Sample records for diploidy

  1. SirA enforces diploidy by inhibiting the replication initiator DnaA during spore formation in Bacillus subtilis.

    Wagner, Jennifer K; Marquis, Kathleen A; Rudner, David Z


    How cells maintain their ploidy is relevant to cellular development and disease. Here, we investigate the mechanism by which the bacterium Bacillus subtilis enforces diploidy as it differentiates into a dormant spore. We demonstrate that a sporulation-induced protein SirA (originally annotated YneE) blocks new rounds of replication by targeting the highly conserved replication initiation factor DnaA. We show that SirA interacts with DnaA and displaces it from the replication origin. As a result, expression of SirA during growth rapidly blocks replication and causes cell death in a DnaA-dependent manner. Finally, cells lacking SirA over-replicate during sporulation. These results support a model in which induction of SirA enforces diploidy by inhibiting replication initiation as B. subtilis cells develop into spores.

  2. SirA enforces diploidy by inhibiting the replication initiator DnaA during spore formation in Bacillus subtilis

    Jennifer K. Wagner; Marquis, Kathleen A.; Rudner, David Z.


    How cells maintain their ploidy is relevant to cellular development and disease. Here, we investigate the mechanism by which the bacterium Bacillus subtilis enforces diploidy as it differentiates into a dormant spore. We demonstrate that a sporulation-induced protein SirA (originally annotated YneE) blocks new rounds of replication by targeting the highly conserved replication initiation factor DnaA. We show that SirA interacts with DnaA and displaces it from the replication origin. As a resu...

  3. Porcine Cloned Embryos Reconstructed with the Cell Nuclei of Tetraploid M-phase Fibroblast Cells Can Restore Normal Diploidy at the Blastocyst Stage.

    Zhao, Q; Qiu, Y G; Tian, J T; Wang, C S; An, T Z


    The cell cycle of donor cells as a major factor that affects cloning efficiency remains debatable. G2/M phase cells as a donor can successfully produce cloned animals, but a minimal amount is known regarding nuclear remodeling events. In this study, porcine fetal fibroblasts (PFFs) were carefully synchronized at G1 or M phase as donor cells. Most of the cloned embryos reconstructed from PFFs at G1 (G1-embryos) or M (M-embryos) phase formed a pronucleus-like nucleus (PN) within 6-h post fusion (hpf), but the M-embryos formed PN earlier than the G1-embryos did. Moreover, 77.4% of the M-embryos formed two PNs, whereas the G1-embryos formed a single PN. The rate of extrusion of polar body-like structures by the M-embryos was significantly lower than that extruded by the G1-embryos (26.3% vs. 37.1%, P M-embryos were octoploid before the first cleavage. Furthermore, 81.25% of the blastomeres of blastocysts developed from the M-embryos showed abnormal ploidy compared with those developed from the G1-embryos (22.55%). However, some of the blastomeres remained diploid in all the M-embryos tested. A portion of the blastomeres restored normal diploidy in some of the M-embryos at the blastocyst stage. This finding provides an explanation for M-embryos developing to term.

  4. 蜂群中的二倍体雄蜂%Research on Diploidy Drone in Bees Colony

    颜伟玉; 吴小波; 王子龙; 曾志将


    @@ 在正常的蜂群中,受精卵发育成雌性蜂(蜂王和工蜂),未受精卵发育成雄性蜂(雄蜂).蜂王可根据工蜂和雄蜂的巢房大小"随意"产下受精卵和未受精卵.但高度近亲交配的蜂王产生的受精卵,在人工培育下也能发育成雄蜂(即二倍体雄蜂).

  5. Meiotic Observations of the Microspore Mother Cells and Unreduced Diploidy Polleninduction in Manihot esculenta%木薯小孢子母细胞减数分裂观察及花粉加倍技术研究

    赖杭桂; 陈霞; 李开棉; 欧文军; 叶建秋; 庄南生; 覃秋林


    利用2n配子途径实现有性多倍化是植物遗传改良的一种有效途径,人工诱导植物2n配子是克服天然2n配子比率低及难于利用的有效方法.本研究对木薯花序发育过程的小孢子母细胞减数分裂进行观察,以掌握木薯小孢子母细胞分裂过程中加倍的有效时期与花序发育及花蕾的外部形态特征的相关性,采用秋水仙素溶液棉浸法对木薯花序进行诱导,获得了加倍2n花粉.结果表明:当幼嫩花序长度约1.5~2.5cm时,侧生小花梗开始出现,雄花蕾直径约1.0~1.5mm时,木薯小孢子母细胞进入减数分裂前期Ⅰ至中期Ⅰ;该期采用0.3%秋水仙素+1%二甲基亚砜(DMSO)处理花序4~5d,可获得2n雄配子,最高诱导率可达12.56%.%Chromosome polyploidization through 2n gamate is one of the efficient approaches for plant genetic improvement.The rate of 2n gamete formation by artificial induction is much higher than that from natural formation.In this research,to find the efficient 2n gamate inductive period during the meiotic process of the microspore mother cells in cassava,different meiotic phases of the microspore mother cells closely correlated with the inflorescent and floral development and morphology were investigated.2n pollen were obtained through colchicine dip the flowers.The results showed that when the inflorence was 1.5~2.5 cm in length,the axilla inflorence initiated to grow,and the male flower bud was 1.0~1.5 mm in diameter,the microspore mother cells of cassava were in the meiotic stages of prophase Ⅰ and metaphase Ⅰ,in which the 2n gamete was efficient induction by dipping the flower buds with 0.3% colchicine +1%DMSO for 4~5 days.The rate of 2n pollen formation was up to 12.56% in SC5.

  6. Analyses of pollen viability and stigma receptivity of tetraploidy and diploidy of Dioscorea zingiberensis%四倍体和二倍体盾叶薯蓣的花粉活力及柱头可受性分析

    赵猛; 徐增莱; 汪琼; 史云云; 贾成森


    盾叶薯蓣(Dioscorea zingiberens C.H.Wright)叶片呈盾形,叶面有不规则的黄白色斑块;蒴果干燥后呈蓝黑色,表面常附有白色粉状物;染色体数20;花粉两端略尖,外壁具有明显条纹。盾叶薯蓣根茎横生,俗称黄姜,可用于治疗皮肤感染、软组织损伤、蜂蜇虫咬及各种外科炎症。盾叶薯蓣根茎所含的薯蓣皂苷元(皂素,diosgenin)是合成甾体激素药物的主要原料,除具有抗炎、避孕等作用外,还可月季于合成镇痛药、杀虫剂及治疗冠心病的药物。

  7. Cytology of Wolbachia-induced parthenogenesis in Leptopilina clavipes (Hymenoptera : Figitidae)

    Pannebakker, BA; Pijnacker, LP; Zwaan, BJ; Beukeboom, LW; Zwaan, Bas J.; Traut, W.


    Parthenogenesis induced by cytoplasmatically inherited Wolbachia bacteria has been found in a number of arthropod species, mainly Hymenoptera. Previously, two different forms of diploidy restoration have been reported to underlie parthenogenesis induction in Hymenoptera by Wolbachia. Both are a form

  8. Morphological systematics of Serapias L. (Orchidaceae) in Southwest Europe

    Venhuis, C.; Venhuis, P.; Oostermeijer, J.G.B.; van Tienderen, P.H.


    Abstract. We measured morphological characters and relative DNA contents to assess variation and phylogenetic relationships among Serapias species in three populations of each of the 10 putative taxa that occur in Southwest Europe. DNA contents indicated diploidy for most species, except for tetrapl

  9. Impact of different patterns of sperm chromosomal abnormalities on the chromosomal constitution of preimplantation embryos.

    Rodrigo, Lorena; Peinado, Vanessa; Mateu, Emilia; Remohí, José; Pellicer, Antonio; Simón, Carlos; Gil-Salom, Manuel; Rubio, Carmen


    To evaluate the effect of sperm chromosome abnormalities--disomy for sex chromosomes and diploidy--in the chromosomal constitution of preimplantation embryos. Retrospective cohort study. Infertility clinic. Three groups: 46,XY infertile men with increased incidence of sex chromosome disomy in sperm; 46,XY infertile men with increased diploidy rates in sperm; 47,XYY infertile men with increased sex chromosome disomy and diploidy rates in sperm. Sperm collection for fluorescence in situ hybridization analysis. Embryo biopsy for preimplantation genetic screening. Frequencies of numerical abnormalities in sperm for chromosomes 13, 18, 21, X, and Y, and in embryos for chromosomes 13, 16, 18, 21, 22, X, and Y. A significant increase of chromosomally abnormal and mosaic embryos was observed in the three study groups compared with controls. Those sperm samples with increased sex chromosome disomy rates produced significantly higher percentages of aneuploid embryos, with a threefold increase for sex chromosomes. Sperm samples with increased diploidy rates were mainly associated to the production of triploid embryos. A strong correlation between sperm and embryo chromosomal constitution has been shown in infertile men with 46,XY and 47,XYY karyotypes. Copyright (c) 2010 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

  10. Development of Special Biological Products


    5 TOTAL GAPS - 4 DICENTRICS - 1 MARKER CHROMOSOMES - 98 Hyperdiploidy was 89% while only 10% was diploidy. This indicates CV-1 (P41) is no longer...cells observed (98%) had chromosome aberration(s), which included breaks, gaps, dicentric and stable marker chromosomes . Most of the marker chromosomes ...9 No. 4 Chromosome Analysis on Three Lots of FRhL-2 (P17) ------------- 10 No. 5 Certification and Testing Status of Three Lots

  11. Acentrosomal Spindle Assembly & Chromosome Segregation During Oocyte Meiosis

    Dumont, Julien; Desai, Arshad


    The ability to reproduce relies in most eukaryotes on specialized cells called gametes. Gametes are formed by the process of meiosis in which, after a single round of replication, two successive cell divisions reduce the ploidy of the genome. Fusion of gametes at fertilization reconstitutes diploidy. In most animal species, chromosome segregation during female meiosis occurs on spindles assembled in the absence of the major microtubule-organizing center, the centrosome. In mammals, oocyte mei...

  12. A clonal theory of parasitic protozoa : the population structures of Entamoeba, Giardia, Leishmania, Naegleria, Plasmodium, Trichomonas, and Trypanosoma and their medical and taxonomical consequences


    We propose a general theory of clonal reproduction for parasitic protozoa, which has important medical and biological consequences. Many parasitic protozoa have been assumed to reproduce sexually, because of diploidy and occasional sexuality in the laboratory. However, a population genetic analysis of extensive data on biochemical polymorphisms indicates that the two fundamental consequences of sexual reproduction (i.e. segregation and recombination) are apparently rare or absent in natural p...

  13. Mating system and gene flow in the red seaweed Gracilaria gracilis: effect of haploid-diploid life history and intertidal rocky shore landscape on fine-scale genetic structure.

    Engel, C R; Destombe, C; Valero, M


    The impact of haploid-diploidy and the intertidal landscape on a fine-scale genetic structure was explored in a red seaweed Gracilaria gracilis. The pattern of genetic structure was compared in haploid and diploid stages at a microgeographic scale (shore. In this intertidal species, biased spore dispersal may occur during the transport of spores and gametes at low tide when small streams flow from high- to lower-shore pools. The longevity of both haploid and diploid free-living stages and the long generation times typical of G. gracilis populations may promote the observed pattern of high genetic diversity within populations relative to that among populations.

  14. Chromosome numbers and pollen stainability of three species of Pacific Island breadfruit (Artocarpus, Moraceae).

    Ragone, D


    Chromosome numbers were determined for 48 accessions of breadfruit (Artocarpus altilis, A. mariannensis, and A. camansi [Moraceae]) from 16 Pacific Island groups, Indonesia, and the Philippines. Artocarpus camansi and A. mariannensis exhibit counts of 2n = 56; 2n = 56 (diploidy) and 2n = 84 (triploidy) were observed for A. altilis. Most diploid cultivars of A. altilis were seeded, but two cultivars with reduced seed number were observed. Micronesian accessions included putative interspecific hybrids between A. altilis and A. mariannensis. The majority of these accessions were seedless diploids, but triploid putative hybrids were also observed. Pollen stainablility was shown to correlate with the degree of seediness.

  15. Acentrosomal spindle assembly and chromosome segregation during oocyte meiosis.

    Dumont, Julien; Desai, Arshad


    The ability to reproduce relies in most eukaryotes on specialized cells called gametes. Gametes are formed by the process of meiosis in which, after a single round of replication, two successive cell divisions reduce the ploidy of the genome. Fusion of gametes at fertilization reconstitutes diploidy. In most animal species, chromosome segregation during female meiosis occurs on spindles assembled in the absence of the major microtubule-organizing center, the centrosome. In mammals, oocyte meiosis is error prone and underlies most birth aneuploidies. Here, we review recent work on acentrosomal spindle formation and chromosome alignment/separation during oocyte meiosis in different animal models.

  16. Parthenogenetic reproduction demonstrated in the diploid Spasalus puncticollis (Le Peletier & Serville 1825), n. stat., from the Antilles (Coleoptera, Scarabaeoidea, Passalidae).

    Boucher, Stéphane; Dutrillaux, Anne-Marie; Dutrillaux, Bernard


    Only females were observed in Spasalus crenatus (Mac Leay 1819) in the Antilles, from Puerto Rico to Saint-Vincent, whereas both sexes are in Trinidad and on the continent. No difference in endo- and ectodermic female genitalia could be noticed between the two populations. Chromosomes of specimens from Guadeloupe reveal a 26,XX karyotype, as in females of various sexual species of Passalini, which demonstrates its diploidy. Breedings were developed with isolated immature stages. After nine years, descendants from a single female are demonstrating their parthenogenetic reproduction. This is the first recorded parthenogenesis in Passalidae and a rare telytoky in diploid insects. Relationships between parthenogenesis, diploidy and insularity are discussed in the scheme of geographical parthenogenesis. No discriminant morphological character on adults could be found between the two populations, except the total length. The modes of reproduction distinguishing the two geographically separated populations suggest the presence of two taxa: S. crenatus on the continent and Trinidad; the parthenote S. puncticollis (Le Peletier & Serville 1825), n. stat., on the Arc of the Antilles.

  17. Onset of Quiescence Following p53 Mediated Down-Regulation of H2AX in Normal Cells

    Inase, Aki; Shinohe, Keitaro; Yoshioka, Yoshiko; Shikanai, Mima; Ichijima, Yosuke; Unno, Junya; Mizutani, Shuki; Tsuchiya, Naoto; Hippo, Yoshitaka; Nakagama, Hitoshi; Masutani, Mitsuko; Teraoka, Hirobumi; Yoshioka, Ken-ichi


    Normal cells, both in vivo and in vitro, become quiescent after serial cell proliferation. During this process, cells can develop immortality with genomic instability, although the mechanisms by which this is regulated are unclear. Here, we show that a growth-arrested cellular status is produced by the down-regulation of histone H2AX in normal cells. Normal mouse embryonic fibroblast cells preserve an H2AX diminished quiescent status through p53 regulation and stable-diploidy maintenance. However, such quiescence is abrogated under continuous growth stimulation, inducing DNA replication stress. Because DNA replication stress-associated lesions are cryptogenic and capable of mediating chromosome-bridge formation and cytokinesis failure, this results in tetraploidization. Arf/p53 module-mutation is induced during tetraploidization with the resulting H2AX recovery and immortality acquisition. Thus, although cellular homeostasis is preserved under quiescence with stable diploidy, tetraploidization induced under growth stimulation disrupts the homeostasis and triggers immortality acquisition. PMID:21858116

  18. Onset of quiescence following p53 mediated down-regulation of H2AX in normal cells.

    Yuko Atsumi

    Full Text Available Normal cells, both in vivo and in vitro, become quiescent after serial cell proliferation. During this process, cells can develop immortality with genomic instability, although the mechanisms by which this is regulated are unclear. Here, we show that a growth-arrested cellular status is produced by the down-regulation of histone H2AX in normal cells. Normal mouse embryonic fibroblast cells preserve an H2AX diminished quiescent status through p53 regulation and stable-diploidy maintenance. However, such quiescence is abrogated under continuous growth stimulation, inducing DNA replication stress. Because DNA replication stress-associated lesions are cryptogenic and capable of mediating chromosome-bridge formation and cytokinesis failure, this results in tetraploidization. Arf/p53 module-mutation is induced during tetraploidization with the resulting H2AX recovery and immortality acquisition. Thus, although cellular homeostasis is preserved under quiescence with stable diploidy, tetraploidization induced under growth stimulation disrupts the homeostasis and triggers immortality acquisition.

  19. Effects of ploidy and recombination on evolution of robustness in a model of the segment polarity network.

    Kerry J Kim


    Full Text Available Many genetic networks are astonishingly robust to quantitative variation, allowing these networks to continue functioning in the face of mutation and environmental perturbation. However, the evolution of such robustness remains poorly understood for real genetic networks. Here we explore whether and how ploidy and recombination affect the evolution of robustness in a detailed computational model of the segment polarity network. We introduce a novel computational method that predicts the quantitative values of biochemical parameters from bit sequences representing genotype, allowing our model to bridge genotype to phenotype. Using this, we simulate 2,000 generations of evolution in a population of individuals under stabilizing and truncation selection, selecting for individuals that could sharpen the initial pattern of engrailed and wingless expression. Robustness was measured by simulating a mutation in the network and measuring the effect on the engrailed and wingless patterns; higher robustness corresponded to insensitivity of this pattern to perturbation. We compared robustness in diploid and haploid populations, with either asexual or sexual reproduction. In all cases, robustness increased, and the greatest increase was in diploid sexual populations; diploidy and sex synergized to evolve greater robustness than either acting alone. Diploidy conferred increased robustness by allowing most deleterious mutations to be rescued by a working allele. Sex (recombination conferred a robustness advantage through "survival of the compatible": those alleles that can work with a wide variety of genetically diverse partners persist, and this selects for robust alleles.

  20. Detection of aneuploidy in sperm of an ataxia telangiectasia patient using three-chromosome fluorescence in situ hybridization

    Lowe, X.R.; Baulch, J.E. [Lawrence Livermore National Lab., CA (United States); Arnheim, N. [USC, Los Angeles, CA (United States)] [and others


    Ataxia telangiectasia (A-T) is an inherited, recessive, cancer-prone disorder. Fluorescence in situ hybridization (FISH) with DNA probes specific for three chromosomes was applied to sperm of an A-T patient to determine if there may be an increased germinal risk for aneuploidy. Air-dried sperm smears were treated with proteinase K and were decondensed with DTT and LIS. The slides were then hybridized with fluorescently labeled repetitive DNA probes specific for chromosomes X, Y and 8, and a total of 11,825 sperm cells were scored. The ratio of sperm bearing X-8 and Y-8 was 1:1, as predicted. The frequencies of hyperhaploidy were 3.9, 1.0, 17.6 and 7.8 per 10,000 cells for categories X-X-8, Y-Y-8, X-Y-8 and 8-8-(X or Y), respectively, In addition, the frequency of diploidy (X-Y-8-8) was 18.6 and auto-diploidies (X-X-8-8 and Y-Y-8-8) were 1.0 and 2.0, respectively. These frequencies were not significantly different when compared with levels in healthy men (p > 0.1). Our finding suggests that chromosome X, Y and 8 aneuploidies are not elevated in the sperm of A-T patients, but studies with additional patients and chromosomes are needed.

  1. Investigating Correlation of DNA Ploidy and Prognosis of Malignant Tumors in Central Nervous System

    ChenRuoping; WangBingyu; DingMeixiu; ShiGuiying; ShiXuegeng


    Objective Investigating the correlation between DNA ploidy and prognosis of malignant tumors in central nervous system (CNS) .Methods 44 cases,including tumors of neuroepithelium, meninges, metastasis, and germioma, were investigated, which divided into two groups, the research and the control (tissue around the tumor). Pancreatin digestion was applied to change all tissue into monocellular suspension. Then DNA ploidy was detected by flow cytometer after propidium iodide (PI) staining. Results Of total 44 cases,61.36% were diploidy tumors(27 cases),31.82% were heterodiploidy(14cases),and 6.82% were tetraploidy(3cases).F-test showed out that the average survival time free of neoplasm of diploidy cases [ (6.19 + 3.37)months] was much longer than that of heterodiploidy and tetraploidy cases [(4.35+4.03)months, P=0.0076]. Conclusion The research suggests positive correlation between DNA ploidy and prognosis, which means that DNA ploidy is possible to be one of predicting indexes.

  2. Chromosomal aneuploidies and DNA fragmentation of human spermatozoa from patients exposed to perfluorinated compounds.

    Governini, L; Guerranti, C; De Leo, V; Boschi, L; Luddi, A; Gori, M; Orvieto, R; Piomboni, P


    This study investigated chromosomal aneuploidies and DNA damage in spermatozoa from male patients contaminated by perfluorinated compounds (PFCs) in whole blood and seminal plasma. Sperm aneuploidy and diploidy rate for chromosomes 18, X and Y were evaluated by FISH; sperm DNA fragmentation was assessed by terminal deoxynucleotidyl transferase-mediated dUTP nick end-labelling technique coupled to flow cytometry. Our results indicated that PFC contamination was present in 58% of subjects included in the study. A significant increase in alterations of sperm parameters was observed in PFC-positive subjects compared to PFC-negative subjects. As regards the sperm aneuploidy, both disomy and diploidy rates resulted significantly increased in subjects positive for PFC contamination compared to PFC-negative samples. In addition, sperm DNA fragmentation index resulted significantly increased in PFC-contaminated subjects compared to PFC-non-contaminated subjects, with a significant increased level of dimmer DNA fragmentation index. Our results clearly indicate that PFC contamination may detrimentally affect spermatogenesis, disturbing both meiotic segregation and DNA integrity. We could therefore suggest cautions to reduce or eliminate any contact with these compounds because the long-term effects of PFC accumulation in the body are not predictable. © 2014 Blackwell Verlag GmbH.

  3. Reciprocal uniparental disomy in yeast.

    Andersen, Sabrina L; Petes, Thomas D


    In the diploid cells of most organisms, including humans, each chromosome is usually distinguishable from its partner homolog by multiple single-nucleotide polymorphisms. One common type of genetic alteration observed in tumor cells is uniparental disomy (UPD), in which a pair of homologous chromosomes are derived from a single parent, resulting in loss of heterozygosity for all single-nucleotide polymorphisms while maintaining diploidy. Somatic UPD events are usually explained as reflecting two consecutive nondisjunction events. Here we report a previously undescribed mode of chromosome segregation in Saccharomyces cerevisiae in which one cell division produces daughter cells with reciprocal UPD for the same pair of chromosomes without an aneuploid intermediate. One pair of sister chromatids is segregated into one daughter cell and the other pair is segregated into the other daughter cell, mimicking a meiotic chromosome segregation pattern. We term this process "reciprocal uniparental disomy."

  4. Expression of placental protein 14 by the new endometrial cancer cell line MFE-280 in vitro and by endometrial carcinomas in vivo.

    Hackenberg, R; Loos, S; Nia, A H; Kunzmann, R; Schulz, K D


    MFE-280 endometrial cancer cells express PP14 (placental protein 14) in vitro. PP14 is normally found in the secretory endometrium and in placental tissue. MFE-280 cells, which are tumorigenic in nude mice, were derived from a recurrent, poorly differentiated endometrial carcinoma. The cells were initially grown in suspension culture and later transferred to monolayer cultures. Karyotyping revealed near-diploidy with a complex heterogeneous aberration pattern. MFE-280 cells were positive for the cytokeratins 7, 8, 18 and 19 as well as for vimentin. The expression of PP14 in MFE-280 cells was demonstrated by immunochemistry and reverse transcriptase--polymerase chain reaction. PP14-mRNA was also detected in one out of five endometrial cancer specimen. In tumor tissue the expression of PP14 was not dependent on progestins.

  5. Suppression of first cleavage in the Mexican axolotl (Ambystoma mexicanum) by heat shock or hydrostatic pressure

    Gillespie, L.L.; Armstrong, J.B.


    Androgenetic diploid axolotls were produced by ultraviolet inactivation of the egg pronucleus shortly after fertilization, followed by suppression of the first cleavage division by hydrostatic pressure or heat shock. After treatment at 14,000 psi for 8 minutes, diploidy was restored in 74% of the embryos, but only 0.8% survived to hatching. A 36-37 degrees C heat shock of 10-minutes duration, applied 5.5 hours after the eggs were collected, yielded a slightly lower percentage of diploids. However, the proportion surviving to hatching was significantly greater (up to 4.6%). A second generation of androgenetic diploids was produced from one of the oldest of the first generation males with a similar degree of success. The lack of significant improvement suggests that the low survival is due to the heat shock per se and not to the uncovering of recessive lethal genes carried by the parent.

  6. Multipolar mitosis of tetraploid cells: inhibition by p53 and dependency on Mos.

    Vitale, Ilio; Senovilla, Laura; Jemaà, Mohamed; Michaud, Mickaël; Galluzzi, Lorenzo; Kepp, Oliver; Nanty, Lisa; Criollo, Alfredo; Rello-Varona, Santiago; Manic, Gwenola; Métivier, Didier; Vivet, Sonia; Tajeddine, Nicolas; Joza, Nicholas; Valent, Alexander; Castedo, Maria; Kroemer, Guido


    Tetraploidy can constitute a metastable intermediate between normal diploidy and oncogenic aneuploidy. Here, we show that the absence of p53 is not only permissive for the survival but also for multipolar asymmetric divisions of tetraploid cells, which lead to the generation of aneuploid cells with a near-to-diploid chromosome content. Multipolar mitoses (which reduce the tetraploid genome to a sub-tetraploid state) are more frequent when p53 is downregulated and the product of the Mos oncogene is upregulated. Mos inhibits the coalescence of supernumerary centrosomes that allow for normal bipolar mitoses of tetraploid cells. In the absence of p53, Mos knockdown prevents multipolar mitoses and exerts genome-stabilizing effects. These results elucidate the mechanisms through which asymmetric cell division drives chromosomal instability in tetraploid cells.

  7. Chromosome counts of some Zingiberaceous species from Thailand

    Ladda Eksomtramage


    Full Text Available The data on chromosome numbers of 22 species belonging to 10 genera of Zingiberaceae distributed in Thailand were investigated. The somatic numbers range from 20 to 48 showing diploidy and polyploidy. Ten of these species are firstly reported here, i.e. Alpinia purpurata (Vielli K. Schum. (2n = 48, Boesenbergia aff. rotunda (2n = 20, Cornukaempferia aurantiflora J. Mood & K. Larsen (2n = 46, Curcuma aff. oligantha Trimen (2n = 42, C. rhabdota Sirirugsa & M.F. Newman (2n = 24, Etlingera elatior (Jack. R.M. Smith (white form (2n = 48, E. hemisphaerica (Bl. R.M. Smith (2n = 48, Hedychium gomezianum Wall. (2n = 34, H. longicornutum Bak. (2n = 34 and Zingiber aff. wrayi (2n = 22



    Triploidy red sea bream were induced by cold-shock techniques (0-3℃) in Qingdao in April 1994, May 1995 and May 1996. Normal diploidy and triploidy chromosome metaphases were produced by chromosome spreads from the gastrula. Counts of 104 chromosome metaphases of normal diploid showed each of them consisted of 2 acrocentric (st) and 46 telocentric (t) chromosomes. Based on the relative lengths and arm ratios, the 48 chromosomes were matched into 24 pairs. Counts of 107 chromosome metaphases of induced triploid showed that each metaphase consisted of 3 acrocentric (st) and 69 telocentric (t) chromosomes. The 72 chromosomes were easily matched into three sets of chromosomes, based on the relative lengths and arm ratios.

  9. A clonal theory of parasitic protozoa: the population structures of Entamoeba, Giardia, Leishmania, Naegleria, Plasmodium, Trichomonas, and Trypanosoma and their medical and taxonomical consequences.

    Tibayrenc, M; Kjellberg, F; Ayala, F J


    We propose a general theory of clonal reproduction for parasitic protozoa, which has important medical and biological consequences. Many parasitic protozoa have been assumed to reproduce sexually, because of diploidy and occasional sexuality in the laboratory. However, a population genetic analysis of extensive data on biochemical polymorphisms indicates that the two fundamental consequences of sexual reproduction (i.e., segregation and recombination) are apparently rare or absent in natural populations of the parasitic protozoa. Moreover, the clones recorded appear to be stable over large geographical areas and long periods of time. A clonal population structure demands that the medical attributes of clones be separately characterized; ubiquitous clones call for priority characterization. Uniparental reproduction renders unsatisfactory Linnean taxonomy; this needs to be supplemented by the "natural clone" as an additional taxonomic unit, which is best defined by means of genetic markers.

  10. Relationship between histologic grade and cytofluorometric cellular DNA and RNA content in primary bone tumors.

    Takeshita, H; Kusuzaki, K; Kuzuhara, A; Tsuji, Y; Ashihara, T; Gebhardt, M C; Mankin, H J; Springfield, D S; Hirasawa, Y


    The diagnosis and grading of bone tumors remains a challenging problem. We studied the relationship between histologic grade and cytofluorometric cellular DNA and RNA content in 108 primary bone tumors. The data included DNA ploidy, mean DNA content (MDC), S-phase fraction (SPF), mean RNA content (MRC) and RNA/DNA ratio (RDR; MRC/MDC) which represents the RNA content normalized for the DNA content. Benign tumors had a diploid stem line with low MDC (mean; 1.04), low SPF (0.9), high MRC (2.41) and high RDR (2.31). Giant cell tumors of bone, which are locally aggressive benign tumors, showed diploidy with relatively higher MDC (1.07, p osteosarcomas revealed aneuploidy with remarkably higher MDC (1.70 in osteosarcomas, p < 0.01) and SPF (6.5, p < 0.01), but lower RDR (1.70, p < 0.01). In contrast, small cell sarcomas, such as Ewing's sarcomas, showed diploidy with low MDC (1.11 in Ewing's sarcomas, N.S.) and SPF (2.5, p < 0.01) and extremely low RDR (1.34, p < 0.01). The RDR value was higher in well-differentiated tumors than in primitive tumors, rendering it useful in grading bone tumors with a diploid stem line. By combining the RDR value with the MDC value, 96% of diploid sarcomas could be distinguished from benign tumors. These results indicate that cellular DNA and RNA content analysis may be of value in assessing the malignant potential of diploid as well as aneuploid bone sarcomas.

  11. Ancient evolutionary trade-offs between yeast ploidy states.

    Enikö Zörgö


    Full Text Available The number of chromosome sets contained within the nucleus of eukaryotic organisms is a fundamental yet evolutionarily poorly characterized genetic variable of life. Here, we mapped the impact of ploidy on the mitotic fitness of baker's yeast and its never domesticated relative Saccharomyces paradoxus across wide swaths of their natural genotypic and phenotypic space. Surprisingly, environment-specific influences of ploidy on reproduction were found to be the rule rather than the exception. These ploidy-environment interactions were well conserved across the 2 billion generations separating the two species, suggesting that they are the products of strong selection. Previous hypotheses of generalizable advantages of haploidy or diploidy in ecological contexts imposing nutrient restriction, toxin exposure, and elevated mutational loads were rejected in favor of more fine-grained models of the interplay between ecology and ploidy. On a molecular level, cell size and mating type locus composition had equal, but limited, explanatory power, each explaining 12.5%-17% of ploidy-environment interactions. The mechanism of the cell size-based superior reproductive efficiency of haploids during Li(+ exposure was traced to the Li(+ exporter ENA. Removal of the Ena transporters, forcing dependence on the Nha1 extrusion system, completely altered the effects of ploidy on Li(+ tolerance and evoked a strong diploid superiority, demonstrating how genetic variation at a single locus can completely reverse the relative merits of haploidy and diploidy. Taken together, our findings unmasked a dynamic interplay between ploidy and ecology that was of unpredicted evolutionary importance and had multiple molecular roots.

  12. Cryptic fitness advantage: diploids invade haploid populations despite lacking any apparent advantage as measured by standard fitness assays.

    Aleeza C Gerstein

    Full Text Available Ploidy varies tremendously within and between species, yet the factors that influence when or why ploidy variants are adaptive remains poorly understood. Our previous work found that diploid individuals repeatedly arose within ten replicate haploid populations of Saccharomyces cerevisiae, and in each case we witnessed diploid takeover within ~1800 asexual generations of batch culture evolution in the lab. The character that allowed diploids to rise in frequency within haploid populations remains unknown. Here we present a number of experiments conducted with the goal to determine what this trait (or traits might have been. Experiments were conducted both by sampling a small number of colonies from the stocks frozen every two weeks (~ 93 generations during the original experiment, as well through sampling a larger number of colonies at the two time points where polymorphism for ploidy was most prevalent. Surprisingly, none of our fitness component measures (lag phase, growth rate, biomass production indicated an advantage to diploidy. Similarly, competition assays against a common competitor and direct competition between haploid and diploid colonies isolated from the same time point failed to indicate a diploid advantage. Furthermore, we uncovered a tremendous amount of trait variation among colonies of the same ploidy level. Only late-appearing diploids showed a competitive advantage over haploids, indicating that the fitness advantage that allowed eventual takeover was not diploidy per se but an attribute of a subset of diploid lineages. Nevertheless, the initial rise in diploids to intermediate frequency cannot be explained by any of the fitness measures used; we suggest that the resolution to this mystery is negative frequency-dependent selection, which is ignored in the standard fitness measures used.

  13. Is there an advantage to be gained from adding digital image cytometry of brush cytology to a standard biopsy protocol in patients with Barrett's esophagus?

    Borovicka, J; Schönegg, R; Hell, M; Kradolfer, D; Bauerfeind, P; Dorta, G; Netzer, P; Binek, J; Meyenberger, C; Fischer, J E; Spieler, P


    The current gold standard in Barrett's esophagus monitoring consists of four-quadrant biopsies every 1-2 cm in accordance with the Seattle protocol. Adding brush cytology processed by digital image cytometry (DICM) may further increase the detection of patients with Barrett's esophagus who are at risk of neoplasia. The aim of the present study was to assess the additional diagnostic value and accuracy of DICM when added to the standard histological analysis in a cross-sectional multicenter study of patients with Barrett's esophagus in Switzerland. One hundred sixty-four patients with Barrett's esophagus underwent 239 endoscopies with biopsy and brush cytology. DICM was carried out on 239 cytology specimens. Measures of the test accuracy of DICM (relative risk, sensitivity, specificity, likelihood ratios) were obtained by dichotomizing the histopathology results (high-grade dysplasia or adenocarcinoma vs. all others) and DICM results (aneuploidy/intermediate pattern vs. diploidy). DICM revealed diploidy in 83% of 239 endoscopies, an intermediate pattern in 8.8%, and aneuploidy in 8.4%. An intermediate DICM result carried a relative risk (RR) of 12 and aneuploidy a RR of 27 for high-grade dysplasia/adenocarcinoma. Adding DICM to the standard biopsy protocol, a pathological cytometry result (aneuploid or intermediate) was found in 25 of 239 endoscopies (11%; 18 patients) with low-risk histology (no high-grade dysplasia or adenocarcinoma). During follow-up of 14 of these 18 patients, histological deterioration was seen in 3 (21%). DICM from brush cytology may add important information to a standard biopsy protocol by identifying a subgroup of BE-patients with high-risk cellular abnormalities.


    林梅绥; 金嘉平; 陈颖; 花井淳


    Objective To study DNA ploidy and genetic changes in the different stages of neoplastic growth in the vocal cord, as well as their biological behavior, for further recognition of the lesions of carcinoma in situ and early carcinoma. Methods 18 tumor lesions of the vocal cord were DNA analyzed by laser scanning cytometry and followed up, and 62 lesions were immunohistochemically investigated for p53, Ki67 and Bcl-X, and with main observation on carcinomas in situ (CISs) and early microinvasive carcinomas (EMICs) which were compared with invasive carcinomas and polyps. Results DNA analysis showed that almost all the CISs and EMICs were diploidy, while 90% invasive carcinomas were aneuploidy. Follow-up data displayed that no one died of the tumor in CIS and EMIC, as well as in the patients with diploidy tumor, and all the patients died of the tumors were with anueploidy tumor. Immunohistochemically, 86% of CIS and EMIC and 91% of invasive carcinoma expressed p53 protein, and the positivities for Ki67 in them were respectively 29% and 27%, which were very significantly different from those of polyps of the vocal cord(P<0. 001). In contrast, expression of Bcl-X were decreasing from benign to malignant lesions, and it was lowest in the invasive carcinomas, significantly different from that of polyp(P=0. 002). Conclusion The present study showed that there were differences of DNA ploidy and genetic expressions among benign lesions, CISs and EMICs, and invasive carcinomas of the vocal cord, indicating that they might be different in biological entities. CIS of the vocal cord could be considered as a borderline lesion, and is better to receive conservative treatment. Moreover, p53 protein determination combined with Ki67 would be helpful in diagnosis of the carcinomas of the vocal cord.

  15. Correlation study between sperm concentration, hyaluronic acid-binding capacity and sperm aneuploidy in Hungarian patients.

    Mokánszki, Attila; Molnár, Zsuzsanna; Ujfalusi, Anikó; Balogh, Erzsébet; Bazsáné, Zsuzsa Kassai; Varga, Attila; Jakab, Attila; Oláh, Éva


    Infertile men with low sperm concentration and/or less motile spermatozoa have an increased risk of producing aneuploid spermatozoa. Selecting spermatozoa by hyaluronic acid (HA) binding may reduce genetic risks such as chromosomal rearrangements and numerical aberrations. Fluorescence in-situ hybridization (FISH) has been used to evaluate the presence of aneuploidies. This study examined spermatozoa of 10 oligozoospermic, 9 asthenozoospermic, 9 oligoasthenozoospermic and 17 normozoospermic men by HA binding and FISH. Mean percentage of HA-bound spermatozoa in the normozoospermic group was 81%, which was significantly higher than in the oligozoospermic (Psex chromosomes (P=0.014) and chromosome 17 (P=0.0019), diploidy (P=0.03) and estimated numerical chromosome aberrations (P=0.004) were significantly higher in the oligoasthenozoospermic group compared with the other groups. There were statistically significant relationships (Pchromosome aberrations (r=-0.668) and between HA binding and estimated numerical chromosome aberrations (r=-0.682). HA binding and aneuploidy studies of spermatozoa in individual cases allow prediction of reproductive prognosis and provision of appropriate genetic counselling. Infertile men with normal karyotypes and low sperm concentrations and/or less motile spermatozoa have significantly increased risks of producing aneuploid (diminished mature) spermatozoa. Selecting spermatozoa by hyaluronic acid (HA) binding, based on a binding between sperm receptors for zona pellucida and HA, may reduce the potential genetic risks such as chromosomal rearrangements and numerical aberrations. In the present study we examined sperm samples of 45 men with different sperm parameters by HA-binding assay and fluorescence in-situ hybridization (FISH). Mean percentage of HA-bound spermatozoa in the normozoospermic group was significantly higher than the oligozoospermic, the asthenozoospermic and the oligoasthenozoospermic groups. Using FISH, disomy of sex

  16. Preferential Bivalent Formation in Tetraploid Male of Pacific Oyster Crassostrea gigas Thunberg

    ZHANG Zhengrui; WANG Xinglian; ZHANG Quanqi; Standish Allen Jr


    Artificially induced tetraploid Pacific oyster, Crassostrea gigas Thunberg, produces more aneuploid gametes than nor-mal diploid one, although they showed a comparable fecundity to diploidy. The meiotic chromosome configuration of 3 tetraploid and 1 tetraploid/triploid mosaic males were analyzed through direct chromosome observation. A majority of metaphase I spermato-cytes contained both bivalents and quadrivalents. The chromosome configuration of these males was characterized by preferential formation of bivalents to quadrivalents. Bivalents appeared in all spermatocytes and consisted of 86%of all chromosome aggregates. In comparison, quadrivalents occurred in 91%spermatocytes and consisted of only 12.6%of all chromosome aggregates. The mean bivalent frequency per spermatocyte varied between 14.4 and 17.2; while that of quadrivalents varied between 2.2 and 2.7. Most quadrivalents were tandemly chained (58%) or circled (39%). The total number of chromosome aggregates per spermatocyte ranged from 13 to 20 with an average of 17.6;while 18 (16 bivalents and 2 quadrivalents) was the most frequent. Univalents and trivalents appeared in very low frequency. Aneuploid (hypotetraploid) spermatocytes were observed in a low frequency. The chromosome con-figuration of in the mosaic individual was similar to that of tetraploid individuals. The percentage of triploid spermatocytes (2%) of the mosaic individual was significantly lower (χ2=30, P<0.01) than that of triploid cells (46%) in its somatic tissue.

  17. Parthenogenesis in Insects: The Centriole Renaissance.

    Riparbelli, Maria Giovanna; Gottardo, Marco; Callaini, Giuliano


    Building a new organism usually requires the contribution of two differently shaped haploid cells, the male and female gametes, each providing its genetic material to restore diploidy of the new born zygote. The successful execution of this process requires defined sequential steps that must be completed in space and time. Otherwise, development fails. Relevant among the earlier steps are pronuclear migration and formation of the first mitotic spindle that promote the mixing of parental chromosomes and the formation of the zygotic nucleus. A complex microtubule network ensures the proper execution of these processes. Instrumental to microtubule organization and bipolar spindle assembly is a distinct non-membranous organelle, the centrosome. Centrosome inheritance during fertilization is biparental, since both gametes provide essential components to build a functional centrosome. This model does not explain, however, centrosome formation during parthenogenetic development, a special mode of sexual reproduction in which the unfertilized egg develops without the contribution of the male gamete. Moreover, whereas fertilization is a relevant example in which the cells actively check the presence of only one centrosome, to avoid multipolar spindle formation, the development of parthenogenetic eggs is ensured, at least in insects, by the de novo assembly of multiple centrosomes.Here, we will focus our attention on the assembly of functional centrosomes following fertilization and during parthenogenetic development in insects. Parthenogenetic development in which unfertilized eggs are naturally depleted of centrosomes would provide a useful experimental system to investigate centriole assembly and duplication together with centrosome formation and maturation.

  18. The Chromosomal Constitution of Embryos Arising from Monopronuclear Oocytes in Programmes of Assisted Reproduction

    Bernd Rosenbusch


    Full Text Available The assessment of oocytes showing only one pronucleus during assisted reproduction is associated with uncertainty. A compilation of data on the genetic constitution of different developmental stages shows that affected oocytes are able to develop into haploid, diploid, and mosaic embryos with more or less complex chromosomal compositions. In the majority of cases (~80%, haploidy appears to be caused by gynogenesis, whereas parthenogenesis or androgenesis is less common. Most of the diploid embryos result from a fertilization event involving asynchronous formation of the two pronuclei or pronuclear fusion at a very early stage. Uniparental diploidy may sometimes occur if one pronucleus fails to develop and the other pronucleus already contains a diploid genome or alternatively a haploid genome undergoes endoreduplication. In general, the chance of obtaining a biparental diploid embryo appears higher after conventional in vitro fertilization than after intracytoplasmic sperm injection. If a transfer of embryos obtained from monopronuclear oocytes is envisaged, it should be tried to culture them up to the blastocyst since most haploid embryos are not able to reach this stage. Comprehensive counselling of patients on potential risks is advisable before transfer and a preimplantation genetic diagnosis could be offered if available.

  19. Cytogeography of essential oil chemotypes of Eremophila longifolia F. Muell (Scrophulariaceae).

    Sadgrove, Nicholas John; Jones, Graham Lloyd


    Previous studies have demonstrated that the widely distributed desert plant Eremophila longifolia has at least six geographically defined essential oil chemotypes. The focus of the present study is to extend and enhance information concerning known chemotypes and to investigate the involvement of cell nuclei ploidy in this variation. Forty field collected specimens of E. longifolia were taken from most of the mainland states of Australia then subjected to hydrodistillation to produce essential oils, which were then chemically characterised. Ploidy was determined using relative fluorescence of cell nuclei stained with propidium iodide, measured in a flow cytometer. Using principal component analysis (PCA), at least three essential oil chemotypes, in addition to the six already described, were identified in the present study. Previously described high yielding essential oil chemotypes were also characterised in terms of diploidy. For the first time diploid populations were identified in New South Wales, correlating with high yielding isomenthone/menthone and karahanaenone chemotypes. Furthermore, the separate diploid population previously described from Western Australia was demonstrated to be the safrole/methyl eugenol type, which is restricted to a small geographic range in far north-west Western Australia (Murchison District). All other chemotypes were shown to be tetraploid, including apparently randomly emerging individuals, representative of chemotypes producing low yields of isomenthone/menthone and karahanaenone similar in composition to the high yielding diploid types.

  20. An Andean radiation: polyploidy in the tree genus Polylepis (Rosaceae, Sanguisorbeae).

    Schmidt-Lebuhn, A N; Fuchs, J; Hertel, D; Hirsch, H; Toivonen, J; Kessler, M


    The Andean tree genus Polylepis (Rosaceae) is notorious for the high morphological plasticity of its species and the difficulty in their circumscription. The evolutionary mechanisms that have driven diversification of the genus are still poorly understood, with factors as diverse as ecological specialisation, reticulate evolution, polyploidisation and apomixis being proposed to contribute. In the present study, chromosome counts, flow cytometry and stomata guard cell size measurements were employed to document for the first time the presence of polyploidy in the genus and to infer ploidy levels for most species. Inferred ploidy levels show a clear progression from diploidy in cloud forest species to polyploidy (tetra- to octoploidy) in the morphologically and ecologically specialised incana group, indicating that polyploidisation may have played a major role in speciation processes and the colonisation of novel habitats during the Andean uplift. At least two species of Polylepis comprise populations with varying degrees of ploidy. More extensive studies are needed to obtain a better understanding of the prevalence and effects of intraspecific polyploidy in the genus.

  1. The nucleoporin Mlp2 is involved in chromosomal distribution during mitosis in trypanosomatids.

    Morelle, Christelle; Sterkers, Yvon; Crobu, Lucien; MBang-Benet, Diane-Ethna; Kuk, Nada; Portalès, Pierre; Bastien, Patrick; Pagès, Michel; Lachaud, Laurence


    Nucleoporins are evolutionary conserved proteins mainly involved in the constitution of the nuclear pores and trafficking between the nucleus and cytoplasm, but are also increasingly viewed as main actors in chromatin dynamics and intra-nuclear mitotic events. Here, we determined the cellular localization of the nucleoporin Mlp2 in the 'divergent' eukaryotes Leishmania major and Trypanosoma brucei. In both protozoa, Mlp2 displayed an atypical localization for a nucleoporin, essentially intranuclear, and preferentially in the periphery of the nucleolus during interphase; moreover, it relocated at the mitotic spindle poles during mitosis. In T. brucei, where most centromeres have been identified, TbMlp2 was found adjacent to the centromeric sequences, as well as to a recently described unconventional kinetochore protein, in the periphery of the nucleolus, during interphase and from the end of anaphase onwards. TbMlp2 and the centromeres/kinetochores exhibited a differential migration towards the poles during mitosis. RNAi knockdown of TbMlp2 disrupted the mitotic distribution of chromosomes, leading to a surprisingly well-tolerated aneuploidy. In addition, diploidy was restored in a complementation assay where LmMlp2, the orthologue of TbMlp2 in Leishmania, was expressed in TbMlp2-RNAi-knockdown parasites. Taken together, our results demonstrate that Mlp2 is involved in the distribution of chromosomes during mitosis in trypanosomatids.

  2. Flow cytometric studies of human osteosarcoma.

    Mankin, H J; Gebhardt, M C; Springfield, D S; Litwak, G J; Kusazaki, K; Rosenberg, A E


    A number of recent studies have emphasized the potential value of flow cytometry as a "marker" to assess the malignity and therefore to help predict the biologic behavior of neoplasms, including bone tumors. Using propidium iodide and a home-built flow cytometer, the authors have studied the DNA distribution in 95 patients with osteosarcoma and determined the percentage of cells in diploidy, S-phase, tetraploidy, and aneuploidy. Using these values and a derived one, mean DNA concentration, it was possible to demonstrate the extent of the abnormalities observed in this group of neoplasms and show their severity as compared with the normal pattern. When the data are compared against disease-free survival and total survival, correlations were noted that, although weak, suggested that some patterns were predictive of increased risk of metastasis and death. The effect of treatment could also be assessed by evaluating the pattern before and after chemotherapy and correlating these with survival. It seems likely that with some improvement in technology, flow cytometry will be of value in the future in assessing the prognosis for osteosarcoma and predicting whether treatment has been effective.

  3. Could sperm aneuploidy rate determination be used as a predictive test before intracytoplasmic sperm injection?

    Petit, François M; Frydman, Nelly; Benkhalifa, Moncef; Le Du, Anne; Aboura, Azzedine; Fanchin, Renato; Frydman, Rene; Tachdjian, Gerard


    Chromosome abnormalities in embryos are a major cause of implantation and development failures. Some couples with normal karyotypes have repeated implantation failures after intracytoplasmic sperm injection (ICSI). In order to value patients at risk for genetic ICSI failures and the validity of sperm aneuploidy analysis, we have studied cytogenetic abnormalities in sperm from ICSI patients. Twenty-nine patients with normal karyotypes were included. Ten patients had at least 4 ICSI treatments without pregnancy (group A). Nine patients had a pregnancy after 1 to 3 ICSI treatments (group B). Ten fertile men with normal semen parameters were studied as controls (group C). Fluorescent in situ hybridization (FISH) was used for sperm nucleus cytogenetic analysis using chromosomes 8, 9, 13, 18, 21, X, and Y specific probes. Aneuploidy for each chromosome and diploidy rates were significantly higher in group A than in group B and in group B than in group C (P < .05). Considering each patient in groups A and B, aneuploidy rate for each chromosome was too variable to be considered as a significant test. We proposed analysis of the total sperm aneuploidy. Chromosomal sperm nuclei profile could be used as a predictive biological test before ICSI in order to improve genetic counseling for oligoasthenoteratozoospermia patients.

  4. Genome-wide analysis of neuroblastomas using high-density single nucleotide polymorphism arrays.

    Rani E George

    Full Text Available BACKGROUND: Neuroblastomas are characterized by chromosomal alterations with biological and clinical significance. We analyzed paired blood and primary tumor samples from 22 children with high-risk neuroblastoma for loss of heterozygosity (LOH and DNA copy number change using the Affymetrix 10K single nucleotide polymorphism (SNP array. FINDINGS: Multiple areas of LOH and copy number gain were seen. The most commonly observed area of LOH was on chromosome arm 11q (15/22 samples; 68%. Chromosome 11q LOH was highly associated with occurrence of chromosome 3p LOH: 9 of the 15 samples with 11q LOH had concomitant 3p LOH (P = 0.016. Chromosome 1p LOH was seen in one-third of cases. LOH events on chromosomes 11q and 1p were generally accompanied by copy number loss, indicating hemizygous deletion within these regions. The one exception was on chromosome 11p, where LOH in all four cases was accompanied by normal copy number or diploidy, implying uniparental disomy. Gain of copy number was most frequently observed on chromosome arm 17q (21/22 samples; 95% and was associated with allelic imbalance in six samples. Amplification of MYCN was also noted, and also amplification of a second gene, ALK, in a single case. CONCLUSIONS: This analysis demonstrates the power of SNP arrays for high-resolution determination of LOH and DNA copy number change in neuroblastoma, a tumor in which specific allelic changes drive clinical outcome and selection of therapy.

  5. Detection of structural and numerical chomosomal abnormalities by ACM-FISH analysis in sperm of oligozoospermic infertility patients

    Schmid, T E; Brinkworth, M H; Hill, F; Sloter, E; Kamischke, A; Marchetti, F; Nieschlag, E; Wyrobek, A J


    Modern reproductive technologies are enabling the treatment of infertile men with severe disturbances of spermatogenesis. The possibility of elevated frequencies of genetically and chromosomally defective sperm has become an issue of concern with the increased usage of intracytoplasmic sperm injection (ICSI), which can enable men with severely impaired sperm production to father children. Several papers have been published about aneuploidy in oligozoospermic patients, but relatively little is known about chromosome structural aberrations in the sperm of these patients. We examined sperm from infertile, oligozoospermic individuals for structural and numerical chromosomal abnormalities using a multicolor ACM FISH assay that utilizes DNA probes specific for three regions of chromosome 1 to detect human sperm that carry numerical chromosomal abnormalities plus two categories of structural aberrations: duplications and deletions of 1pter and 1cen, and chromosomal breaks within the 1cen-1q12 region. There was a significant increase in the average frequencies of sperm with duplications and deletions in the infertility patients compared with the healthy concurrent controls. There was also a significantly elevated level of breaks within the 1cen-1q12 region. There was no evidence for an increase in chromosome-1 disomy, or in diploidy. Our data reveal that oligozoospermia is associated with chromosomal structural abnormalities suggesting that, oligozoospermic men carry a higher burden of transmissible, chromosome damage. The findings raise the possibility of elevated levels of transmissible chromosomal defects following ICSI treatment.

  6. Defining fitness in an uncertain world.

    Crewe, Paul; Gratwick, Richard; Grafen, Alan


    The recently elucidated definition of fitness employed by Fisher in his fundamental theorem of natural selection is combined with reproductive values as appropriately defined in the context of both random environments and continuing fluctuations in the distribution over classes in a class-structured population. We obtain astonishingly simple results, generalisations of the Price Equation and the fundamental theorem, that show natural selection acting only through the arithmetic expectation of fitness over all uncertainties, in contrast to previous studies with fluctuating demography, in which natural selection looks rather complicated. Furthermore, our setting permits each class to have its characteristic ploidy, thus covering haploidy, diploidy and haplodiploidy at the same time; and allows arbitrary classes, including continuous variables such as condition. The simplicity is achieved by focussing just on the effects of natural selection on genotype frequencies: while other causes are present in the model, and the effect of natural selection is assessed in their presence, these causes will have their own further effects on genoytpe frequencies that are not assessed here. Also, Fisher's uses of reproductive value are shown to have two ambivalences, and a new axiomatic foundation for reproductive value is endorsed. The results continue the formal darwinism project, and extend support for the individual-as-maximising-agent analogy to finite populations with random environments and fluctuating class-distributions. The model may also lead to improved ways to measure fitness in real populations.

  7. Meiotic behaviour of sex chromosomes investigated by three-colour FISH on 35,142 sperm nuclei from two 47,XYY males.

    Chevret, E; Rousseaux, S; Monteil, M; Usson, Y; Cozzi, J; Pelletier, R; Sele, B


    Meiotic segregation of sex chromosomes from two fertile 47,XYY men was analysed by a three-colour fluorescence in situ hybridisation procedure. This method allows the identification of hyperhaploidies (spermatozoa with 24 chromosomes) and diploidies (spermatozoa with 46 chromosomes), and their meiotic origin (meiosis I or II). Alpha-satellite probes specific for chromosomes X, Y and 1 were observed simultaneously in 35,142 sperm nuclei. For both 47,XYY men (24,315 sperm nuclei analysed from one male and 10,827 from the other one) the sex ratio differs from the expected 1:1 ratio (P XYY men compared with control sperm (142,050 sperm nuclei analysed from five control men), whereas the rates of hyperhaploidy XY, disomy X and disomy 1 were not significantly different from those of control sperm. These results support the hypothesis that the extra Y chromosome is lost before meiosis with a proliferative advantage of the resulting 46,XY germ cells. Our observations also suggest that a few primary spermatocytes with two Y chromosomes are able to progress through meiosis and to produce Y-bearing sperm cells. A theoretical pairing of the three gonosomes in primary spermatocytes with an extra sex chromosome, compatible with active spermatogenesis, is proposed.

  8. Patterns of homoeologous gene expression shown by RNA sequencing in hexaploid bread wheat.

    Leach, Lindsey J


    BACKGROUND: Bread wheat (Triticum aestivum) has a large, complex and hexaploid genome consisting of A, B and D homoeologous chromosome sets. Therefore each wheat gene potentially exists as a trio of A, B and D homoeoloci, each of which may contribute differentially to wheat phenotypes. We describe a novel approach combining wheat cytogenetic resources (chromosome substitution \\'nullisomic-tetrasomic\\' lines) with next generation deep sequencing of gene transcripts (RNA-Seq), to directly and accurately identify homoeologue-specific single nucleotide variants and quantify the relative contribution of individual homoeoloci to gene expression. RESULTS: We discover, based on a sample comprising ~5-10% of the total wheat gene content, that at least 45% of wheat genes are expressed from all three distinct homoeoloci. Most of these genes show strikingly biased expression patterns in which expression is dominated by a single homoeolocus. The remaining ~55% of wheat genes are expressed from either one or two homoeoloci only, through a combination of extensive transcriptional silencing and homoeolocus loss. CONCLUSIONS: We conclude that wheat is tending towards functional diploidy, through a variety of mechanisms causing single homoeoloci to become the predominant source of gene transcripts. This discovery has profound consequences for wheat breeding and our understanding of wheat evolution.

  9. Infant acute lymphoblastic leukemia: a 20-year children's hospital experience.

    Murray, Rebecca A F; Thom, Giddel; Gardner, Renee V; Craver, Randall D


    We reviewed our 20-year experience with infant acute lymphoblastic leukemia (ALL). Nine infants (4.2% of all ALL) were identified; all were < 6 months of age. White blood cell counts ranged from 42,000-1.6 million/microL, 6 of 8 had hepatosplenomegaly, and 6 of 9 (66.6%) had central nervous system disease. Of 7 with cytogenetic information, 6 (85.7%) had diploidy; the remaining child was 47, XY,+8,del(21)(q22). Four had the MLL-11q23 abnormality. All received chemotherapy. Four underwent stem cell transplantation. Survival was 67%, (15 months-21 years). Deaths occurred at 9 months, 15 months (graft vs. host), and 7 years (complications of small bowel transplantation). Only 1 undergoing stem cell transplantation died. There were no late recurrences or second malignancies. Despite extensive disease and age < 6 months at diagnosis (a poor prognostic feature), for ALL patients our 67% survival is at least as good as reported, although it is less favorable than childhood ALL.

  10. Diploid Male Production of Two Amazonian Melipona Bees (Hymenoptera: Apidae

    Izaura Bezerra Francini


    Full Text Available The diploid male has already been recorded for Melipona Illger, and herein, in Melipona seminigra merrillae Cockerell and Melipona interrupta manaosensis Schwarz. This paper was carried out at the Instituto Nacional de Pesquisas da Amazônia (INPA, Manaus, AM, Brazil. We produced and monitored 31 new colonies of M. s. merrillae and 32 new colonies of M. i. manaosensis. We sampled 2,995 pupae of M. s. merrillae and 2,020 of M. i. manaosensis. In colonies with a 1 : 1 sex ratio, male diploidy was confirmed by cytogenetic analysis and workers’ behavior. We estimated 16 sex-determining alleles in M. s. merrillae and 22 in M. i. manaosensis. In colonies of M. i. manaosensis in a 1 : 1 sex ratio, workers killed the males and the queen that produced them soon after they emerged, as predicted. This behavior was not registered for M. s. merrillae, and sex ratios did not stay 1 : 1, indicating polyandry for this species.

  11. Establishment of Cell Lines from Both Myeloma Bone Marrow and Plasmacytoma: SNU_MM1393_BM and SNU_MM1393_SC from a Single Patient

    Youngil Koh


    Full Text Available Purpose. We tried to establish clinically relevant human myeloma cell lines that can contribute to the understanding of multiple myeloma (MM. Materials and Methods. Mononuclear cells obtained from MM patient’s bone marrow were injected via tail vein in an NRG/SCID mouse. Fourteen weeks after the injection, tumor developed at subcutis of the mouse. The engraftment of MM cells into mouse bone marrow (BM was also observed. We separated and cultured cells from subcutis and BM. Results. After the separation and culture of cells from subcutis and BM, we established two cell lines originating from a single patient (SNU_MM1393_BM and SNU_MM1393_SC. Karyotype of the two newly established MM cell lines showed tetraploidy which is different from the karyotype of the patient (diploidy indicating clonal evolution. In contrast to SNU_MM1393_BM, cell proliferation of SNU_MM1393_SC was IL-6 independent. SNU_MM1393_BM and SNU_MM1393_SC showed high degree of resistance against bortezomib compared to U266 cell line. SNU_MM1393_BM had the greater lethality compared to SNU_MM1393_SC. Conclusion. Two cell lines harboring different site tropisms established from a single patient showed differences in cytokine response and lethality. Our newly established cell lines could be used as a tool to understand the biology of multiple myeloma.

  12. STRAP Is a Strong Predictive Marker of Adjuvant Chemotherapy Benefit in Colorectal Cancer

    Martin Buess


    Full Text Available BACKGROUND: Molecular predictors for the effectiveness of adjuvant chemotherapy in colorectal cancer are of considerable clinical interest. To this aim, we analyzed the serine threonine receptor-associated protein (STRAP, an inhibitor of TGF-βsignaling, with regard to prognosis and prediction of adjuvant 5-FU chemotherapy benefit. i The gene copy status of STRAP was determined using quantitative realtime polymerase chain reaction in 166 colorectal tumor biopsies, which had been collected from a randomized multicenter trial of 5-fluorouracil (5-FU/mitomycin C (MMC adjuvant chemotherapy of the Swiss Group for Clinical Cancer Research (SAKK. RESULTS: Amplification of STRAP was found in 22.8% of the tumors. When left without adjuvant chemotherapy, patients bearing tumors with a STRAP amplification had a significantly better prognosis (hazard ratio for death: 0.26; P = .004. Interestingly, these patients, when receiving adjuvant treatment, had a worse survival (hazard ratio for death: 3.48; P = .019 than without chemotherapy, whereas patients carrying tumors with diploidy or deletion of STRAP benefited from the treatment (hazard ratio for death: 0.44; P = .052. This suggests the amplification of STRAP as a strong predictor of an unfavorable effect of 5-FU-based adjuvant chemotherapy. CONCLUSION: If confirmed, the STRAP gene copy status might provide a parameter to decide about the use of 5-FU-based adjuvant chemotherapy.

  13. Novel and Recently Evolved MicroRNA Clusters Regulate Expansive F-BOX Gene Networks through Phased Small Interfering RNAs in Wild Diploid Strawberry.

    Xia, Rui; Ye, Songqing; Liu, Zongrang; Meyers, Blake C; Liu, Zhongchi


    The wild strawberry (Fragaria vesca) has recently emerged as an excellent model for cultivated strawberry (Fragaria × ananassa) as well as other Rosaceae fruit crops due to its short seed-to-fruit cycle, diploidy, and sequenced genome. Deep sequencing and parallel analysis of RNA ends were used to identify F. vesca microRNAs (miRNAs) and their target genes, respectively. Thirty-eight novel and 31 known miRNAs were identified. Many known miRNAs targeted not only conserved mRNA targets but also developed new target genes in F. vesca. Significantly, two new clusters of miRNAs were found to collectively target 94 F-BOX (FBX) genes. One of the miRNAs in the new cluster is 22 nucleotides and triggers phased small interfering RNA production from six FBX genes, which amplifies the silencing to additional FBX genes. Comparative genomics revealed that the main novel miRNA cluster evolved from duplications of FBX genes. Finally, conserved trans-acting siRNA pathways were characterized and confirmed with distinct features. Our work identified novel miRNA-FBX networks in F. vesca and shed light on the evolution of miRNAs/phased small interfering RNA networks that regulate large gene families in higher plants. © 2015 American Society of Plant Biologists. All Rights Reserved.

  14. Chemotherapy induces transient sex chromosomal and autosomal aneuploidy in human sperm.

    Robbins, W A; Meistrich, M L; Moore, D; Hagemeister, F B; Weier, H U; Cassel, M J; Wilson, G; Eskenazi, B; Wyrobek, A J


    Each year more than 20,000 children and young persons of reproductive age are exposed to known mutagens in the form of chemo- and/or radiotherapy for cancer in the States. As more of these treatments are effective there is growing concern that genetic defects are introduced in the germ cells of these young patients. It is well documented for male rodents that treatment with chemo- and radio-therapeutic agents before mating can cause genetic damage in the germ line, and the magnitude of heritable effects depends on the spermatogenic cell stage treated. Similar germinal effects are suspected to occur in humans but remain unproven. Hodgkin's disease (HD) is an example of a malignancy which is typically diagnosed during a patient's reproductive years. In our study we observed eight male HD patients who were treated with NOVP (Novanthrone, Oncovin, Vinblastine, Prednisone) chemotherapy. We evaluated sperm aneuploidy using multi-colour fluorescence in situ hybridization (FISH), and found approximately 5-fold increases in sperm with disomies, diploidies and complex genotypes involving chromosome X, Y and 8. Increases in sex chromosome aneuploidies arose from segregation errors at meiosis I as well as meiosis II. The aneuploidy effects were transient, however, declining to pretreatment levels within approximately 100 days after the end of the therapy. When compared with normal men, some HD patients showed higher proportions of certain sperm aneuploidy types even before their first therapy.

  15. DNA and RNA content analysis by flow cytometry in the pathobiologic assessment of bone tumors

    El-Naggar, A.K.; Hurr, K.; Tu, Z.N.; Teague, K.; Raymond, K.A.; Ayala, A.G.; Murray, J. [Univ. of Texas, Houston, TX (United States)


    Studies of simultaneous DNA and RNA contents by flow cytometry in hematologic and some solid neoplasms have been shown to provide information that may be useful in pathobiological evaluation of these neoplasms. We contend that similar analysis may be equally valuable in assessing bone tumors. Our data revealed significant statistical differences in DNA ploidy and proliferative fraction between benign and malignant bone neoplasms. Benign tumors manifested predominantly DNA diploidy and low proliferative activity, whereas the majority of malignant tumors were DNA aneuploid and showed high proliferation rate. No significant difference in the RNA content between different histopathologic categories was found. We observed, however, a distinct and consistently high RNA content pattern in giant cell tumors, aneurysmal bone cysts, and chondroblastomas that may be useful in their differential diagnosis. Analysis of different prognostic factors in malignant tumors indicated that histologic grade and DNA content are significant prognostic factors. Further analysis of malignant tumors showed that a correlation between the proliferative activity and the clinical outcome in the low grade category and between RNA content and patients` survival in osteosarcomas. Our study also showed that preoperative treatment significantly impacted on the extent of the proliferative fraction in malignant tumors. We conclude that DNA/RNA analysis of bone tumor may assist in: (1) the differential diagnosis of certain bone tumors, (2) evaluation of treatment response, and (3) the biological assessment of osteosarcomas. 38 refs., 4 figs., 5 tabs.

  16. Derivation and differentiation of haploid human embryonic stem cells.

    Sagi, Ido; Chia, Gloryn; Golan-Lev, Tamar; Peretz, Mordecai; Weissbein, Uri; Sui, Lina; Sauer, Mark V; Yanuka, Ofra; Egli, Dieter; Benvenisty, Nissim


    Diploidy is a fundamental genetic feature in mammals, in which haploid cells normally arise only as post-meiotic germ cells that serve to ensure a diploid genome upon fertilization. Gamete manipulation has yielded haploid embryonic stem (ES) cells from several mammalian species, but haploid human ES cells have yet to be reported. Here we generated and analysed a collection of human parthenogenetic ES cell lines originating from haploid oocytes, leading to the successful isolation and maintenance of human ES cell lines with a normal haploid karyotype. Haploid human ES cells exhibited typical pluripotent stem cell characteristics, such as self-renewal capacity and a pluripotency-specific molecular signature. Moreover, we demonstrated the utility of these cells as a platform for loss-of-function genetic screening. Although haploid human ES cells resembled their diploid counterparts, they also displayed distinct properties including differential regulation of X chromosome inactivation and of genes involved in oxidative phosphorylation, alongside reduction in absolute gene expression levels and cell size. Surprisingly, we found that a haploid human genome is compatible not only with the undifferentiated pluripotent state, but also with differentiated somatic fates representing all three embryonic germ layers both in vitro and in vivo, despite a persistent dosage imbalance between the autosomes and X chromosome. We expect that haploid human ES cells will provide novel means for studying human functional genomics and development.

  17. Detection of structural and numerical chromosomal abnormalities by ACM-FISH analysis in sperm of oligozoospermic infertility patients.

    Schmid, T E; Brinkworth, M H; Hill, F; Sloter, E; Kamischke, A; Marchetti, F; Nieschlag, E; Wyrobek, A J


    Modern reproductive technologies are enabling the treatment of infertile men with severe disturbances of spermatogenesis. The possibility of elevated frequencies of genetically and chromosomally defective sperm has become an issue of concern with the increased usage of ICSI, which can enable men with severely impaired sperm production to father children. Several papers have been published reporting aneuploidy in oligozoospermic patients, but relatively little is known about chromosome structural aberrations in the sperm of these patients. We examined sperm from infertile, oligozoospermic individuals for structural and numerical chromosomal abnormalities using a multicolour ACM fluorescence in situ hybridization (FISH) assay that utilizes DNA probes specific for three regions of chromosome 1 to detect human sperm that carry numerical chromosomal abnormalities plus two categories of structural aberrations: duplications and deletions of 1pter and 1cen, and chromosomal breaks within the 1cen-1q12 region. There was a significant increase in the average frequencies of sperm with duplications and deletions in the infertility patients compared with the healthy concurrent controls. There was also a significantly elevated level of breaks within the 1cen-1q12 region. There was no evidence for an increase in chromosome 1 disomy, or in diploidy. Our data reveal that oligozoospermia is associated with chromosomal structural abnormalities, suggesting that oligozoospermic men carry a higher burden of transmissible, chromosome damage. The findings raise the possibility of elevated levels of transmissible chromosomal defects following ICSI treatment.

  18. p53 Dependent Centrosome Clustering Prevents Multipolar Mitosis in Tetraploid Cells

    Yi, Qiyi; Zhao, Xiaoyu; Huang, Yun; Ma, Tieliang; Zhang, Yingyin; Hou, Heli; Cooke, Howard J.; Yang, Da-Qing; Wu, Mian; Shi, Qinghua


    Background p53 abnormality and aneuploidy often coexist in human tumors, and tetraploidy is considered as an intermediate between normal diploidy and aneuploidy. The purpose of this study was to investigate whether and how p53 influences the transformation from tetraploidy to aneuploidy. Principal Findings Live cell imaging was performed to determine the fates and mitotic behaviors of several human and mouse tetraploid cells with different p53 status, and centrosome and spindle immunostaining was used to investigate centrosome behaviors. We found that p53 dominant-negative mutation, point mutation, or knockout led to a 2∼ 33-fold increase of multipolar mitosis in N/TERT1, 3T3 and mouse embryonic fibroblasts (MEFs), while mitotic entry and cell death were not significantly affected. In p53-/- tetraploid MEFs, the ability of centrosome clustering was compromised, while centrosome inactivation was not affected. Suppression of RhoA/ROCK activity by specific inhibitors in p53-/- tetraploid MEFs enhanced centrosome clustering, decreased multipolar mitosis from 38% to 20% and 16% for RhoA and ROCK, respectively, while expression of constitutively active RhoA in p53+/+ tetraploid 3T3 cells increased the frequency of multipolar mitosis from 15% to 35%. Conclusions p53 could not prevent tetraploid cells entering mitosis or induce tetraploid cell death. However, p53 abnormality impaired centrosome clustering and lead to multipolar mitosis in tetraploid cells by modulating the RhoA/ROCK signaling pathway. PMID:22076149

  19. The parental origin correlates with the karyotype of human embryos developing from tripronuclear zygotes.

    Joergensen, Mette Warming; Labouriau, Rodrigo; Hindkjaer, Johnny; Stougaard, Magnus; Kolevraa, Steen; Bolund, Lars; Agerholm, Inge Errebo; Sunde, Lone


    It has previously been suggested that embryos developing from intracytoplasmic sperm-injected (ICSI) zygotes with three pronuclei (3PN) are endowed with a mechanism for self-correction of triploidy to diploidy. 3PN are also observed in zygotes after conventional in vitro fertilization (IVF). The parental origin, however, differs between the two fertilization methods. Whereas the vast majority of 3PN IVF zygotes are of dispermic origin and thus more likely to have two centrioles, the 3PN ICSI zygotes are digynic in origin and therefore, more likely to have one centriole. In the present study, we examine whether the parental origin of 3PN embryos correlates with the karyotype. The karyotype of each nucleus was estimated using four sequential fluorescence in situ hybridizations-each with two probes-resulting in quantitative information of 8 different chromosomes. The karyotypes were then compared and correlated to the parental origin. 3PN ICSI embryos displayed a significantly larger and more coordinated reduction from the assumed initial 3 sets of chromosomes than 3PN IVF embryos. The differences in the parental origin-and hence the number of centrioles-between the 3PN IVF and the 3PN ICSI zygotes are likely to be the cause of the differences in karyotypes.

  20. Complete hydatidiform mole and a coexistent fetus following ovulation induction in a patient with Sheehan's syndrome: a first case report and review of literature.

    Huang, Xuekun; Liang, Jingyao; Huang, Yonghan; Huang, Juanhua


    Pregnancy in Sheehan's syndrome (SS) is extremely rare. We present the first reported case of twin pregnancy with complete hydatiform mole (CHM) and a coexistent fetus (CHCF) in a patient with SS. A 29-year-old Chinese patient with SS became pregnant following one cycle of ovulation induction with human menopausal gonadotropin after secondary infertility. A normal live fetus and a low echogenic mass suspected hydatidiform mole (HM) were detected by ultrasound examinations at gestational week 8. The couple highly desired to continue the pregnancy because it is very hard to get pregnant for the patients with SS. However, the pregnancy was terminated for the size of the HM component increased rapidly at gestational week 15. Histological examinations confirmed CHCF. Genetic studies showed that the CHM genome was derived from paternal diploidy, and the normal fetus was from biparental genomes. Furthermore, a literature review on these topics is included. This case highlighted that even in a patient with SS, twin pregnancy with CHCF can still occur after ovulation induction.

  1. Constraints on the evolution of asexual reproduction.

    Engelstädter, Jan


    Sexual reproduction is almost ubiquitous among multicellular organisms even though it entails severe fitness costs. To resolve this apparent paradox, an extensive body of research has been devoted to identifying the selective advantages of recombination that counteract these costs. Yet, how easy is it to make the transition to asexual reproduction once sexual reproduction has been established for a long time? The present review approaches this question by considering factors that impede the evolution of parthenogenesis in animals. Most importantly, eggs need a diploid chromosome set in most species in order to develop normally. Next, eggs may need to be activated by sperm, and sperm may also contribute centrioles and other paternal factors to the zygote. Depending on how diploidy is achieved mechanistically, further problems may arise in offspring that stem from 'inbreeding depression' or inappropriate sex determination systems. Finally, genomic imprinting is another well-known barrier to the evolution of asexuality in mammals. Studies on species with occasional, deficient parthenogenesis indicate that the relative importance of these constraints may vary widely. The intimate evolutionary relations between haplodiploidy and parthenogenesis as well as implications for the clade selection hypothesis of the maintenance of sexual reproduction are also discussed.

  2. The population structure of Trypanosoma cruzi: expanded analysis of 54 strains using eight polymorphic CA-repeat microsatellites

    Riva P Oliveira


    Full Text Available Recently we cloned and sequenced the first eight Trypanosoma cruzi polymorphic microsatellite loci and studied 31 clones and strains to obtain valuable information about the population structure of the parasite. We have now studied 23 further strains, increasing from 11 to 31 the number of strains obtained from patients with chronic Chagas disease. This expanded set of 54 strains and clones analyzed with the eight microsatellites markers confirmed the previously observed diploidy, clonal population organization and very high polymorphism of T. cruzi. Moreover, this new study disclosed two new features of the population genetic structure of T. cruzi. The first was the discovery that, similarly to what we had previously shown for strains isolated from insect vectors, mammals and humans with acute disease, isolates from patients in the chronic phase of Chagas disease could also be multiclonal, albeit at a reduced proportion. Second, when we used parsimony to display the genetic relationship among the clonal lineages in an unrooted Wagner network we observed, like before, a good correlation of the tree topography with the classification in three clusters on the basis of single locus analysis of the ribosomal RNA genes. However, a significant new finding was that now the strains belonging to cluster 2 split in two distant sub-clusters. This observation suggests that the evolutionary history of T. cruzi may be more complex than we previously thought.

  3. Correlation of cell kinetic findings with morphology of non-Hodgkin's malignant lymphomas.

    Silvestrini, R; Piazza, R; Riccardi, A; Rilke, F


    Kinetic studies were carried out on 6 benign and 37 malignant lymph nodes from patients with non-Hodgkin's malignant lymphomas (ML). The labeling index, DNA content, and cell distribution through the cell cycle were analyzed in the ML, which were classified according to the Kiel classification. Approximately 90% of the ML studied showed a clear diploidy; the only cases of polyploidy were limited to some centroblastic-centrocytic ML with more than 30% malignant centroblasts and to be single centroblastic ML. The labeling indexes ranged from 0.05 to 33%. No correlation was found between the proliferative rate and the degree of ploidy, while a grading of labeling index was found in relation to the three main DNA distribution patterns observed (i.e., G1 peak, S accumulation, and bimodal distribution through the cell cycle). From a kinetic point of view, the most heterogeneous groups were the lymphoplasmacytoid (subtype polymorphous) and centroblastic-centrocytic ML, where the degree of proliferation increased as the mixture of cell type (relative to the former group) and the malignant centroblastic component (relative to the latter group) increased.

  4. A simple non-invasive protocol to establish primary cell lines from tail and toe explants for cytogenetic studies in Australian dragon lizards (Squamata: Agamidae)

    O’Meally, Denis; Quinn, Alexander E.; Sarre, Stephen D.; Georges, Arthur; Marshall Graves, Jennifer A.


    Primary cell lines were established from cultures of tail and toe clips of five species of Australian dragon lizards: Tympanocryptis pinguicolla, Tympanocryptis sp., Ctenophorus fordi, Amphibolurus norrisi and Pogona vitticeps. The start of exponential cell growth ranged from 1 to 5 weeks. Cultures from all specimens had fibroblastic morphology. Cell lines were propagated continuously up to ten passages, cryopreserved and recovered successfully. We found no reduction in cell viability after short term (<6 months) storage at −80 °C. Mitotic metaphase chromosomes were harvested from these cell lines and used in differential staining, banding and fluorescent in situ hybridisation. Cell lines maintained normal diploidy in all species. This study reports a simple non-invasive method for establishing primary cell lines from Australian dragon lizards without sacrifice. The method is likely to be applicable to a range of species. Such cell lines provide a virtually unlimited source of material for cytogenetic, evolutionary and genomic studies. PMID:19199067

  5. A study of aneuploidy and DNA fragmentation in spermatozoa of three men with sex chromosome mosaicism including a 45,X cell line.

    Nguyen, Minh Huong; Morel, Frederic; Bujan, Louis; May-Panloup, Pascale; De Braekeleer, Marc; Perrin, Aurore


    Meiotic segregation of mosaic males with a 45,X cell line has been little examined. In this study, we evaluated the risk of aneuploid gametes using fluorescence in situ hybridization (FISH) and DNA fragmentation in ejaculated spermatozoa of three men with sex chromosome mosaicism including a 45,X cell line. Triple- and dual-color FISH were performed. Sperm DNA fragmentation was detected using the TUNEL assay. A significantly increased frequency of XY disomic spermatozoa was observed for patients (P)1 and P2. A significant increase in diploidy and autosomal aneuploidy was found in P2 and P3, respectively. The rate of DNA fragmentation was not different from that observed in a control group. Data from the literature are scarce (only 3 cases reported), making comparison of the present data difficult, especially as the frequencies of the cell lines comprising the mosaicism differed between patients. Furthermore, the proportion of the different cell lines can differ from one tissue to another in the same patient. Whether the relative levels of the several cell lines present in the mosaicism can influence the rate of aneuploid spermatozoa remains unknown.

  6. Parasitic Cape honeybee workers, Apis mellifera capensis, evade policing

    Martin, Stephen J.; Beekman, Madeleine; Wossler, Theresa C.; Ratnieks, Francis L. W.


    Relocation of the Cape honeybee, Apis mellifera capensis, by bee-keepers from southern to northern South Africa in 1990 has caused widespread death of managed African honeybee, A. m. scutellata, colonies. Apis mellifera capensis worker bees are able to lay diploid, female eggs without mating by means of automictic thelytoky (meiosis followed by fusion of two meiotic products to restore egg diploidy), whereas workers of other honeybee subspecies are able to lay only haploid, male eggs. The A. m. capensis workers, which are parasitizing and killing A. m. scutellata colonies in northern South Africa, are the asexual offspring of a single, original worker in which the small amount of genetic variation observed is due to crossing over during meiosis (P. Kryger, personal communication). Here we elucidate two principal mechanisms underlying this parasitism. Parasitic A. m. capensis workers activate their ovaries in host colonies that have a queen present (queenright colonies), and they lay eggs that evade being killed by other workers (worker policing)-the normal fate of worker-laid eggs in colonies with a queen. This unique parasitism by workers is an instance in which a society is unable to control the selfish actions of its members.

  7. Comprehensive meiotic segregation analysis of a 4-breakpoint t(1;3;6) complex chromosome rearrangement using single sperm array comparative genomic hybridization and FISH.

    Hornak, Miroslav; Vozdova, Miluse; Musilova, Petra; Prinosilova, Petra; Oracova, Eva; Linkova, Vlasta; Vesela, Katerina; Rubes, Jiri


    Complex chromosomal rearrangements (CCR) represent rare structural chromosome abnormalities frequently associated with infertility. In this study, meiotic segregation in spermatozoa of an infertile normospermic carrier of a 4-breakpoint t(1;3;6) CCR was analysed. A newly developed array comparative genomic hybridization protocol was used, and all chromosomes in 50 single sperm cells were simultaneously examined. Three-colour FISH was used to analyse chromosome segregation in 1557 other single sperm cells. It was also used to measure an interchromosomal effect; sperm chromatin structure assay was used to measure chromatin integrity. A high-frequency of unbalanced spermatozoa (84%) was observed, mostly arising from the 3:3 symmetrical segregation mode. Array comparative genomic hybridization was used to detect additional aneuploidies in two out of 50 spermatozoa (4%) in chromosomes not involved in the complex chromosome rearrangement. Significantly increased rates of diploidy and XY disomy were found in the CCR carrier compared with the control group (P < 0.001). Defective condensation of sperm chromatin was also found in 22.7% of spermatozoa by sperm chromatin structure assay. The results indicate that the infertility in the man with CCR and normal spermatozoa was caused by a production of chromosomally unbalanced, XY disomic and diploid spermatozoa and spermatozoa with defective chromatin condensation.

  8. Influence of spermatogenic profile and meiotic abnormalities on reproductive outcome of infertile patients.

    Barri, Pedro N; Vendrell, Jose M; Martinez, Francisca; Coroleu, Buenaventura; Arán, Begoña; Veiga, Anna


    Genetic aspects of male infertility and the possible risks of new assisted reproduction and their influence on the development of zygotes and children born after intracytoplasmic sperm injection (ICSI) need further research. These patients have an increased risk of diploidy, and disomies are frequent in their spermatozoa. Meiotic disorders are more common in testicular biopsies of patients with severe oligoasthenozoospermia. For these reasons, a detailed andrological study is absolutely mandatory before accepting a couple with these characteristics into an IVF-ICSI programme. When an andrological patient has plasma FSH values >10 IU/l and/or very low total motile sperm count meiotic study in order to rule out meiotic arrest or synaptic anomalies. Another important aspect to be considered is the possible benefit of applying preimplantation genetic diagnosis in these cases because they normally have a high percentage of chromosomally abnormal embryos, although in the present study this was not evident. All studies agree on the necessity of conducting follow-up studies in the population of children born after IVF-ICSI. In this way, it will be possible to find out if these infertile patients and their offspring have a higher risk of suffering epigenetic errors and imprinting disorders.

  9. New and emerging prognostic and predictive genetic biomarkers in B-cell precursor acute lymphoblastic leukemia

    Moorman, Anthony V.


    Acute lymphoblastic leukemia (ALL) is a heterogeneous disease at the genetic level. Chromosomal abnormalities are used as diagnostic, prognostic and predictive biomarkers to provide subtype, outcome and drug response information. t(12;21)/ETV6-RUNX1 and high hyper-diploidy are good-risk prognostic biomarkers whereas KMT2A (MLL) translocations, t(17;19)/TCF3-HLF, haploidy or low hypodiploidy are high-risk biomarkers. t(9;22)/BCR-ABL1 patients require targeted treatment (imatinib/dasatinib), whereas iAMP21 patients achieve better outcomes when treated intensively. High-risk genetic biomarkers are four times more prevalent in adults compared to children. The application of genomic technologies to cases without an established abnormality (B-other) reveals copy number alterations which can be used either individually or in combination as prognostic biomarkers. Transcriptome sequencing studies have identified a network of fusion genes involving kinase genes - ABL1, ABL2, PDGFRB, CSF1R, CRLF2, JAK2 and EPOR. In vitro and in vivo studies along with emerging clinical observations indicate that patients with a kinase-activating aberration may respond to treatment with small molecular inhibitors like imatinib/dasatinib and ruxolitinib. Further work is required to determine the true frequency of these abnormalities across the age spectrum and the optimal way to incorporate such inhibitors into protocols. In conclusion, genetic biomarkers are playing an increasingly important role in the management of patients with ALL. PMID:27033238

  10. Study of the Sperm Chromosomal Aneuploidies of Teratozoospermic Men%畸形精子症患者精子染色体非整倍体的研究

    朱元; 伍琼芳; 辛才林; 赵琰; 林戈; 谭跃球; 程德华; 卢光琇


    Objective: To explore the sperm chromosomal aneuploidies with teratozoospermic men. Methods: Sperm obtained from isolated teratozoospermic men (n=18) and control men with normal fertility (n=5) were analyzed using fluorescent in situ hybridization (FISH) to detect aneuplody of chromosomes 18, X and Y. Results: Totally 58 178 spermatozoa were counted from the teratozoospermia group and 16 369 spermatozoa were counted from the control, with the hybridization rates of 97.5% and 98.3%, respectively. The major types of chromosomal aneuploidies were disomy (YY18, XX18, XY18, Y1818 and X1818) and diploidy (1818XX, 1818YY, 1818XY). In the teratozoospermic group and the control, the disomy rates of 18 chromosome were 0.29 ± 0.16% and 0.03 ± 0.02%, the disomy rates of sex chromosome were 0.65 ± 0.24% and 0.05 ± 0.02%, the diploidy rates were 0.14 ± 0.12% and 0.04 ± 0.03%, respectively. All the differences between the 2 groups were significant (P<0.05). Conclusion: Sperm of isolated teratozoospermic men have higher rates of 18, X and Y chromosomal aneuploidies than those of the fertile controls.%目的:研究畸形精子症患者精子染色体的非整倍体率.方法:应用18号、X和Y染色体着丝粒探针,采用荧光原位杂交(FISH)技术比较畸形精子症患者(畸精组,n=18)和生育力正常且精子正常形态率、浓度、活力等均正常男性(对照组,n=5)精子中18号、X和Y染色体的非整倍体率.结果:畸精组共计数精子58178条,对照组共计数精子16369条.畸精组和对照组杂交效率分别为97.5%和98.3%;染色体非整倍体类型主要有二体(XX 18、YY18、XY18、Y1818和X1818)和二倍体(1818XX、1818YY、1818XY).畸精组和对照组的18号染色体二体率分别为0.29±0.16%和0.03±0.02%,性染色体二体率分别为0.65±0.24%和0.05±0.02%,二倍体率分别为0.14±0.12%和0.04±0.03%.18号、X和Y染色体非整倍体率组间均有统计学差异(P<0.05).结论:与生育力和

  11. 35. Detection of numerical chromosome aberrations in sperm of workers exposed to benzene series by two-color fluorescence in situ hybridization


    To study the frequency of numerical chromosome aberrations in sperm of workers exposed to benzene series, the aneuploidy frequencies of 9 and 18 chromosomes were detected in sperm of workers by two color fluorescence in situ hybridization with digoxingenin labeled 9 chromosome probe (D9zl) and biotin labeled 18 chromosome probe (D18zl). The timeweight average air concentration (TWA ) of benzene in the workplace was 86.49 mg/m3, it was two fold higher than the national maximum allowable concentration. The concentration of urinary trans,trans-muconic acid (ttMA) in exposed group was significantly higher than that of control group. The sperms counted in 14 exposed workers and 16 control workers were 136 401 and 156 955 respectively. The results showed that the disomy frequencies of sperm for chromosome 9, 18 in (0.168%±0.063% and 0.055%±0.031% respectively) and the frequency of diploidy sperm (0.073%±0.045%) were statistically increased over that of the control group(0.050%±0.030%; 0.033%±0.025% and 0.040%±0.036%, respectively). The nullisomic frequencies of 9,18 chromosome in the exposed group (0.206%±0.047%, 0.068%±0.044%) were statistically increased over that of the control group too (0.067%±0.037%, 0.048%±0.034%). The frequency of overall numerical chromosome aberrations in the exposed group was 0.570%±0.144%, when this value was compared with that of control (0.218%±0.071%), a statistically significance was presented. Our experiments showed that exposed to benzene at higher concentration may induce increase in aneuploidy frequency of sperm autosomal chromosome in exposed workers.

  12. 18,X,Y aneuploidies and transmission electron microscopy studies in spermatozoa from five carriers of different reciprocal translocations

    Elena Moretti; Nicola Antonio Pascarelli; Valentina Giannerini; Michela Geminiani; Cecilia Anichini; Oiulia Collodel


    We analysed ejaculated spermatozoa from five infertile men with different balanced reciprocal translocations to contribute to the study of meiotic segregation of chromosomes 18, X and Y and also to evaluate sperm morphology by transmission electron microscopy (TEM) analysis. Conventional lymphocyte karyotype analyses highlighted dif-ferent reciprocal balanced translocations: t(12; 13), t(4;9), t(X;8), t(8; 10) and t(3; 16). Semen analysis was performed by light and TEM. Fluorescence in situ hybridization was performed directly on sperm nuclei using centromeric probes for chromosomes 18, X and Y. The carriers of the balanced reciprocal translocations considered in the pres-ent study showed a very similar pattern of sperm pathologies: diffused presence of apoptosis and immaturity. All patients showed meiotic segregation derangements, highlighted by the presence of sperm diploidies and sex chro-mosome disomies particularly related to the failure of the first meiotic division. However, an increased incidence of chromosome 18 aneuploidy was detected in spermatozoa from t(X;8) and t(8;10) carriers. We have also reported values from sex chromosomes such as t(X;8), although the X chromosome was involved in translocation. Since pa-tients with reciprocal translocations and spermatogenetic impairment are candidates for intracytoplasmic sperm in-jection cycles, the study of sperm parameters, and particularly of the level of aneuploidy rates, would provide better information for couples at risk and would contribute to the data in the literature for a better understanding of the ef-fects of chromosomal rearrangement on the whole meiotic process and, in particular, on chromosomes not involved in translocation.

  13. Induction of aneuploidy in male mouse germ cells detected by the sperm-FISH assay: a review of the present data base.

    Adler, I-D; Schmid, T E; Baumgartner, A


    Multicolour fluorescence in situ hybridization (FISH) with chromosome-specific DNA-probes can be used to assess aneuploidy (disomy) and diploidy in sperm of any species provided the DNA-probes are available. In the present EU research project, DNA-probes for mouse chromosomes 8, X and Y were employed each labelled with different colours. Male mice were treated with the test chemicals and sperm were sampled from the Caudae epididymes 22-24 days later to allow spermatocytes exposed during meiosis to develop into mature sperm. At present, the data base comprises 10 chemicals: acrylamide (AA), carbendazim (CB), colchicine (COL), diazepam (DZ), griseofulvin (GF), omeprazole (OM), taxol (TX), thiobendazole (TB), trichlorfon (TF) and vinblastine (VBL). Of these, COL and TF induced disomic sperm only. DZ and GF induced disomic and diploid sperm, while CB and TB induced diploid sperm only. VBL gave contradictory results in repeated experiments in an inter-laboratory comparison. AA, OM and TX did not induce an increase in disomic or diploid sperm at the doses used. The induction of aneuploidy by DZ was also tested in humans. Sperm samples from patients after attempted suicide and from patients with chronic Valium((R)) abuse were evaluated using human DNA-probes specific for chromosomes 1,16, 21, X and Y. A quantitative comparison between mouse and man indicates that male meiosis in humans is 10-100 times more sensitive than in mice to aneuploidy induction by DZ. The positive response of mice to TF supports the hypothesis by Czeizel et al. [Lancet 341 (1993) 539] that TF may be causally related to the occurrence of congenital abnormality clusters in a Hungarian village.

  14. Yield and quality analysis of variety Ji2003-8 as ovule mutagenesis cross combination%胚珠诱变杂交组合吉2003-8产质量分析

    杨国; 卞桂杰; 黄淑兰; 王绍伦; 潘亚丽


    The ovule mutagenesis cross combination Ji 2008 -8 variety was bred in 2003 by taking the polyembryonic sugar beet diploidy male sterile line Ji75 -08 MS as female parent and the ovule mutagenesis tetraploid sugar beet pure line 9488 - 15 - 1 - 1 as male parent, hybrid seed 3: 1. Through the plot trials in 2007 and 2008, the root yield got 44925.4kg/hm2 ,which increased 24.4% compared with the CK ( Jitian 301 ) ;sugar content was 14.42% ,0.57 degrees higher than the CK; sugar yield got 6480.7kg/hm2 ,which increased by 30.2%compared with the CK.%胚珠诱变杂交组合吉2003-8是2003年以本所自育的多胚二倍体雄性不育系吉75-08MS为毋本,胚珠诱变四倍体纯系9488-15-1-1为父本,按母、父比例3:1自然杂交育成.经2007和2008两年小区试验:根产量44 925.4kg/hm2,比对照(吉甜301)提高24.4%;含糖率14.42%,比对照提高0.57度;产糖量6 480.7kg/hm2,比对照提高30.2%

  15. Inhibitory effects of Curcuma aromatica oil on proliferation of hepatoma in mice

    Wan Yin Wu; Qin Xu; Ling Chun Shi; Wei Bin Zhang


    AIM To reveal the inhibitory effects of Curcuma aromatica oil ( CAO ) on cell proliferation of hepatoma in mice. METHODS Two tumor inhibitory experiments of CAO on hepatoma in mice were conducted.The inhibitory effects of CAO on proliferation of hepatoma in mice were evaluated by DNA image cytometry and immunohistochemical staining of proliferating cell nuclear antigen (PCNA).RESULTS The tumor inhibitory rates of CAO were 52% and 51% in two experiments,respectively. Compared with those of the salinetreated control groups, both differences were statistically significant (P < 0.01). In the group of mice treated with CAO, the cellular nuclear DNA OD value (249 ± 70), areas (623μnm2 ±228 μm2) and DNA (2.38 ± 0.67) index of hepatic carcinomas were significantly lower than those of the control group (430 ± 160, 1073μm2 ± 101 um2 and 4.48 ± 0.71 ). CAO also could increase diploidy cell rates (29.00% ± 9.34% vs 2.97% ± 5.69%, P<0.01 ) and decrease pentaploidy cell exceeding rate (30.04% ± 15.10% vs 70.89%±14.94%, P<0.01). In the group of mice treated with CAO, the labeling indexes of proliferating cell nuclear antigen (PCNA-LI) were 30% ± 4%, which were significantly lower than 40% ± 6% of the control group (P<0.01). CONCLUSION The inhibition of CAO on the growth of hepatoma in mice might be associated with its depression on cellular proliferative activity.

  16. Investigation of the frequency of chromosomal aneuploidy using triple fluorescence in situ hybridization in 12 Chinese infertile men

    张群芳; 卢光琇


    Background Chromosomal aberrations are the major cause of pre-and post-implantation embryo wastage and some studies suggest that half of all human conceptions have a chromosomal abnormality. A chromosomal aberration in human sperms is also one of the causes of failure of in vitro fertilization. This study was designed to ascertain whether chromosomal aneuploidy in spermatozoa is a risk factor for male infertility.Methods Twelve infertile men were divided into two groups: 10 with oligoasthenoteratozoospermia (OAT, Group A) and two with a normal semen analysis (Group B). Two normal healthy sperm donors acted as controls (Group C). We used fluorescence in situ hybridization (FISH) and probes for chromosomes X, Y and 18 to determine the frequency of aneuploidy.Results The frequencies of spermatozoa disomy for chromosomes X, Y and 18 were 0.30% and 0.30%, respectively, in Group B. The percentages were not significantly different from those of Group C (0.15% and 0. 16%). The frequencies of nullisomy for chromosomes X, Y and 18 were 0.15%and 0 for Group B, and 0 and 0.15% for Group C (P>0.05). In Group A, the incidences of disomy were 1.13% and 0. 96% and the frequencies of nullisomy were 1.13% and 1.60%. In these three groups, the incidences of diploidy were 0.60%, 1.00%, and 0.30%, respectively. Both the frequencies of disomic and nullisomic spermatozoa for chromosomes X, Y, and 18 and of diploid spermatozoa were significantly higher in Group A than in Groups B and C. The estimated total aneuploidy rates in the sperm from the three groups were 42.44%, 6.05%, and 2.59%,respectively.Conclusion These results indicate that chromosomal aneuploidy in spermatozoa may be a risk factor for infertility.

  17. Effects of oocyte quality, incubation time and maturation environment on the number of chromosomal abnormalities in IVF-derived early bovine embryos.

    Demyda-Peyrás, Sebastian; Dorado, Jesus; Hidalgo, Manuel; Anter, Jaouad; De Luca, Leonardo; Genero, Enrique; Moreno-Millán, Miguel


    Chromosomal aberrations are one of the major causes of embryo developmental failures in mammals. The occurrence of these types of abnormalities is higher in in vitro-produced (IVP) embryos. The aim of the present study was to investigate the effect of oocyte morphology and maturation conditions on the rate of chromosomal abnormalities in bovine preimplantational embryos. To this end, 790 early cattle embryos derived from oocytes with different morphologies and matured under different conditions, including maturation period (24 v. 36h) and maturation media (five different serum supplements in TCM-199), were evaluated cytogenetically in three sequential experiments. The rates of normal diploidy and abnormal haploidy, polyploidy and aneuploidy were determined in each embryo. Throughout all the experiments, the rate of chromosomal abnormalities was significantly (P<0.05) affected by oocyte morphology and maturation conditions (maturation time and culture medium). Lower morphological quality was associated with a high rate of chromosome abnormalities (P<0.05). Moreover, polyploidy was associated with increased maturation time (P<0.01), whereas the maturation medium significantly (P<0.05) affected the rates of haploidy and polyploidy. In general, supplementing the maturation medium with oestrous cow serum or fetal calf serum resulted in higher rates of chromosomal aberrations (P<0.05) compared with the other serum supplements tested (bovine steer serum, anoestroues cow serum, bovine amniotic fluid and bovine serum albumin). On the basis of the results of the present study, we conclude that the morphological quality of oocytes and the maturation conditions affect the rate of chromosomal abnormalities in IVP bovine embryos.

  18. Cell cycle regulatory factors in juxta-tumoral renal parenchyma.

    Petruşcă, Daniela Nicoleta; Petrescu, Amelia; Vrabie, Camelia; Niculescu, L; Jinga, V; Diaconu, Carmen; Braşoveanu, Lorelei


    The aim of this study was to evaluate regulatory cell cycle factors in juxta-tumoral renal parenchyma in order to obtain information regarding early primary changes occurred in normal renal cells. Specimens of juxta-tumoral renal parenchyma were harvested from the tumoral kidney in 10 patients with no history of treatment before surgery. The expression of p53, Bcl-2, Rb and PCNA was studied by immunohistochemical methods in paraffin-embedded tissues. The apoptotic status was evaluated by flow-cytometry analysis following propidium iodide incorporation. The p53 protein expression was recognized in most of the cases (80%) with different intensities. High intensity apoptotic process detected in juxta-tumoral parenchyma seemed to be p53 dependent and well correlated with the low Bcl-2 expression. 70% of cases were Rb positive. In this type of tissue Rb has only an anti-proliferative and anti-tumoral role. PCNA was present in half of the cases being low expressed due to the tissue regenerating mechanism. Our data suggest that the high intensity of programmed cell death in this type of tissue is supported by the status of cell regulatory factors that control this process. Previous studies have demonstrated that healthy renal tissue has neither apoptosis nor mitotic activity. Juxta-tumoral renal tissue is also displaying normal morphology and DNA content (diploidy) but the microenvironmental status induced by the tumor presence prompts cells to choose death rather than malignant transformation. Further studies are necessary to emphasize if these results have a clinical relevance for the outcome of therapeutical approaches in renal carcinomas.

  19. Multicolor fluorescence in situ hybridization analysis of meiotic chromosome segregation in a 47,XYY male and a review of the literature.

    Shi, Q; Martin, R H


    The frequencies of aneuploid and diploid sperm were determined in a 47,XYY male using multi-color fluorescence in situ hybridization (FISH) analysis, and compared with those from 10 control donors. A total of 30,078 sperm from the patient was scored, 15,044 by two-color FISH for chromosomes 13 and 21, and 15,034 by three-color FISH for the sex chromosomes using chromosome 1 as an internal autosomal control for diploidy and lack of hybridization. The frequencies of X-bearing (49.73%) and Y-bearing sperm (49.46%) in control males were not significantly different from the expected 50% (chi(2)-test for goodness of fit). The ratio of 24,X (50.60%) to 24, Y sperm (48.35%) in the patient, however, was significantly different from the controls (P = 0.0144, chi(2)-test for independence) and from the expected 1:1 ratio (P = 0.0055, chi(2)-test for goodness of fit). There was no significant increase in the frequency of diploid sperm when compared with the controls (chi(2)-test for independence). Significantly increased frequencies were found for 24,YY (0.07% vs. 0.02%, P = 0.0009) and 24,XY (0.44% vs. 0.29%, P = 0.0025), but not for 24,XX (0.05% vs. 0.05%, P > 0. 05), 24,+13 (0.07% vs. 0.07%, P > 0.05) or 24,+21 sperm (0.21% vs. 0. 18%, P > 0.05) in the 47,XYY male when compared with control donors (chi(2)-test for independence). Our results support the theory that loss of the extra Y chromosome occurs during spermatogenesis in most cells. In this XYY patient there was a significant increase in the frequency of sperm with sex chromosomal abnormalities but no suggestion of an inter-chromosomal effect on autosomes. All 3-color FISH studies in the literature demonstrate a significantly increased risk of gonosomal aneuploidy in XYY males, with the risk being on the order of 1%.

  20. 32. Study the aneugenic effect of benzene on germ cell of animal and workers


    Objective: To study the aneupoidy effect of benzene on germ cells of animal and humans. Method: The NIH adult female mice were treated with varies doses of benzene (942, 1881 and 3762mg/kg respectively) by single gavage and (706, 1922 and 4864mg/m3 respectively) by inhalation (2h/d, 15d), the ovulated oocytes were collected for conventional cytogenetic analyses, and the frequencies of aneuploidy were detected. The frequencies of aneuploidy in sperm of benzene exposed workers were detected by two color fluorescence in situ hybridization with digoxingenin labeled 9 chromosome probe(D9Z1) and biotin labeled 18 chromosome probe (D18Z1). Result: The frequencies of aneuploidy in MII oocytes were significantly increased over the control in three groups treated by inhalation (7.06%,7.50% and 7.76% respectively control 1.30%, P<0.05 ), a dose-dependent response was present, But in gavage groups only the high dose group was increased over that of control. P<0.05. The timeweight average air concentration (TWA) of benzene in the workplace was 86.49mg/m3, it was two fold higher than the national maximum allowable concentration. The concentration of urinary trans, trans-muconic acid (ttMA) in exposed group was significantly higher than that of control group. A total of 136 401 sperm nuclei in 14 exposed workers and 156 955 sperm nuclei in 16 control workers were counted. The results showed that the frequencies of disomic sperm for chromosome 9 and 18, and diploidy sperm in exposed-workers (0.168%, 0.055%, 0.073%, respectively) were statistically increased over that (0.050%、 0. 033% and 0.040%, respectively) of controls. P<0.05. The frequencies of nullisomic sperm for chromosome 9,18 in the exposed group (0.206%,0.068%) were statistically increased also over that (0.067%,0.048% respectively) of control. The frequency of overall numerical chromosome aberrations (0.570%) in tbe exposed group was increased over that of control(0.218%)P<0.05.Conclusion:An increased aneuploidy

  1. Complete mole in a dichorionic twin pregnancy after intracytoplasmic sperm injection Mola hidatiforme completa em gravidez bicoriônica após injecção intracitoplasmática de espermatozoides

    Tiago José Santos de Matos Ferraz


    Full Text Available A dichorionic twin pregnancy with complete hydatidiform mole and coexistent fetus is a rare and challenging situation, whose pathogenesis has not been yet fully understood. We present a case of a 39-year-old woman who underwent intracytoplasmic sperm injection with two embryos transfer. The 12-week gestation ultrasound examination revealed normal fetus and placenta with features of hydatidiform mole, leading to pregnancy termination. Autopsy and histological examinations diagnosed a complete mole coexisting with a normal fetus, and the genetic analysis showed a diploid fetus with biparental genome and molar tissue with paternal diploidy. This case highlighted that complete molar pregnancies may still occur even though pregnancy is achieved after intracytoplasmic sperm injection. A review of the literature was performed by collecting data from the few similar reported cases and by commenting on the pathogenesis of this rare condition.Uma gravidez bicoriônica com mola hidatiforme completa e feto normal é uma situação rara e desafiadora, cuja patogênese não foi ainda totalmente compreendida. Apresenta-se o caso de uma mulher de 39 anos submetida à injeção intracitoplasmática de espermatozoides com transferência de dois embriões. Na ecografia pré-natal realizada na 12ª semana de gestação, foi identificado um embrião morfologicamente normal e uma placenta com características molares. Esta situação resultou na terminação eletiva da gravidez. A autópsia e o estudo histológico permitiram o diagnóstico definitivo de uma mola hidatiforme completa coexistindo com feto normal. A análise genética mostrou feto diploide com genoma biparental e tecido molar com diploidia paterna. Este caso ressaltou que as gestações com mola hidatiforme completa poderão ainda ocorrer, mesmo que a gravidez seja realizada após uma injeção intracitoplasmática de espermatozoides. Foram realizadas uma revisão dos raros casos descritos na literatura

  2. A CpG island hypermethylation profile of primary colorectal carcinomas and colon cancer cell lines

    Rognum Torleiv O


    Full Text Available Abstract Background Tumor cell lines are commonly used as experimental tools in cancer research, but their relevance for the in vivo situation is debated. In a series of 11 microsatellite stable (MSS and 9 microsatellite unstable (MSI colon cancer cell lines and primary colon carcinomas (25 MSS and 28 MSI with known ploidy stem line and APC, KRAS, and TP53 mutation status, we analyzed the promoter methylation of the following genes: hMLH1, MGMT, p16INK4a (CDKN2A α-transcript, p14ARF (CDKN2A β-transcript, APC, and E-cadherin (CDH1. We compared the DNA methylation profiles of the cell lines with those of the primary tumors. Finally, we examined if the epigenetic changes were associated with known genetic markers and/or clinicopathological variables. Results The cell lines and primary tumors generally showed similar overall distribution and frequencies of gene methylation. Among the cell lines, 15%, 50%, 75%, 65%, 20% and 15% showed promoter methylation for hMLH1, MGMT, p16INK4a, p14ARF, APC, and E-cadherin, respectively, whereas 21%, 40%, 32%, 38%, 32%, and 40% of the primary tumors were methylated for the same genes. hMLH1 and p14ARF were significantly more often methylated in MSI than in MSS primary tumors, whereas the remaining four genes showed similar methylation frequencies in the two groups. Methylation of p14ARF, which indirectly inactivates TP53, was seen more frequently in tumors with normal TP53 than in mutated samples, but the difference was not statistically significant. Methylation of p14ARF and p16INK4a was often present in the same primary tumors, but association to diploidy, MSI, right-sided location and female gender was only significant for p14ARF. E-cadherin was methylated in 14/34 tumors with altered APC further stimulating WNT signaling. Conclusions The present study shows that colon cancer cell lines are in general relevant in vitro models, comparable with the in vivo situation, as the cell lines display many of the same

  3. 早期口腔癌检查及诊断方法的研究进展%Advances in detection and diagnosis of early oral cancers

    李金忠; 李鑫; 郑家伟


    过去几十年,口腔癌的治疗方法取得了很大进步,但中晚期口腔癌的预后并未碍到明显改善,早期诊断、早期治疗仍然是提高治愈率的关键措施.早期确诊的主要方法是切取活检,但该方法有创,且异常增生与早期口腔癌的区分过于依赖病理医师的临床经验.为诊断口腔异常增生和早期口腔癌,迫切需要可在门诊条件下操作简便、实用且无创的诊断工具.目前发展迅速的早期口腔癌检测方法包括光学系统检查、活组织染色、唾液检测、刷取活检、DNA分析等.本文对这些方法诊断口腔鳞癌及癌前病损的价值进行回顾与评价.%The therapeutic modalities have accomplished important advances in last several decades, while the prognosis of oral cancers remains unchanged. Since oral cancers have a much better prognosis in early stage than in advanced stage, the detection and diagnosis of early oral cancers is the key in the overall control of oral cancers. The essential method in identification of early cancers is still scalpel biopsy. However, this method is invasive and restricted to highly suspicious lesions. Furthermore, scalpel biopsy depends heavily on the experiences of pathologists in the differential diagnosis of dysplasia and early oral cancers. There is urgent need to devise easy, practical, and non-invasive methods for the detection and diagnosis of early oral cancers that can be performed easily in out-patient set-up. This paper reviewed the current methods, including light-based oral cancer screening system, vital staining, saliva testing, brush biopsy, DNA diploidy analysis, et al, and evaluated them in the detection and diagnosis of early oral cancers and precancerous lesions.

  4. Uniparental disomy (UPD). Genomic imprinting and a case for new genetics (prenatal and clinical implications: the "Likon" concept).

    Engel, E


    Uniparental disomy (UPD) is often the result of an aneuploid event masquerading under the features of diploidy. As such, it may never be recognized, being at 2 opposite phenotypic poles, harmless to the bearer, or, if harmful, eventually responsible for uncharacteristic although perhaps serious conditions. UPD can also be associated with problems such as recessiveness or mosaicism. This article considers the chances of unmasking UPD, in the course of CVS or AC prenatal diagnosis, by reviewing the main cytogenetic signals and major familial or personal antecedents raising its suspicion. Once suspected, the lead toward UPD may or may not be followed through appropriate molecular studies. UPD for either maternal or paternal chromosomes 13, 21 and 22 may not have consistent, common deleterious effects, while other identified UPD's are too rare to call. Unconditionally, main, consistent or near consistent damages to the phenotype have been traced to specific chromosome pairs such as 15 mat (Prader-Willi syndrome), 15 pat (Angelman syndrome), 11 pat (Wiedemann-Beck with syndrome), 14 mat and pat (multiple cogenital and developmental anomalies [MCDA]-several rather constant) and 7 mat (Russel-Silver [RS] and Growth-failure [GF]). The above problems all stem from an alteration of the normal, developmentally important genomic imprinting processes and most of them may recognize several etiopathogenic paths, other than UPD, none of which abides by straight Mendelian rules. In this very area, therefore, a new, non-traditional type of inheritance is confronting genetic counselling. In this paper, for want of appropriate semantic language, the neologism "likon" (or "laïkon") is coined to make reference to the hemizygously expressed sequences of the genomic parts imprinted in the somatic tissues. Broadening the definition, the word is then applied to the 4 possible epigenotypes of imprinted domains, which depend on the parental sex-of-origin: germinally "resting" (R), or "acting

  5. Establishment of a rapid technical assessment of sex chromosome numerical abnormality in spermatozoa%快速检测精子性染色体分析技术的建立

    乐威; 钱君海; 赵华颖; 袁涛; 吴登龙; 章劲夫


    Objective To establish a rapid method to analyze the numerical abnormality of chromosome X and Y in spermatozoa from the healthy males by using two-color fluorescence in situ hybridization( FISH) .Methods Fluorescence in situ hybridization was carried out by using centromeric probes targeting chromosomes, including X and Y upon the sperm samples harvested from 7 healthy men with normal semen quality.The frequency of numerical abnormality ( aneuploidy, diploidy) was rated in each sample.Results A total of 10 078 sperm cells were counted.The total hybridization rate was 99.000% with ( 49.789 ± 1.346 ) % for chromosome X and (48.814 ± 1.296)% for chromosome Y.Abnormality in these samples included XX-, XY-, and YY-typed chromosomes, respectively with incidence of (0.110 ±0.053 ) % , (0.222 ±0.077)% , and(0.094 ±0.038) % .Conclusion A rapid and efficient method has been established to analyze the numerical abnormality in sperm sex chromosomes by using fluorescence in situ hybridization.This method may be adopted to examining genetically the quality of spermatozoa and to estimate male's infertility.%目的 建立一种应用双色荧光原位杂交技术快速分析正常男性精子XY染色体数目及异常率的方法.方法 采用X、Y号染色体着丝粒探针对7例健康男性精子样本进行荧光原位杂交实验,检测精子XY染色体数目异常情况(非整倍体,双倍体).结果 计数10078个精子,杂交率99.00%.其中X染色体杂交率为(49.789±1.346)%,Y染色体杂交率为(48.814±1.296)%,精子中双体类型为XX、XY、YY,其中XX杂交率为(0.110±0.053)%,XY杂交率为(0.222±0.077)%,YY杂交率为(0.094±0.038)%.结论 采用荧光原位杂交技术建立了一种能够快速高效分析精子染色体数目异常的检测方法,可作为精子质量遗传学检测手段,应用于男性不育症的临床检测.

  6. Sperm chromosome analysis and preimplantation genetic diagnosis in an infertile male with mosaic trisomy 18%一例嵌合型18三体少精子症患者精染色体分析及植入前遗传学诊断

    罗玉琴; 钱羽力; 朱瑞建; 叶英辉; 朱宇宁; 金帆


    Objective To analyze the numerical aberration rate of X, Y and chromosome 18 in sperms from an oligozoospermic male with mosaic trisomy 18 and to perform preimplantation genetic diagnosis (PGD) for the couple. Methods G-banding and fluorescence in situ hybridization (FISH) were performed on metaphase chromosome. Sperm was analyzed in three-color FISH with a probe mixture containing CEP18, CEPY and Tel Xq/Yq. A healthy man with normal semen parameters was used as control. Results Significant difference in the rates of disomy for chromosome 18 (0. 63% vs. 0. 16%) and the gonosomes (0. 94% vs. 0. 35%) and diploidy (0. 87% vs. 0. 31%) was found in the spermatozoa between the patient and the control. After four embryos were biopsied in one PGD cycle, two embryos with XY1818 and XX1818 were selected for implanting and clinical pregnancy was ongoing. Conclusion SpermFISH allows further understanding of aneuploidy rate and accurate genetic counseling. FISH-PGD was effective for patient with mosaic trisomy 18.%目的 分析1例嵌合型18三体少精子患者精子18、X、Y染色体数目畸变并进行植入前遗传学诊断(preimplantation genetic djagnosis,PGD).方法 采用G带及荧光原位杂交(fluorescence in situ hybridjzation,FISH)对中期分裂相进行分析,应用三色探针CEP18、CEPY、Tel Xq/Yq对患者精子进行FISH分析,同时以1名染色体正常男性的正常精液作为对照,并对嵌合型18三体患者进行PGD.结果 患者精子18二体率、性染色体二体率和二倍体率分别为0.63%、0.94%和0.87%,与对照组相比(0.16%、0.35%、0.31%)差异有统计学意义.患者进行1个PGD周期的治疗、活检4个胚胎,移植正常的XY1818、XX1818各1胚胎后获得临床妊娠.结论 精子FISH分析可为其提供更准确的遗传咨询及指导植入前遗传学诊断,FISH-PGD可有效地应用于嵌合型18三体的植入前遗传学诊断.

  7. p57K1P2和PHLDA2蛋白表达对葡萄胎的诊断意义%The value of p57K1P2 and PHLDA2 in the differential diagnosis of placental hydropic diseases

    方芳; 万希润; 向阳; 冯风芝; 任彤


    Objective To investigate the value of combined use of p57K1P2 and PHLDA2 immunohistochemis-try and flow cytometry in the difierential diagnosis of placental hydropic diseases. Methods Specimens of 29 cases of mole pregnancy, formerly diagnosed as complete hydatidiform moles (CHM, n=13), and partial hydatidiform moles (PHMn=16), were reviewed by a senior pathologist. p57K1P2 and PHLDA2 immunohistochemical staining and flow cytometry DNA ploidy analysis were performed in all 29 cases. Results Flow cytometry in all 29 molar cases suggested that 14 cases were triploidy or tetraploid (means these cases are PHM) and 15 cases were diploidy (means these cases are CHM) . All flow cytometrically confirmed partial moles were both p57K1P2 and PHLDA2 positive. There was strong PHLDA2 staining of the cytoplasm in virtually all cells of the villous cytotrophoblast, while p57K1P2 was localized to the nucleus in a subset of those cells. All flow cytometrically showed that complete moles cases were both p57K1P2 and PHLDA2 negative. Conclusion Immunohistochemistry for p57K1P2 and PHL-DA2 may be serves as a practical and reliable diagnostic marker for the diagnosis of complete mole from partial mole.%目的 研究母源表达的印记基因p57K1P2和PHLDA2(IPL/TSSC3)蛋白表达对葡萄胎的辅助诊断意义.方法 收集北京协和医院刮宫组织存档的石蜡包埋标本,其中病理组织学诊断完全性葡萄胎( completehydatidiform mole,CHM) 13例,部分性葡萄胎(partial hydatidiform mole,PHM) 16例.全部病例行流式细胞术DNA倍体分析,采用免疫组化二步法检测p57K1p2及PHLDA2在病理组织中表达.结果 29例病例DNA倍体分析:二倍体15例,三倍体13例,四倍体1例,诊断为CHM 15例,PHM 14例,病理诊断的符合率为86.2%.部分性葡萄胎p57K1P2和PHLDA2绒毛滋养细胞层免疫组织化学全部阳性(100%,14/14).PHLDA2染色阳性位于胞质和胞膜,表达为强阳性.p57K1P2染色阳性位于