Asynchrony of DNA replication and mitotic spiralization along heterochromatic portions of Chinese hamster chromosomes

Sequence of DNA synthesis and mitotic chromosome spiralization along heterochromatic portions of the sex (X₁X₂) and of some marker chromosomes in cultured Chinese hamster cells were studied, employing two methods: study of segmentation pattern caused in chromosomes with colcemid, and autoradiography with tritiated thymidine. The heterochromatic portions of all chromosomes studied were characterized by striking internal asynchrony of DNA replication. In particular, they had segments that replicated relatively early. The short arm of the X₂ chromosome, heterochromatic in female somatic cells, had at least three such segments. Replication patterns of the long arms of the X₁ and X₂ chromosomes were different. In X₁ this arm contains several segments showing relatively early replication. The long arm of X₂ had no similar segments. The possible significance of the data obtained is discussed with regard to the problem of genetic inertness of heterochromatin. At the terminal stage of the S period, H³-thymidine seems to be incorporated into condensed chromatin of interphase nuclei. On the basis of the data obtained, it is proposed that during replication of heterochromatin consecutive despiralization of parts of it takes place.     

CHROMOSOMAL BASIS OF DOSAGE COMPENSATION IN DROSOPHILA : III. Early Completion of Replication by the Polytene X-Chromosome in Male: Further Evidence and Its Implications

Thymidine-³H labeling patterns on the X (section 1 A to 12 E of Bridges' map) and 2 R (section 56 F to 60 F of Bridges' map) segments in the salivary gland chromosomes of Drosophila melanogaster have been analyzed in male and female separately. The observed patterns fit, with a few exceptions, in a continuous to discontinuous labeling sequence. In nuclei with similar labeling patterns on the 2R segment in both sexes, the number of labeled sites on the X in male is always less than in female X's. The labeling frequency of the different sites on the male X is considerably lower than those on the female X's, while the sites on the 2R segment have very similar frequency in the two sexes. The rate of thymidine-³H incorporation (as judged by visual grain counting) is relatively higher in male X than in female X's. It is concluded that the model sequence of replication in polytene chromosomes follows a continuous to discontinuous labeling sequence, and that the single X in male completes its replication earlier than either the autosomes in male or the X's in female. This asynchronous and faster rate of replication by the polytene X-chromosome in male substantiates the hypothesis of hyperactivity of the single X in male as the chromosomal basis of dosage compensation in Drosophila.     

Conserved autonomy of replication of the X-chromosomes in hybrids of Drosophila miranda and Drosophila persimilis

The chromosome complement of hybrid males from the cross between Drosophila miranda female and D. persimilis male provides an interesting chromosomal situation where an autosome, the 3rd chromosome of D. persimilis, coexists with a homologue that developed into a sex chromosome, the X₂ in D. miranda. Except for certain inversions and a few minor translocations, these two chromosomes (X₂ and the 3rd) still look alike as polytene elements. However, in hybrid males pairing of the two chromosomes, the X₂ and 3rd, is rare, while in female hybrids it occurs frequently. — ³H-TdR labeling shows that while the X₂ and 3rd chromosomes replicate synchronously in hybrid female, in the hybrid male the former completes its replication earlier than the 3rd chromosome, as do the two arms of the X₁ (XL and XR). The frequency and relative intensity of ³H-TdR labeling of each site of the X₂ and that of the 3rd chromosome in hybrid males closely agree with those of the corresponding sites in the X₂ of the miranda male and the 3rd chromosome of the persimilis male (or female), respectively. The results suggest that timing and rate of replication of the X₂ are determined autonomously and follow the pattern in the respective parental species.     

Studies of multipolar mitoses in euploid tissue cultures

In tissue cultures of male Microtus agrestis, diploid mitoses with two X or two Y chromosomes were found. For identifiying the sex chromosomes in nonhypotonioally treated mitoses, the asynchrony of DNA replication of the sex chromosomes of both sexes was used. The constitutive heterochromatin of Y replicates later in the S period than X, and X₂ of the female replicates later than X₁. Autoradiographic studies of tetraploid tripolar mitoses showed that the diploid daughter nuclei contain either XX or YY in the male; in the female, X₁X₂ daughter nuclei were found less frequently than X₁X₁ and X₂X₂ cells.     

X1X1X2X2/X1X2Y sex chromosome systems in the Neotropical Gymnotiformes electric fish of the genus Brachyhypopomus

Several types of sex chromosome systems have been recorded among Gymnotiformes, including male and female heterogamety, simple and multiple sex chromosomes, and different mechanisms of origin and evolution. The X₁X₁X₂X₂/X₁X₂Y systems identified in three species of this order are considered homoplasic for the group. In the genus Brachyhypopomus, only B. gauderio presented this type of system. Herein we describe the karyotypes of Brachyhypopomus pinnicaudatus and B. n. sp. FLAV, which have an X₁X₁X₂X₂/X₁X₂Y sex chromosome system that evolved via fusion between an autosome and the Y chromosome. The morphology of the chromosomes and the meiotic pairing suggest that the sex chromosomes of B. gauderio and B. pinnicaudatus have a common origin, whereas in B . n. sp. FLAV the sex chromosome system evolved independently. However, we cannot discard the possibility of common origin followed by distinct processes of differentiation. The identification of two new karyotypes with an X₁X₁X₂X₂/X₁X₂Y sex chromosome system in Gymnotiformes makes it the most common among the karyotyped species of the group. Comparisons of these karyotypes and the evolutionary history of the taxa indicate independent origins for their sex chromosomes systems. The recurrent emergence of the X₁X₁X₂X₂/X₁X₂Y system may represent sex chromosomes turnover events in Gymnotiformes.     

DNA-Replikation und Chromosomenstruktur von Mesostoma (Turbellaria)

During meiosis in M. ehrenbergi (2n=10) and M. lingua (2n=8) male certain chromosomes never pair completely. In these bivalents only terminal pairing appears, crossing over could not be proved by ³H-thymidine autoradiography. DNA amounts of the M. ehrenbergi and M. lingua genomes are in a proportion of 10∶1. The mitotic S-phase of spermatogonia in M. ehrenbergi is twice as long as in M. lingua. In metaphase of spermatogonia a differentiated DNA replication pattern can be identified in M. ehrenbergi as late-pulse-replicating segments. After incorporation of ³H-thymidine X₂-metaphase chromosomes can be found, which show single chromatid labeling, terminal and intercalary isolabeling as well as kinds of chromosome labeling, which can only result from sister strand exchange. After treating the chromosomes with low temperature, colchicine or by hydrolysis (60° C) substructures of the chromatin become visible in both spezies which however are evaluated as artefacts. — Formation of the different isolabeling types is discussed on the basis of a two-strand model of the chromosome fibril. A hypothesis is formulated that the surplusage of DNA in M. ehrenbergi is distributed over all the length of the chromatids as small parts of heterochromatin. This hypothesis is supported by investigations of the DNA replication and the contractility of the chromosomes. Furthermore, a pattern of small DNA particles can be demonstrated after partial destruction of the DNA in metaphase chromosomes of M. ehrenbergi, which could represent this intercalary heterochromatin.     

Karyological analysis of Cylindera trisignata (Latreille & Dejean, 1822) from Portugal (Coleoptera,Cicindelidae)

Cellular suspensions of germinal tissues of Cylindera trisignata provided the definition of its karyotype: 2n=23 and 2n=26 for the males and females respectively. This Palearctic species has a sex chromosome system of the X₁X₂X₃X₄Y / X₁X₁X₂X₂X₃X₃X₄X₄ type, only found until now in Cicindela maroccana pseudomaroccana. The heterosomes are not well-differentiated from the general morphological standpoint. To explain the origin of the 4X condition, a mechanism of dissociation of the X chromosomes rather than an incorporation of autosomal segments is proposed. However, based on the occurrence of distinct ploidy levels, both in male and female cells, with a relatively high incidence for the tetraploid condition with regular autosomal bivalents, the hypothesis of a possible evolutionary role of polyploidy is suggested.     

Genetic variation in the serotonin transporter gene (5-HTTLPR, rs25531) influences the analgesic response to the short acting opioid Remifentanil in humans

A growing body of evidence indicates that P2X receptors (P2XRs), a family of ligand-gated cation channels activated by extracellular ATP, play an important role in pain signaling. In contrast to the role of the P2X₃R subtype that has been extensively studied, the precise roles of others among the seven P2XR subtypes (P2X₁R-P2X₇R) remain to be determined because of a lack of sufficiently powerful tools to specifically block P2XR signaling in vivo. In the present study, we investigated the behavioral phenotypes of a line of mice in which the p2rx4 gene was disrupted in a series of acute and chronic pain assays. While p2rx4 ^-/- mice showed no major defects in pain responses evoked by acute noxious stimuli and local tissue damage or in motor function as compared with wild-type mice, these mice displayed reduced pain responses in two models of chronic pain (inflammatory and neuropathic pain). In a model of chronic inflammatory pain developed by intraplantar injection of complete Freund's adjuvant (CFA), p2rx4 ^-/- mice exhibited attenuations of pain hypersensitivity to innocuous mechanical stimuli (tactile allodynia) and also of the CFA-induced swelling of the hindpaw. A most striking phenotype was observed in a test of neuropathic pain: tactile allodynia caused by an injury to spinal nerve was markedly blunted in p2rx4 ^-/- mice. By contrast, pain hypersensitivity to a cold stimulus (cold allodynia) after the injury was comparable in wild-type and p2rx4 ^-/- mice. Together, these findings reveal a predominant contribution of P2X₄R to nerve injury-induced tactile allodynia and, to the lesser extent, peripheral inflammation. Loss of P2X₄R produced no defects in acute physiological pain or tissue damaged-induced pain, highlighting the possibility of a therapeutic benefit of blocking P2X₄R in the treatment of chronic pain, especially tactile allodynia after nerve injury.     

Die Cytogenetik zweier norddeutscher Populationen von Nosopsyllus fasciatus Bosc (Aphaniptera)

Specimens of the populations Hamburg and Wilhelmshaven of the ratflea N. fasciatus exhibit variation of the chromosome number in the range of 2n=20–23 and 2n=20–27 respectively, resulting from individual differences in the number of supernumerary chromosomes beyond the basic chromosome complement of 2n=20. The supernumerary chromosomes are mostly euchromatic and partly or completely homologous to each other and to the 10. pair of the basic complement. The numerical variation in the population Wilhelmshaven is produced by recurrent mitotic non-disjunction of the supernumerary chromosomes in anaphase II of spermatogenesis. Constant mitotic non-disjunction and preferential segregation of the supernumerary chromosomes towards the pronucleus leads to their accumulation in the population.—A multiple sex-chromosome mechanism of the type X₁ X₂ Y₁ Y₂ (male): X₁ X₁ X₂ X₂ (female) has been demonstrated for the population Wilhelmshaven of N. fasciatus. The X₁ X₂ Y₁ Y₂-chain of four is restricted to the male meiosis, in oogenesis two sex bivalents (X₁ X₁ and X₂ X₂) are formed. — The cytogenetic data presented do not support the concept of a closer phylogenetic relationship between the Aphaniptera and Nematocera, but do not preclude the possibility of a kinship of Aphaniptera and Neomecoptera.     

Quantification of Cell Proliferation and Alpha-Toxin Gene Expression of Clostridium perfringens in the Development of Necrotic Enteritis in Broiler Chickens

Cell proliferation and alpha-toxin gene expression of Clostridium perfringens in relation to the development of necrotic enteritis (NE) were investigated. Unlike bacitracin-treated chickens, non-bacitracin-treated birds exhibited typical NE symptoms and reduced growth performance. They also demonstrated increased C. perfringens proliferation and alpha-toxin gene expression that were positively correlated and progressed according to the regression model y = b₀ + b₁X − b₂X². The average C. perfringens count of 5 log₁₀ CFU/g in the ileal digesta appears to be a threshold for developing NE with a lesion score of 2.     

Evolutionary breakpoint regions and chromosomal remodeling in Harttia (Siluriformes: Loricariidae) species diversification

The Neotropical armored catfish genus Harttia presents a wide variation of chromosomal rearrangements among its representatives. Studies indicate that translocation and Robertsonian rearrangements have triggered the karyotype evolution in the genus, including differentiation of sex chromosome systems. However, few studies used powerful tools, such as comparative whole chromosome painting, to clarify this highly diversified scenario. Here, we isolated probes from the X₁ (a 5S rDNA carrier) and the X₂ (a 45S rDNA carrier) chromosomes of Harttia punctata, which displays an X₁X₁X₂X₂/X₁X₂Y multiple sex chromosome system. Those probes were applied in other Harttia species to evidence homeologous chromosome blocks. The resulting data reinforce that translocation events played a role in the origin of the X₁X₂Y sex chromosome system in H. punctata. The repositioning of homologous chromosomal blocks carrying rDNA sites among ten Harttia species has also been demonstrated. Anchored to phylogenetic data it was possible to evidence some events of the karyotype diversification of the studied species and to prove an independent origin for the two types of multiple sex chromosomes, XX/XY₁Y₂ and X₁X₁X₂X₂/X₁X₂Y, that occur in Harttia species. The results point to evolutionary breakpoint regions in the genomes within or adjacent to rDNA sites that were widely reused in Harttia chromosome remodeling.     

Spatial Structure of the Transmembrane Domain Heterodimer of ErbB1 and ErbB2 Receptor Tyrosine Kinases

Growth factor receptor tyrosine kinases of the ErbB family play a significant role in vital cellular processes and various cancers. During signal transduction across plasma membrane, ErbB receptors are involved in lateral homodimerization and heterodimerization with proper assembly of their extracellular single-span transmembrane (TM) and cytoplasmic domains. The ErbB1/ErbB2 heterodimer appears to be the strongest and most potent inducer of cellular transformation and mitogenic signaling compared to other ErbB homodimers and heterodimers. Spatial structure of the heterodimeric complex formed by TM domains of ErbB1 and ErbB2 receptors embedded into lipid bicelles was obtained by solution NMR. The ErbB1 and ErbB2 TM domains associate in a right-handed α-helical bundle through their N-terminal double GG4-like motif T⁶⁴⁸G⁶⁴⁹X₂G⁶⁵²A⁶⁵³ and glycine zipper motif T⁶⁵²X₃S⁶⁵⁶X₃G⁶⁶⁰, respectively. The described heterodimer conformation is believed to support the juxtamembrane and kinase domain configuration corresponding to the receptor active state. The capability for multiple polar interactions, along with hydrogen bonding between TM segments, correlates with the observed highest affinity of the ErbB1/ErbB2 heterodimer, implying an important contribution of the TM helix-helix interaction to signal transduction.     

End-to-End Association of <Emphasis Type="Italic">X</Emphasis> and <Emphasis Type="Italic">Y</Emphasis> Chromosomes in Mouse Meiosis

ACCORDING to the hypothesis of Crew and Koller¹ and Koller and Darlington², there are homologous segments in the X and Y chromosomes of the mouse and other mammals. The homologous regions in the mouse were believed to be localized in the extremely short arms proximal to the kinetochores. The end-to-end association at meiosis was thought to be the result of the formation of a chiasma between these homologous regions³. Electron microscopy revealed a short synaptonemal complex in mouse meiotic cells⁴. However, partial sex linkage has never been demonstrated in the mouse⁵ and other authors^6–10 believe that the X and Y chromosomes associate only by connexion between the chromosome ends furthest from the centromeres.     

A unique cytogenetic system in monotremes

All 3 extant genera of monotremes show a unique kind of cytogenetic system involving the formation of a structurally heterozygous chain multiple apparently coupled with a system of complementary gametic elimination. In the echidna Tachyglossus there are 63 chromosomes in the male and 64 in the female. This is associated with an X₁X₂Y/X₁X₁X₂X₂ sex chromosome system. Additionally in both sexes there are 6 mitotic chromosomes (a-f) which have no obvious homologous partners. At male meiosis these are included with the 3 sex chromosomes in a chain multiple of nine which has the constitution X₁·Y·X₂·f·e·d·c·b·a. This shows convergent orientation at first metaphase leading to the production of two kinds of sperm, namely X₁X₂ eca and Yfdb. Since no individual of either sex has been found homozygous for any of the a-f elements it follows that only gametes carrying different combinations of the three unpaired elements give rise to viable offspring. Whether this depends on gametic selection or on zygotic lethality is not known. An apparently identical system operates in Zaglossus. In the platypus Ornithorhynchus, on the other hand, there are 52 chromosomes in both males and females associated with an XY/XX sex chromosome mechanism and the presence of 4 consistently unpaired elements (a-d) at mitosis. A chain multiple of 10 forms at male meiosis involving the 2 sex chromosomes, the 4 unpaired elements and two of the small pairs of autosomes. Additionally the six longest autosome pairs in Tachyglossus and the X₁ show a polymorphism for size which in heterozygous combination leads to the formation of unequal bivalents at male meiosis.     

Cytogenetic characterization of ten araneomorph spiders (Araneae): Karyotypes and meiotic features

Karyotypes, sex chromosome systems and meiotic characteristics are reported for ten spider species belonging to the families Gnaphosidae, Philodromidae, Salticidae, Oxyopidae and Sicariidae by using standard Giemsa staining. The male diploid numbers (2n) and sex chromosome systems are as follows: Berinda hakani 2n = 22 (X₁X₂), Berinda ensigera 2n = 22 (X₁X₂), Trachyzelotes lyonneti 2n = 22 (X₁X₂), Trachyzelotes malkini 2n = 22 (X₁X₂), Zelotes caucasius 2n = 22 (X₁X₂) (Gnaphosidae); Thanatus pictus 2n = 28 (X₁ X₂), Tibellus macellus 2n = 24 (X₁ X₂) (Philodromidae); Neon reticulatus 2n = 21 (X₀) (Salticidae); Peucetia virescens 2n = 28 (X₁X₂) (Oxyopidae) and Loxosceles rufescens 2n = 21 (X₁ X₂Y) (Sicariidae). All species have monoarmed chromosomes with the exception of L. rufescens that has biarmed (metacentric and submetacentric) chromosomes. The obtained data are the first results for the genera Berinda, Trachyzelotes and Neon. Additionally, with the exception of L. rufescens, all species are being chromosomally analyzed for the first time.     

The Clinical Significance and Risk Factors of Solitary Lymph Node Metastasis in Gastric Cancer

Aims

To assess the clinical significance and risk factors of solitary lymph node metastasis (SLM) in gastric carcinoma and establish a more accurate method to evaluate the possibility of lymph node metastasis (LM).

Methods

A total of 385 patients with gastric carcinoma who underwent D2 lymphadenectomy at the Cancer Center of Sun Yat-Sen University were included in this research. Then we used a group of data from Sun Yat-sen University Gastrointestinal Hospital (SYSUGIH) to validate the accuracy of our developed method. The χ² test, Kaplan–Meier analysis, log-rank test, COX model, and discriminate analysis were used to analyze the data with SPSS13.0.

Results

We found that the LM number and pathological T staging were independent prognostic risk factors. CEA grading, LN status by CT, and T staging by CT were independent risk factors for LM in gastric carcinoma. In addition, we developed the equation Y = -5.0 + X ₁ + 1.8X ₃ + 0.7X ₄ (X ₁ = CEA grading, X ₃ = LN status by CT, X ₄ = T staging by CT) to evaluate the situation of LM. The data from SYSUGIH shows this equation has a better accuracy compared with CT.

Conclusions

SLM is an independent risk factor in gastric cancer. And there was no survival difference between the skip metastasis group and the other SLM group (P = 0.659). It is inappropriate for the patient with SLM doing a standard D2 lymphadenectomy, due to the fact that LM rarely occurs in the splenic artery, splenic hilum. The risk factors for LM include CEA grading, LN status by CT, and T staging by CT. And we can use Y = -5.0 + X ₁ + 1.8X ₃ + 0.7X ₄ (X ₁, CEA grading, X ₃ = LN status by CT, X ₄ = T staging by CT, the critical value is 0.3) to estimate the possibility of LM, which has a better accuracy compared with CT.     

High catalytic activities in the norbornene polymerization with neutral palladium complexes containing N4-type tetradentate chelating ligands

Norbornene polymerization catalyzed by new Pd(II) complexes bearing N4-type tetradentate ligands obtained from the reaction between a 6-methyl-2-picolinic acid or picolinic acid and appropriate diamines has been studied. A class of new palladium complexes, [Pd(X¹X²bpb)] and [Pd(X¹X²-6-Me₂bpb)] (X¹ = Me, X² = Me (1 and 4); X¹ = H, X² = H (2 and 5); X¹ = H, X² = NO₂ (3 and 6); bpb = N,N′-(o-phenylene)bis(pyridine-2-carboxamidate); 6-Me₂bpb = N,N′-(o-phenylene)bis(6-methylpyridine-2-carbox-amidate)) were synthesized and characterized. The molecular structure of Pd complex 5 was determined by X-ray crystallography, showing distorted square planar configurations. Using modified methylaluminoxanes (MMAO) as an activator, the palladium complexes exhibited high catalytic activities for the polymerization of norbornene. The catalytic activities up to 4.0 × 10⁶ g of PNBEs/mol_Pd·h and M_w up to 8.34 × 10⁵ g/mol with PDI < 2.53 were observed. Amorphous polynorbornenes (PNBEs) were obtained with good solubility in halogenated aromatic solvents. Interestingly, the structural modification with the methyl groups of pyridyl rings and the strong electron-withdrawing substituents induced improvement in solubility, thermal stability and catalytic activity. FT-IR, ¹H, and ¹³C NMR analyses of the polymers suggest that the catalytic polymerization occurs via vinyl addition mechanism.     

The Evolutionary History of DROSOPHILA BUZZATII. Xii. the Genetic Basis of Sterility in Hybrids between D. BUZZATII and Its Sibling D. SERIDO from Argentina

The genetic basis of hybrid sterility has been investigated in backcross segmental hybrids between two sibling species, Drosophila buzzatii and D. serido. Asynapsis of homologous bands in hybrid polytene chromosomes has been used to identify the D. serido chromosome segments introgressed into the D. buzzatti genome. All the investigated chromosomes contain male sterility factors. For autosomes, sterility is produced when an introgressed D. serido chromosome segment, or combination of segments, reaches a minimum size. On the other hand, any introgressed X chromosome segment from D. serido, irrespective of its size, produces either male hybrid sterility or inviability.