Indentification of three different chromosomal additions by chromosome painting using fluorescence in situ hybridization [FISH] technique

Fluorescence in situ hybridization (FISH) is a very useful method for assessing chromosome rearrangements. When neither banding pattern nor clinical symptoms are sufficient to determine the origin of additional chromosomal fragment, FISH with multiple chromosome-specific libraries (chromosome painting), allows to solve this diagnostic problem rapidly. Three chromosomal additions, 7q+, 13p+ and 22q+, found in routine cytogenetic studies performed in children with phenotypic abnormalities were analysed using FISH. This technique documented the origin of the extra material to be derived from chromosome 16[der(7)t(7; 16)(q36.3;p 13.11)], 18[der(13)t(13; 18)(p12;q 12.2)] and 22[dup(22)(q11.2q13.1)], respectively. In two cases the abnormality arose de novo, while in the third case the product of translocation t(13;18) was maternal by origin. It was present in 30% of mother's lymphocytes, and in 70% of them a balanced Robertsonian translocation t(13q;15q) was found. In the presented cases the chromosome analysis with both traditional banding and chromosome painting techniques, allowed to establish final clinical diagnosis

Not Ideal, But Still Acknowledged: A 10-Country Survey on Empathy for Victims of Anti-LGBT Violence

Using data from a cross-national survey conducted on representative samples of populations from 10 European countries (n = 10,766), the present study is the first one to empirically measure the validity of Christie’s influential ideal victim model. We use a range of scenarios built around common types of anti-LGBT violence to verify the extent to which the public’s empathy for victims is contingent on the victim’s identity and the circumstances of the crime. The results provide strong evidence that, when applied to this group of victims, the rules of the ideal victim work, adequately moderating the public’s emotional reactions. We found that all victims receive relatively high levels of empathy, but the further the victim is from the ideal, the less support they can count on. Thus, even though no victim is “rejected,” a clear hierarchy of victimization emerges. As a group, LGBT people suffer from an empathy deficit, but there also are considerable variations within this group, with a lesbian attacked by extremists receiving the most, and a drunk transgender person receiving the least empathy from the public. The study contributes to the development of theory by embedding the ideal victim model in a broader sociological paradigm of dramaturgical analysis. Since our research shows that the victim’s LGBT status decreases the levels of empathy (being seen as a type of stigma), we call for more attention to be paid to the actor’s identity in Goffman’s framework. Implications for practice and further research are offered.</p

Impact of selected factors on extraction yield and composition of fatty acids in microbial oil produced from Yarrowia lipolytica yeast cells

Drożdże Yarrowia lipolytica to modelowy gatunek mikroorganizmów olejogennych, zdolny do kumulacji lipidów wewnątrz komórki w ilości powyżej 20 % suchej masy. Oprócz prac nad zwiększaniem wydajności biosyntezy oleju mikrobiologicznego ważnym zagadnieniem jest także dobór warunków ekstrakcji tłuszczu z biomasy drożdży. Celem pracy była ocena wpływu wybranych parametrów (rodzaju rozpuszczalnika, sposobu przygotowania próbki oraz czasu trwania procesu) na wydajność ekstrakcji tłuszczu zawartego w komórkach dzikiego szczepu drożdży Y. lipolytica KKP 379, prowadzonej w aparacie Soxhleta oraz na skład kwasów tłuszczowych w uzyskanym oleju. Hodowle okresowe drożdży prowadzono w bioreaktorze laboratoryjnym w bogatym podłożu YPO oraz w podłożach mineralnych z limitowaną zawartością azotu, co umożliwiło otrzymanie biomasy komórkowej o zróżnicowanej zawartości oleju mikrobiologicznego. W badaniach zastosowano plany kwadratów łacińskich, co pozwoliło na odmaskowanie efektu wpływu sposobu przygotowania próbki na zmienną zależną, jaką była zawartość tłuszczu w biomasie drożdży. Najlepsze rezultaty osiągnięto po przygotowaniu biomasy metodą lityczną z wykorzystaniem komercyjnego preparatu Y-PER. Zastosowany czas ekstrakcji oraz rodzaj rozpuszczalnika (eter naftowy lub heksan) nie wpłynęły istotnie na wydajność procesu. Rodzaj rozpuszczalnika miał z kolei wpływ na zawartość trzech kwasów tłuszczowych spośród czternastu zidentyfikowanych w lipidach ekstrahowanych z biomasy drożdży, tj. kwasu palmitooleinowego, eikozapentaenowego i dokozatetraenowego. Większą zawartość wymienionych kwasów oznaczono w próbkach ekstrahowanych heksanem.Yarrowia lipolytica yeasts are a model species of oleaginous microorganisms capable of accumulating lipids inside the cell in an amount exceeding 20 % of dry matter. In addition to studies on how to increase the efficiency of oil biosynthesis, an important issue is, also, the selection of conditions for lipid extraction from yeast biomass. The objective of the research study was to assess the impact of some selected parameters (type of solvent, sample treatment method, and the duration of the process) on the yield of extraction, run in an Soxhlet extractor, of lipids contained in cells of wild strain of Y. lipolytica KKP 379 yeast, and on the composition of fatty acids in the microbial oil produced. The yeasts were cultured in a batch process, in a laboratory bioreactor in a rich YPO medium, as well as in nitrogen restricted mineral media; thus, it was possible to obtain a cell biomass with varying contents of microbial oil. In the research, Latin Square Designs were applied so that the effect of biomass treatment method could be unmasked on the dependent variable, i.e. the content of lipids in the yeast biomass. The best results were achieved when the biomass was prepared using a commercial preparation of Y-PER. The applied duration time of extraction and the used type of solvent (petroleum ether or hexane) did not significantly impact the process yield. However, the type of solvent impacted the content of three fatty acids of the fourteen identified in the lipids extracted from the yeast biomass, ie. the content of palmitoleic, eicosapentaenoic, and docosatetraenoic acids. Higher contents of the above mentioned compounds were determined in the hexane extracted samples

Supernumerary marker chromosomes characterized by fluorescence in situ hybridization [FISH]

Until recently marker chromosomes have presented a difficult diagnostic problem for cytogeneticists as well as for clinicians. Introduction of FISH to cytogenetic analysis has enabled identification of their origin giving possibility to outline specific phenotypic effects of defined marker chromosomes. Nine marker chromosomes were analysed with FISH using centromeric probes, chromosome- specific libraries and unique DNA sequences probes for PWS/AS critical region. The origin from acrocentric chromosomes was established in 6 cases. One marker was a product of maternal 11;22 translocation and two others were pericentromeric regions of chromosome 2 and 4. Among 6 markers, derived from acrocentric chromosomes, 2 consisted of pericentromeric part of chromosome 15, one was identified as mar (21) and in 3 other cases the origin could not be differentiated between chromosomes 13 and 21 or 14 and 22. Clinical consequences of marker chromosomes including the risk for chromosomal nondisjunction and trisomy 21 as well as the risk for uniparental disomy (UPD) are discussed

The attempt to apply Yarrowia Lipolytica KKP379 yeast in fishery industry wastes management

Celem pracy przedstawionej w artykule była ocena możliwości zastosowania kilku produktów odpadowych pochodzących z przetwórni ryb (solanki, dwóch rodzajów odpadów po procesie wędzenia ryb oraz szlamu) jako źródła węgla do wzrostu dzikiego szczepu drożdży Y. lipolytica KKP 379. Najwyższy średni plon biomasy drożdży uzyskano w podłożu zawierającym szlam jako źródło węgla, natomiast najwyższą średnią aktywność lipolityczną oznaczono w płynie pohodowlanym z podłoża zawierającego olej odpadowy po procesie wędzenia. Wprawdzie średnia zewnątrzkomórkowa aktywność lipolityczna drożdży była niższa w porównaniu z uzyskaną w podłożu z oliwą z oliwek (znanym aktywatorem syntezy lipaz), ale zaletą przedstawionego rozwiązania mogą być względy ekonomiczne i ekologiczne.The aim of the study presented in the paper was to evaluate the possibility of application some fishery wastes (oily wastes from fish smoking process, fish leach and sludge) as a carbon sources in the culture medium for wild strain of yeast Y. lipolytica KKP 379. The highest average yeast biomass yield was achieved in the presence of sludge, but the highest lipase activity was observed in the presence of oily wastes from fish smoking process. The extracellular lipase activity in the culture medium was lower than that obtained in the presence of oil olive (the common activator of lipase synthesis), however this approach may be recommended due to the economic and environmental reasons

Utilization of waste from food and fuel industries by lipolytic yeast of Yarrowia lipolytica

Przemysł spożywczy i paliwowy wytwarzają znaczne ilości trudnych w utylizacji hydrofobowych odpadów, stąd istnieje potrzeba poszukiwania nowych metod ich zagospodarowania. Celem badań była ocena możliwości zastosowania wybranych hydrofobowych odpadów przemysłu spożywczego i paliwowego jako głównego źródła węgla oraz induktora syntezy enzymów lipolitycznych w hodowli szczepu drożdży Yarrowia lipolytica W29. Hodowle wstrząsane prowadzono przez 65 h w 28°C w zmodyfikowanym podłożu YPG, w którym glukoza została zastąpiona olejem po procesie wędzenia ryb, tłuszczem po procesie wędzenia wędlin wieprzowych, tłuszczem po pieczeniu kaczej tuszki, zjełczałym masłem klarowanym lub zużytym olejem silnikowym. Stwierdzono, że odpady te mogą być wykorzystywane jako źródło węgla w hodowli drożdży Y. lipolytica. Produkcję enzymów lipolitycznych zaobserwowano w podłożach zawierających tłuszczowe substraty, a aktywność enzymów korelowano ze składem kwasów tłuszczowych. Wykazano, że istnieje możliwość utylizacji zastosowanych substratów odpadowych w procesach mikrobiologicznych do syntezy enzymów o aktywności lipolitycznej.Waste disposal and by-product management in many branches of industry pose problems in the areas of environmental protection and sustainability. Hydrophobic waste substrates of food and fuel origin stands for one of the continuously gaining ground for waste management fields. The aim of the study was to evaluate the possibility to use food origin wastes and fuel industry waste as a carbon sources in the culture medium for Y. lipolytica W29 with their simultaneous valorization. Culture media contained 2% of waste substrates. In the study there were evaluated yeast biomass yield, number of yeast cells in 1 cm³ of medium and extracellular lipase activity after 65 h of yeast growth on a rotary shaker at 28°C. Five wastes were estimated: oily waste from duck roasting process, oily waste from sausages smoking process, rancid ghee, oily waste from fish smoking process and waste engine oil. Additionally fatty acid composition of lipid waste was analyzed using gas chromatography. It was shown the possibility of using these wastes in cultivation of yeast with their simultaneous valorization by obtaining valuable products, e.g. enzymes such as extracellular lipases as well as biomass intended for feed. Yeast biomass yielded from 19.77 g DM·dm⁻³ for oily waste from duck roasting process to 12.28 g DM·dm⁻³ for oily waste from fish smoking process. It has been found that waste substrates stimulate the synthesis of extracellular lipases with different efficiency. The highest activity was obtained in medium containing smoked fish oil (0.313 U·cm⁻³). Furthermore, in waste engine oil medium no lipase activity of Y. lipolytica yeast was observed, but cells did grow and formed a biofilm. The analysis of fatty acid compositions showed the highest oleic acid content in oily waste from duck roasting process (41.9%), slightly lower waste from sausages smoking process (37.8%) and two-fold lower in fish oil (17.3%). This is very important information, because some authors believe that lipolytic enzymes synthesized by the yeast Y. lipolytica show substrate specificity as compared to that oil. Furthermore, the waste oil from fish smoking process was characterized by the presence of polyunsaturated fatty acids containing more than 20 carbons in chain length (EPA and DHA). There was no correlation between lipolytic activity and oleic acid content in waste fat used as the carbon source in medium, but it can be concluded that Y. lipolytica yeast prefered unsaturated rather than saturated fatty acids in extracellular lipase production

Molecular versus classical cytogenetics - evaluation of 20 Prader-Willi syndrome patients

Prader-Willi syndrome (PWS) is a developmental disorder caused by a deficiency of paternal contribution of the chromosome region 15q11.2-q13 arising from differently sized deletions, maternal disomy, or rarely imprinting mutations. We have analyzed 20 PWS patients using combined cytogenetic high resolution technique (HRT), fluorescence in situ hybridization (FISH) and molecular studies to identify parental origin (uniparental disomy) or molecular defect (deletion) of the Prader-Willi region. Lack of a paternal copy of 15q11.2-q13 resulting from its deletion was found in 16 patients. Using high resolution GTG banding on prometaphase chromosomes, deletion in the 15q11.2-q13 region was detected in only 8 patients. Application of FISH with different sets of PWS specific unique sequence probes (D15S11, SNRPN, D15S10, GABRß3) revealed microdeletions in 12 patients. In 12 out of 20 cases FISH confirmed HRT studies, while in 8 cases inconsistent results were obtained. No discrepancies between results of FISH and molecular studies were found, although the latter had a higher sensitivity. We conclude that FISH appears to be a rapid and reliable method of microdeletion identification and should be performed as a method of choice in cytogenetic diagnosis of Prader-Willi syndrome

Identification of supernumerary marker chromosomes derived from chromosomes 5, 6, 19, and 20 using FISH

A large number of cases with supernumerary marker chromosomes (SMCs) should be compared to achieve a better delineation of karyotype-phenotype correlations. Here we present four phenotypically abnormal patients with autosomal marker chromosomes analysed by fluorescence in situ hybridisation using centromeric, telomeric, and unique sequence probes, as well as forward and reverse painting. We also report the first case, to the best of our knowledge, of an SMC derived from chromosome 5. Furthermore, a marker chromosome 20 in a patient with sex differentiation abnormalities, a double mar(6) in a boy with psychomotor retardation, and the association of r(19) with dup(21q21.2q22.12) are described. Although the mar(6) was very small, the presence of euchromatin was shown, suggesting that the partial trisomy of pericentric region derived sequences is implicated in the aetiology of the abnormal phenotypes.


Keywords: supernumerary marker chromosomes; fluorescence in situ hybridisation; phenotype-genotype correlatio