EVOLUTION, Issue 9 2002
Karin S. Pfennig
Abstract., The fitness consequences of hybridization critically affect the speciation process. When hybridization is costly, selection favors the evolution of prezygotic isolating mechanisms (e.g., mating behaviors) that reduce heter-ospecific matings and, consequently, enhance reproductive isolation between species (a process termed reinforcement). If, however, selection to avoid hybridization differs between species, reinforcement may be impeded. Here, we examined both the frequency and fitness effects of hybridization between plains spadefoot toads (Spea bombifrons) and New Mexico spadefoot toads (S. multiplicata). Hybridization was most frequent in smaller breeding ponds that tend to be ephemeral, and heterospecific pairs consisted almost entirely of S. bombifrons females and S. multiplicata males. Moreover, in controlled experimental crosses, hybrid offspring from crosses in which S. multiplicata was maternal had significantly lower survival and longer development time than pure S. multiplicata offspring. By contrast, hybrid offspring from crosses in which S. bombifrons was maternal outperformed pure S. bombifrons offspring by reaching metamorphosis faster. These data suggest that, although S. multiplicata females are under selection to avoid hybridization, selection might favor those S. bombifrons females that hybridize with S. multiplicata if their breeding pond is highly ephemeral. Generally, the strength of selection to avoid hybridization may differ for hybridizing species, possibly impeding reinforcement. [source]

DETECTION OF SALMONELLA TYPHIMURIUM IN OYSTERS BY PCR AND MOLECULAR HYBRIDIZATION

JOURNAL OF FOOD QUALITY, Issue 5 2006
A.A. CORRÊA
ABSTRACT Because shellfish (oysters, clams and mussels) are filter feeders, i.e., able to concentrate pathogens from the surrounding waters within their tissues, they have been widely associated with outbreaks illness. The incidence of salmonellosis caused by the consumption of raw or undercooked shellfish, is a primary concern of public health agencies. Then, in recent years, more rapid and specific methods based on the DNA sequence of salmonella genes have been developed to detect low levels of pathogens in environmental and food samples. In this study, we developed a sensitive method to detect low levels of Salmonella typhimurium in oyster tissues (0.1 cfu/g). This methodology consisted of dissection of the gastrointestinal oyster tract, pre-enrichment of the samples in nonselective medium, DNA extraction and polymerase chain reaction followed by molecular hybridization using a digoxygenin-labeled amplicon-derived probe. These results can benefit the public health agencies and shellfish producers concerning microbiological and quality aspects of the commercial oyster production. [source]

Typological thinking and the conservation of subspecies: the case of the San Clemente Island loggerhead shrike

DIVERSITY AND DISTRIBUTIONS, Issue 4 2000
Michael A. Patten
Abstract. ,Hybridization with closely related taxa poses a significant threat to endangered subspecies (e.g. outbreeding depression, inbreeding) and confounds efforts to manage and conserve these taxa through a loss of taxonomic identity, in part because of the practical necessity of defining subspecies in a typological manner. We examined nine morphological characters in 167 post-juvenile museum specimens to determine if loggerhead shrikes Lanius ludovicianus Linnaeus 1766 on San Clemente Island (off the coast of California) remain diagnosable as L. l. mearnsi Ridgway (1903); an island endemic listed as endangered by the United States Fish and Wildlife Service. Four recent shrike specimens from the island were compared to historical specimens using a bivariate scatter plot and a discriminant function (the latter was used to classify recent specimens post hoc). The few recent specimens were not diagnosable as L. l. mearnsi, but instead appear to be intergrades between L. l. mearnsi and L. l. anthonyi Mearns 1898 (the subspecies endemic to Santa Cruz, Santa Catalina, Santa Rosa and Anacapa islands), and are perhaps closer to pure anthonyi. Our data and the species' natural history and distribution suggest that shrikes currently on San Clemente Island are the result of genetic ,swamping' of mearnsi by anthonyi. Under a necessarily typological definition of a subspecies, it is evident that mearnsi is probably no longer diagnosable. However, we conclude that protection of the entire Channel Islands population of the loggerhead shrike would be the best management strategy, as the species has declined drastically throughout the islands. [source]

Avoidance by grazers facilitates spread of an invasive hybrid plant

ECOLOGY LETTERS, Issue 2 2010
E. Grosholz
Ecology Letters (2010) 13: 145,153 Abstract Biological invasions greatly increase the potential for hybridization among native and non-native species. Hybridization may influence the palatability of novel hybrids to consumers potentially influencing invasion success; however, the palatability of non-native hybrids relative to the parent species is poorly known. In contrast, studies of native-only hybrids find they are nearly always more palatable to consumers than the parent species. Here, I experimentally demonstrate that an invasive hybrid cordgrass (Spartina) is dramatically less palatable to grazing geese than the native parent species. Using field and aviary experiments, I show that grazing geese ignore the hybrid cordgrass and preferentially consume native Spartina. I also experimentally demonstrate that reduced herbivory of the invasive hybrid may contribute to faster spread in a California estuary. These results suggest that biological invasions may increase future opportunities for creating novel hybrids that may pose a greater risk to natural systems than the parent species. [source]

Flow Injection Analysis of Sulfide Using a Cinder/Tetracyano Nikelate Modified Screen-Printed Electrode

ELECTROANALYSIS, Issue 9 2005
Jyh-Myng Zen
Abstract Flow injection analysis (FIA) of sulfide is presented using a screen-printed carbon electrode modified with a cinder/tetracyano nickelate hybrid (designated as cinder/NiTcSPE). Hybridization of NiTc was achieved in iron-enriched industrial waste cinder material through the bimetallic formation of FeIII[NiII(CN)4]. The electrocatalytic oxidation of sulfide is mediated by the higher oxidation state of Ni in this hybrid-bimetallic complex. The system shows a detection limit (S/N=3) of 0.06,,M and a linear working range up to 1,mM in pH,10, 0.1,M KCl solution. Taking into account the relatively low volatility of the analyte in alkaline conditions, the system is ideally suited for the accurate detection of sulfide. The response of the electrode to sulfide is highly reproducible, thereby offering the potential development of a disposable amperometric sensor for sulfide. Selective detection of sulfide in cigarette smoke is presented in this study as an example of a real sample application. [source]

DNA Hybridization at Magnetic Nanoparticles with Electrochemical Stripping Detection

ELECTROANALYSIS, Issue 23 2004
Ningning Zhu
Abstract A simple and practical method for electrochemical DNA hybridization assay has been developed to take advantage of magnetic nanoparticles for ssDNA immobilization and zinc sulfide nanoparticle as oligonucleotide label. Magnetic nanoparticles were prepared by coprecipitation of Fe2+ and Fe3+ with NH4OH, and then amino silane was coated onto the surface of magnetite nanoparticles. The magnetic nanoparticles have the advantages of easy preparation, easy surface modification and low cost. The target ssDNA with the phosphate group at the 5, end was then covalently immobilized to the amino group of magnetite nanoparticles by forming a phosphoramidate bond in the presence of 1-ethyl-3-(3-dimeth-ylaminopropyl)carbodiimide (EDAC). The zinc sulfide (ZnS) nanoparticle-labeled oligonucleotides probe was used to identify the target ssDNA immobilized on the magnetic nanoparticles based on a specific hybridization reaction. The hybridization events were assessed by the dissolution of the zinc sulfide nanoparticles anchored on the hybrids and the indirect determination of the dissolved zinc ions by anodic stripping voltammetry (ASV) at a mercury film glassy carbon electrode (GCE). The proposed method couples the high sensitivity of anodic stripping analysis for zinc ions with effective magnetic separation for eliminating nonspecific adsorption effects and offers great promise for DNA hybridization analysis. [source]

Label-Free and Label Based Electrochemical Detection of Hybridization by Using Methylene Blue and Peptide Nucleic Acid Probes at Chitosan Modified Carbon Paste Electrodes

ELECTROANALYSIS, Issue 24 2002
Pinar Kara
Abstract A chitosan modified carbon paste electrode (ChiCPE) based DNA biosensor for the recognition of calf thymus double stranded DNA (dsDNA), single stranded DNA (ssDNA) and hybridization detection between complementary DNA oligonucleotides is presented. DNA and oligonucleotides were electrostatically attached by using chitosan onto CPE. The amino groups of chitosan formed a strong complex with the phosphate backbone of DNA. The immobilized probe could selectively hybridize with the target DNA to form hybrid on the CPE surface. The detection of hybridization was observed by using the label-free and label based protocols. The oxidation signals of guanine and adenine greatly decreased when a hybrid was formed on the ChiCPE surface. The changes in the peak currents of methylene blue (MB), an electroactive label, were observed upon hybridization of probe with target. The signals of MB were investigated at dsDNA modified ChiCPE and ssDNA modified ChiCPE and the increased peak currents were observed, in respect to the order of electrodes. The hybridization of peptide nucleic acid (PNA) probes with the DNA target sequences at ChiCPE was also investigated. Performance characteristics of the sensor were described, along with future prospects. [source]

Metal-Assisted Hybridization of Oligonucleotides, Evaluation of Circular 2,- O -Me RNA as Ligands for the TAR RNA Target

EUROPEAN JOURNAL OF ORGANIC CHEMISTRY, Issue 6 2003
Laurence Zapata
Abstract Two complementary oligonucleotides were conjugated with terpyridine ligands at their nearby 5,- and 3,-ends. Addition of a stoichiometric amount of a transition metal (Zn2+, Fe2+) resulted in a large increase in the melting temperature of the duplex. The conjugation of TPY to stem-loop oligomers provided an efficient procedure for the cyclisation of the oligomer after the addition of metal ions. Such a short stem-loop oligomer was designed to target the HIV-1 TAR RNA through loop,loop interactions. The addition of Zn2+ ions yielded a good ligand (Kd = 30 nM) for this RNA structural element. (© Wiley-VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2003) [source]

ENVIRONMENT-DEPENDENT ADMIXTURE DYNAMICS IN A TIGER SALAMANDER HYBRID ZONE

EVOLUTION, Issue 6 2004
Benjamin M. Fitzpatrick
Abstract After an estimated five million years of independent evolution, the barred tiger salamander (Ambystoma tigrinum mavortium) was introduced by bait dealers into the native range of the California tiger salamander (A. californiense). Hybridization and backcrossing have been occurring in central California for 50,xs60 years, or an estimated 15,30 generations. We studied genetic and ecological factors influencing admixture of these two divergent gene pools by analyzing frequencies of hybrid genotypes in three kinds of breeding habitats: natural vernal pools, ephemeral man-made cattle ponds, and perennial man-made ponds. Perennial ponds tended to have higher frequencies of nonnative alleles than either type of seasonal pond, even in cases where perennial and seasonal ponds are within a few hundred meters. Thus, the hybrid zone has a mosaic structure that depends on pond hydrology or ecology. The presence of some broadly acting constraints on admixture is suggested by linkage disequilibria between physically unlinked molecular markers within ponds. In addition, we found several marker-specific deviations from Hardy-Weinberg equilibrium. One marker showed a consistent deficit of heterozygotes across pond types. Another showed heterozygote deficits only in vernal pools. A third was more likely to have heterozygote excess in ephemeral cattle ponds. These patterns indicate that admixture is influenced by complex genotype-by-environment interactions. [source]

DIFFERENTIAL SELECTION TO AVOID HYBRIDIZATION IN TWO TOAD SPECIES

Latitudinal variation in cold hardiness in introduced Tamarix and native Populus

EVOLUTIONARY APPLICATIONS (ELECTRONIC), Issue 4 2008
Jonathan M. Friedman
Abstract To investigate the evolution of clinal variation in an invasive plant, we compared cold hardiness in the introduced saltcedar (Tamarix ramosissima, Tamarix chinensis, and hybrids) and the native plains cottonwood (Populus deltoides subsp. monilifera). In a shadehouse in Colorado (41°N), we grew plants collected along a latitudinal gradient in the central United States (29,48°N). On 17 occasions between September 2005 and June 2006, we determined killing temperatures using freeze-induced electrolyte leakage and direct observation. In midwinter, cottonwood survived cooling to ,70°C, while saltcedar was killed at ,33 to ,47°C. Frost sensitivity, therefore, may limit northward expansion of saltcedar in North America. Both species demonstrated inherited latitudinal variation in cold hardiness. For example, from September through January killing temperatures for saltcedar from 29.18°N were 5,21°C higher than those for saltcedar from 47.60°N, and on September 26 and October 11, killing temperatures for cottonwood from 33.06°N were >43°C higher than those for cottonwood from 47.60°N. Analysis of nine microsatellite loci showed that southern saltcedars are more closely related to T. chinensis while northern plants are more closely related to T. ramosissima. Hybridization may have introduced the genetic variability necessary for rapid evolution of the cline in saltcedar cold hardiness. [source]

Quantum-Dot-Functionalized Poly(styrene- co -acrylic acid) Microbeads: Step-Wise Self-Assembly, Characterization, and Applications for Sub-femtomolar Electrochemical Detection of DNA Hybridization

ADVANCED FUNCTIONAL MATERIALS, Issue 7 2010
Haifeng Dong
Abstract A novel nanoparticle label capable of amplifying the electrochemical signal of DNA hybridization is fabricated by functionalizing poly(styrene- co -acrylic acid) microbeads with CdTe quantum dots. CdTe-tagged polybeads are prepared by a layer-by-layer self-assembly of the CdTe quantum dots (diameter,=,3.07,nm) and polyelectrolyte on the polybeads (diameter,=,323,nm). The self-assembly procedure is characterized using scanning and transmission electron microscopy, and X-ray photoelectron, infrared and photoluminescence spectroscopy. The mean quantum-dot coverage is (9.54,±,1.2),×,103 per polybead. The enormous coverage and the unique properties of the quantum dots make the polybeads an effective candidate as a functionalized amplification platform for labelling of DNA or protein. Herein, as an example, the CdTe-tagged polybeads are attached to DNA probes specific to breast cancer by streptavidin,biotin binding to construct a DNA biosensor. The detection of the DNA hybridization process is achieved by the square-wave voltammetry of Cd2+ after the dissolution of the CdTe tags with HNO3. The efficient carrier-bead amplification platform, coupled with the highly sensitive stripping voltammetric measurement, gives rise to a detection limit of 0.52 fmol L,1 and a dynamic range spanning 5 orders of magnitude. This proposed nanoparticle label is promising, exhibits an efficient amplification performance, and opens new opportunities for ultrasensitive detection of other biorecognition events. [source]

Photovoltaics Based on Hybridization of Effective Dye-Sensitized Titanium Oxide and Hole-Conductive Polymer P3HT

ADVANCED FUNCTIONAL MATERIALS, Issue 15 2009
Ke-Jian Jiang
Abstract Here, the fabrication of quasi-solid-state TiO2/dye/poly(3-hexylthiophene) (P3HT) solar cells is reported, in which the dyes with oleophilic thienyl groups were employed and ionic liquid (IL), 1-ethyl-3-methylimidazolium (EMIm) containing lithium bis(trifluromethanesulfone)amide (Li-TFSI) and 4- tert -butylpyridine (t -BP) are assembled with dyed TiO2 surfaces. One of the devices gave a high conversion efficiency of up to 2.70% under 1 sun illumination. The excellent performance is ascribed to successful molecular self-organization at interface of the dye molecules and P3HT, and to the efficient charge separation and diffusion acquired by introduction of the IL coupled with Li-TFSI and t-BP. [source]

Stage-specific gene expression in early differentiating oligodendrocytes

GLIA, Issue 2 2002
Francesca Blasi
Abstract The screening of a differential library from precursor and differentiated oligodendrocytes, obtained through the representational difference analysis (RDA) technique, has generated a number of cDNA recombinants corresponding to mRNA coding for known and unknown proteins: (1) mRNA coding for proteins involved in protein synthesis, (2) mRNA coding for proteins involved in the organization of the cytoskeleton, and (3) mRNA coding for proteins of unknown function. The expression profile of the mRNA was studied by Northern blot hybridization to the poly-A+ mRNA from primary rat progenitor and differentiated oligodendrocytes. In most cases, hybridization to the precursor was higher than hybridization to the differentiated mRNA, supporting the validity of the differential screening. Hybridization of the cDNA to rat cerebral hemisphere and brain stem poly-A+ mRNA, isolated from 1- to 90-day-old rats, confirms the results obtained with the mRNA from differentiating oligodendrocytes. The intensity of the hybridization bands decreases as differentiation proceeds. The pattern of expression observed in oligodendrocytes is different from that found in the brain only in the case of the nexin-1 mRNA, the level of which remains essentially constant throughout differentiation both in the brain stem and in the cerebral hemispheres, in agreement with the published data. In contrast, the intensity of hybridization to the oligodendrocyte mRNA is dramatically lower in the differentiated cells compared with the progenitor oligodendrocyte cells. Some of the recombinant cDNA represent mRNA sequences present at high frequency distribution in the cells, while others belong to the rare sequences group. Six recombinants code for proteins of the ribosomal family, suggesting that of approximately 70 known ribosomal proteins, only a few are upregulated during oligodendrocyte differentiation. The third category of open reading frame (ORF) is represented by rare messengers coding for proteins of unknown functions and includes six clones: RDA 279, 11, 95, 96, 254, and 288. GLIA 39:114,123, 2002. © 2002 Wiley-Liss, Inc. [source]

Electrodeposition of Inorganic/Organic Hybrid Thin Films

ADVANCED FUNCTIONAL MATERIALS, Issue 1 2009
Tsukasa Yoshida
Abstract Electrodeposition of inorganic compound thin films in the presence of certain organic molecules results in self-assembly of various hybrid thin films with new properties. Examples of new discoveries by the authors are reviewed, taking cathodic formation of a ZnO/dye hybrid as the leading example. Hybridization of eosinY leads to the formation of highly oriented porous crystalline ZnO as the consequence of dye loading. The hybrid formation is a highly complicated process involving complex chemistry of many molecular and ionic constituents. However, electrochemical analyses of the relevant phenomena indicate the possibility of reaching a comprehensive understanding of the mechanism, giving us the chance to further develop them into industrial technologies. The porous crystals are ideal for photoelectrodes in dye-sensitized solar cells. As the process also permits the use of non-heat-resistant substrates, the technology can be applied for the development of colorful and light-weight plastic solar cells. [source]

Hybridization between perennial ryegrass and Italian ryegrass in naturalized Japanese populations

GRASSLAND SCIENCE, Issue 2 2008
Hiroyuki Tobina
Abstract Introduced Lolium species, including perennial ryegrass (Lolium perenne) and Italian ryegrass (Lolium multiflorum), have been widely utilized in Japan for forage, turf and soil conservation. These ryegrasses have escaped from cultivated areas and become naturalized, and this has become a serious issue in recent years. Interspecific hybrids between perennial ryegrass and Italian ryegrass have often been found in naturalized populations. It has also been suggested that hybridization between plant species might serve as a stimulus for the evolution of invasiveness. We surveyed the genetic structure of naturalized ryegrass populations in Japan using genetic markers that distinguished perennial ryegrass and Italian ryegrass. Of the 55 naturalized populations surveyed, 41 exhibited morphological traits of Italian ryegrass. DNA analysis using simple sequence repeat and chloroplast DNA markers characterized 20 of these 41 populations as Italian ryegrass, with the remaining populations as interspecific hybrid derivatives. Approximately half of the naturalized ryegrasses populations in Japan were inferred to include interspecific hybrids. [source]

Visualization of Helicobacter Species Within the Murine Cecal Mucosa Using Specific Fluorescence In Situ Hybridization

HELICOBACTER, Issue 2 2005
Vivian Chan
ABSTRACT Background., Members of the genus Helicobacter have been associated with colitis development in a number of immunodeficient animal models. While it is known that these organisms can initiate colitis development, the location and spatial distribution of these bacteria within the intestinal tract is currently unknown. In this study, we developed and optimized fluorescence in situ hybridization (FISH) probes specifically for Helicobacter species. Materials and Methods., Based on 16S-RNA gene alignments, two probes specific for the entire family Helicobacteraceae and two probes specific for Helicobacter ganmani and Helicobacter hepaticus were designed. Evaluation of these probes was determined using ATCC reference strains and cecum samples from ten IL-10 knockout mice. The presence of Helicobacter species was determined using FISH and verified using PCR-DGGE and microscopic examination of silver stained sections. Results., Analysis of the ATCC reference strains revealed that the probes HEL274/HEL717 were specific for the family Helicobacteraceae, while HEP642 was specific for H. hepaticus and GAN1237 for H. ganmani. Using these probes, a pattern of spatial localization of the two different Helicobacter species was observed in the cecum tissues of IL-10 knockout mice. This consistently showed that H. ganmani was localized to the lower regions and H. hepaticus to the mid-upper regions of the crypts. Conclusion., We have developed FISH probes specific for the family Helicobacteraceae as well as two individual Helicobacter species. This study will allow the future use of the FISH to better understand host-pathogen interactions in vitro. [source]

Sarcoglycanopathies and the risk of undetected deletion alleles in diagnosis,,

HUMAN MUTATION, Issue 1 2005
Stefan J. White
Abstract We have designed Multiplex Amplifiable Probe Hybridization (MAPH) probes for 28 exons of the sarcoglycan genes SGCA, SGCB, SGCG, and SGCD. The set was used to screen DNA from limb-girdle muscular dystrophy (LGMD) patients for the presence of pathogenic deletion or duplication mutations. An unexpected heterozygous deletion of SGCG exon 7 was detected in a patient from a consanguineous family in which a known c.525delT mutation segregates. The exon 7 deletion was inherited from the father, who was part of the consanguineous c.525delT branch of the family but who did not carry the c.525delT mutation. A similar, homozygous deletion had been identified in two unrelated LGMD patients from southern Italy. The deletion breakpoints were mapped, isolated, and sequenced, and were identical in all cases. Haplotype analysis showed the same alleles segregating with the mutation in all three patients, suggesting a common ancestor. Exonic deletions in sarcoglycanopathies appear to be rare events. However, we recommend screening for exonic deletions/duplications in patients where a mutation has not been identified in both alleles, as well as in seemingly homozygous cases where segregation of the mutations can not be confirmed in the parents. © 2005 Wiley-Liss, Inc. [source]

BUB1 infrequently mutated in human breast carcinomas,,

HUMAN MUTATION, Issue 5 2003
Anita Langerød
Abstract The BUB1 gene is a key player in the mitotic spindle checkpoint machinery that monitors proper segregation of sister chromatides during mitosis. It has been suggested that mutations in BUB1 may disrupt the spindle checkpoint and thereby cause chromosomal instability, which is a hallmark of solid tumors including those from the breast. From a series of breast carcinomas we selected 20 cases with genomic instability, as scored by Comparative Genome Hybridization (CGH), and without somatic TP53 (p53) mutations, and sequenced the entire coding region of the BUB1 gene. Two different constitutional sequence variants were found; a base substitution in exon 5, c.481G>A (CAG>CAA, a synonymous change encoding Gln144) in two samples, and a base substitution 8 bp upstream of exon 10, c.1007-8T>C in two other samples. No somatic mutations were detected. These results indicate that genomic instability scored as copy number alterations by CGH in TP53 wild type breast carcinomas is not caused by somatic mutations in the BUB1 gene. © 2003 Wiley-Liss, Inc. [source]

Gene structure and expression of nanos (nos) and oskar (osk) orthologues of the vector mosquito, Culex quinquefasciatus

INSECT MOLECULAR BIOLOGY, Issue 5 2008
J. Juhn
Abstract The products of the maternal-effect genes, nanos (nos) and oskar (osk), are important for the development of germ cells in insects. Furthermore, these genes have been proposed as candidates for donating functional DNA regulatory sequences for use in gene drive systems to control transmission of mosquito-borne pathogens. The nos and osk genes of the cosmopolitan vector mosquito, Culex quinquefasciatus, encode proteins with domains common to orthologues found in other mosquitoes. Expression analyses support the conclusion that the role of these genes is conserved generally among members of the nematocera. Hybridization in situ analyses reveal differences in mRNA distribution in early embryos in comparison with the cyclorraphan, Drosophila melanogaster, highlighting a possible feature in the divergence of the clades each insect represents. [source]

Real-time PCR quantification of haematopoietic chimerism after transplantation: a comparison between TaqMan and hybridization probes technologies

INTERNATIONAL JOURNAL OF LABORATORY HEMATOLOGY, Issue 1p1 2010
J. MARTINEZ-LOPEZ
Summary This study aimed to compare the sensitivity and accuracy of two methods of quantitative real-time polymerase chain reaction (qrt-PCR), in order to determine haematopoietic chimerism (CH): single nucleotide polymorphisms using TaqMan (TM) probes and insertion/deletion polymorphisms using Hybridization (Hyb) probes. A total of 106 samples from 20 patients who underwent allogenic stem cell transplantation (n = 14) or live-donor liver transplantation (n = 6) were studied. The mean level of chimerism was 8.37% for the TM method and 7.73% in the Hyb method, which was not significantly different (P = 0.69). The Pearson correlation coefficient between the two methods was r = 0.91 (P < 0.001). The estimation of the regression line, using the Passing and Balbock method was Intercept A ,0.0381 [95% confidence interval (CI) ,0.1265 to 0.0296) and Slope B: 1.04609(95% CI 0.9349,1.161). Bland,Altman data showed that the standard deviations, which differed between the two methods (%Hyb,%TM), were 0.98 and ,1.28. The accuracy and sensitivity of qrt-PCR chimerism is independent of the method used if the optimization is adequate and satisfies the criteria for adequate study. Real-time PCR, independent of the method adopted, is a very good tool for study levels of CH. [source]

Characterization of human fetal osteoblasts by microarray analysis following stimulation with 58S bioactive gel-glass ionic dissolution products

JOURNAL OF BIOMEDICAL MATERIALS RESEARCH, Issue 2 2006
Ioannis Christodoulou
Abstract Bioactive glasses dissolve upon immersion in culture medium, releasing their constitutive ions in solution. There is evidence suggesting that these ionic dissolution products influence osteoblast-specific processes. Here, we investigated the effect of 58S sol,gel-derived bioactive glass (60 mol % SiO2, 36 mol % CaO, 4 mol % P2O5) dissolution products on primary osteoblasts derived from human fetal long bone explant cultures (hFOBs). We used U133A human genome GeneChip® oligonucleotide arrays to examine 22,283 transcripts and variants, which represent over 18,000 well-substantiated human genes. Hybridization of samples (biotinylated cRNA) derived from monolayer cultures of hFOBs on the arrays revealed that 10,571 transcripts were expressed by these cells, with high confidence. These included transcripts representing osteoblast-related genes coding for growth factors and their associated molecules or receptors, protein components of the extracellular matrix (ECM), enzymes involved in degradation of the ECM, transcription factors, and other important osteoblast-associated markers. A 24-h treatment with a single dosage of ionic products of sol,gel 58S dissolution induced the differential expression of a number of genes, including IL-6 signal transducer/gp130, ISGF-3/STAT1, HIF-1 responsive RTP801, ERK1 p44 MAPK (MAPK3), MAPKAPK2, IGF-I and IGFBP-5. The over 2-fold up-regulation of gp130 and MAPK3 and down-regulation of IGF-I were confirmed by real-time RT-PCR analysis. These data suggest that 58S ionic dissolution products possibly mediate the bioactive effect of 58S through components of the IGF system and MAPK signaling pathways. © 2005 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2006 [source]

Rcl is a novel ETV1/ER81 target gene upregulated in breast tumors

JOURNAL OF CELLULAR BIOCHEMISTRY, Issue 3 2008
Sook Shin
Abstract ETV1 (ER81) is a transcription factor that can be activated by HER2/Neu, a proto-oncoprotein often overexpressed in metastatic breast tumors. Here, we demonstrate that ETV1 downregulation suppresses proliferation of HER2/Neu-positive MDA-MB-231 breast cancer cells in vitro and tumor formation in vivo, proving for the first time the existence of a critical role of ETV1 in breast cancer cell physiology. A screen for novel ETV1 target genes hinted at Rcl, an enzyme involved in nucleotide metabolism. To characterize the human Rcl gene, we cloned its promoter and found that ETV1 and HER2/Neu cooperated in activating the Rcl promoter, whereas a dominant-negative ETV1 molecule suppressed the Rcl promoter. Moreover, ETV1 and HER2/Neu synergized to upregulate the endogenous Rcl gene. ETV1 also bound to the Rcl promoter in vivo, indicating that Rcl is a bona fide target gene of ETV1. Hybridization of Rcl cDNA to a breast cancer array revealed that Rcl is overexpressed in ,40% of all breast tumors. Importantly, its expression significantly escalates with increasing tumor grade, strongly implicating that Rcl contributes to breast tumorigenesis. Since joint overexpression of Rcl with vascular endothelial growth factor, another target gene of ETV1, has been shown to induce tumor formation, Rcl may be one crucial effector of ETV1 and HER2/Neu in breast tumors. Furthermore, given its expression pattern and enzymatic function in nucleotide metabolism, Rcl presents itself as a novel target in breast cancer therapy via modulation of its activity by small molecule drugs. J. Cell. Biochem. 105: 866,874, 2008. © 2008 Wiley-Liss, Inc. [source]

Identification of Differentially Expressed Genes During Anther Abortion of Taigu Genic Male Sterile Wheat by Combining Suppression Subtractive Hybridization and cDNA Array

JOURNAL OF INTEGRATIVE PLANT BIOLOGY, Issue 11 2006
Qing-Shan Chang
Abstract Taigu Genic Male Sterile Wheat (TGMSW; Triticum aestivum L.), a dominant genic male sterile germplasm, is of considerable value in the genetic improvement of wheat because of its stable inherence, complete male abortion, and high cross-fertilization rate. To identify specially transcribed genes in sterile anther, a suppression subtractive hybridization (SSH) library was constructed with sterile anther as the tester and fertile anther as the driver. A total of 2 304 SSH inserts amplified by polymerase chain reaction were arrayed using robotic printing. The cDNA arrays were hybridized with 32P-labeled probes prepared from the RNA of forward- and reverse-subtracted anthers. Ninety-six clones were scored as upregulated in sterile anthers compared with the corresponding fertile anthers and some clones were selected for sequencing and analysis in GenBank. Based on their putative functions, 87 non-redundant clones were classified into the following groups: (i) eight genes involved in metabolic processes; (ii) four material transportation genes; (iii) three signal transduction-associated genes; (iv) four stress response and senescence-associated protein genes; (v) seven other functional protein genes; (vi) five genes with no known function; and (vii) another 56 genes with no match to the databases. To test the hybridization efficiency, eight genes were selected and analyzed by Northern blot. The results of the present study provide a comprehensive overview of the genes and gene products involved in anther abortion in TGMSW. (Managing editor: Li-Hui Zhao) [source]

Variations of 18S rDNA Loci Among Six Populations of Paeonia obovata Maxim. (Paeoniaceae) Revealed by Fluorescence In Situ Hybridization

JOURNAL OF INTEGRATIVE PLANT BIOLOGY, Issue 5 2006
Rui Luo
Abstract The localization of 18S ribosomal RNA genes (rDNA) by fluorescence in situ hybridization (FISH) had been performed for some species of Paeonia. However, the pattern of 18S rDNA loci among populations is indistinct. In the present study, we localized 18S rDNA loci on meiotic or mitotic chromosomes of six populations of Paeonia obovata Maxim. (Paeoniaceae). Different numbers of rDNA loci were found with different diploid (2n=10) populations, namely eight (Lushi and Mt. Jiuhua populations), 10 (Mt. Taibai population), and seven (Mt. Guandi population), whereas tetraploid (2n=20) populations were all found with 16 loci. All rDNA loci were mapped near telomeres of mitotic chromosomes and there was no chromosome with two loci. The present results show that molecular cytological polymorphism exists among P. obovata diploid populations, indicating that structural variations occurred frequently during the evolutionary history of this species, accompanied with differentiation among populations. (Managing editor: Wei Wang) [source]

Sensitive and Specific Digoxigenin-labelled RNA Probes for Routine Detection of Citrus tristeza virus by Dot-blot Hybridization

JOURNAL OF PHYTOPATHOLOGY, Issue 6 2006
L. Barbarossa
Abstract A non-radioactive dot-blot hybridization assay for the successful detection of Citrus tristeza virus (CTV) RNA in total nucleic acid extracts of infected citrus was developed. Two digoxigenin (DIG)-labelled minus-sense riboprobes, complementary to the coat protein gene sequence of a Chinese and an Apulian CTV isolate were synthesized. Several citrus tissues were evaluated as optimal virus source and leaf petioles were found appropriate material for reliable detection. The hybridization assay showed a detection limit corresponding to 0.2 mg of fresh infected tissue. The riboprobes allowed CTV detection in isolates from different geographical areas, grown in the screenhouse or in the field, resulting in similar hybridization patterns. The infected trees were tested during different seasons with positive results, although from July to August most of the samples gave a weaker hybridization signal, compared to other seasons. The high sensitivity and reliability of the molecular hybridization assay described make it a good alternative to serological methods for CTV detection. [source]

Hyperbranched Fluoropolymers and their Hybridization into Complex Amphiphilic Crosslinked Copolymer Networks

MACROMOLECULAR CHEMISTRY AND PHYSICS, Issue 15 2007
Jeremy W. Bartels
Abstract This feature article highlights three types of hyperbranched fluoropolymers (HBFPs) with different structural features, which were synthesized by either polycondensation of fluorinated ABx monomers or self-condensing vinyl (co)polymerization of fluorinated inimers and/or fluorinated comonomers. Amphiphilic crosslinked networks with hybridization of these hydrophobic HBFPs and linear hydrophilic poly(ethylene glycol)s are also discussed. As microphase-segregated materials with nanoscale surface heterogeneities, these networks possessed unusual anti-biofouling abilities, atypical sequestration and release behaviors for guest molecules, and special mechanical properties. [source]

Does hybridization between divergent progenitors drive whole-genome duplication?

MOLECULAR ECOLOGY, Issue 16 2009
RICHARD J. A. BUGGS
Abstract Hybridization and whole-genome duplication are both potential mechanisms of rapid speciation which sometimes act in concert. Recent surveys, showing that homoploid hybrid species tend to be derived from parents that are less evolutionarily divergent than parents of polyploid hybrid species (allopolyploids), have been interpreted as supporting a hypothesis that high divergence between hybridizing species drives whole-genome duplication. Here, we argue that such conclusions stem from problems in sampling (especially the omission of autopolyploids) and null model selection, and underestimate the importance of selection. The data simply demonstrate that hybridization between divergent parents has a higher probability of successfully producing a species if followed by polyploidization. [source]

Extreme changes to gene expression associated with homoploid hybrid speciation

MOLECULAR ECOLOGY, Issue 5 2009
MATTHEW J. HEGARTY
Abstract Hybridization is an important cause of abrupt speciation. Hybrid speciation without a change in ploidy (homoploid hybrid speciation) is well-established in plants but has also been reported in animals and fungi. A notable example of recent homoploid hybrid speciation is Senecio squalidus (Oxford ragwort), which originated in the UK in the 18th Century following introduction of hybrid material from a hybrid zone between S. chrysanthemifolius and S. aethnensis on Mount Etna, Sicily. To investigate genetic divergence between these taxa, we used complementary DNA microarrays to compare patterns of floral gene expression. These analyses revealed major differences in gene expression between the parent species and wild and resynthesized S. squalidus. Comparisons of gene expression between S. aethnensis, S. chrysanthemifolius and natural S. squalidus identified genes potentially involved in local environmental adaptation. The analysis also revealed non-additive patterns of gene expression in the hybrid relative to its progenitors. These expression changes were more dramatic and widespread in resynthesized hybrids than in natural S. squalidus, suggesting that a unique expression pattern may have been fixed during the allopatric divergence of British S. squalidus. We speculate that hybridization-induced gene-expression change may provide an immediate source of novel phenotypic variation upon which selection can act to facilitate homoploid hybrid speciation in plants. [source]

Introgressive hybridization of human and rodent schistosome parasites in western Kenya

MOLECULAR ECOLOGY, Issue 23 2008
MICHELLE L. STEINAUER
Abstract Hybridization and introgression can have important consequences for the evolution, ecology and epidemiology of pathogenic organisms. We examined the dynamics of hybridization between a trematode parasite of humans, Schistosoma mansoni, and its sister species, S. rodhaini, a rodent parasite, in a natural hybrid zone in western Kenya. Using microsatellite markers, rDNA and mtDNA, we showed that hybrids between the two species occur in nature, are fertile and produce viable offspring through backcrosses with S. mansoni. Averaged across collection sites, individuals of hybrid ancestry comprised 7.2% of all schistosomes collected, which is a large proportion given that one of the parental species, S. rodhaini, comprised only 9.1% of the specimens. No F1 individuals were collected and all hybrids represented backcrosses with S. mansoni that were of the first or successive generations. The direction of introgression appears highly asymmetric, causing unidirectional gene flow from the rodent parasite, S. rodhaini, to the human parasite, S. mansoni. Hybrid occurrence was seasonal and most hybrids were collected during the month of September over a 2-year period, a time when S. rodhaini was also abundant. We also examined the sex ratios and phenotypic differences between the hybrids and parental species, including the number of infective stages produced in the snail host and the time of day the infective stages emerge. No statistical differences were found in any of these characteristics, and most of the hybrids showed an emergence pattern similar to that of S. mansoni. One individual, however, showed a bimodal emergence pattern that was characteristic of both parental species. In conclusion, these species maintain their identity despite hybridization, although introgression may cause important alterations of the biology and epidemiology of schistosomiasis in this region. [source]