Literatura académica sobre el tema ""cytogenetic markers""

Mixed phenotypic acute leukemia (MPAL) comprises of both lymphoid and myeloid markers or blasts in a single population. Diagnostic criteria hinges on classifications provided by identifying these lineages based on cytogenetic markers taken throughout the disease course. We describe an interesting presentation of a patient who had first presented with Acute Myeloid leukemia (AML) but 8 weeks later transformed into Early precursor T cell ALL (ETP-ALL). Cytogenetics were taken throughout the course of the cancer and confirmed the presence of a CD34 precursor cell marker. This transformation and cytogenic markers indicated a pluripotent progenitor cell origin confirming the diagnosis of MAPL. This case highlights a pluripotent progenitor origin with initial presentation as AML (myeloid clone) and later as ALL after initial partial response to AML therapy due to clonal evolution.

Abstract Background: One of the utilities of molecular markers in hematological malignancies is their potential to predict response to drug therapy. Accordingly, we inquired about the effect of both cytogenetic profile and JAK2V617F mutational status on drug therapy response in myelofibrosis with myeloid metaplasia (MMM). Methods: Mutation analysis for JAK2V617 was performed in DNA derived from peripheral blood mononuclear cells, granulocytes, or both. Genomic DNA was amplified by PCR and fluorescent dye chemistry sequencing was performed using the same primers used for amplification (Levin et al. Cancer Cell2005;7:387). Abnormal cytogenetic findings were considered to be either favorable (sole 13q- or 20q- abnormalities) or unfavorable (other abnormalities) (Tefferi et al. BJH2001;113:763). Results: i. Patients and treatment: A total of 69 patients with MMM (median age 62 years, range 38–75; 47 males) received the following drugs as first-line therapy for either anemia or symptomatic splenomegaly; erythropoietin (Epo) alone or in combination with hydroxyurea (n=25), hydroxyurea alone (HU; n=17), interferon alpha (IFN; n=11), combination of thalidomide and prednisone (ThalPred; n=7), androgen preparations (Andro; n=5), and etanercept (n=4). ii. Results of cytogenetic studies and JAK2 mutation screening: Cytogenetic findings were abnormal in 35 patients (51%); favorable in 9 and unfavorable in 26. JAK2V617 mutation analysis revealed wild-type allele in 31 patients (45%) and either heterozygous (n=31) or homozygous (n=7) mutation in the remaining 38 patients (55%). iii. Correlation of response to cytogenetic/molecular markers: Overall, 17 patients (25%) achieved either a major (n=8) or minor (n=9) response in anemia from treatment with one of the aforementioned drugs. Regardless of either JAK2V617 mutational status or cytogenetic profile, treatment-induced responses in anemia were poor for IFN (0%) and HU (12%). In contrast, the best anemia responses were documented for ThalPred (57%), Andro (40%), and Epo-based therapy (32%). Among the 25 patients that received Epo-based therapy, all 3 patients with favorable cytogenetic abnormalities achieved major responses in anemia whereas such responses were seen in none of the 9 patients with unfavorable cytogenetic abnormalities and only 1 of 13 patients with normal cytogenetics (p=0.004). Furthermore, 3 of the 4 major responders were heterozygous for JAK2V617 and one carried the wild-type allele. In contrast, none of the 3 JAK2V617 homozygotes showed any type of response but all 3 also displayed unfavorable cytogenetics. None of the 7 ThalPred-treated patients carried favorable cytogenetics and yet 3 achieved major responses including one with unfavorable cytogenetics. JAK2V617 mutational status was heterozygous in 1 and wild-type in the other two. There were no major responses among the 5 Andro-treated patients despite the presence of favorable cytogenetic abnormalities in 2 patients. In order to investigate the relationship between cytogenetic profile and JAK2V617 mutational status further, we referred to an expanded database of 116 patients and found the incidence of unfavorable cytogenetics to be higher in JAK2V617 homozygotes compared to non-homozygotes (57% vs. 31%; p=0.16) Conclusion: The current study suggests clustering of favorable cytogenetic abnormalities in MMM with anemia response to Epo therapy and unfavorable cytogenetic abnormalities with homozygosity for JAK2V617 mutation.

Astyanax Baird et Girard, 1854, is one of the largest genera in the family Characidae and comprises 177 valid species. This genus has been the focus of cytogenetic studies primarily owing to the presence of B chromosomes and high karyotypic diversity among different populations. The intense genetic variability in Astyanax is one of the factors responsible for the occurrence of species complexes, which are groups (1) with certain difficulties in establishing common genetic pools or (2) belonging to different cryptic species. To evaluate cytogenetic marker inheritance and the possibility of the identification of these hybrids, this study aimed to describe cytogenetic hybrids from three strains of species of the genera Astyanax and Hyphessobrycon Eigenmann, 1908. A. lacustris Lütken, 1875, A. schubarti Britski, 1964, A. fasciatus Cuvier, 1819, and H. anisitsi Eigenmann, 1907 were used to generate three hybrid lineages. The diploid number, heterochromatin sites, and ribosomal genes (18S and 5S rDNA) of the parental strains and the hybrids were analyzed. The results indicated that the three hybrid lineages had cytogenetic markers of both parents, presenting Mendelian inheritance. However, differences in distribution of heterochromatic blocks were observed between the hybrids and the parent strains. Our results allowed the identification of the hybrid strains based on the cytogenetic markers applied, reinforcing the efficiency of cytogenetic markers as tools for identification and indicating that such events may increase the karyotypic diversity in the genera Astyanax and Hyphessobrycon.

Astyanax Baird et Girard, 1854, is one of the largest genera in the family Characidae and comprises 177 valid species. This genus has been the focus of cytogenetic studies primarily owing to the presence of B chromosomes and high karyotypic diversity among different populations. The intense genetic variability in Astyanax is one of the factors responsible for the occurrence of species complexes, which are groups (1) with certain difficulties in establishing common genetic pools or (2) belonging to different cryptic species. To evaluate cytogenetic marker inheritance and the possibility of the identification of these hybrids, this study aimed to describe cytogenetic hybrids from three strains of species of the genera Astyanax and Hyphessobrycon Eigenmann, 1908. A. lacustris Lütken, 1875, A. schubarti Britski, 1964, A. fasciatus Cuvier, 1819, and H. anisitsi Eigenmann, 1907 were used to generate three hybrid lineages. The diploid number, heterochromatin sites, and ribosomal genes (18S and 5S rDNA) of the parental strains and the hybrids were analyzed. The results indicated that the three hybrid lineages had cytogenetic markers of both parents, presenting Mendelian inheritance. However, differences in distribution of heterochromatic blocks were observed between the hybrids and the parent strains. Our results allowed the identification of the hybrid strains based on the cytogenetic markers applied, reinforcing the efficiency of cytogenetic markers as tools for identification and indicating that such events may increase the karyotypic diversity in the genera Astyanax and Hyphessobrycon.

Alpaca is a camelid species of broad economic, biological and biomedical interest, and an essential part of the cultural and historical heritage of Peru. Recently, efforts have been made to improve knowledge of the alpaca genome, and its genetics and cytogenetics, to develop molecular tools for selection and breeding. Here, we report cytogenetic mapping of 35 new markers to 19 alpaca autosomes and the X chromosome. Twenty-eight markers represent alpaca SNPs, of which 17 are located inside or near protein-coding genes, two are in ncRNA genes and nine are intergenic. The remaining seven markers correspond to candidate genes for fiber characteristics (BMP4, COL1A2, GLI1, SFRP4), coat color (TYR) and development (CHD7, PAX7). The results take the tally of cytogenetically mapped markers in alpaca to 281, covering all 36 autosomes and the sex chromosomes. The new map assignments overall agree with human–camelid conserved synteny data, except for mapping BMP4 to VPA3, suggesting a hitherto unknown homology with HSA14. The findings validate, refine and correct the current alpaca assembly VicPac3.1 by anchoring unassigned sequence scaffolds, and ordering and orienting assigned scaffolds. The study contributes to the improvement in the alpaca reference genome and advances camelid molecular cytogenetics.

UVOD Prenatalna dijagnostika predstavlja skup metoda i postupaka čiji je cilj da potvrde ili isključe postojanje kongenitalnih anomalija ploda. Prenatalni skrining može biti ne invazivni i invazivni. Ne invazivni skrining treba da ima visku senzitivnost i da omogući adekvatnu selekciju trudnica kojima će se predložiti genetsko ispitivanje ploda iz uzoraka dobijenih invazivnim metodama prenatalne dijagnostike. Prenatalni skrining prvog trimestra trudnoće obuhvata ultrazvučni pregled debljine nuhalne translucencije i laboratorijsku analizu dva biohemijska markera od 11 do 14 nedelje trudnoće, Prenatalni skrining drugog trimestra trudnoće koji se zasniva na biohemijskom skriningu i tripl testu iako je jedini koji se primenjuje zbog niske senzitivnosti od 20% do 40%, ne može se smatrati validnim. Integrativni biohemijski test prvog i drugog trimestra imaj veću senzitivnost (od 40 do 60%) ali ni on nije dao očekivane rezultate u adekvatnoj selekciji trudnica za genetsku analizu ploda zbog visoke stope lažno pozitivnih rezultata. Drugi trimestar trudnoće omogućava sonografskim pregledima i biohemijskim analizama dopunski način a u nekim slučajevima i jedini u proceni postojanja rizika Daunovog sindroma ili nekih drugih hromozomskih aberacija ploda Zato je primena integrativnih prenatalnih ne invazivnih metoda prvog i drugog trimestra trudnoće veoma značaja u poboljšanju dijagnostičkih vrednosti prenatalnih skrining testova i ima za cilj da smanji procenat invazivnih procedura zbog mogućih komplikacija i ne potrebnih finansijskih troškova. Daunov sindrom(trizomija 21 para hromozoma) je najčešća hromozomska numerička aberacija praćena mentalnom retardacijom dece (I.Q<70. ) Deca sa Daunovim sindromom su karakterističnog fenotipskog izgleda i sa čestim kongenitalnim anomailjama koje im onemogućavaju normalan život a često su i uzrok njihove prerane smrtnosti. Kongenitalne anomalije su zastupljene kod 2% do 5% živo rođene dece, predstavljaju 25 % perinatalne smrtnosti, četvrtina su uslovljnene hromozomskim aberacijama ili naslednom osnovom, od čega 0, 2%-0, 4% su sa Daunovim sindromom. CILJEVI Ciljevi četrorogodišnjeg istraživanja su bili da se poboljša dijagnostička vrednost postojećih prenatalnih testova, da se potvrdi značaj ultrazvučnog skrininga drugog trimestra trudnoće analizom debljine vratne brazde i dužine butne kosti ploda te da se poboljša njegova senzitivnost korporativnom sonografskom analizom cefaličnog indexa, intraorbitalnog rastojanja i dužine fronto-talamične distance. MATERIJAL I METODE Ukupan broj trudnica obuhvaćen četvorogodišnjim ispitivanjem koje su ultrazvučno pregledane i kojima je savetovano genetsko ispitivanje ploda blio je 4552. Tokom Retrospektivnog dvogodišnjieg ( 2010.2011)bila je 2169 dok je prospektivnom dvogodišnjom analizom (2012, 2013)je bilo obuhvaćeno 2383 trudnica. Ispitivana grupa su bile trudnice kod kojih je genetskom analizom otkriven patološki kariotip ploda, kontrolna grupa je obuhvatila sve ostale trudnice kod je kariotip ploda bio normalan od kojih su 124 trudnice odabrane metodom slučajnog izbora. Retrospektivnom studijom ultrazvučna je pregledana dužina vratne brazde(>6mm i dužina butne kosti<0, 6 od 14 do 22 nedelje trudnoće. Analizirana je cirkulacija fetalne krvi kroz duktus venosus ploda( a talas) i postojanje nosne kosti ploda(+, -). Prospektivnom analizom je ultrazvučnim pregledom ploda dodatno analiziran cefalični index(>85%), i intraorbitalna distanca i duzina fronto-talamične distance(<80%) ploda. Korišćene su metode deskriptivne statističke analize, aritmetička sredina, standardna devijacija, najmanja i najveća vrednost kod parametrijskih obeležja dok su za ne parametrijska postojanje nosne kosti i alfa talasa duktusu venozusu korišćene druge statističke metode, a komparativnim statističkim metodama kod normalnih, patoloških i kariotipova sa Daunovim sindromom ploda. Statistička značajnost je dokazana t testom a definisana nivoom p<0, 05 i p<0, 001 odnosom kod normalnih, patoloških kariotipova i Daunovog sindroma. Multifaktorskom regresivnom logističkom analizom je urađena procena senzitvnosti prenatalnog ultrazvučnog skrininga sa ispitivanim obeležjima drugog trimestra trudnoće REZULTATI I DISKUSIJA Od ukupnog broja ultrazvučno pregledaninh trudnica 4552 kojima je savetovano genetska analiza ploda citogenetskom analizom je otkriveno 66 patoloških kariotipova 1, 49%, sa Dunovim sindromom 31 0, 68%. Deskriptivnom statističkom obradom ultrazvučno ispitivanih obeležja od 14 do 22 nedelje trudnoće, uočeno je odstupanje i potvrđen značaj starije životne dobi trudnica, debljine vratne brazde i dužine frontotalamične distance u odnosu na normalne nalaze katiotipova ploda u predikciji Daunovog sindroma.Vrednosti dužine butne kosti, cefaličnog indeksa i intraorbitalnog rastojanja nisu imala veća odstupanja u poređenju patoškokih i normalnih nalaza kariotipova.Studentovim t testom je i dokazano p<0, 001 za debljinu vratne brazde i dužinu fronto-talamične distance, dok je za stariju životnu dob trudnice potvrđeno a;0, 001. Senzitivnost prenatalnog skrininga drugog trimestra analizom debljine vratne brazde i dužine butne kosti je veća u odnosu na standardno primenjivan biohemijski skrining drugog trimestra tripl testa (senzitivnost 40%-60) sa velikom stopom lažno pozitivnih rezultata.Dokazan je značaj poboljšanja senzitivnosti prenatalnih skrining testova dopunskom analizom tri ultrazvučna parametra, dužine fronto-talamične distance, cefaličnog indeksa i intraorbitalnog rastojanja u predikciji Daunovog sindroma, ali i kod ostalih hromozomskih aberacija ploda u periodu od 14 do 22 nedelje trudnoće primenomi multifaktorske logističke regresivne analize senzitivnost preko 93% sa 7% lažno pozitivnih rezultata. Postojanje korelacije između debljine vratne brazde i dužine fronto-talamične distance ploda poboljšavai senzitivnost prenatalnih ultrazvučnog skrininga. Integrativnim pristupom ultrazvučnog i biohemijskog skrininga drugog trimesra trudnoće, tripl testa očekuje se poboljšati dijagnostičkih vrednosti prenatalnog skrininga senzitivnost ne invazivnog skrininga u predikciji Daunovog sindroma i ostalih hromozomskih aberacija ploda. ZAKLJUČCI 1Potvrđen je značaj starije životne dobi trudnice u povećanju rizika Daunovog sindoma, i ostalih hromozomskih aberacija ploda ( p<0, 001) Potvrđen je značaj zadebljanja vratne brazde ploda >6mm(p<0, 001) i skraćenja butne kosti kod Daunovog sindroma ploda od 14 do 22 nedelje trudnoće u prenatalnom otkrivanju Dunovog sindroma i ostalih hromozomskih aberacija ploda i selekciji trudnica kojima će se predložiti genetsko ispitivanje ploda.Potvrđena je hipoteza da skraćenje fronto-talamične distance poboljšava senzitivnost ultrasonografskog skrininga, jer češće postoji kod Daunovog sindroma ploda ali i ostalih numeričkih hromozomskih aberacija tipa, nego kod normalnih nalaza kariotipa ploda( p<0, 001).Komparativnom analzom ultrazvučnim pregledom fronto-talamična distance debljine vratne brazde i dužine butne kosti ploda od 14 do 22 nedelje trudnoće može se značajno poboljšati vrednost dijagnostičkih prenatalnih testova u predikciji Daunovog sindroma. Postojanje korelacija između fronto-talamične distance i debljine vratne brazde dopunjuje ultrazvučni skrining i povećava njegovu senzitivnost na preko 90%, što je multifaktorskom regresivnom logaritamskom analizom i potvrđeno. Značaj multidisciplinarnog pristupa pogotovo je izražen u predikciji Daunovog sindroma, obzirom na različite specijalnosti koje u njemu učestvuju. Cost – benefit analiza. Visoka senzitivnost ne invazivnog prenatalnog skrininga u predikciji Daunovog sindroma, smanjuje troškove za pojedince i državu jer je njihova cena i do dest puta manja od cene citogenetskih analiza, a i trudnice se ne izlažu riziku mogućih komplikacija prilikom izvođenja invazivnih metoda
INTRODUCTIONS Prenatal diagnostic procedure represent a set of methods and techniques with the aim to afirmate or eliminate the presence of Down’s syndrome and other congenital anomalies Can be non-invasive and invasive methods. Non-invasive methods (laboratory or ultrasonographic) have the aim to make possible the most valid assessment of the risk of presence of an affected fetus in the pregnancy, selected pregnancy for invasive diagnostics procedures and citogenetics analisseskariotipingfoeti. Down’s syndrome, aneuploidy with trisomy 21 chromosomal, is the most common chromosomal numerical aberration associated with mental retardation of children (IQ< 70). Children with Down’s syndrome have characteristic phenotypic appearance with high frequent congenital anomalies that preclude a normal life and are frequently the cause of their earlier death. AIM The aim of the four year long investigation was to confirm the importance of ultrasound screening by the analyses of the basic ultrasound parameters for the second trimester, the thickness of the nuchal fold and the length of the femur of the fetus in the prediction of Down’s syndrome and other chromosomal aberrations of the fetus, as well as to improve other existing ultrasonic screenings of the first and second trimester of pregnancy by ultrasonic examination and analyses of the cephalic index and intraorbital space and the length of the fronto-thalamic distance. MATERIAL AND METODS Retrospective investigation (2010. 2011) and prospective investigation (2012.2013) includes 4655 pregnant women. For all pregnant women the genetic investigation of the fetus was performed. A total of 68 were found with chromosomal aberrations, 38 with Down’s syndrome. The method of haphazard choice in retrospective study and in prospective study ultrasound markers are examined. In retrospective analyses of the nuchal fold (<6mm and the length of femur <0.6, that represent basic ultrasound screening of the second trimester and are analyzed as parametric signs of the second trimester, and are analyzed as parametric markers, and analyses of the circulation of fetal blood through ductus venosus of the fetus. In the retrospective study the length of the nuchal fold (>6mm in length, that represent a basic ultrasound screening of the second trimester, and are analyzed as parametric markers in the prediction of Down’s syndrome and other chromosomal aberrations. RESULTS AND DISCUSION Cytogenetic analyses revealed 66 (1, 49%) pathologic karyotypes and Down syndrome were present in 31 (0, 68%) cases. All pathologic karyotypes were obtained after ultrasound examinations of 4552 pregnant women. Ultrasound markers for period 14th-22nd GW were analyzed with descriptive statistical methods and importance of pregnancy in older women, thickness of nuchal fold and lengths frontal thalamic distance were proofed in case of Down syndrome. Femoral bone lengths, cephalic index and intraorbital distances were similar for both groups, normal and pathologic karyotypes. Student’s t test revealed statistical significance with p<0, 001 values for nuchal fold thickness, frontal thalamic distance and older ages.Three additional ultrasound markers (frontal thalamic distance, cephalic index, intraorbital distance) improve prediction of Down syndrome and other chromosomal aberrations between 14th and 22nd GW as well. Multifactorial logistic regressive analyses revealed 93% sensitivity with 7% false positive results. Corelation between nuchal fold thickness and frontal thalamic distance improve prenatal ultrasound screening sensitivity. Using both ultrasound and biochemical screening (triple test) is way to improve sensitivity of non invasive screening in prediction of Down syndrome and other chromosomal aberrations. CONCLUSIONS Importance of pregnant women ages and higher risk for Down syndrome and other chromosomal aberrations was proofed (p<0, 001).Importance of nuchal fold thickness above 6mm (p<0, 001) and shorter femoral bone marker in period from 14th to 22nd GW in prediction of Down syndrome and other chromosomal aberrations are proofed (p<0, 001). Hypothesses that frontal thalamic distance improve ultrasound screening sensitivity was proofed was proofed (p<0, 001) since it is significantly shorter in Down syndrome and other chromosomal aberrations in comparison with fetuses with normal karyotypes. Comparative analyses of frontothalamic distance, nuchal fold thickness and femoral bone length in period from 14th to 22nd GW can signifi cantly improve prenatal diagnostic testing in Down syndrome prediction. Correlation between frontothalamic distance and nuchal fold thickness improve ultrasound screening sensitivity on 93% that is proofed with multifactorial logistic regressive analyses. Significance of multidisciplinary approach is high in Down syndrome prediction. Cost-benefit: High sensitivity of non invasive prenatal screening in Down syndrome prediction reduces costs for families and government since it costs ten time less than cytogenetic analyses and risk with invasive procedures is avoided.

Genetic resistance to Wheat streak mosaic virus (WSMV) offers the most attractive and environmentally safe strategy for disease control. While effective resistance in hexaploid bread wheat (Triticum aestivum) has recently been described in only one case, the Wsm2 gene, more successful resistance has been introgressed from the related hexaploid wheatgrass, Thinopyrum intermedium, as the Wsm1 and Wsm3 genes. In the current study, fluorescent in situ hybridization (FISH) with genomic DNA from Th. intermedium, Aegilops tauschii and repetitive DNA probes was applied to four breeding populations of wheat-Th. intermedium, previously tested for WSMV-resistance. Three different wheat-Th. intermedium recombinant chromosomes, the well-known 4Ai#2S.4DL and two novel, 1B and 3D, were identified to be associated with WSMV-resistance. These novel introgressed genes from Th. intermedium were designated as Wsm4 and Wsm5 respectively. The Wsm4 gene was pinpointed to a 6% interstitial map region on the 1BS flanked by Xgwm4144 and Xgwm1100 markers. Six new PCRmarkers, five linked to Wsm1 and one to Wsm4 were identified. Molecular markers now provide a good coverage of the 4Ai#2S arm for effective marker assisted selection and the new genes increase our arsenal to combat the disease. Two highly repetitive satellite DNA families, Afa and pSc119.2, were isolated for the first time from Th. intermedium and their diversity in respect to copies from other Triticeae species were investigated. They showed contrasting evolutionary dynamics leading to time dependent or independent homogenization of Afa and pSc119.2 sequences. Both repeats are excellent cytological markers and characterized the 4Ai#2S chromosomal arm, in the alien wheat lines and the Th. intermedium genome. Southern hybridization, with methylation sensitive and insensitive restriction enzymes and immunostaining with anti-5-methyl-cytosine antibodies were employed to assess DNA methylation. Overall, no massive changes were evident in the wheat genome, however the alien arm showed reduced cytosine methylation which is characteristic for actively transcribing chromatin.

DNA markers tightly-linked to nuclear fertility restorer genes for cytoplasmic male sterility (CMS) are valuable tools for breeders and researchers working with these genes. Two different targeting approaches were used to identify markers linked to the Rfp1 restorer gene for the pol CMS of canola (Brassica napus L.): nearly isogenic line (NIL) comparison and bulked segregant analysis. These methods were equally efficient in identifying markers linked to Rfp1; combining them allowed a targeting efficiency of 100% to be achieved. The efficiency of bulked segregant analysis was found to be limited by the inadvertent occurrence of shared homozygosity at specific chromosomal regions in the bulks, in contrast with the efficiency of NIL comparison which was limited by the occurrence of residual DNA from the donor cultivar at scattered sites around the genome of the NILs. Eleven DNA markers linked to the Rfp1 gene were identified, one of which perfectly co-segregates with Rfp1. The linkage group on which Rfp1 is localized contains 17 DNA markers. Two restorer genes of the pol CMS, Rfp1 and Rfp2, and a Rfn restorer gene of the nap CMS were found to be at least tightly linked to one another and may all reside at the same locus. A fourth restorer gene, the Rfo restorer for the ogu CMS, was, however, found to be unlinked to the other restorer genes. Different restorer genes for the nap CMS were found in the lines 'Westar-Rf and 'Karat'. A linkage map of the B. napus genome containing 146 markers organized into 23 linkage groups covering a total length of 850.2 cM was constructed from a BC$ sb1$ population. This map contains 63 loci previously localized on the B. napus genome through analysis of an F$ sb2$ population. Comparative analysis indicates that the total length of the BC$ sb1$-derived map is smaller than that of the F$ sb2$-derived map, which suggests that a reduction in recombination frequency is occurring in male gametes. The preferential use of two or three probe-

A two-component transposable element system consisting of a stabilized Activator (Acst) and a chimeric Dissociation (Ds) element has been introduced into the genome of Brassica napus. This Acst/ Ds system incorporates the use of several highly effective screenable and selectable markers. One of these markers is the maize Lc gene, a transcriptional regulator of flavonoid biosynthetic genes. This substrate-independent screenable marker was tested for the first time in B. napus and I show that when overexpressed, there is augmented trichome production and a light-dependent, enhanced accumulation of anthocyanins in B. napus plants. The phenotypes are expressed under a wide range of conditions, are visually distinct, and are observed throughout plant development. When used as a visual marker for the Acst element, Lc B. napus plants were rapidly identified among F2 segregating populations. As part of my goal to develop a very efficient Acst/Ds system for use in B. napus, a conditional negative selectable marker, the E. coli codA gene, was also tested for the first time in B. napus. This was done because use of a substrate-dependent negative selectable marker can facilitate the rapid and reliable identification of stable Ds transposition events when used as a marker for the Acst T-DNA. The enzyme cytosine deaminase, encoded by the codA gene, catalyzes the deamination of the non-toxic compound 5-fluorocytosine (5-FC) to the highly toxic compound 5-fluorouracil. In codA transformed B. napus seedlings, expression of cytosine deaminase results in a severe suppression of growth and this phenotype is dependent on the presence of the 5-FC substrate. Wild-type seedlings, however, lack endogenous cytosine deaminase activity and appear unaffected by the presence of 5-FC in the growth media. These results indicate that codA has the potential to be used effectively in B. napus as a substrate-dependent negative selectable marker for the Acst T-DNA. To determine if Ac transposase cou

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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
A hibridação artificial interespecífica de peixes é utilizada em diversos estabelecimentos voltados à piscicultura no país, com a finalidade de produzir indivíduos mais vantajosos e favoráveis para o cultivo. Porém, devido principalmente às dificuldades encontradas na identificação dos híbridos, esta prática pode determinar o surgimento de sérios problemas como a contaminação genética dos estoques de cultivo, a comercialização de produtos híbridos como espécies puras, a introdução de espécies exóticas e escapes de produtos de piscicultura para o ambiente natural e até eventos de introgressão genética e extinção das espécies nativas. Neste contexto, o presente trabalho teve por objetivo analisar geneticamente exemplares das linhagens parentais de Pseudoplatystoma corruscans (pintado) e Pseudoplatystoma reticulatum (cachara) e seus híbridos interespecíficos pintachara (macho de pintado x fêmea de cachara) e cachapinta (fêmea de cachara x macho de pintado), provenientes dos estoques de cultivo do CEPTAlICMBio, Pirassununga, SP, a fim de identificar e estabelecer marcadores genéticos para possibilitar sua identificação e diferenciação. Também foram analisadas geneticamente amostras de P. corruscans e P. reticulatum coletadas no Rio Paraguai, MS (bacia do Paraguai) e de P. corruscans capturados do rio Mogi- Guaçu, SP (bacia do Alto Paraná), a fim de identificar a possível ocorrência de híbridos entre estas espécies na natureza. As análises citogenéticas revelaram um número diploide de 2n=56 cromossomos para ambas as espécies parentais, com cariótipos caracterizados por cromossomos dos tipos 20m+12am+12st+12a e número fundamental (NF) igual a 100, indicando uma fórmula cariotípica conservada entre estas espécies. A análise dos padrões de heterocromatina pelo bandamento C revelou blocos heterocromáticos localizados nas porções...
Artificial interspecific hybridization of fish is used in various establishments linked to fish farming in the country, in order to produce individuais more advantageous and favorable for cultivation. However, due mainly to difficulties in the hybrid products identification, this practice may determine the onset of serious problems such as genetic contamination of the breeder stocks, marketing of hybrids as pure species, introduction of exotic species and escapes of farmed products to the natural environment and sometimes leading to events of genetic introgression and extinction of native species. In such context, the present study aimed to analyze genetically copies of the parental lines of Pseudoplatystoma corruscans (pintado) and Pseudoplatystoma reticulatum (cachara) and their interspecific hybrids pintachara (female of pintado x male of cachara) and cachapinta (female of cachara x male of pintado), from the stocks manteined in CEPTAlICMBio, Pirassununga, SP, in order to characterize and establish genetic markers to allow their identification and differentiation. Samples of P. corruscans and P. reticulatum collected in the Paraguay river, MS (Paraguay basin) and P. corruscans captured in Mogi-Guaçu river, SP (Alto Paraná basin), also were genetically analyzed in order to identify the possible occurrence of hybrids between these species in nature. The cytogenetic analysis revealed a diploid number of 2n = 56 chromosomes for both parental species with karyotypes characterized by the formula 20m+12am+12st+12a and fundamental number (NF) of 100, indicating a karyotypic formula conserved among these species. The analysis of the heterochromatin C-banding patterns revealed heterochromatic blocks located in the pericentromeric and terminal portions of some chromosomes, the NOR was sim pie and located in one chromosome pai r and 5S ribosomal genes located on two different subtelocentric... (Complete abstract click electronic access below)

Orientador: Fábio Porto Foresti
Banca: José Augusto Senhorini
Banca: Celso Benites
Resumo: A hibridação artificial interespecífica de peixes é utilizada em diversos estabelecimentos voltados à piscicultura no país, com a finalidade de produzir indivíduos mais vantajosos e favoráveis para o cultivo. Porém, devido principalmente às dificuldades encontradas na identificação dos híbridos, esta prática pode determinar o surgimento de sérios problemas como a contaminação genética dos estoques de cultivo, a comercialização de produtos híbridos como espécies puras, a introdução de espécies exóticas e escapes de produtos de piscicultura para o ambiente natural e até eventos de introgressão genética e extinção das espécies nativas. Neste contexto, o presente trabalho teve por objetivo analisar geneticamente exemplares das linhagens parentais de Pseudoplatystoma corruscans (pintado) e Pseudoplatystoma reticulatum (cachara) e seus híbridos interespecíficos "pintachara" (macho de pintado x fêmea de cachara) e "cachapinta" (fêmea de cachara x macho de pintado), provenientes dos estoques de cultivo do CEPTAlICMBio, Pirassununga, SP, a fim de identificar e estabelecer marcadores genéticos para possibilitar sua identificação e diferenciação. Também foram analisadas geneticamente amostras de P. corruscans e P. reticulatum coletadas no Rio Paraguai, MS (bacia do Paraguai) e de P. corruscans capturados do rio Mogi- Guaçu, SP (bacia do Alto Paraná), a fim de identificar a possível ocorrência de híbridos entre estas espécies na natureza. As análises citogenéticas revelaram um número diploide de 2n=56 cromossomos para ambas as espécies parentais, com cariótipos caracterizados por cromossomos dos tipos 20m+12am+12st+12a e número fundamental (NF) igual a 100, indicando uma fórmula cariotípica conservada entre estas espécies. A análise dos padrões de heterocromatina pelo bandamento C revelou blocos heterocromáticos localizados nas porções... (Resumo completo, clicar acesso eletrônico abaixo)
Abstract: Artificial interspecific hybridization of fish is used in various establishments linked to fish farming in the country, in order to produce individuais more advantageous and favorable for cultivation. However, due mainly to difficulties in the hybrid products identification, this practice may determine the onset of serious problems such as genetic contamination of the breeder stocks, marketing of hybrids as pure species, introduction of exotic species and escapes of farmed products to the natural environment and sometimes leading to events of genetic introgression and extinction of native species. In such context, the present study aimed to analyze genetically copies of the parental lines of Pseudoplatystoma corruscans (pintado) and Pseudoplatystoma reticulatum (cachara) and their interspecific hybrids "pintachara" (female of pintado x male of cachara) and "cachapinta" (female of cachara x male of pintado), from the stocks manteined in CEPTAlICMBio, Pirassununga, SP, in order to characterize and establish genetic markers to allow their identification and differentiation. Samples of P. corruscans and P. reticulatum collected in the Paraguay river, MS (Paraguay basin) and P. corruscans captured in Mogi-Guaçu river, SP (Alto Paraná basin), also were genetically analyzed in order to identify the possible occurrence of hybrids between these species in nature. The cytogenetic analysis revealed a diploid number of 2n = 56 chromosomes for both parental species with karyotypes characterized by the formula 20m+12am+12st+12a and fundamental number (NF) of 100, indicating a karyotypic formula conserved among these species. The analysis of the heterochromatin C-banding patterns revealed heterochromatic blocks located in the pericentromeric and terminal portions of some chromosomes, the NOR was sim pie and located in one chromosome pai r and 5S ribosomal genes located on two different subtelocentric... (Complete abstract click electronic access below)
Mestre

Soil pollution with industrial leftovers is of real danger to living organisms since harmful effects can arise after exposure to the contaminants in the soil. In our study, we applied a plant bioassay battery to monitor soil genotoxicity after short-term exposure to the soil. The soil was collected in 3 rounds: at the central part of the brownfield before (S-I) and after (S-III) topsoil removal, and at the brownfield periphery (S-II). The permissible value of the total contamination index is &amp;amp;lt;16 and the corresponding values were 780 in S-I, 69 in S-II and 133 in S-III soil showing that whole brownfield territory is extremely polluted with heavy metals. Cytogenetic markers were recorded in Allium and Tradescantia test-systems and two types of molecular markers, RAPD and ISSR, were analysed in Allium. Our results revealed that the most polluted soil sample has induced an alarming increase of apoptotic cells in onion roots. Chromosome aberration and micronuclei frequency in Allium decreased inconsistently along with the pollution reduction in the soil. Increased frequencies of all cytogenetic markers were revealed in Tradescantia cuttings after exposure to the S-I soil extracts. Cluster analysis of Allium RAPD and ISSR markers showed that the most polluted soil samples induced genetic changes in onions different from those induced by the least polluted soil. Both plant test-systems in this study confirm that soil from the brownfield is harmful to plants and is potentially hazardous to humans.

As part of the prenatal screening of the first trimester of pregnancy, biochemical parameters of blood were analyzed in 1214 women. The level of serum markers β-hCG and PAPP-A in pregnant women are sensitive indicators of chromosomal pathology of the fetus. With trisomy on the 21st chromosome, an increase in the content of β-hCG and a decrease in the level of PAPP-A are often observed in the patient's blood. With fetal trisomy on chromosome 18, both indicators show a tendency to a significant decrease relative to the norm. It should be taken in account that both with fetal euploidy and with chromosomal abnormalities, atypical values of indicators can be established, which requires the involvement of additional markers in the analysis. In any case, biochemical analysis allows only to establish the likelihood of the presence of pathology of the fetus and attribute pregnancy to the risk group. The final conclusion about the presence or absence of chromosomal pathology can only be given by a cytogenetic analysis of the fetal material.

Lung cancer (LC) is leading oncological pathology, posing a serious threat for patient’s lives. Accordingly to World Health Organization (WHO) 2,1 million of new cases and 1,8 of deaths are annually registered. It was accumulated a lot of information about significant influence of smoking on increased risk of LC development. 80-90% of patients with LC are namely smokers. However at present time it was registered increased level of mortality from this pathology among non-smoking patients [1]. LC formation in non-smoking individuals can occur due to environmental pollution by industrial and household cancerogens and also because of molecular and genetical and cytogenetical dissimilarities. Since LC development can be associated with anomalous immunological response, immune genes can be considered as potential biological markers [2]. Objective: To assess the influence of polymorphic variants of innate immunity genes on LC development in non-smoking patients.