pod multispektralnim belim (W) i monohromatskim plavim (B) LED svetiljkama (tri kontejnera po LED svetlu i po genotipu), dok su tri kontejnera sa biljkama ostavljena u stakleniku i služila su kao kontrola. Biljke su gajene u komorama za rast biljaka 25 dana, sa 9h/15h i 10h/14h (svetlost/mrak) fotoperiodom, čime su simulirani spoljašnji uslovi. Morfološki parametri rasta (broj listova i težina biljaka) mereni su 11. i 25. dana. Sadržaj hlorofila meren je 4., 11., 18. i 25. dana. Biljke gajene pod belim i plavim LED svetiljkama imale su značajno veći broj listova, težinu i sadržaj hlorofila u poređenju sa biljkama gajenim u stakleniku. Upotreba energetski efikasnih belih i plavih LED svetiljki u proizvodnji rasada zelene salate tokom 18-25 dana pozitivno utiče na kvalitet rasada, a biljke dobijene iz ovakvog rasada mogu da ostvare visoke prinose i kvalitet