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At least 145 records · Page 8Linked to original sources
PubMed1977

Tumor x host cell hybrids in the mouse: chromosomes from the normal cell parent maintained in malignant hybrid tumors.

We investigated whether the malignancy of hybrids between normal and malignant cells could be correlated with the loss of specific genes borne by specific chromosomes from the normal parent cells. Tumors produced in mice by the inoculation of Cl.1D cells (an L cell derivative) contained tumor x host cell hybrids. Hybrid cell populations isolated from 14 tumors were injected into 123 mice, of which 108 (87%) developed tumors. Metaphases of growing hybrid cell tumors were analyzed by use of a trypsin-Giemsa banding technique. The chromosomes contributed by the host (normal) parent cell could be distinguished from Cl.1D chromosomes, since the latter exhibited morphologic differences due to rearrangements. In the 14 hybrid tumors analyzed, we found that any one of the chromosomes of the host cell might be present, which indicated that none of the chromosomes from the normal cell bore genetic information capable of suppressing the malignancy of Cl.1D cells. Absence of complimentation in the hybrids suggested that, even if the accumulation of several mutations were necessary for malignant tumor growth of Cl.D1 cells, none of these mutations is recessive.

Animals↗
PubMedDate not supplied

Hybridization among the ancient mariners: characterization of marine turtle hybrids with molecular genetic assays.

Reports of hybridization between marine turtle species (family Cheloniidae) have been difficult to authenticate based solely on morphological evidence. Here we employ molecular genetic assays to document the sporadic, natural occurrence of viable interspecific hybrids between species representing four of the five genera of cheloniid sea turtles. Using multiple DNA markers from single-copy nuclear loci, eight suspected hybrids (based on morphology) were confirmed to be the products of matings involving the loggerhead turtle (Caretta caretta) x Kemp's ridley (Lepidochelys kempii) (N = 1 specimen), loggerhead turtle x hawksbill (Eretmochelys imbricata) (N = 2), loggerhead turtle x green turtle (Chelonia mydas) (N = 4), and green turtle x hawksbill (N = 1). Molecular markers from mitochondrial DNA permitted identification of the maternal parental species in each cross. The species involved in these hybridization events represent evolutionary lineages thought to have separated 10-75 million years ago (mya) and thus may be among the oldest vertebrate lineages capable of producing viable hybrids in nature. In some cases, human intervention with the life cycles of marine turtles (e.g., through habitat alteration, captive rearing, or attempts to establish new breeding sites) may have increased the opportunities for interspecific hybridization.

Animals↗
PubMed1991

Biotinylated probes to detect Leptospira interrogans on dot blot hybridization or by in situ hybridization.

Total genomic biotinylated probes which can identify leptospires by hybridization on filters or by in situ hybridization are described in this study. According to the weak G + C content of the strains studied (35-39%) and owing to the decreasing melting temperature (Tm) due to overbiotinylation, hybridization and wash temperatures were optimized at 33 degrees C and at 42 degrees C respectively. Fourteen serovars of Leptospira interrogans belonging to 11 different serogroups and three serovars of Leptospira biflexa were used in this study. Cross-hybridization results show that it is possible, by means of such probes, specifically to recognize pathogenic strains. These probes did not hybridize with the three saprophytic strains: L. buenos-aires, L. patoc and L. andamana. We also ran a total genomic probe, specific to the serovar buenos-aires which hybridizes only with homologous DNA.

Biotin↗
PubMed1981

The suppression of tumorigenicity in human X mouse cell hybrids. I. Derivation of hybrid clones, chromosome analysis and tumorigenicity studies.

The nature of genetic determinants associated with the regulation of malignancy was investigated in interspecies (human X mouse) cell hybrids. An apparently transformed diploid human cell strain (ME) was fused with a malignant, HGPRT-deficient, mouse cell line (A9). A primary hybrid with an unusual stellate appearance that contained the isozymic forms of lactate dehydrogenase from both parent cells was selected. Through further cloning, four tertiary hybrid cell clones were obtained in which malignancy was substantially inhibited and in which the human chromosome content was reduced or completely absent. Two of the hybrid clones whose malignant phenotype was stably suppressed contained no apparent human chromosomes, whereas the other two, whose phenotype was unstable, contained a part of a human chromosome. Our results are consistent with the view that suppression of malignancy occurs in interspecies hybrids consisting of a malignant mouse and a transformed human parent cell. At present, however, any assignment of a specific human chromosome, or specific fragments thereof, that may suppress the expression of malignancy in these hybrids, is a matter of conjecture.

Adenoma↗
PubMed1987

Construction and characterisation of a hybrid-hybrid monoclonal antibody recognising both carcinoembryonic antigen (CEA) and vinca alkaloids.

Recent developments of hybridoma technology have allowed us to prepare a bispecific monoclonal antibody recognising both the tumour-associated antigen carcinoembryonic antigen (CEA) and the cytostatic vinca alkaloids. The yields of the hybrid-hybrid 28.19.8 monoclonal after affinity chromatography purification are close to 50% of the total Ig produced. The hybrid-hybrid has a molecular weight ca. 150,000 daltons. The heavy chains of the hybrid-hybrid are a gamma 1 heavy chain from the parental anti-CEA monoclonal and a gamma 2a heavy chain from the anti-vinca alkaloid donor lymphocytes.

Animals↗
PubMed1987

Specific in vitro and in vivo drug localisation to tumour cells using a hybrid-hybrid monoclonal antibody recognising both carcinoembryonic antigen (CEA) and vinca alkaloids.

By using a bispecific monoclonal antibody recognising both carcinoembryonic antigen (CEA) and the cytostatic vinca alkaloid drugs we have been able to show specific tumour localisation of vinca alkaloids. In vitro studies with sections of human colorectal tumours have demonstrated that the hybrid-hybrid 28.19.8 monoclonal is able to specifically localise vindesine to cells expressing CEA. Furthermore, the hybrid-hybrid 28.19.8 localises in vivo preferentially to tumour tissues in nude mice bearing the MAWI human xenograft tumour. This tumour-bound hybrid-hybrid monoclonal antibody induces profound changes in the bio-distribution of vinca alkaloid drugs, targeting them specifically to tumour tissues.

Animals↗
PubMed1996

Generation of a panel of radiation-reduced hybrids containing human 11q22-23 fragments bearing a HPRT selective marker: identification of hybrids carrying various subregions around the ataxia-telangiectasia locus.

A human-mouse monochromosomal hybrid that contains a human t(X;11) translocated chromosome carrying pter-->q23 segment of chromosome 11 was used to construct a panel of radiation-reduced hybrids. The hypoxhanthine phosphoribosyltransferase (HPRT) gene located close to the translocation breakpoint was used as a marker to select for the hybrids that preferentially retain the 11q22-23 region. Twenty-three HAT-resistant hybrids were isolated and screened by polymerase chain reaction (PCR) for the retention of 31 loci on 11q22-23 region. Among the 14 hybrids that had breakpoints within the 11q22-23 region, 6 hybrids contained fragments that extend either from centromere or telomere to the 5-Mb region spanned by GRIA4 and FDX, carrying various breakpoints within the region. This subpanel could be a potential resource to analyze the ataxia-telangiectasia disease locus and its neighboring region.

Animals↗
PubMed1978

Pharmacologic responses of cells of a neuroblastoma X glioma hybrid clone and modulation of synapses between hybrid cells and mouse myotubes.

Cells of the hybrid clone NG108-15 responded to 5-hydroxytryptamine (5-HT), dopamine or acetylcholine with graded depolarizations involving membrane conductance increases. Responses desensitized during continuous application of the neurotransmitters, and responses to 5-HT and dopamine cross-desensitized: a desensitizing application of one neurotransmitter also desensitized the hybrd cell to the other neurotransmitter. 5-HT and acetylcholine did not cross-desensitize. The hybrid cell 5-HT response was not attenuated by D-LSD, and was blocked by 10(-5) M morphine, although not via binding to naloxone-sensitive opiate receptors. 5-HT or the prostaglandin PGF2alpha caused the release of acetylcholine at the synapses of hybrid cells with mouse myotubes. Application of 5-HT or PGF2alpha also facilitated the synaptic release elicited by hybrid cell action potentials. Following treatment with the antimitotic agent cytosine arabinoside, co-cultures of hybrid cells and mouse myotubes exhibited plentiful synaptic connections only if maintained in medium containing 1 mM dibutyryl cAMP (dBcAMP). After X-irradiation, co-cultures were synaptically active even in the absence of dBcAMP. Thus, methods have been found to regulate both the short-term and long-term synaptic activity of NG108-15 hybrid cells.

Acetylcholine↗
PubMed1984

Evidence for the presence of a hybrid of phosphoglyceromutase/bisphosphoglyceromutase in the red cells: partial characterization of the hybrid.

Purified phosphoglyceromutase was hybridized in vitro with pure biphosphoglyceromutase . The hybrid showed an electrophoretic mobility identical to that of the intermediate band of red cell phosphoglyceromutase activity in the hemolysate patterns. Electrophoretic tests showed that the partially purified hybrid displayed both bisphosphoglyceromutase and phosphoglyceromutase activities and that the sample contained also a small portion of non-hybrid bisphosphoglyceromutase . By constrast to a non-hybrid mixture of the two purified mutases the hybrid exhibited heat instability of bisphosphoglyceromutase activity and neutralization of phosphoglyceromutase activity by anti- bisphosphoglyceromutase antibody.

Bisphosphoglycerate Mutase↗
PubMed2003

Hybrid global optimization algorithms for protein structure prediction: alternating hybrids.

Hybrid global optimization methods attempt to combine the beneficial features of two or more algorithms, and can be powerful methods for solving challenging nonconvex optimization problems. In this paper, novel classes of hybrid global optimization methods, termed alternating hybrids, are introduced for application as a tool in treating the peptide and protein structure prediction problems. In particular, these new optimization methods take the form of hybrids between a deterministic global optimization algorithm, the alphaBB, and a stochastically based method, conformational space annealing (CSA). The alphaBB method, as a theoretically proven global optimization approach, exhibits consistency, as it guarantees convergence to the global minimum for twice-continuously differentiable constrained nonlinear programming problems, but can benefit from computationally related enhancements. On the other hand, the independent CSA algorithm is highly efficient, though the method lacks theoretical guarantees of convergence. Furthermore, both the alphaBB method and the CSA method are found to identify ensembles of low-energy conformers, an important feature for determining the true free energy minimum of the system. The proposed hybrid methods combine the desirable features of efficiency and consistency, thus enabling the accurate prediction of the structures of larger peptides. Computational studies for met-enkephalin and melittin, employing sequential and parallel computing frameworks, demonstrate the promise for these proposed hybrid methods.

Algorithms↗
PubMed1995

A hybrid sigma subunit directs RNA polymerase to a hybrid promoter in Escherichia coli.

Most of the sigma (transcriptional initiation specificity) subunits of RNA polymerase, from a wide range of eubacteria, show strong elements of amino acid sequence similarity. There is evidence that two of the "conserved" regions, 2.4 and 4.2, are involved in recognition of the consensus DNA sequences centred near -10 and -35, respectively, which define promoter sites for the initiation of transcription. Since all the alternative sigma subunits of the above type function by binding to a common core polymerase enzyme in a given bacterium, it can be predicted that a hybrid sigma might be functional, and if so should permit RNA polymerase to initiate only at a correspondingly hybrid promoter. To test these predictions, a hybrid gene encoding the amino-proximal 529 amino acids of the major Escherichia coli sigma protein, sigma 70 (including region 2.4) followed by the last 82 amino acids of the heat-shock sigma protein, sigma 32 (including region 4.2) was constructed and fused to Plac on a plasmid. Major-consensus, heat-shock and hybrid promoters were fused to a chloramphenicol acetyl transferase (CAT) reporter gene on a compatible plasmid. CAT assays showed that, as predicted, a promoter with a "heat-shock" -35 consensus and a "major" -10 consensus sequence (PHM) required Plac-dependent production of the hybrid sigma (sigma 70-32) for activity in vivo. PHM then became a strong promoter. The hybrid sigma gene has potential advantages over its parents for structure-function studies.

Amino Acid Sequence↗
PubMed1992

Recombinant human adenoviruses containing hybrid adenovirus type 5 (Ad5)/Ad12 E1A genes: characterization of hybrid E1A proteins and analysis of transforming activity and host range.

Hybrid adenovirus type 12 (Ad12)/Ad5 E1A genes were constructed by homologous recombination in Escherichia coli, a technique which offers several advantages over conventional mutagenesis for genetic analysis of proteins. In particular, functional differences between the proteins can be mapped by correlating the replacement of specific sequences with the acquisition of new properties, and there is no requirement for common unique restriction sites or polymerase chain reaction strategies to construct the hybrids. Recombinant adenoviruses expressing these hybrid E1A proteins were capable of replicating efficiently in HeLa cells, with the exception of one construct which contained a hybrid transactivation domain. The transforming activity of the hybrid E1A constructs was assayed by DNA transfection of primary baby rat kidney cells. Plasmids containing Ad12 E1 were approximately 20-fold less efficient at transformation than those with E1 of Ad5, and it was found that two regions in exon 1 of E1A mediate this difference. No differences were found in the abilities of any hybrid E1A proteins to bind to cellular proteins previously determined to be important for transformation by E1A.

Adenovirus Early Proteins↗
PubMed1986

Carotid bifurcation: results of spliced hybrid subtraction angiography compared with standard temporal and hybrid images.

Hybrid subtraction angiography can provide a higher percentage of two-view examinations of the carotid bifurcation. Unfortunately, hybrid images are lower in signal-to-noise ratio (S/N) than temporal subtraction angiographic images and are often described as "noisy." The present study was designed to determine if the lower S/N of hybrid subtraction angiography could be restricted to the contralateral carotid bifurcation by "splicing" a small hybrid region of interest into a larger temporal image. Twenty-two selected images were examined on four image quality scales by seven readers. Similarity of reader scoring was characterized by cluster analysis and was good. Ipsilateral carotid arteries were preferred as temporal images, and contralateral carotid arteries were preferred as hybrid images. The splice image was preferred over both. The splice image retained the superiority of temporal subtraction angiography in the region of the ipsilateral carotid artery and incorporated the superiority of hybrid subtraction angiography in the region of the contralateral carotid artery. The splice image incorporated the best features of both image modalities.

Aged↗
PubMed1989

Amino acid sequence of an alpha-delta-glycophorin hybrid. A structure reciprocal to Sta delta-alpha-glycophorin hybrid.

In this report we examine the primary sequence of a variant glycophorin obtained from erythrocytes of an individual who exhibits an unusual MNSs blood group phenotype. We show that this protein is a hybrid molecule constructed from sequences of alpha- and delta-glycophorins (glycophorins A and B) in a alpha-delta arrangement. Serological typing revealed that the donor's phenotype was M+N+S+s+U+; yet his erythrocytes reacted with some but not all examples of anti-S antisera. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed a variant glycophorin band, and immunoblotting and reaction with N-glycanase suggested that its amino terminus resembled that of M-alpha-glycophorin but that its carboxyl terminus did not. A preparation highly enriched in the variant was obtained and used to generate peptide fragments for sequencing. The sequence revealed that the variant was a hybrid molecule whose amino terminus corresponded to M-alpha-glycophorin and whose carboxyl terminus corresponded to S-delta-glycophorin. CNBr cleavage of the variant glycophorin yielded four peptides. The sequence of the amino-terminal CNBr peptide (residues 1-8) was identical to the amino-terminal octapeptide of M-alpha-glycophorin. The proceeding peptide (residues 9-61) contained a segment identical to residues 9-58 of alpha glycophorin, but its carboxyl-terminal sequence had the Gly-Glu-Met sequence from S-delta-glycophorin (residues 27-29). The other two peptides, insoluble in aqueous solutions, contained highly hydrophobic sequences, identical to residues 30-52 and 53-68 of delta-glycophorin. Sequences of overlapping peptides generated by trypsin and V8 protease confirmed the hybrid nature of the variant glycophorin: residues 1-58 were identical to residues 1-58 of M-alpha-glycophorin, and residues 59-100 were entirely identical to residues 27-68 of S-delta-glycophorin. The variant glycophorin is expected to have 4 additional residues at its carboxyl terminus that correspond to the carboxyl-terminal residues 69-72 of delta-glycophorin. The amino acid sequence arrangement of the variant alpha-delta-glycophorin is an exact reciprocal of that found in another hybrid glycophorin, Sta, that is a delta-alpha hybrid. We propose that the two hybrid glycophorins represent the two possible products resulting from a reciprocal recombination event.

Amino Acid Sequence↗
PubMedDate not supplied

[Characteristics of the segregation of a new remote hybrid compared to an earlier remote hybrid obtained from crossing Saccharomyces cerevisiae with Schizosaccharomyces pombe].

The variability of two remote hybrids between Saccharomyces cerevisiae and Schizosaccharomyces pombe was compared in the course of their vegetative splitting. Hybrid 69 was obtained upon fusion of germinating spores, hybrid 92 was produced by copulation of vegetative cells. The variability of hybrid 92 was more diverse in terms of the size and shape of its cells; moreover, the hybrid yielded adenine-dependent haploid cells typical of S. cerevisiae with red colonies as well as dividing cells very similar to S. pombe with cross septa. Hybrid 69 did not produce such cultures.

Ascomycota↗
PubMed1990

Direct nonradioactive in situ hybridization of somatic cell hybrid DNA to human lymphocyte chromosomes.

Biotinylated DNA from various human-rodent hybrids was hybridized to human lymphocyte spreads after preannealing of the repeated sequences with sonicated total human DNA. Fluorescent labeling was achieved by successive treatments with fluorescein-labeled avidin and biotinylated antiavidin antibody. The use of labeled total DNA from hybrids with known chromosome composition permits the fluorescent staining-("painting") of specific chromosomes, or parts thereof, in human lymphocyte metaphases. Alternatively, the human chromosome content of cell hybrids with unknown chromosome composition is directly assessed from the labeling pattern of human lymphocyte spreads using the total hybrid DNA as probe.

Animals↗
PubMed1992

Isolation and characterization of radiation-reduced hybrids containing portions of the proximal long arm of the human X chromosome: identification of hybrids containing the Menkes' disease locus.

The proximal long arm of the human X chromosome (Xcen----Xq13) encompasses an estimated 23 megabases of DNA and contains numerous identified genetic loci. In order to generate a highly enriched source of DNA from this region, radiation-reduced human-hamster hybrids were constructed and screened to identify those that contained at least part of proximal Xq. Eight such hybrids were identified and characterized by Southern blot and fluorescence in situ hybridization analyses to determine more precisely the human DNA complement in each. One hybrid contains the entire proximal long arm and will be useful for mapping Xcen----Xq13 in its entirety and for localizing genes within this region. Another hybrid contains a smaller portion of the proximal long arm that includes the region reported to contain the gene for Menkes' disease.

Base Sequence↗