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At least 451 records · Page 25Linked to original sources
PubMed2006

Stage-specific expression of Caenorhabditis elegans ribonuclease H1 enzymes with different substrate specificities and bivalent cation requirements.

Ribonuclease H1 (RNase H1) is a widespread enzyme found in a range of organisms from viruses to humans. It is capable of degrading the RNA moiety of DNA-RNA hybrids and requires a bivalent ion for activity. In contrast with most eukaryotes, which have one gene encoding RNase H1, the activity of which depends on Mg(2+) ions, Caenorhabditis elegans has four RNase H1-related genes, and one of them has an isoform produced by alternative splicing. However, little is known about the enzymatic features of the proteins encoded by these genes. To determine the differences between these enzymes, we compared the expression patterns of each RNase H1-related gene throughout the development of the nematode and the RNase H activities of their recombinant proteins. We found gene-specific expression patterns and different enzymatic features. In particular, besides the enzyme that displays the highest activity in the presence of Mg(2+) ions, C. elegans has another enzyme that shows preference for Mn(2+) ion as a cofactor. We characterized this Mn(2+)-dependent RNase H1 for the first time in eukaryotes. These results suggest that there are at least two types of RNase H1 in C. elegans depending on the developmental stage of the organism.

Alternative Splicing↗
PubMed1999

Comparative immunogenicity of two bivalent botulinum vaccines.

OBJECTIVE: To compare the ability of a new single-dose botulinum vaccine containing a non-mineral oil adjuvant with a single dose of a conventional botulinum vaccine product to produce antibody to Clostridium botulinum types C and D in cattle in Northern Australia. DESIGN AND PROCEDURE: One hundred and fifty Brahman steer weaners were randomly divided into two groups receiving either a single dose of CSL Bivalent Botulinum vaccine or Websters Singvac. Blood samples were collected at 0, 8 and 24 weeks and tested by antibody ELISA. The final samples were also tested by the toxin neutralisation test, to test titres of neutralising antibody. RESULTS: Six months after inoculation, cattle vaccinated with Websters Singvac had ELISA antibody response twice that of CSL conventional product. However, this difference was only evident for neutralising antibody to type C botulinum toxin. Both products produced similar titres of type D neutralising antibody after a single dose. CONCLUSION: Websters' Singvac produces a greater neutralising antibody response to type C botulism upon single inoculation than a conventional vaccine. The product produces an equivalent neutralising antibody response to type D.

Animals↗
PubMed1991

[Immunization of healthy children with mumps-rubella bivalent live vaccine and simultaneous vaccination with mumps-rubella and varicella vaccines].

Bivalent virus vaccine, containing rubella TCRB-19 strain and mumps NK-M46 strain (MR vaccine), was administered to a total of 95 healthy children who had already received measles vaccine or had been infected with wild measles virus. The seroconversion rates for rubella and mumps viruses in subjects having no antibody to rubella or to mumps virus were 99% (75/76) and 97% (63/65), respectively, at 6-8 weeks after vaccination. The seroconversion rates for both rubella and mumps in vaccinees initially seronegative to both viruses were 95% (56/59). Immune responses after MR vaccine injection were comparable to those after administration of monovalent rubella or mumps vaccine. Clinical reactions observed in some subjects who received MR vaccine were mild fever (3.6%), exanthem (8%), lymphadenopathy (1.8%), and swelling of the parotis region (1.8%). MR vaccine could be simultaneously injected with varicella vaccine at the opposite site producing no adverse effect on immune response. Our results indicate that MR vaccine is a safe and effective vaccine, especially for children who have had wild measles or who have received measles vaccine.

Adolescent↗
PubMed2005

Characterization of a novel bivalent morphinan possessing kappa agonist and micro agonist/antagonist properties.

Previous research has shown that compounds with mixed kappa and mu activity may have utility for the treatment of cocaine abuse and dependence. The present study characterizes the pharmacological profile of a bivalent morphinan that was shown to be a kappa opioid receptor agonist and a mu opioid receptor agonist/antagonist. MCL-145 [bis(N-cyclobutylmethylmorphinan) fumarate] is related to the morphinan cyclorphan and its N-cyclobutylmethyl derivative MCL-101 [3-hydroxy-N-cyclobutylmethyl morphinan S-(+)-mandelate]. MCL-145 consists of two morphinans connected by a spacer at the 3-hydroxy position. This compound had K(i) values of 0.078 and 0.20 nM for the kappa and mu opioid receptors, respectively, using radioligand binding assays as shown by Neumeyer et al. in 2003. In the guanosine 5'-O -(3-[(35) S]thiotriphosphate) binding assay, MCL-145 produced an E(max) value of 80% for the kappa opioid receptor and 42% for the mu opioid receptor. The EC(50) values obtained for this compound were 4.3 and 3.1 nM for the kappa and mu opioid receptors, respectively. In vivo MCL-145 produced a full dose-response curve in the 55 degrees C warm water tail-flick test and was equipotent to morphine. The agonist properties of MCL-145 were antagonized by the mu-selective antagonist beta-funaltrexamine and the kappa-selective antagonist nor-binaltorphimine. MCL-145 also acted as a mu antagonist, as measured by the inhibition of morphine-induced antinociception.

Analgesics, Opioid↗
PubMed2005

Interaction of bivalent ligand KDN21 with heterodimeric delta-kappa opioid receptors in human embryonic kidney 293 cells.

KDN21 is a bivalent ligand that contains delta and kappa opioid antagonist pharmacophores linked through a 21-atom spacer. It has been reported that KDN21 bridges delta and kappa receptors that are organized as heterodimers. We have shown previously that when using [(3)H]diprenorphine as radioligand, KDN21 displayed greatly enhanced affinity in this series for coexpressed delta and kappa opioid receptors (CDK). The present study used in vitro expression systems to investigate interactions of members of the KDN series with delta-kappa heterodimers through competition binding using selective ligands and the mitogen-activated protein kinase (MAPK) assay. In this regard, the use of the selective radioligands [(3)H]naltrindole and [(3)H]norbinaltorphimine (nor-BNI) in competition binding studies revealed that KDN21 has much higher affinity than other KDN members for CDK and bound to CDK more selectively relative to mixed delta and kappa opioid receptors or singly expressed delta and kappa opioid receptors. Other experiments revealed that the binding of naltrindole to delta opioid receptors could increase the binding of nor-BNI to kappa opioid receptors and vice versa, suggesting reciprocal allosteric modulation of receptors in the heterodimer. Regarding the selectivity of KDN21 for phenotypic delta and kappa opioid receptors, we investigated the effect of KDN21 on the activation of MAPKs [extracellular signal-regulated kinases 1 and 2 (ERK1/2)] by delta- or kappa-selective agonists. KDN21 inhibited the activation of ERK1/2 by [D-Pen(2),D-Pen(5)]-enkephalin (delta(1)) and bremazocine (kappa(2)) but had no effect on the activation by deltorphin II (delta(2)) and (+)-(5alpha,7alpha,8beta)-N-methyl-N-[7-(1-pyrrolidinyl)-1-oxaspiro[4.5]dec-8-yl]benzeneacetamide (U69593, kappa(1)). 7-Benzylidenenaltrexone (delta(1)) and bremazocine (kappa(2)) significantly reduced the binding of KDN21 to CDK, whereas naltriben (delta(2)) and U69593 produced no such change. Taken together, these data support the idea that the organization of delta and kappa receptors as heterodimers gives rise to delta(1) and kappa(2) phenotypes.

Amino Acid Sequence↗
PubMed2004

Increasing secretion of a bivalent anti-T-cell immunotoxin by Pichia pastoris.

The bivalent anti-T-cell immunotoxin A-dmDT390-bisFv(G(4)S) was developed for treatment of T-cell leukemia and autoimmune diseases and for tolerance induction for transplantation. This immunotoxin was produced extracellularly in toxin-sensitive Pichia pastoris JW102 (Mut(+)) under control of the AOX1 promoter. There were two major barriers to efficient immunotoxin production, the toxicity of the immunotoxin for P. pastoris and the limited capacity of P. pastoris to secrete the immunotoxin. The immunotoxin toxicity resulted in a decrease in the methanol consumption rate, cessation of cell growth, and low immunotoxin productivity after the first 22 h of methanol induction. Continuous cell growth and continuous immunotoxin secretion after the first 22 h of methanol induction were obtained by adding glycerol to the methanol feed by using a 4:1 methanol-glycerol mixed feed as an energy source and by continuously adding a yeast extract solution during methanol induction. The secretory capacity was increased from 22.5 to 37 mg/liter by lowering the induction temperature. A low temperature reduced the methanol consumption rate and protease activity in the supernatant but not cell growth. The effects of adding glycerol and yeast extract to the methanol feed were synergistic. Adding yeast extract primarily enhanced methanol utilization and cell growth, while adding glycerol primarily enhanced immunotoxin production. The synergy was further enhanced by decreasing the induction temperature from 23 to 15 degrees C, which resulted in a robust process with a yield of 37 mg/liter, which was sevenfold greater than the yield previously reported for a toxin-resistant CHO cell expression system. This methodology should be applicable to other toxin-related recombinant proteins in toxin-sensitive P. pastoris.

Bioreactors↗
PubMed1984

Construction of a potential live oral bivalent vaccine for typhoid fever and cholera-Escherichia coli-related diarrheas.

We used the Salmonella typhi galactose epimerase (galE) mutant strain Ty21a, shown to be a safe, effective, living, attenuated oral typhoid vaccine, as a recipient for a recombinant plasmid containing the gene for production of the nontoxic B subunit of the heat-labile enterotoxin of Escherichia coli. The S. typhi derivative, strain SE12, produced heat-labile enterotoxin subunit B that was structurally and immunologically indistinguishable from heat-labile enterotoxin subunit B produced by strains of E. coli harboring the same plasmid. Tests in mice and guinea pigs showed that strain SE12 was safe when given orally and was capable of inducing a significant antitoxic antibody response when injected parenterally. Moreover, it retained the galactose sensitivity of the parent strain, preserving its utility as a typhoid vaccine. This strain may prove to be a useful live oral bivalent vaccine strain for typhoid fever and cholera-E. coli-related diarrheas.

Administration, Oral↗
PubMed1990

Specific immunoglobulin A-secreting cells in peripheral blood of humans following oral immunization with a bivalent Salmonella typhi-Shigella sonnei vaccine or infection by pathogenic S. sonnei.

The ability of bivalent Salmonella typhi-Shigella sonnei vaccine strain 5076-1C to stimulate an intestinal immunoglobulin A response in humans was evaluated by detecting gut-derived, trafficking antibody-secreting cells (ASC) in peripheral blood. Following vaccination, an immunoglobulin A-ASC response to O antigens of S. typhi and S. sonnei was observed in 10 of 13 and 13 of 13 vaccine recipients, respectively. Experimental challenge with pathogenic S. sonnei stimulated an ASC response to the S. sonnei O antigen in all subjects who developed clinical illness. The magnitude of the ASC response to challenge was significantly greater than that resulting from vaccination. Furthermore, compared with the response of the unimmunized controls, individuals previously immunized with 5076-1C demonstrated a significantly greater ASC response following challenge with S. sonnei.

Administration, Oral↗
PubMed1983

Bivalent attachment of antibody onto poliovirus leads to conformational alteration and neutralization.

The treatment of nonsaturating, neutralizing antibody-poliovirus complexes with papain generally led to the loss of viral neutralization and to the loss of the neutralization-associated change in the isoelectric point (pI) of the virion. Subsequent treatment with anti-immunoglobulin G antibodies restored the neutralization of the virus and the alteration of the viral pI. It appears that, under nonsaturating conditions, poliovirus neutralization by an antibody is dependent upon the ability of the antibody to bivalently attach to the virion. Exceptions are monospecific neutralizing antibodies with an affinity for capsid protein VP3.

Antibodies, Monoclonal↗
PubMed2001

Individual and bivalent vaccines based on alphavirus replicons protect guinea pigs against infection with Lassa and Ebola viruses.

Lassa and Ebola viruses cause acute, often fatal, hemorrhagic fever diseases, for which no effective vaccines are currently available. Although lethal human disease outbreaks have been confined so far to sub-Saharan Africa, they also pose significant epidemiological concern worldwide as demonstrated by several instances of accidental importation of the viruses into North America and Europe. In the present study, we developed experimental individual vaccines for Lassa virus and bivalent vaccines for Lassa and Ebola viruses that are based on an RNA replicon vector derived from an attenuated strain of Venezuelan equine encephalitis virus. The Lassa and Ebola virus genes were expressed from recombinant replicon RNAs that also encoded the replicase function and were capable of efficient intracellular self-amplification. For vaccinations, the recombinant replicons were incorporated into virus-like replicon particles. Guinea pigs vaccinated with particles expressing Lassa virus nucleoprotein or glycoprotein genes were protected from lethal challenge with Lassa virus. Vaccination with particles expressing Ebola virus glycoprotein gene also protected the animals from lethal challenge with Ebola virus. In order to evaluate a single vaccine protecting against both Lassa and Ebola viruses, we developed dual-expression particles that expressed glycoprotein genes of both Ebola and Lassa viruses. Vaccination of guinea pigs with either dual-expression particles or with a mixture of particles expressing Ebola and Lassa virus glycoprotein genes protected the animals against challenges with Ebola and Lassa viruses. The results showed that immune responses can be induced against multiple vaccine antigens coexpressed from an alphavirus replicon and suggested the possibility of engineering multivalent vaccines based upon alphavirus vectors for arenaviruses, filoviruses, and possibly other emerging pathogens.

Alphavirus↗
PubMed2005

Live bivalent vaccine for parainfluenza and influenza virus infections.

Influenza and human parainfluenza virus infections are of both medical and economical importance. Currently, inactivated vaccines provide suboptimal protection against influenza, and vaccines for human parainfluenza virus infection are not available, underscoring the need for new vaccines against these respiratory diseases. Furthermore, to reduce the burden of vaccination, the development of multivalent vaccines is highly desirable. Thus, to devise a single vaccine that would elicit immune responses against both influenza and parainfluenza viruses, we used reverse genetics to generate an influenza A virus that possesses the coding region for the hemagglutinin/neuraminidase ectodomain of parainfluenza virus instead of the influenza virus neuraminidase. The recombinant virus grew efficiently in eggs but was attenuated in mice. When intranasally immunized with the recombinant vaccine, all mice developed antibodies against both influenza and parainfluenza viruses and survived an otherwise lethal challenge with either of these viruses. This live bivalent vaccine has obvious advantages over combination vaccines, and its method of generation could, in principle, be applied in the development of a "cocktail" vaccine with efficacy against several different infectious diseases.

Animals↗
PubMed1987

Ring XY bivalent: a new phenomenon at metaphase I of meiosis in man.

The unusual appearance of a ring XY bivalent at metaphase I of meiosis is reported in some cells of an oligospermic human male. Higher than usual frequencies of ring configuration in the XY pair were also observed during prophase I. The defect could be attributable to loss of some DNA sequences from the distal heterochromatic tip of the Y chromosome long arm.

Adult↗
PubMed1985

Passive K+-Cl- fluxes in low-K+ sheep erythrocytes: modulation by A23187 and bivalent cations.

A fraction of the ouabain-resistant (OR) K+ flux of low-K+ (LK) sheep erythrocytes is Cl- dependent (K+-Cl- transport) and is activated reversibly by cell swelling or irreversibly by treatment with N-ethylmaleimide (NEM). The effect of the ionophore A23187 plus bivalent cations (Me2+) or ethyleneglycol-bis(beta-aminoethylether)-N,N'-tetraacetic acid (EGTA) was studied on K+-Cl- transport in control or NEM-treated LK cells. The following observations were made. 1) A23187 (6 microM), at a hematocrit of 10% (vol/vol) and in the presence of 1 mM EGTA, activated severalfold OR K+-Cl- transport in shrunken or swollen cells but failed to stimulate further K+-Cl- flux in NEM-treated cells. 2) In the absence of EGTA, but at very low external Ca2+ concentrations [( Ca2+]o = 10(-7) M), A23187 stimulated OR K+-Cl- flux in controls less than with EGTA and inhibited it slightly in NEM-treated cells. 3) When [Ca2+]o was raised to 10(-3) M, an almost complete inhibition of OR K+-Cl- fluxes occurred in shrunken, swollen, or NEM-treated cells. 4) Other Me2+ inhibited OR K+-Cl- flux in the presence of A23187 in the following order of decreasing potency: Mn2+ much greater than Ca2+ greater than Mg2+ greater than Sr2+ much much greater than Ba2+. 5) Stimulation of OR K+-Cl- flux by A23187 +/- EGTA and inhibition by A23187 + Ca2+ were reversible and did not alter significantly cellular ATP. 6) The stimulatory effect of A23187 plus EGTA, perhaps by Me2+ removal, on K+-Cl- flux and its inhibition by Ca2+ were reversibly abolished in metabolically depleted cells.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1983

Heterochromatin heterogeneity in Chinese hamster sex bivalents.

The heterochromatin of the Chinese hamster sex chromosomes was analyzed by different banding techniques. Combined results obtained after differential Ba(OH)2 treatment, BrdU incorporation, Giemsa 11, and staining with quinacrine permitted the characterization of different regions in the heterochromatic portions of the X and Y chromosomes. In the light of these observations, the chiasma observed in the sex bivalent of Chinese hamster spermatocytes was localized within specific heterochromatic regions. The homologous segments consist of the entire short arm of the Y and the distal end of the long arm of the X up to band q21. These regions are probably not rich in highly repetitive DNA sequences, which are more resistant to alkali denaturation, or in satellite DNA, which is stained by Giemsa 11. Thus the heterochromatin in the homologous regions of the sex chromosomes allows the formation of a chiasma. The heterogeneity found in these heterochromatic regions may help to establish a more precise relationship between heterochromatin and recombination.

Animals↗
PubMed1997

Targeting medullary thyroid carcinomas with bispecific antibodies and bivalent haptens. Results and clinical perspectives.

The present article reviews the clinical trials that have been performed in recurrent medullary thyroid carcinoma patients with the Affinity Enhancement System. This technique uses bispecific antibodies to target radiolabelled bivalent haptens to tumour cells. Its sensitivity in the detection of known tumour sites is high (90%) and this technique also achieves good sensitivity (61%) in the detection of occult disease as revealed by abnormal thyrocalcitonin blood levels. Due to its high targeting capacity, this technique is now considered for use as a therapeutic agent in medullary thyroid carcinoma patients.

Antibodies, Anti-Idiotypic↗
PubMed1977

Extensive pairing of the XY bivalent in mouse spermatocytes as visualized by whole-mount electron microscopy.

Autosomes and sex chromosomes of mouse spermatocytes were examined during zygotene, pachytene, and diplotene by a whole-mount electron-microscope technique after cell dispersion in a detergent solution (Nonidet-P40). Zygotene, pachytene, and diplotene stages can be adequately identified in the preparations. Thus, asynchronous side-by-side pairing of homologous autosomes, some of them displaying attached nucleoli, defines zygotene. Pachytene is identified by complete pairing of homologues. Diplotene is characterized by disjunction of bivalents (autosomes and sex chromosomes), lack of autosomal-attached nucleoli, divergent expansions observed at lateral element endings of disassembled synaptonemal complexes, end-to-end association of the XY pair and well defined outward deformations ('bulges') along sex chromosomal axial cores. X and Y chromosomes display at pachytene an extensive side-by-side pairing segment which decreases in length as meiotic prophase advances. Each sex chromosomal axial core appears double and is formed by close apposition of 2 nearly parallel elements displayed separately along the entire length of the chromosomal core. This double structural feature suggests that each sex chromosomal axial core is presumably composed of 2 chromatid axial cores, each of which, in turn, constitutes the respective lateral elements of short synaptonemal complexes observed at the unpaired segment.

Animals↗
PubMed1979

An experimental analysis of bivalent interlocking in spermatocytes of the new Triturus vulgaris.

Interlocked bivalents at 1st meiotic metaphase are relatively uncommon in spermatocytes of the newt Triturus vulgaris, but their frequency of occurrence can be significantly increased by subjecting newts to a 24-h heat shock. Newt spermatocytes are sensitive to a heart shock at any stage between the end of premeiotic S and mid to late pachytene. The heat shock does not cause evidence desynapsis, nor does it significantly affect chiasma frequency; therefore the interlocked condition induced in spermatocytes which were subjected to a heat shock when they were in zygotene or pachytene is unlikely to be a consequence of synaptic trapping. By way of explanation it is suggested that a heat shock may cause telomers to detach from the nuclear membrane, or from the synaptonemal complex where the latter is attached to the membrane, thus allowing non-homologous chromonemata to become interwined before chiasmata have formed. If this explanation is valid, it is then further suggested that the recombination process which results in chiasma formation probably takes place in chromosomal regions lying outside the synaptonemal complex, rather than inside, between its 2 lateral elements.

Animals↗