Genome sensitivity in patients with chromosomal instability disorders

U cilju ispitivanja senzitivnosti genoma i potvrde klinički postavljene dijagnoze bolesti hromozomske nestabilnosti, korišćeni su uzorci dve grupe pacijenata sa kliničkom slikom: Fankonijeve anemije (FA) i Nijmegenovog sindroma nestabilnosti (NSN) (eng. Nijmegen Breakage syndrome, NBS). Specifična hipersenzitivnost ćelija obolelih od FA na diepoksibutan (DEB), odnosno kod obolelih od NSN na bleomicin (BLC), iskorišćena je u diferencijalno dijagnostičke svrhe. Studija je obuhvatila ukupno 100 pacijenata: 90/100 sa kliničkom slikom FA i 10/100 pacijenata sa kliničkim znacima NSN, koji su dijagnostikovani na Institutu za zdravstvenu zaštitu majke i deteta Srbije „Dr Vukan Čupić“. U detekciji DEB-om indukovane hromozomske nestabilnosti obolelih od FA primenjene su standardne citogenetičke metode. Za potvrdu ili isključenje mozaičnog fenotipa FA korišćene su dodatne metode: analiza oštećenja DNK komet testom i protočno citometrijska analiza zastoja ćelija u G2 fazi ćelijskog ciklusa. Hromozomska osetljivost ćelija obolelih od NSN je analizirana primenom bleomicinovog testa (BLC test). Molekularna analiza prisustva ili odsustva c.657_661del5 mutacije u NBN genu obolelih od NSN, urađena je korišćenjem modifikovane PCR metode uz analizu heterodupleksa. Kod 11,11% (10/90) pacijenata pod sumnjom da boluju od FA (FA grupa), otkrivena je povećana hromozomska osetljivost na DEB u odnosu na kontrolnu grupu (Mann-Whitneyev test: p < 0,05), dok su kod 3,33% (3/90) pacijenata dobijene granične vrednosti iste analize (FA* grupa). U cilju preciznijeg klasifikovanja pacijenata u FA grupi, primenjen je indeks hromozomske fragilnosti. Na ovaj način je u FA grupi, kod 40% (4/10) pacijenata otkriven ćelijski fenotip FA nemozaičnog tipa, dok je kod 60% (6/10) pacijenata dobijen mozaični odgovor na DEB. Kod svih pacijenata sa mozaičnim fenotipom FA, kao i kod dva pacijenta sa graničnim vrednostima diepoksibutanovog testa (DEB testa), daljim analizama primenom komet testa i analizom zastoja ćelija u G2 fazi, potvrđena je dijagnoza FA. U drugoj grupi od 10 bolesnika sa sumnjom da boluju od NSN analizirana je BLC-om indukovana hromozomska nestabilnost, koja je bila povećana kod 40% (4/10) pacijenata (NBS grupa). Kod 30% (3/10) pacijenata dobijeni rezultati su se preklapali sa rezultatima kontrolne grupe (ne-NBS grupa), dok je kod preostala tri pacijenta analiza bila neuspešna zbog slabog mitotskog indeksa. Molekularnim analizama je otkriveno homozigotno prisustvo c.657_661del5 mutacije u NBN genu kod sva četiri pacijenta iz NBS grupe, kao i kod tri pacijenta kod kojih je citogenetička analiza bila neuspešna, i tako potvrđena konačna dijagnoza NSN. Na osnovu dobijenih rezultata može se zaključiti da su analize korišćene u ovoj studiji omogućile potvrdu klinički postavljene dijagnoze FA i NSN, što opravdava njihovu dalju primenu u diferencijalnoj dijagnostici ovih oboljenja.In order to determine the sensitivity of the genome and to confirm a clinical diagnosis of chromosomal instability disorders, blood samples from patients with clinical suspicion of Fanconi’s anemia (FA) and Nijmegen Breakage syndrome (NBS), were collected for chromosome fragility evaluation by the diepoxybutane (DEB) and bleomycin (BLC) tests. The study considered a total of 100 patients: 90/100 with the hematological and/or congenital phenotypic symptoms reminiscent of FA and 10/100 patients with clinical features of NBS, all diagnosed at the Mother and Child Health Care Institute of Serbia "Dr Vukan Čupić”. The DEB-induced chromosomal fragility analysis in patients with FA symptoms was carried out by using standard cytogenetic methods. The additional methods, such as: DNA damage analysis by comet assay and flow cytometric analysis of G2 phase cell cycle arrest, were used in order to confirm FA phenotype in patients with mosaic cytogenetic response to DEB. Chromosomal sensitivity of cells in patients with NBS symptoms was analyzed using a BLC test. Molecular analysis for the presence of the c.657_661del5 mutation in NBN gene in patients with NBS was carried out using modified PCR method and heteroduplex analysis on PAGE gel. In this study 11.11% (10/90) of patients with clinical suspicion of FA were found to have an increased chromosomal sensitivity to DEB (FA group), comparing to healthy controls (Mann-Whitney test: p < 0.05), while 3.33% (3/90) of patients showed borderline results of the same analysis (FA* group). The chromosome fragility index was used in order to provide a clear cut-off diagnostic level distinguishing FA mosaic from other patients in the FA group. In this group, 40% (4/10) of FA patients revealed non-mosaic FA phenotype, while the remaining 60% (6/10) of FA patients showed a mosaic response to DEB. In all of FA mosaic patients, and two patients with borderline DEB-sensitivity, further analyses were performed using comet assay and analysis of G2 phase cell cycle arrest, in order to confirm the diagnosis of FA. In the second group of 10 patients with clinical suspicion of NBS, the cytogenetic analysis revealed an increased chromosomal sensitivity to BLC in 40% (4/10) of them (NBS group). The BLC-induced chromosome fragility values in 30% (3/10) of these patients were overlapping those in the control group (non-NBS group). In the remaining of three patients the cytogenetic analysis was unsuccessful, due to the low mitotic index. Molecular analysis confirmed the presence of homozygous c.657_661del5 mutation in all seven NBS patients, including three patients with unsuccessful cytogenetic analysis, which confirmed the final diagnosis of NBS. Based on these results it can be concluded that the analyses used in this study are useful in confirmation of clinical diagnosis of FA and NBS, which justifies their further application in the differential diagnosis of these two diseases

Analiza deformacionih karakteristika šavova u zavisnosti od strukturnih parametara primenjenih konaca i tkanina

Production of clothes of appropriate quality, low prices and increased productivity is conditioned by properly defined various parameters of the production process. By knowing the connections between structural and mechanical properties of sewing thread, fabric and seams, would be provided conditions for defining the optimal parameters of the production of clothing. During exploitation, clothing is exposed to various influences and particularly stresses. These stresses are greatly affected by the movement of the body, cut off the garment, the method of production, materials and others. Too much stress can lead to damage or deformation of clothing which most often appear on the materials in the area of the seam, on the seam and rarely on the material itself. Mechanical characteristics of sewn seams depends on their structural solutions, sewing thread and fabric, as well as the technological conditions of the process of sewing. Methods for testing are based on the examination and analysis of breaking characteristics of seams, shear of threads in fabric or thread tearing in the seam area (at constant one way load). Based on the experimental results and theoretical assumptions were set realistic mathematical dependencies, by which can be predicted the tensile forces and forces on the limit of elasticity of sewn seams. The breaking force of the seam (relative) and the force on the limit of elasticity of seam are shown as a function of the applied tensile forces of sewing thread, density of stitch and coefficients of correction that takes into account the changes that occur due to dynamic loading of materials involved in the process of sewing. The proposed equations can be used for predicting breaking force of seams and elastic forces on the limit of elasticity , thus simplifying and improving the technical preparation of clothes, saving materials and energy

Filogenetska analiza molekularne evolucije hantavirusa u različitim vrstama glodara

Hantaviruses, members of the order Bunyavirales, family Hantaviridae, are enveloped viruses with a negative sense three-segmented RNA genome, consisting of large – L (6.5–6.6 kb), medium – M (3.6–3.7 kb) and small – S (1.7–2.0 kb) segments, coding for viral polymerase, viral glycoprotein precursor further processed into two separate envelope glycoproteins (Gn and Gc) and viral nucleocapsid protein, respectively. Hantaviruses are unique among the bunyaviruses in not being transmitted by an arthropod vector. These viruses are persistently active in their natural reservoirs including rodents, insectivores and bats. In nature, hantaviruses are circulating via horizontal transmission among infected natural reservoirs. Their primary natural hosts are rodents belonging to four different subfamilies, including Sigmodontinae, Arvicolinae, Murinae and Neotominae. Newly published data have extended hantavirus hosts to insectivores (families Soricidae and Talpidae) and bats (order Chiroptera). Hantaviruses are commonly divided into two groups: old world hantaviruses, present in Europe and Far East, mostly including pathogenic hantavirus species that cause hemorrhagic fever with renal syndrome (HFRS), and new world hantaviruses, present in the Americas, defined as etiological agents of hantavirus pulmonary syndrome (HPS). Mortality rates of these zoonotic diseases, caused by pathogenic hantaviruses, vary with up to 12%-18% for HFRS and 60% for HPS. Human infection occurs through respiratory exposure to contaminated secreta and excreta including urine, saliva, and feces. Humans are not among the natural hosts of hantaviruses and they are thought to be dead-end hosts, without further virus transmission, with the exception of few reported cases of human-to-human transmission of Andes virus (ANDV) in Argentina and Chile...Hantavirusi, taksonomski svrstani u red Bunyavirales, familija Hantaviridae, su virusi sa omotačem i negativnim jednolančanim RNK genomom koji je segmentiran i sastoji se od velikog – L (6.5–6.6 kb), srednjeg – M (3.6–3.7 kb) i malog – S (1.7–2.0 kb) segmenta. L segment kodira virusnu polimerazu, M segment kodira glikoproteinski prekursor od koga nastaju dva glikoproteina (Gn i Gc) i S segment kodira nukleokapsidni protein. Za razliku od ostalih bunjavirusa, transmisija hantavirusa se ne odvija putem artropodnog vektora. Ovi virusi trajno cirkulišu u svojim prirodnim rezervoarima koji obuhvataju glodare, insektivore i slepe miševe. U prirodi, hantavirusi se prenose putem horizontalne i vertikalne transmisije između zaraženih životinja. Najznačajniji rezervoari hantavirusa su glodari koji pripadaju različitim potfamilijama (Sigmodontinae, Arvicolinae, Murinae and Neotominae). Hantavirusi su podeljeni u dve grupe: hantavirusi starog sveta (prisutni u Evropi i na dalekom istoku) koji obuhvataju patogene vrste hantavirusa koji izazivaju hemoragijsku groznicu sa bubrežnim sindromom (HGBS) i hantavirusi novog sveta (prisutni u Americi) koji obuhvataju patogene vrste hantavirusa koji izazivaju hantavirusni pulmonarni sindrom (HPS). Stopa smrtnosti kod ovih zoonoza, uzrokovanih patogenim vrstama hantavirusa, varira od 12-18% za HGBS i do 60% za HPS. Čovek se inficira resporatornim putem u kontaktu sa izlučevinama zaraženih životinja (urin, feces, pljuvačka). S obzirom da čovek nije prirodni domaćin hantavirusa, dalji interhumani prenos hantsvirusa nije moguć, sa izuzetkom nekoliko opisanih slučajeva prenosa Andes virusa (ANDV) u Čileu i Argentini. Hantavirusi, kao virusi sa segmentiranim RNK genomom , spadaju u visoko varijabilne mikroorganizme sa supstitucionom stopom od 10-2 - 10-4 izmena/mestu/godini. Genetički drift, homologa rekombinacija i izmena genskih segmenata su glavni mehanizmi koji leže u osnovi evolucije hantavirusa. U ovom istraživanju smo se fokusirali na analizu evolucije hantavirusa..

CHEMICAL EVALUATION OF AUTOCHTHONOUS VARIETY 'PROKUPAC' RED WINE WITH THE ADDITION OF SELECTED AROMATIC HERBS

The quality parameters and chemical composition (total phenolics, total flavonoids and total anthocyanins) of the red wine obtained from the autochthonous variety 'Prokupac' (Vitis vinifera L.) with the addition of the selected aromatic herbs (Pimpinella anisum L., Cinnamomum zeylanicum, Artemisia absinthium and Glycyrrhiza glabra) were examined in two consequtive vintages (2013-2014). The differences in contents of total extract, reducing sugars and ash between the studied samples were observed, but with no statistical significance between both examined years and analyzed wines. 'Prokupac' red wines with the addition of aromatic herbs stood out with significantly higher contents of total phenolics (371.4-594.3 mg GAE L-1) and total flavonoids (136.2-243.2 mg CTE L-1) while lower total anthocyanins content was noticed, when compared to 'Prokupac' red wine used as control (p&lt;0.05). The unfavourable weather conditions in the vintages 2013 and 2014, reflected notably on the quality of the obtained wine. To analyze the correlations between wine quality parameters and the similarity of individual wine samples, Principal Component Analysis (PCA) was used

A contribution to the data on tench (Tinca tinca L., Cyprinidae, pisces) distribution in Serbia

The studies on the ichthyofauna of Serbia that have been conducted so far indicate the sporadic tench (Tinca tinca L.) presence in typical habitats. The hydroenvironmental conditions necessary for the survival of this fish species are highly variable and, generally, are more favourable in the northern part of the country (the Vojvodina Province). Although the tench presence has been registered at over 30 sites, there is a tendency of a decrease in population abundance. This is due to the reduction of the surface area of the macrophyte-covered flood zones induced by the damming and regulation of rivers, high water level fluctuations, and water amount reduction in the majority of watercourses, a massive increase in the number of allochthonous phytofagous species, reservoir sediment deposition, water quality deterioration and other factors. The tench conservation status in Serbia is officially designated as Low Risk - Least Concern. It may be transferred to a higher category if the tendency towards deterioration of survival conditions for the species should continue

Cosmological constant and the fate of the DDM theory

We investigate the impact of the non-zero cosmological constant on the classical decaying dark matter theory developed by the late Dennis Sciama. In particular, we concentrate on the change in relevant values of cosmological parameters in comparison to the high-precision estimates given by Sciama (1997). It is shown that the appropriate changes in resulting parameter values are such to make the DDM concept less plausible. This is in complete agreement with recently reported observational results detrimental to this theory.Comment: 4 pages, 2 eps figures, uses AA latex styl

Factors affecting elimination of polycyclic aromatic hydrocarbons from traditional smoked common carp meat

Smoking techniques have been progressively improved and different procedures have been developed in different regions for treating fish. In these times, the technology is mainly used for enrichment of fish with specific taste and odour, to extend the shelf-life of these perishable products and appearance required widely on the market. A lot of chemical contaminants such as polycyclic aromatic hydrocarbons (PAHs) are formed during the combustion of fuel in the smoking process. PAHs are a group of compounds that have been the subject of great concern in the recent years due to their toxic, mutagenic and/or carcinogenic potentials to humans. These fact can have a significant impact on the acceptance of these products by consumers. In this review article, the objective is to describe factors affecting elimination of polycyclic aromatic hydrocarbons from traditional smoked common carp meat

Kvantitativno i kvalitativno određivanje enrofloksacina u tkivima riba

Presence of enrofloxacin residues in fish liver, kidney and muscle tissue was investigated after per os application of the drug. For the purpose of determination of enrofloxacin, the following analytical methods were used: microbiological method - plate pH 8 with Escherichia coli ATCC 11303 and HPLC method with fluorescence detection. After a 5-day oral treatment of carps, enrofloxacin residues in tissues were determined up to the 10th day after the end of the drug application. Enrofloxacin content determined by the HPLC method was lower than MRL; drug residues were determined in liver on the 6th day after treatment, in kidney on the 7th day and in muscle on the 9th day after treatment. The results of enrofloxacin residues determination by screening method on the medium with E. Coli ATCC 11303, pH 8 show that this procedure can be used for qualitative determination of enrofloxacin. The screening method allows determination of enrofloxacin in fish tissues below the MRL. Cyprofloxacin was not detected in fish liver, kidney and muscle tissue.Prisustvo rezidua enrofloksacina u jetri, bubregu i mesu riba ispitano je posle njegovog peroralnog aplikovanja. Za ispitivanje rezidua su korišćene: mikrobiološ ka metoda - ploča pH 8 sa Escherichia coli ATTC 11303 i HPLC metoda sa flurescentnom detekcijom. Posle petodnevne oralne terapije šarana rezidue enrofloksacina u tkivima riba su dokazane i devetog dana po prestanku terapije. Sadržaj enrofloksacina dokazan HPLC postupkom, niži od MRL vrednosti, u jetri je dokazan šestog dana po prestanku terapije, u bubregu sedmog dana a u mišićnom tkivu devetog dana po prestanku terapije. Rezultati utvrđivanja rezidua enrofloksacina skrining postupkom na podlozi pH 8 E.coli ATCC 11303 pokazuju da se ovaj postupak može koristiti za kvantitativno dokazivanje enrofloksacina. Skrining postupkom u tkivima riba mogu da se dokažu količine enrofloksacina ispod MRL vrednosti. Ciprofloksacin nije utvrđen u jetri, bubrezima i mesu riba

Astrobiological Complexity with Probabilistic Cellular Automata

Search for extraterrestrial life and intelligence constitutes one of the major endeavors in science, but has yet been quantitatively modeled only rarely and in a cursory and superficial fashion. We argue that probabilistic cellular automata (PCA) represent the best quantitative framework for modeling astrobiological history of the Milky Way and its Galactic Habitable Zone. The relevant astrobiological parameters are to be modeled as the elements of the input probability matrix for the PCA kernel. With the underlying simplicity of the cellular automata constructs, this approach enables a quick analysis of large and ambiguous input parameters' space. We perform a simple clustering analysis of typical astrobiological histories and discuss the relevant boundary conditions of practical importance for planning and guiding actual empirical astrobiological and SETI projects. In addition to showing how the present framework is adaptable to more complex situations and updated observational databases from current and near-future space missions, we demonstrate how numerical results could offer a cautious rationale for continuation of practical SETI searches.Comment: 37 pages, 11 figures, 2 tables; added journal reference belo