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Studies on caprine responsiveness to nematodiasis: segregation of male goats into responders and non-responders.

Eighty-three 2-4-year-old intact male goats exposed to a combination of artificial and natural challenge were segregated into responders and non-responders by ranking of weekly faecal egg counts (FECs). Retrospective analysis of samples over a 15-week-period showed responders had a statistically lower mean FEC than non-responders. Estimates of repeatability between consecutive egg counts were significant in both groups. The 6 top responders and bottom non-responders were subsequently given an artificial trickle challenge with Teladorsagia circumcincta and Trichostrongylus vitrinus. Mean faecal egg output was significantly lower in responders than non-responders. Peripheral eosinophil numbers following challenge were significantly greater in responder than non-responder goats. Abomasal and intestinal worm burdens were considerably lower in responders, with evidence of retardation of worm development compared to non-responders. Both abomasal and jejunal tissue eosinophil numbers were significantly higher in responders, although there was no difference in mucosal mast cell or globule leucocyte numbers. These results suggest that under temperate climatic conditions, it is possible to segregate male goats into responders and non-responders on the basis of simple parasitological criteria.

Abomasum↗

Structure of replicating simian virus 40 minichromosomes. The replication fork, core histone segregation and terminal structures.

The structure of replicating simian virus 40 (SV40) minichromosomes was studied by DNA crosslinking with trimethyl-psoralen. The procedure was used both in vitro with extracted SV40 minichromosomes as well as in vivo with SV40-infected cells. Both procedures gave essentially the same results. Mature SV40 minichromosomes are estimated to contain about 27 nucleosomes (error +/- 2), except for those molecules with a nucleosome-free gap, which are interpreted to contain 25 nucleosomes (error +/- 2). In replicative intermediates, nucleosomes are present in the unreplicated parental stem with the replication fork possibly penetrating into the nucleosomal DNA before the histone octamer is removed. Nucleosomes reassociate on the newly replicated DNA branches at distances from the branch point of 225 ( +/- 145) nucleotides on the leading strand and of 285( +/- 120) nucleotides on the lagging strand. In the presence of cycloheximide, daughter duplexes contained unequal numbers of nucleosomes, supporting dispersive and random segregation of parental nucleosomes. These were arranged in clusters with normal nucleosome spacing. We detected a novel type of interlocked dimer comprising two fully replicated molecules connected by a single-stranded DNA bridge. We cannot decide whether these dimers represent hemicatenanes or whether the two circles are joined by a Holliday-type structure. The joining site maps within the replication terminus. We propose that these dimers represent molecules engaged in strand segregation.

Cross-Linking Reagents↗

The mcm2 mutation of yeast affects replication, rather than segregation or amplification of the two micron plasmid.

We have studied the maintenance of the endogenous two micron (2 mu) plasmid in a strain of yeast carrying the nuclear mutation mcm2. This mutation, earlier shown to affect the maintenance of yeast minichromosomes in an ARS-dependent manner, also affected the copy number of the 2 mu plasmid. The effect was more pronounced at 35 degrees C leading to the elimination of the plasmid from the cells cultured at this temperature. The mutant cells could be efficiently cured of the circle by transformation with 2 mu ORI-carrying hybrid vectors, an observation consistent with the low copy number of the endogenous plasmid. A chromosomal revertant of this mutant for another ARS(ARS1) was found also to confer stability on the 2 mu ORI-carrying minichromosomes and had elevated levels of the endogenous plasmid. The mutation neither affected the segregation nor the amplification process mediated by site-specific recombination at FRT sites requiring the FLP gene-encoded protein action. ARS131C, an ARS that was unaffected in the mutant at 25 degrees C, could elevate the copy number of a 2 mu hybrid vector in the mutant cells. In view of these results, some aspects of segregation and copy number control of the endogeneous plasmid have been discussed. We propose that the mutation impairs the 2 mu ORI function, leading to its loss.

Cell Division↗

Mechanism of post-segregational killing by the hok/sok system of plasmid R1. Sok antisense RNA regulates hok gene expression indirectly through the overlapping mok gene.

The hok/sok locus of plasmid R1, which mediates plasmid stabilization by killing of plasmid-free segregants, codes for two RNAs, Hok mRNA and Sok antisense RNA. Hok mRNA encodes the Hok killer protein of 52 amino acid residues. Expression of hok is regulated post-transcriptionally by Sok antisense RNA. Killing of plasmid-free daughter-cells by the hok/sok system is accomplished through differential decay of the Hok and Sok-RNAs: Hok mRNA is very stable while Sok-RNA decays rapidly, thus leading to derepression of Hok mRNA translation in plasmid-free segregants, ensuring a rapid and selective killing of these cells. Sok antisense RNA is complementary to the leader region of the Hok mRNA. However, the region of complementarity does not overlap with the hok Shine-Dalgarno sequence. Thus, Sok-RNA regulates hok translation indirectly by an as yet unknown mechanism. We show here that Sok antisense RNA regulates the translation of another reading frame located in the hok/sok locus. This new reading frame, which overlaps with almost the entire hok gene, was denoted mok (mediation of killing). Point-mutations that prevent mok translation through the hok translational initiation region abolish efficient expression of hok. Furthermore, these mutations abolish the Sok-RNA-mediated control of hok gene expression. Hence, the antisense-RNA-mediated regulation of the hok gene seems to occur via translational coupling between the hok and mok reading-frames.

Base Sequence↗

Including the environment in models for genetic segregation.

Current models for linkage and segregation analysis do not allow for many of the different ways in which the environment may affect risk for a psychiatric disorder. A variety of mechanisms for the contribution of the environment and the correlation and interaction of environment effects with genetic differences are described. The approach to including these effects in models for segregation and linkage is outlined.

Genotype↗

The effect of exposure fractionation on the yield of nondisjunctional segregation involving compound X and free 4-chromosomes in Drosophila.

This paper describes the effect of fractionation of an exposure of 2000-R X-rays into two 1000-R exposures separated by a 3-h interval on the frequency of various segregation products following chromatoid interchange between a compound X and the 4th chromosome. It was observed that fractionation leads to a decrease of all exceptional progeny, the result being significant at the 0.05-probability level for both classes of detachments; i.e., with or without a 4th chromosome. The finding that the same effect of fractionation was observed for numerical aberration (non-disjunction) and detachment supports the idea that these reflect different consequences of the same primary effect, radiation-induced rearrangement. Furthermore, alternative recoveries were observed to occur at equal frequencies. Following C(1)-4 interchange, the univalent 4 does not segregate at random, but assorts apart from its homologue 1.5 as frequently as it moves to the same pole.

Animals↗

Amplification of the provirus region in Rous sarcoma virus-transformed Chinese hamster cells and segregation of the amplified copies in somatic cell hybrids.

Four independent clones of RSV-transformed Chinese hamster fibroblasts were isolated. Southern blots and dot hybridization studies showed that in three out of four clones there were four to eight times as many integrated proviruses as in the fourth clone which contained at least one complete provirus. Restriction mapping studies showed that although the integration site varied from clone to clone, all the proviral copies in the same clone shared the same flanking cellular sequences. In one clone there are at least two polymorphic proviral variants A and B, one with and one without a BglI site. Further experiments were performed to see if the variants could be physically separated. RSV-Transformed Chinese hamster cells resistant to thioguanine were fused with mouse cells to give somatic hybrids which preferentially segregate Chinese hamster chromosomes. Ten out of eleven hybrids positive for virus rescue have lost up to 90% of the parental provirus copies. Four of these hybrids were found to contain the provirus variant A alone, one the variant B alone, and the rest contained both variants. All the proviruses retained in somatic hybrids shared the flanking cellular sequences of the parental provirus. Provirus segregation in somatic hybrids confirms that multiple (about ten) copies of the provirus region are present in the karyotype of parental RSV-transformed cells and, furthermore, suggests that the amplified copies of this region are translocated to different chromosomes.

Animals↗

Segregation of cauliflower mosaic virus symptom genetic determinants.

We have created a series of hybrid cauliflower mosaic virus (CaMV) genomes between a severe virus strain (Cabb BJI) and a mild strain (Bari 1) to map the virus genetic loci responsible for specific systemic symptom characters produced in infected turnip plants. Recombinants were generated in vivo by recombinational rescue and in vitro by restriction enzyme fragment exchange. On infection, hybrids induced either parental (wild-type) symptoms or segregated parental characters. Some of the engineered hybrid genomes produced novel symptomatic effects not observed in either of the parental strains whilst others reverted to express parental symptom characters following passaging. Determinants defining differences between the two CaMV strains in respect of four specific symptom characters were delimited to separate genome regions. A locus involved in determining the rate of spread of systemic vein clearing symptoms mapped to a region containing part of gene VII and gene I (nts 109-780). This phenomenon is consistent with the putative involvement of the CaMV gene I product in mediating virus movement within infected plants. Determinants influencing the degree of leaf chlorosis were located in a separate genome domain encompassing part of gene VI together with the large intergenic region and part of gene VII (nts 6103-90). Determinants controlling timing of initial systemic symptom appearance were mapped to a region between nts 2150 and 4438 containing part of gene III, gene IV, and part of gene V. Plant stunting was influenced by loci in at least two separate regions, one containing parts of gene I and II, and a second within the reverse transcriptase gene (V). We conclude that symptoms produced by CaMV infection can be subdivided into individual characters, the genetic determinants of which segregate to different virus genetic loci and are not restricted to a single gene product.

Brassica↗

Texture segregation based on two-dimensional relative phase differences in composite sine-wave grating patterns.

Experiments examined the visual processing of relative phase relations between differently oriented components of a textured pattern. The textures were a super position of three 1.3 c/deg sine-wave gratings rotated 60 deg relative to one another. Global orientation and relative phase were varied. Subjects rated the segregation of pairs of textures presented in a figure/ground configuration at 10 deg retinal eccentricity. Both orientation and phase differences were used in making ratings. It was not simply the difference in relative phase that mattered. The results also depended on the values of the relative two-dimensional (2-D) phases in figure and ground. Mirror-image texture pairs (which may have a large relative phase difference) segregated poorly compared to other pairs with similar phase differences. This suggests that the peripheral visual system does not completely encode 2-D relative phase. Secondary issues include spatial frequency effects, the relevance of figure/ground borders, and better performance in the lower visual field.

Contrast Sensitivity↗

The role of the background: texture segregation and figure-ground segmentation.

The effects of a texture surround composed of line elements on a stimulus within which a target line element segregates, were studied. Detection and discrimination of the target when it had the same orientation as the surround were impaired at short presentation time; on the other hand, no effect was present when they were reciprocally orthogonal. These results are interpreted as background completion in texture segregation; a texture made up of similar elements is represented as a continuous surface with contour and contrast of an embedded element inhibited. This interpretation is further confirmed with a simple line protruding from an annulus. Generally, the results are taken as evidence that local features are prevented from segmenting when they are parts of a global entity.

Discrimination, Psychological↗

The effect of local cortical microfilament disorganization on ooplasmic segregation in the loach (Misgurnus fossilis) egg.

Injections of cytochalasin D (CD) or DNase I under the surface of fertilized loach egg result in local disorganization of microfilamentous cortex (MC) as revealed by transmission electron microscopy. This effect correlates with the loss of the cortex ability to contract in vitro. The disorganization of MC in the vegetal hemisphere of the egg does not affect the ooplasm segregation or blastodisk cleavage. Injection under the animal pole suppresses blastodisk formation and results in the autonomous separation of ooplasm in the central part of the egg. The experiments suggest that (1) autonomous separation of ooplasm from the yolk granules can proceed in the central part of the egg without the participation of MC; (2) normal segregation of ooplasm at the animal pole requires that the structures of microfilaments in the animal hemisphere (but not in the vegetal one) be preserved.

Actin Cytoskeleton↗

Membrane-induced segregation of motile and stable domains in cytoplasm: possible role in morphological differentiation of tissue cells.

Many morphological differentiations and modulations involve alterations of degree of segregation in two types of cytoplasmic domains: lamellar-pseudopod-forming motile structures, actinoplasts, and non-motile cylindrical processes, tubuloplasts. Pronounced reversible segregation of actinoplasts and tubuloplasts can be induced in certain fibroblastic cells by phorbol ester. Analysis of this model reorganization suggests that formation of tubuloplasts from actinoplasts may involve local collapse of actin network and microtubule-directed translocation of actin material from collapsing zone; these cytoskeletal alterations may be controlled by membrane-activated regulatory enzymes.

Actins↗

Conservative assembly and segregation of nucleosomal histones.

The assembly of new histones into nucleosomes and the segregation of old histones during replication were investigated using a density gradient, sedimentation equilibrium analysis of histones labeled in vivo with dense amino acids. After a 1 hr pulse of dense amino acids and 3H-lysine, nucleosomes were isolated from chick myoblast organ cultures, and the histones were cross-linked to octamers. The octamers were purified from DNA and then banded to equilibrium in cesium-formate guanidinium-HCI density gradients. The cross-linked dense octamers have the same density as the noncross-linked dense histones, and both were significantly heavier than histones synthesized in the presence of light amino acids. This experiment shows that new histone does not mix with old histone in the new nucleosomes, since the labeling protocol allows density labeling of only one histone for every seven preexisting unlabeled histones. Thus the assembly of new histone octamers is conservative. Using essentially the same experimental design, but varying the details of the labeling procedures, we also show that the dense histone octamer is stable over 3-4 generations, that neighboring octamers tend to be synthesized at the same time, and that old and new histone octamers segregate conservatively over 2-3 generations.

Animals↗

Histone H2B repression causes cell-cycle-specific arrest in yeast: effects on chromosomal segregation, replication, and transcription.

To determine which cellular processes are dependent on histones, we blocked histone H2B mRNA synthesis in asynchronously growing yeast after fusing the H2B gene to a repressible GAL10 promoter. Chromosomal segregation, replication, and transcription were then examined. We found that the cells arrested in mitosis, with a cell division cycle (cdc) phenotype. Chromatin structure and nuclear segregation were disrupted. A full round of DNA replication took place after the repression of histone H2B mRNA synthesis. Active transcription and the induction of new transcripts also continued in the arrested cells.

Cell Cycle↗

Parental DNA strands segregate randomly during embryonic development of Caenorhabditis elegans.

The fate of gamete DNA was followed in the next generation embryos of the nematode C. elegans. Either male worms or spermless hermaphrodites were grown on bromodeoxyuridine-containing E. coli in order to label germ-line DNA. Matings then produced embryos in which only the DNA strands provided by the gametes contained label. This original gamete DNA could be detected during embryonic development by using a fluorescently labeled monoclonal antibody specific to bromodeoxyuridine. Both the number and position of fluorescent spots in the embryo indicate that gamete DNA strands segregate randomly during development. Random segregation of parental DNA strands rules out models of development that invoke chromosome imprinting or immortal DNA strands.

Animals↗

The expression of three members of the achaete-scute gene complex correlates with neuroblast segregation in Drosophila.

The expression of a subset of homologous genes of the AS-C is required during embryogenesis and metamorphosis for proper neural development. Here we study the expression of three of these genes (T3, T4, and T5) and show that their transcripts accumulate at the blastoderm stage in periodic patterns coincident with the dorsoventral extent of the neuroectoderm. Subsequent expression is in partially overlapping patterns that correlate with the segregation of neuroblasts. These genes are not transcribed in neurons. We also show that a deficiency for the AS-C prevents the appearance of at least one identified class of neuroblasts. We propose that the AS-C is part of a network of genes responsible for the segregation of neuroblasts.

Animals↗

Segregation of recombined chromosomes in meiosis I requires DNA topoisomerase II.

To understand better the similarities and differences between meiosis and mitosis, we examined the meiotic role of DNA topoisomerase II, an enzyme that is required mitotically to disentangle sister chromatids at the time of chromosome segregation. In meiosis, we found that topoisomerase II is required only at the time of nuclear division. When cold-sensitive top2 mutants are induced to sporulate at the restrictive temperature, they undergo premeiotic DNA synthesis and commitment to meiotic levels of recombination but fail to complete the first meiotic nuclear division. The introduction of a mutation blocking recombination relieves the requirement for topoisomerase II in meiosis I. These results suggest that topoisomerase II is required at the time of chromosome segregation in meiosis I for the resolution of recombined chromosomes.

Cell Nucleus↗

A kinesin-like protein required for distributive chromosome segregation in Drosophila.

The nod gene is required for the distributive segregation of nonexchange chromosomes during meiosis in D. melanogaster. Loss-of-function nod mutations cause nondisjunction and loss of nonrecombinant chromosomes both at meiosis I and during subsequent mitotic divisions. We have cloned the nod locus, examined its expression patterns, and determined its coding sequence. In adults the nod transcript is only present in females, consistent with the observation that males do not use the distributive segregation system. However, the nod locus is also transcribed in the embryonic, larval, and pupal stages of development, and possibly in all dividing cells. Finally, the N-terminal domain of the predicted nod protein has amino acid similarity to the mechanochemical domain of kinesin heavy chain; however, the C-terminal domain is unlike that of kinesin heavy chain or of any previously reported protein. Thus, the nod protein is a member of the kinesin superfamily and may be a microtubule motor.

Adenosine Triphosphatases↗