EVOLUTION, Issue 3 2006
Charles D. Criscione
Abstract Little is known about actual mating systems in natural populations of parasites or about what constitutes the limits of a parasite deme. These parameters are interesting because they affect levels of genetic diversity, opportunities for local adaptation, and other evolutionary processes. We expect that transmission dynamics and the distribution of parasites among hosts should have a large effect on mating systems and demic structure, but currently we have mostly speculation and very few data. For example, infrapopulations (all the parasites in a single host) should behave as demes if parasite offspring are transmitted as a clump from host to host over several generations. However, if offspring are well mixed, then the parasite component population (all the parasites among a host population) would function as the deme. Similarly, low mean intensities or a high proportion of worms in single infections should increase the selfing rate. For species having an asexual amplification stage, transmission between intermediate and definitive (final) hosts will control the variance in clonal reproductive success, which in turn could have a large influence on effective sizes and rates of inbreeding. We examined demic structure, selfing rates, and the variance in clonal reproductive success in natural populations of Plagioporus shawi, a hermaphroditic trematode that parasitizes salmon. Overall levels of genetic diversity were very high. An a posteriori inference of population structure overwhelmingly supports the component population as the deme, rather than individual infrapopulations. Only a single pair of 597 adult individuals was identified as clones. Thus, the variance in clonal reproductive success was almost zero. Despite being hermaphroditic, P. shawi appears to be almost entirely outcrossing. Genetic estimates of selfing (<5%) were in accordance with the proportion of parasites from single infections. Thus, it appears that individual flukes outcross whenever possible and only resort to selfing when alone. Finally, our data support the hypothesis that aquatic transmission and the use of several intermediate hosts promotes high genetic diversity and well-mixed infrapopulations. [source]

REGULAR(IZED) HEDGE FUND CLONES

THE JOURNAL OF FINANCIAL RESEARCH, Issue 3 2010
Daniel Giamouridis
Abstract This article addresses the problem of portfolio construction in the context of efficient hedge fund investments replication. We propose a modification to the standard Sharpe "style analysis" by introducing a constraint on the asset weights 1-norm and 2-norm. This constraint regularizes the optimization problem, allows efficient selection of relevant factor's and has significant effects on the stability of the resulting asset mix and the risk,return characteristics of the replicating portfolio. The empirical results suggest that the norm-constrained replicating portfolios exhibit significant correlations with their benchmarks, often higher than 0.9; have a fraction, which is about half to two-thirds, of active positions relative to those determined through the standard method; and are obtained with turnover, which is in some instances about one-fourth of that for the standard method. [source]

Post-gall induction performance of Adelges Abietis (L.) (Homoptera: Adelgidae) is influenced by clone, shoot length, and density of colonising gallicolae

ECOLOGICAL ENTOMOLOGY, Issue 1 2010
LEAH FLAHERTY
1. We evaluated the effect of clone (one susceptible and one resistant clone), shoot length, crown level, and gallicola density on post-gall induction performance of Adelges abietis. Galls that had been successfully induced by one fundatrix on a range of shoot sizes were selected, and the number of gallicolae that could colonise the gall was manipulated. 2. Post-induction gall development success was inversely related to shoot length and was higher on the susceptible clone than on the resistant clone. As gallicola density did not influence the proportion of galls that successfully completed development, reduced post-induction gall development on large shoots was not likely to be result of an insufficient stimulus from gallicolae. 3. Clone was the only factor that significantly influenced gall volume and galls were larger on the susceptible clone than on the resistant clone. As gall volume did not increase when more gallicolae attempted to colonise a gall, competition within a gall increased. Gallicola survival was inversely related to the number of colonising gallicolae. Our results suggest that gall size may be limiting at natural densities. 4. Previous studies report positive relationships between gall induction success and fundatrix density, and between gall size and fundatrix density. As each fundatrix produces one egg mass of gallicolae, this study suggests that there may be a trade-off between the successful induction of a large gall and subsequent survival of gallicolae. 5. In the present study, clone influenced all measures of post-gall induction performance. Performance was always higher on the susceptible than on the resistant clone. [source]

Vitamin D Receptor Expression in Human Muscle Tissue Decreases With Age,

JOURNAL OF BONE AND MINERAL RESEARCH, Issue 2 2004
HA Bischoff-Ferrari
Abstract Intracellular 1,25-dihydroxyvitamin D receptor (VDR) is expressed in human skeletal muscle tissue. However, it is unknown whether VDR expression in vivo is related to age or vitamin D status, or whether VDR expression differs between skeletal muscle groups. Introduction: We investigated these factors and their relation to 1,25-dihydroxyvitamin D receptor (VDR) expression in freshly removed human muscle tissue. Materials and Methods: We investigated biopsy specimens of the gluteus medius taken at surgery from 20 female patients undergoing total hip arthroplasty (mean age, 71.6 ± 14.5; 72% > 65 years) and biopsy specimens of the transversospinalis muscle taken at surgery from 12 female patients with spinal operations (mean age, 55.2 ± 19.6; 28% > 65 years). The specimens were obtained by immunohistological staining of the VDR using a monoclonal rat antibody to the VDR (Clone no. 9A7). Quantitative VDR expression (number of VDR positive nuclei) was assessed by counting 500 nuclei per specimen and person. Serum concentrations of 25-hydroxyvitamin D and 1,25-dihydroxyvitamin D were assessed at day of admission to surgery. Results: All muscle biopsy specimens stained positive for VDR. In the univariate analyses, increased age was associated with decreased VDR expression (r = 0.5: p = 0.004), whereas there were no significant correlations between VDR expression and 25-hydroxyvitamin D or 1,25-dihydroxyvitamin D levels. VDR expression did not differ between patients with hip and spinal surgery. In the multivariate analysis, older age was a significant predictor of decreased VDR expression after controlling biopsy location (gluteus medius or the transversospinalis muscle), and 25-hydroxyvitamin D levels (linear regression analysis: ,-estimate = ,2.56; p = 0.047). Conclusions: Intranuclear immunostaining of the VDR was present in muscle biopsy specimens of all orthopedic patients. Older age was significantly associated with decreased VDR expression, independent of biopsy location and serum 25-hydroxyvitamin D levels. [source]

Genome Organization of an Infectious Clone of Tomato Leaf Curl Virus (Philippines), a New Monopartite Begomovirus*

JOURNAL OF PHYTOPATHOLOGY, Issue 11-12 2002
Tatsuya Kon
Abstract Complete nucleotide sequence of infectious cloned DNA of Tomato leaf curl virus from Philippines (ToLCV-Ph) was determined. The single circular DNA molecule comprises 2755 nucleotides. ToLCV-Ph DNA contains six open reading frames (ORFs) each capable of encoding proteins with a molecular weight greater than 10 kDa. A partial dimeric ToLCV-Ph DNA clone was constructed in a binary vector and used to agroinoculate tomato plants (Lycopersicon esculentum Mill. cv. Zuikou 102). Typical leaf curl symptoms were observed, showing that the single DNA component is sufficient for infectivity. In total nucleotide sequence comparisons with other geminiviruses, ToLCV-Ph was most closely related to Ageratum yellow vein virus (AYVV) (79% identity), ToLCV-Laos (78%), Soyabean crinkle leaf virus -Thailand (78%) and ToLCV-Taiwan (77%). The significant but relatively low sequence identity in the genomic DNA between ToLCV-Ph and other geminiviruses suggests that it is a distinct geminivirus in the genus Begomovirus. [source]

Dealing with Saddam or His Clone

MIDDLE EAST POLICY, Issue 4 2000
Ghassan Atiyyah
[source]

Reciprocal Relationship Between a Ph-Negative Clone With Trisomy 8 Associated With Severe Myelodysplasia and a Ph-Positive Clone Following Imatinib Treatment in a Patient With Accelerated-Phase Chronic Myelogenous Leukemia (CML)

AMERICAN JOURNAL OF HEMATOLOGY, Issue 4 2004
Paulina Patchenko
No abstract is available for this article. [source]

Differential gene expression in senescing leaves of two silver birch genotypes in response to elevated CO2 and tropospheric ozone

PLANT CELL & ENVIRONMENT, Issue 6 2010
SARI KONTUNEN-SOPPELA
ABSTRACT Long-term effects of elevated CO2 and O3 concentrations on gene expression in silver birch (Betula pendula Roth) leaves were studied during the end of the growing season. Two birch genotypes, clones 4 and 80, with different ozone growth responses, were exposed to 2× ambient CO2 and/or O3 in open-top chambers (OTCs). Microarray analyses were performed after 2 years of exposure, and the transcriptional profiles were compared to key physiological characteristics during leaf senescence. There were genotypic differences in the responses to CO2 and O3. Clone 80 exhibited greater transcriptional response and capacity to alter metabolism, resulting in better stress tolerance. The gene expression patterns of birch leaves indicated contrasting responses of senescence-related genes to elevated CO2 and O3. Elevated CO2 delayed leaf senescence and reduced associated transcriptional changes, whereas elevated O3 advanced leaf senescence because of increased oxidative stress. The combined treatment demonstrated that elevated CO2 only temporarily alleviated the negative effects of O3. Gene expression data alone were insufficient to explain the O3 response in birch, and additional physiological and biochemical data were required to understand the true O3 sensitivity of these clones. [source]

The motility of glioblastoma tumour cells is modulated by intracellular cofilin expression in a concentration-dependent manner

CYTOSKELETON, Issue 3 2005
Celestial T. Yap
Abstract The invasive behaviour of tumour cells has been attributed in part to dysregulated cell motility. Members of the ADF/Cofilin family of actin-binding proteins are known to increase microfilament dynamics by increasing the rate at which actin monomers leave the pointed end of the filament and by a filament-severing activity. As depolymerisation is a rate-limiting step in actin dynamics, ADF/Cofilins are suspected to facilitate the motility of cells. To test this, we investigated the influence of cofilin on tumour motility by transient and stably overexpressing cofilin in the human glioblastoma cell line, U373 MG. Several different methods were used to ascertain the level of cofilin in overexpressing clones and this was correlated with their rate of random locomotion. A biphasic relationship between cofilin level and locomotory rate was found. Clones that displayed a moderate amount of overproduction of cofilin were found to have increased rates of locomotion approximately linear to the overproduction of cofilin up to an optimal cofilin level of about 4.5 times that of wild type cells at which the cells were almost twice as fast. However, clones producing more than this optimal amount were found to locomote at progressively reduced speeds. Cells that overexpress cofilin have reduced stress fibres compared to control cells showing that the excess cofilin affects the actin cytoskeleton. We conclude that overexpression of cofilin enhances the motility of glioblastoma tumour cells in a concentration-dependent fashion, which is likely to contribute to their invasiveness. Cell Motil. Cytoskeleton 60:153,165, 2005. © 2005 Wiley-Liss, Inc. [source]

Sorting nexin-14, a gene expressed in motoneurons trapped by an in vitro preselection method

DEVELOPMENTAL DYNAMICS, Issue 4 2001
Patrick Carroll
Abstract A gene-trap strategy was set up in embryonic stem (ES) cells with the aim of trapping genes expressed in restricted neuronal lineages. The vector used trap genes irrespective of their activity in undifferentiated totipotent ES cells. Clones were subjected individually to differentiation in a system in which ES cells differentiated into neurons. Two ES clones in which the trapped gene was expressed in ES-derived neurons were studied in detail. The corresponding cDNAs were cloned, sequenced, and analysed by in situ hybridisation on wild-type embryo sections. Both genes are expressed in the nervous system. One gene, YR-23, encodes a large intracellular protein of unknown function. The second clone, YR-14, represents a sorting nexin (SNX14) gene whose expression in vivo coincides with that of LIM-homeodomain Islet-1 in several tissues. Sorting nexins are proteins associated with the endoplasmic reticulum (ER) and may play a role in receptor trafficking. Gene trapping followed by screening based on in vitro preselection of differentiated ES recombinant clones, therefore, has the potential to identify integration events in subsets of genes before generation of mouse mutants. © 2001 Wiley-Liss, Inc. [source]

Multispecific responses by T cells expanded by endogenous self-peptide/MHC complexes

EUROPEAN JOURNAL OF IMMUNOLOGY, Issue 3 2007
Guifang Cai
Abstract The paradox of autoreactivity to self-peptides in physiological as opposed to pathological immune responses is not well understood. Here, we directly examined the human T cell response to endogenous self-peptides in a series of healthy subjects. CFSE-labeled T cells were stimulated with unmanipulated antigen-presenting cells containing endogenous self-antigen, and the resulting CD4+ populations entering into cell cycle (CFSElow) or non-proliferating CD4+ cells (CFSEhigh) were single-cell sorted, cloned and screened against a panel of self-antigens and microbial recall antigens to interrogate their antigen reactivity. The percentage of CD4+ T cells entering cell cycle in response to self-peptide/MHC was calculated to be 0.04%, and entry into cell cycle was dependent upon CD28 costimulation. Clones derived from CFSElow T cells exhibited significantly greater cross-reactivity to multiple antigens than CFSEhigh clones or other CD4+ clones generated after microbial antigen stimulation. Sequencing the TCR, chains indicated that CFSElow clones were indeed clonal. These data demonstrate that T cell clones generated on stimulation by endogenous self-peptides exhibit a high degree of multispecificity, and we speculate that their multispecificity is based upon recognition of shared-backbone MHC determinants. [source]

THREE-GENE IDENTITY COEFFICIENTS DEMONSTRATE THAT CLONAL REPRODUCTION PROMOTES INBREEDING AND SPATIAL RELATEDNESS IN YELLOW-CEDAR, CALLITROPSIS NOOTKATENSIS

EVOLUTION, Issue 10 2008
Stacey Lee Thompson
Asexual reproduction has the potential to promote population structuring through matings between clones as well as through limited dispersal of related progeny. Here we present an application of three-gene identity coefficients that tests whether clonal reproduction promotes inbreeding and spatial relatedness within populations. With this method, the first two genes are sampled to estimate pairwise relatedness or inbreeding, whereas the third gene is sampled from either a clone or a sexually derived individual. If three-gene coefficients are significantly greater for clones than nonclones, then clonality contributes excessively to genetic structure. First, we describe an estimator of three-gene identity and briefly evaluate its properties. We then use this estimator to test the effect of clonality on the genetic structure within populations of yellow-cedar (Callitropsis nootkatensis) using a molecular marker survey. Five microsatellite loci were genotyped for 485 trees sampled from nine populations. Our three-gene analyses show that clonal ramets promote inbreeding and spatial structure in most populations. Among-population correlations between clonal extent and genetic structure generally support these trends, yet with less statistical significance. Clones appear to contribute to genetic structure through the limited dispersal of offspring from replicated ramets of the same clonal genet, whereas this structure is likely maintained by mating among these relatives. [source]

Revelationary biology: A review of The Second Tree: Stem Cells, Clones, Chimeras, and Quests for Immortality, by Elaine Dewar

EVOLUTION AND DEVELOPMENT, Issue 5 2005
Yolanda P. Cruz
First page of article [source]

Reduced FAS transcription in clones of U937 cells that have acquired resistance to Fas-induced apoptosis

FEBS JOURNAL, Issue 2 2009
Jeanette Blomberg
Susceptibility to cell death is a prerequisite for the elimination of tumour cells by cytotoxic immune cells, chemotherapy or irradiation. Activation of the death receptor Fas is critical for the regulation of immune cell homeostasis and efficient killing of tumour cells by apoptosis. To define the molecular changes that occur during selection for insensitivity to Fas-induced apoptosis, a resistant variant of the U937 cell line was established. Individual resistant clones were isolated and characterized. The most frequently observed defect in the resistant cells was reduced Fas expression, which correlated with decreased FAS transcription. Clones with such reduced Fas expression also displayed partial cross-resistance to tumour necrosis factor-, stimulation, but the mRNA expression of tumour necrosis factor receptors was not decreased. Reintroduction of Fas conferred susceptibility to Fas but not to tumour necrosis factor-, stimulation, suggesting that several alterations could be present in the clones. The reduced Fas expression could not be explained by mutations in the FAS coding sequence or promoter region, or by silencing through methylations. Protein kinase B and extracellular signal-regulated kinase, components of signalling pathways downstream of Ras, were shown to be activated in some of the resistant clones, but none of the three RAS genes was mutated, and experiments using chemical inhibitors could not establish that the activation of these proteins was the cause of Fas resistance as described in other systems. Taken together, the data illustrate that Fas resistance can be caused by reduced Fas expression, which is a result of an unidentified mode of regulation. [source]

The C-terminal region of CHD3/ZFH interacts with the CIDD region of the Ets transcription factor ERM and represses transcription of the human presenilin 1 gene

FEBS JOURNAL, Issue 6 2007
Martine Pastorcic
Presenilins are required for the function of ,-secretase: a multiprotein complex implicated in the development of Alzheimer's disease (AD). We analyzed expression of the presenilin 1 (PS1) gene. We show that ERM recognizes avian erythroblastosis virus E26 oncogene homolog (Ets) motifs on the PS1 promoter located at ,10, +90, +129 and +165, and activates PS1 transcription with promoter fragments containing or not the ,10 Ets site. Using yeast two-hybrid selection we identified interactions between the chromatin remodeling factor CHD3/ZFH and the C-terminal 415 amino acids of ERM used as bait. Clones contained the C-terminal region of CHD3 starting from amino acid 1676. This C-terminal fragment (amino acids 1676,2000) repressed transcription of the PS1 gene in transfection assays and PS1 protein expression from the endogenous gene in SH-SY5Y cells. In cells transfected with both CHD3 and ERM, activation of PS1 transcription by ERM was eliminated with increasing levels of CHD3. Progressive N-terminal deletions of CHD3 fragment (amino acids 1676,2000) indicated that sequences crucial for repression of PS1 and interactions with ERM in yeast two-hybrid assays are located between amino acids 1862 and 1877. This was correlated by the effect of progressive C-terminal deletions of CHD3, which indicated that sequences required for repression of PS1 lie between amino acids 1955 and 1877. Similarly, deletion to amino acid 1889 eliminated binding in yeast two-hybrid assays. Testing various shorter fragments of ERM as bait indicated that the region essential for binding CHD3/ZFH is within the amino acid region 96,349, which contains the central inhibitory DNA-binding domain (CIDD) of ERM. N-Terminal deletions of ERM showed that residues between amino acids 200 and 343 are required for binding to CHD3 (1676,2000) and C-terminal deletions of ERM indicated that amino acids 279,299 are also required. Furthermore, data from chromatin immunoprecipitation (ChIP) indicate that CHD3/ZFH interacts with the PS1 promoter in vivo. [source]

Secretion of proteases in serglycin transfected Madin,Darby canine kidney cells

FEBS JOURNAL, Issue 3 2006
Lillian Zernichow
Madin,Darby canine kidney (MDCK) cells, which do not normally express the proteoglycan (PG) serglycin, were stably transfected with cDNA for human serglycin fused to a polyhistidine tag (His-tag). Clones with different levels of serglycin mRNA expression were generated. One clone with lower and one with higher serglycin mRNA expression were selected for this study. 35S-labelled serglycin in cell fractions and conditioned media was isolated using HisTrap affinity chromatography. Serglycin could also be detected in conditioned media using western blotting. To investigate the possible importance of serglycin linked to protease secretion, enzyme activities using chromogenic substrates and zymography were measured in cell fractions and serum-free conditioned media of the different clones. Cells were cultured in both the absence and presence of phorbol 12-myristate 13-acetate (PMA). In general, enzyme secretion was strongly enhanced by treatment with PMA. Our analyses revealed that the clone with the highest serglycin mRNA expression, level of HisTrap isolated 35S-labelled serglycin, and amount of serglycin core protein as detected by western blotting, also showed the highest secretion of proteases. Transfection of serglycin into MDCK cells clearly leads to changes in secretion levels of secreted endogenous proteases, and could provide further insight into the biosynthesis and secretion of serglycin and potential partner molecules. [source]

BJ46a, a snake venom metalloproteinase inhibitor

FEBS JOURNAL, Issue 10 2001
Isolation, characterization, cloning, insights into its mechanism of action
Fractionation of the serum of the venomous snake Bothrops jararaca with (NH4)2SO4, followed by phenyl-Sepharose and C4 -reversed phase chromatographies, resulted in the isolation of the anti-hemorrhagic factor BJ46a. BJ46a is a potent inhibitor of the SVMPs atrolysin C (class P-I) and jararhagin (P-III) proteolytic activities and B. jararaca venom hemorrhagic activity. The single-chain, acidic (pI 4.55) glycoprotein has a molecular mass of 46 101 atomic mass units determined by MALDI-TOF MS and 79 kDa by gel filtration and dynamic laser light scattering, suggesting a homodimeric structure. mRNA was isolated from the liver of one specimen and transcribed into cDNA. The cDNA pool was amplified by PCR, cloned into a specific vector and used to transform competent cells. Clones containing the complete coding sequence for BJ46a were isolated. The deduced protein sequence was in complete agreement with peptide sequences obtained by Edman degradation. BJ46a is a 322-amino-acid protein containing four putative N-glycosylation sites. It is homologous to the proteinase inhibitor HSF (member of the fetuin family, cystatin superfamily) isolated from the serum of the snake Trimeresurus flavoviridis, having 85% sequence identity. This is the first report of a complete cDNA sequence for an endogenous inhibitor of snake venom metalloproteinases (SVMPs). The sequence reveals that the only proteolytic processing required to obtain the mature protein is the cleavage of the signal peptide. Gel filtration analyses of the inhibitory complexes indicate that inhibition occurs by formation of a noncovalent complex between BJ46a and the proteinases at their metalloproteinase domains. Furthermore, the data shows that the stoichiometry involved in this interaction is of one inhibitor monomer to two enzyme molecules, suggesting an interesting mechanism of metalloproteinase inhibition. [source]

Absence of residual effects of a defeated resistance gene in poplar

FOREST PATHOLOGY, Issue 2 2003
K.-S. Woo
Summary In a few plant pathosystems, defeated major genes have been shown to contribute to partial resistance to disease. This hypothesis has never been tested before in a forest tree, but pathogenic variation associated with recent hybridization in poplar rust in the Pacific northwest provided an opportunity. An F2 progeny of 256 poplar clones in the field near Corvallis, Oregon, USA, has been monitored for rust severity and infection type since the advent of the new hybrid rust, Melampsora × columbiana, in the mid-1990s. All 256 clones displayed a susceptible infection type in 1997 and again in 2000, and yet variation in uredinial density (i.e. partial resistance) was still observed. To determine which clones possessed a defeated resistance gene, a greenhouse inoculation was performed with an isolate of M. medusae, one of the parents of M. × columbiana. Clones that would have been resistant to M. medusae, prior to the advent of M. × columbiana, were thus identified. The inoculation resulted in a 1 : 1 segregation (,2=0.772; p=0.38) for resistance, indicating the presence of a major gene. However, the F2 clones possessing the defeated resistance gene displayed the same level of partial resistance in the field in both 1997 and 2000 as their full siblings lacking the gene. Résumé Chez quelques pathosystèmes végétaux, il a été montré que le contournement de gènes majeurs de résistance contribue à une résistance partielle envers la maladie. Cette hypothèse n'a encore jamais été testée chez un arbre forestier, mais le changement de pouvoir pathogène associéà l'hybridation récente de la rouille du peuplier dans le nord-ouest des USA en a fourni l'occasion. Une descendance F2 de 256 clones de peuplier a été suivie au champ près de Corvallis, Oregon, USA, pour la gravité de la rouille et le type d'infection, depuis l'apparition du nouvel hybride Melampsora x columbiana, dans les années 1990. Tous les 256 clones se sont montrés sensibles en 1997 et à nouveau en 2000, et une variation dans la densité des urédies (résistance partielle) a aussi été observée. Pour déterminer quels clones présentaient une résistance contournée, des inoculations ont été réalisées en serre avec un isolat de Melampsora medusae originaire du Kentucky. Des clones qui étaient résistants àM. medusae avant l'apparition de M. x columbiana ont ainsi été identifiés. Les inoculations ont abouti à une ségrégation 1 :1 (,2 = 0,772; P = 0,38) pour la résistance, ce qui indique la présence d'un gène majeur. Cependant, les clones F2 possédant le gène de résistance contourné montraient le même niveau de résistance partielle au champ en 1997 et 2000 que leurs plein-frères qui n'avaient pas ce gène. Zusammenfassung Für einige Pflanzen-Pathosysteme wurde gezeigt, dass unwirksam gewordene Haupt-Resistenzgene immer noch zu einer teilweisen Resistenz beitragen. Für Waldbäume wurde diese Hypothese bisher nie überprüft. Dies wurde jetzt im pazifischen Nordwesten möglich, wo der Pappelrost nach einem rezenten Hybridisierungsereignis stark variierte. An den F2-Nachkommenschaften von 256 Pappelklonen, die unter Freilandbedingungen in der Nähe von Corvallis, Oregon, USA wuchsen, wurde nach dem Auftreten des neuen Hybridrostes (Melampsora × columbiana) ab ca. 1990 die Krankheitsintensität und der Infektionstyp registriert. Alle 256 Klone zeigten einen anfälligen Infektionstyp im Jahre 1997 und dann wieder im Jahre 2000. Dabei wurde eine Variation in der Urediendichte (d.h. partielle Resistenz) beobachtet. Um zu bestimmen, welche Klone ein unwirksam gewordenes Resistenzgen besitzen, wurden Inokulationen im Gewächshaus mit einem Isolat von M. medusae, einem Elter von M. × columbiana, durchgeführt. Damit wurden Klone identifiziert, die vor dem Auftreten von M. × columbiana gegen M. medusae resistent waren. Der Infektionsversuch führte zu einer 1:1 Segregation (,2=0,772; P=0,38) für die Resistenz, was auf das Vorliegen eines Hauptgens hinweist. Die F2-Klone, welche dieses überwundene Resistenzgen besitzen, zeigten jedoch unter Feldbedingungen in den Jahren 1997 und 2000 den gleichen Grad einer Teilresistenz wie ihre Vollgeschwister, welchen dieses Gen fehlt. [source]

Activating and inhibitory nature of the murine paired immunoglobulin-like receptor family

IMMUNOLOGICAL REVIEWS, Issue 1 2001
Toshiyuki Takai
Summary: Clones for murine paired immunoglobulin-like receptors (PIR) were first isolated as those coding for type I transmembrane glycoproteins with six immunoglobulin-like domains homologous to human Fc,R, bovine Fc,2R, and other related receptors. However, they turned out to bind neither IgA nor other immunoglobulins in the case of the ectopic expression on COS-1 fibroblastic cells. PIR-A and B are expressed on a wide variety of cells in the murine immune system, such as in B cells, mast cells, macrophages, and dendritic cells, mostly in a pairwise fashion. PIR-A requires homodimeric Fc receptor common , chain, which harbors an immunoreceptor tyrosine-based activation motif, for its efficient cell surface expression and for the delivery of activation signaling. In contrast, PIR-B contains immunoreceptor tyrosine-based inhibitory motifs (ITIMs) in its cytoplasmic portion and inhibits receptor-mediated activation signaling in vitro upon engagement with other activating-type receptors such as the antigen receptor on B cells and the high affinity Fc receptor for IgE on mast cells. ITIMs of PIR-B on macrophages and B cells have been shown to be constitutively phosphorylated in their tyrosine residues. Although the ligand for PIR still remains unknown, the transgenics and the gene-targeted mice will provide us with valuable information on their physiological roles in the immune regulation. We thank Hiromi Kubagawa for discussion. This work is supported by CREST Program of JST, Virtual Research Institute of Aging funded by Boehringer Ingelheim, and by research grants from the Ministry of Education, Science, Sports and Culture of Japan to T. Takai. [source]

Effect of Temperature on Fecundity, Life Span and Morphology of Long- and Short-Spined Clones of Brachionus caudatus f. apsteini (Rotifera)

INTERNATIONAL REVIEW OF HYDROBIOLOGY, Issue 6 2008
Sujiporn Athibai
Abstract We investigated the effect of temperature (20, 25 and 30 °C) on fecundity, life span and morphology of the rotifer Brachionus caudatus f. apsteini. For each temperature, short posterior-spined and long posterior-spined clones of B. caudatus f. apsteini were individually cultured for up to six generations. The rotifers were fed Chlorella sp. at a density of 1 × 106 cells ml,1. Morphometric data (body size and spine length) were collected. Total number of offspring producing by a single female per life cycle at high temperature was higher than at low temperature. The duration of juvenile period, reproductive period, post-reproductive period and life span of both clones of B. caudatus f. apsteini decreased with increasing temperature. All offspring of short posterior-spined clone produce posterior spines at 20 and 25 °C, with an average length of 19.8 ± 6.6 and 11.9 ± 2.6 ,m, respectively. In contrast, they cannot develop posterior spines at 30 °C, at which the average length of the posterior spine remnant was 6.4 ± 1.3 ,m. On the other hand, all offspring of long posterior-spined clone have long posterior spines with average lengths of 36.8 ± 6.1, 36.3 ± 5.2 and 36.6 ± 6.2 ,m at 20, 25 and 30 °C, respectively. This study indicated that the production of posterior spines can be induced by low temperature and that short posterior-spined and long posterior-spined clones are genetically different. (© 2008 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim) [source]

Greater capacity for division of labour in clones of Fragaria chiloensis from patchier habitats

JOURNAL OF ECOLOGY, Issue 3 2007
SERGIO R. ROILOA
Summary 1Unlike non-clonal plants, clonal plants can develop a division of labour in which connected ramets specialize to acquire different, locally abundant resources. This occurs as a plastic response to a patchy environment where two resources tend not to occur together and different ramets experience high availabilities of different resources. We hypothesized that if division of labour is an important advantage of clonal growth in such environments in nature, then clones from habitats where resource availabilities are negatively associated should show a greater capacity for division of labour than clones from habitats where resource availabilities are more uniform. 2To test this, we collected clones of Fragaria chiloensis from sand dune and grassland sites in each of three regions of the central coast of California, grew pairs of connected or severed ramets under low light and high N or under high light and low N, and measured leaf area, chlorophyll content and final dry mass. Given that previous work has indicated that high availabilities of light and N show a stronger tendency not to occur together in the dune than in the grassland sites, we expected that clones from dunes would show greater capacity for division of labour than clones from grasslands. 3Clones from dunes showed a greater capacity than clones from grasslands to specialize for acquisition of abundant N via high proportional mass of roots. Clones from the two types of habitats showed similar capacity to specialize for acquisition of abundant light via high leaf area and chlorophyll content of leaves. Specialization via leaf area and chlorophyll content took place mainly within the first half of the 60-day experiment. 4These results provide evidence that division of labour in a clonal plant has been selected for in natural habitats where high levels of different resources tend to be spatially separated. Results also show that division of labour can occur, not just via allocation of mass, but also via physiological traits, and that both morphological and physiological specialization can take place within a few weeks. 5Clonal plants dominate many habitats and include many highly invasive species. Division of labour is one of the most striking potential advantages of clonal growth, and is a remarkable instance of phenotypic plasticity in plants. This study further suggests that division of labour in clonal plants is an instance of adaptive plasticity and could therefore play a part in their widespread ecological success. [source]

Analyses of amino acid sequences in hypervariable region-1 of hepatitis C virus (HCV) in sera from chimpanzees infected three times with the same HCV strain

JOURNAL OF MEDICAL PRIMATOLOGY, Issue 1 2010
M. Sugitani
Abstract Background, To investigate whether or not the same strain of hepatitis C virus (HCV) can twice re-infect the same chimpanzee, we analyzed nucleic and amino acid sequences in HCV hypervariable region-1 (HVR1). Two chimpanzees were inoculated, three times each, with the same HCV strain during the 1983,1991. After each inoculation, chimpanzees developed acute hepatitis C, and then recovered. Methods, Using sera, HVR1 cloning and antibody to HVR1 major clone measurement were performed. Results, Clones from the first inoculum were divisible into major and minor types. Clones from the second and third inocula, as well as all post-inoculation sera, were essentially identical to the major type. Titers of antibody to HVR1 major clone were consistently low in pre- and post-inoculation sera. Conclusions, Both chimpanzees were re-infected twice with the same strain of HCV. The sequences from the second and third infections were similar to the major sequences in the first inoculum. [source]

Characterization of a chemokine receptor CCR5-negative T cell line and its use in determining human immunodeficiency virus type 1 phenotype

JOURNAL OF MEDICAL VIROLOGY, Issue 2 2008
Dorothea Binninger-Schinzel
Abstract A human CD4-positive T cell line from a donor homozygous negative for the chemokine receptor CCR5 was established, characterized, and used for determining the coreceptor usage of human immunodeficiency virus type 1 (HIV-1) isolates. Clones of this IL-2 dependent human T-cell lymphotropic virus type 1 (HTLV-I) immortalized cell line, named IsnoR5 clones 1 and 2, are susceptible to infection by HIV-1 isolates that use CXCR4 as a coreceptor but resistant to infection by CCR5 tropic HIV-1 viruses. HIV-1 isolates whose replication is inhibited in IsnoR5 cells in the presence of the bicyclam AMD 3100, a CXCR4 specific inhibitor, utilize a coreceptor distinct from CCR5 and CXCR4. Using a panel of primary HIV-1 isolates we have shown that a single T cell line is sufficient to discriminate between use of CCR5, CXCR4 or an alternative coreceptor. As IsnoR5 clone 1 cells revealed the existence of even minor populations of CXCR4-using virus variants, they could be useful for the early identification of changes in coreceptor usage in HIV infected individuals facilitating the timely introduction of appropriate clinical treatments. J. Med. Virol. 80:192,200, 2008. © 2007 Wiley-Liss, Inc. [source]

PRELIMINARY ANALYSIS OF QUANTITATIVE GENETICS AND PHENOTYPIC PLASTICITY IN AULACOSEIRA SUBARCTICA (BACILLAR-IOPHYTA)

JOURNAL OF PHYCOLOGY, Issue 2000
S. M. Edgar
Several clones of Aulacoseira subarctica were isolated from Yellowstone, Lewis, and East Rosebud Lakes (Montana, Wyoming). Two to four clones from each lake were grown in batch cultures under three light intensities, 2, 11.4 and 115 ,E m,2 s,1. Clones were conditioned to their light environment for a three-week period. Inoculants from the conditioned clones taken during log phase of growth, were grown until in log phase, then samples were collected. Five randomly chosen valves for 2 replicates of each clone were examined using a scanning electron microscope and captured on film at a magnification of 20,000x. Each image was digitized and quantitative morphometric characters were measured. A preliminary quantitative genetic analysis was performed on selected characters within each light environment. Plasticity of characters within clones across the three light regimes were also examined. The amount of variability found within characters in A. subarctica will be discussed in terms of environmental, genetic, and microenvironmental sources. [source]

Simulations of the population of Centaurs , II.

MONTHLY NOTICES OF THE ROYAL ASTRONOMICAL SOCIETY, Issue 2 2004
Individual objects
ABSTRACT Detailed orbit integrations of clones of five Centaurs , namely, 1996 AR20, 2060 Chiron, 1995 SN55, 2000 FZ53 and 2002 FY36 , for durations of ,3 Myr are presented. One of our Centaur sample starts with perihelion initially under the control of Jupiter (1996 AR20), two start under the control of Saturn (Chiron and 1995 SN55) and one each starts under the control of Uranus (2000 FZ53) and Neptune (2002 FY36), respectively. A variety of interesting pathways are illustrated with detailed examples including: capture into the Jovian Trojans, repeated bursts of short-period comet behaviour, capture into mean-motion resonances with the giant planets and into Kozai resonances, as well as traversals of the entire Solar system. For each of the Centaurs, we provide statistics on the numbers (i) ejected, (ii) showing short-period comet behaviour and (iii) becoming Earth- and Mars-crossing. For example, Chiron has over 60 per cent of its clones becoming short-period objects, while 1995 SN55 has over 35 per cent. Clones of these two Centaurs typically make numerous close approaches to Jupiter. At the other extreme, 2000 FZ53 has ,2 per cent of its clones becoming short-period objects. In our simulations, typically 20 per cent of the clones which become short-period comets subsequently evolve into Earth-crossers. [source]

Selection for verticillium wilt resistance in potato breeding populations derived from four cross types

PLANT BREEDING, Issue 3 2009
J. Bae
Abstract Verticillium wilt (VW) is one of the important yield-limiting diseases for potato production. To develop resistant clones, the potential for early generation selection was studied using three basic selection methods, individual, family, and within family selection, for two clonal generations. A total of 152 clones were derived from four cross types (2x × 2x, 2x × 4x, 4x × 2x and 4x × 4x). Clones were evaluated for maturity, symptom expression, yield and stem colonization in replicated trials. Heritability and selection response for the traits were estimated for each selection method. Direct selection in the second clonal generation and individual selection showed more gain than that from other methods. Both 2x × 2x and 4x × 2x families were higher yielding and had lower stem colonization scores than 2x × 4x and 4x × 4x crosses. Therefore, 2x × 2x or 4x × 2x crosses between carefully chosen parents with high yield and VW resistance may produce offspring with superior performance. [source]

Interclonal differences, plasticity and trade-offs of life history traits of Cyperus esculentus in relation to water availability

PLANT SPECIES BIOLOGY, Issue 3 2001
BO LI
Abstract Cyperus esculentus is an exotic clonal (or pseudoannual) weed in Japan, and its range is steadily increasing. To investigate its interclonal variation and phenotypic plasticity in response to water availability, five clones of C. esculentus, collected from different sites in Japan, were grown singly in pots placed outdoors under dry and wet conditions. All the traits examined showed considerable variation among the five clones. However, two clones from Tochigi were similar to each other; thus, they might have originated from the same founder population. The clone from Ibaraki was quite different from the others. Therefore, it is suggested that the Japanese populations of C. esculentus might have resulted from multiple introductions of genotypes from geographically separated and, hence, genetically differentiated, source populations. All the clones also showed considerable plasticity in response to water availability. Clones with a larger ramet number had a greater plasticity, whereas tuber size was invariant across water treatments. Highly plastic traits had generally low interclonal variation in plasticity. All the clones had high productivity and produced more ramets and tubers under wet conditions than under dry conditions. Moreover, water availability could partially regulate the mode of its reproduction; wet conditions favored tuber production (vegetative propagation) while dry conditions favored sexual reproduction. A number of trade-offs occurred between the traits of clonal growth, storage and sexual reproduction, indicating that allocation among the competing functions/organs is mutually exclusive in plants. The results obtained here suggest that C. esculentus is more likely to invade wet habitats than dry habitats. [source]

Genetic, Morphological, and Ecological Diversity of Spatially Separated Clones of Meseres corlissi Petz & Foissner, 1992 (Ciliophora, Spirotrichea)

THE JOURNAL OF EUKARYOTIC MICROBIOLOGY, Issue 4 2008
THOMAS WEISSE
ABSTRACT. We investigated the intraspecific variation of the spirotrich freshwater ciliate Meseres corlissi at the level of genes (SSrDNA, ITS), morphology (14 characters), and ecophysiology (response to temperature and pH). Five of the eight clonal M. corlissi cultures isolated from five localities on four continents were studied at all levels. The null hypothesis was that geographic distance plays no role: M. corlissi lacks biogeography. The intraspecific variation was low at the genetic level (0%,4%), moderate at the morphological level (5%,15%), and high at the ecophysiological level (10%,100%). One clone, isolated from subtropical China, differed significantly at all levels from all other clones, suggesting limited dispersal and local adaptation among M. corlissi. However, other clones from distant areas, such as Australia and Austria, were genetically identical and differed only slightly in morphology and temperature response. We speculate that our findings may be typical for rare species; the chances may be equally high for both global dispersal of most and local adaptation of some populations in areas where dispersal has been permanently or temporarily reduced. [source]

Development and characterization of a minimal inducible packaging cell line for simian immunodeficiency virus-based lentiviral vectors

THE JOURNAL OF GENE MEDICINE, Issue 4 2002
Seraphin Kuate
Abstract Background Lentiviral vectors allow gene transfer into non-dividing cells. Further development of these vector systems requires stable packaging cell lines that enable adequate safety testing. Methods To generate a packaging cell line for vectors based on simian immunodeficiency virus (SIV), expression plasmids were constructed that contain the codon-optimized gag-pol gene of SIV and the gene for the G protein of vesicular stomatitis virus (VSV-G) under the control of an ponasterone-inducible promoter. Stable cell lines expressing these packaging constructs were established and characterized. Results The RT activity and vector titers of cell clones stably transfected with the inducible gag-pol expession plasmid could be induced by ponasterone by more than a factor of 1000. One of these clones was subsequently transfected with the ponasterone-inducible VSV-G expression plasmid to generate packaging cells. Clones of the packaging cells were screened for vector production by infection with an SIV vector and subsequent induction by ponasterone. In the supernatant of selected ponasterone-induced producer clones vector titers of more than 1×105 transducing units/ml were obtained. Producer cell clones were stable for at least five months, as tested by vector production. Conclusions The packaging cells described should be suitable for most preclinical applications of SIV-based vectors. By avoiding regions of high homology between the vector and the packaging constructs, the design of the SIV packaging cell line should reduce the risk of transfer of packaging genes to target cells and at the same time provide flexibility with respect to the SIV vector constructs that can be packaged. Copyright © 2002 John Wiley & Sons, Ltd. [source]

A 2.5-Mb contig constructed from Angus, Longhorn and horned Hereford DNA spanning the polled interval on bovine chromosome 1

ANIMAL GENETICS, Issue 6 2006
K. R. Wunderlich
Summary The polled locus has been mapped by genetic linkage analysis to the proximal region of bovine chromosome 1. As an intermediate step in our efforts to identify the polled locus and the underlying causative mutation for the polled phenotype, we have constructed a BAC-based physical map of the interval containing the polled locus. Clones containing genes and markers in the critical interval were isolated from the TAMBT (constructed from Angus and Longhorn genomic DNA) and CHORI-240 (constructed from horned Hereford genomic DNA) BAC libraries and ordered based on fingerprinting and the presence or absence of 80 STS markers. A single contig spanning 2.5 Mb was assembled. Comparison of the physical order of STSs to the corresponding region of human chromosome 21 revealed the same order of genes within the polled critical interval. This contig of overlapping BAC clones from horned and polled breeds is a useful resource for SNP discovery and characterization of positional candidate genes. [source]