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PubMed2002

A demonstration of a 1:1 correspondence between chiasma frequency and recombination using a Lolium perenne/Festuca pratensis substitution.

A single chromosome of the grass species Festuca pratensis has been introgressed into Lolium perenne to produce a diploid monosomic substitution line 2n = 2x = 14. The chromatin of F. pratensis and L. perenne can be distinguished by genomic in situ hybridization (GISH), and it is therefore possible to visualize the substituted F. pratensis chromosome in the L. perenne background and to study chiasma formation in a single marked bivalent. Recombination occurs freely in the F. pratensis/L. perenne bivalent, and chiasma frequency counts give a predicted map length for this bivalent of 76 cM. The substituted F. pratensis chromosome was also mapped with 104 EcoRI/Tru91 and HindIII/Tru91 amplified fragment length polymorphisms (AFLPs), generating a marker map of 81 cM. This map length is almost identical to the map length of 76 cM predicted from the chiasma frequency data. The work demonstrates a 1:1 correspondence between chiasma frequency and recombination and, in addition, the absence of chromatid interference across the Festuca and Lolium centromeres.

Chromosome Mapping↗
PubMed1996

Evaluation of live attenuated influenza vaccines in children 6-18 months of age: safety, immunogenicity, and efficacy. National Institute of Allergy and Infectious Diseases, Vaccine and Treatment Evaluation Program and the Wyeth-Ayerst ca Influenza Vaccine Investigators Group.

Live attenuated, cold-adapted (ca) monovalent and bivalent influenza A vaccines were evaluated in seronegative infants (ages 6-18 months) in a double-blind placebo-controlled trial to assess safety and immunogenicity. A total of 182 seronegative subjects received a single intranasal dose (10(6.2) TCID50) of ca A/Kawasaki/9/86 (H1N1) or ca A/Los Angeles/2/87 (H3N2), both as a bivalent vaccine, or placebo. Respiratory and systemic symptoms did not differ between groups after vaccination. Hemagglutination antibody seroconversions (> or = 1:8) to H3N2 exceeded 90%. In contrast, seroconversions to A/Kawasaki/9/86 (H1N1) were significantly less frequent in bivalent ca vaccine recipients (31%) than in monovalent ca H1N1 recipients (83%) (P < .002). During a subsequent H3N2 epidemic, nasal washes were cultured for viruses from any subject with respiratory illness. H3N2 infections documented by virus isolation were reduced by 65% in ca H3N2 recipients compared with placebo or ca HIM recipients (P = .01).

Adaptation, Physiological↗
PubMed1995

A monovalent C mu 4-specific ligand enhances the activation of human B cells by membrane IgM cross-linking ligands.

The ligand-receptor binding requirements for achieving full B cell activation through the membrane immunoglobulin (mIg) signaling pathway are relatively demanding, and mIg-antigen engagements which fall below these critical thresholds cause, at most, only the partial activation of B cells. In an effort to resolve new means of enhancing the efficacy of mIgM-mediated signal transduction, as well as to further understand the process by which mIgM-mediated signals are initiated, we have explored the mechanism for a previously reported synergy between certain mixtures of murine anti-IgM mAbs in eliciting human B cell DNA synthesis. We here report that striking synergy occurs when any of several relatively high affinity mAbs specific for diverse domains of mIgM are combined in culture with the relatively low affinity C mu 4-specific ligand, mAb IG6. Although B cell activation was dependent upon the bivalency, and hence mIgM cross-linking potential, of the high affinity ligand, low affinity mAb IG6 could enhance the activation process when present as a monovalent Fab' fragment. This did not appear due to F(ab')2 contamination or Fab' aggregation, since IG6 Fab' preparations were notably compromised in several other functions requiring ligand bivalency. Pulsing studies revealed that the C mu 4-specific ligand exhibits its functional effects only when stimulatory mIgM receptor cross-links are being formed by bivalent ligands, and that IG6 Fab' enhancement is most notable during the later interval of the prolonged mIgM signaling process that leads to S phase entry. A unique region of the membrane-proximal IgM domain may be important for Fab'-mediated enhancement, since Fab' fragments that bind with higher affinities to distinct sites on C mu 4 were not as effective at mediating this phenomenon. Several possibilities for the adjuvant effects of this C mu 4-specific Fab' on B cell responses triggered by mIgM crosslinking ligands are discussed, including the possibility that IG6 Fab' influences the potential for mIgM dimer formation or interactions of mIgM with other signal-transducing molecules.

Animals↗
PubMedDate not supplied

Studies of synaptonemal complexes in farm mammals--a review.

For the last 10 years extensive studies of synaptonemal complexes in farm mammals were carried out. In this article a survey of the obtained results is presented. Studies on synaptic behavior of chromosomes at pachytene substage of meiotic prophase I in carriers of centric fusions (cattle, sheep, goat, pig, and blue fox) showed that the trivalent is rarely involved in an association with the sex bivalent. In carriers of reciprocal translocations (pig and cattle) a wide range of synaptic configurations were found. Besides the expected cross-shaped quadrivalent, the following configurations were observed: open quadrivalent, trivalent plus univalent, and two heteromorphic bivalents. The latter configurations were quite frequently involved in the association with XY bivalent. Studies of synaptonemal complexes in interspecies hybrids (cattle x zebu, river x swamp buffalo, blue fox x silver fox) revealed a wide range of synaptic disturbances. Cases of pericentric inversions, aneuploidy, and chromosomal polymorphisms are also reviewed. This survey shows that synaptonemal complex analysis is a very useful tool in studies on causes of altered fertility in carriers of abnormal chromosome complement.

Aneuploidy↗
PubMed1991

Use of hydrophilic diamines for bridging of two opioid peptide pharmacophores. Synthesis and receptor binding of two new analogues.

The bivalent ligand approach, which assumes that two pharmacophores are connected by a spacer, was used to design receptor type-selective ligands for opioid receptors. The first two opioid peptide bivalent ligands with different spacer lengths containing different numbers of hydroxyl groups, (Tyr-D-Ala-Gly-Phe-NH-CH2-CHOH-)2 (Tyr-D-Ala-Gly-Phe-NH-CH2-CHOH-CHOH-)2, were synthesized and their binding to mu, delta, and kappa opioid receptors was characterized. Both analogues were found to possess high opioid in vitro activities. The length of the hydrophilic spacer does not affect the affinity for delta receptors, whereas shorter spacer length increases affinity for mu and even more so for kappa receptors. Thus receptor type-selective peptides for opioid receptors can be designed using the bivalent approach.

Amino Acid Sequence↗
PubMed1995

Synaptonemal complex analysis in a boar with tertiary, trisomy, product of a rcp(7;17)(q26;q11) translocation.

Synaptonemal complex analysis by electron microscopy showed a trivalent formed by pairing of chromosome 17(7) with chromosome pair 7 in two zygotene cells. In 89 pachytene cells (75.4%) the chromosome 17(7) occurred as a univalent with the axis being unpaired, in the shape of a ring (60.6%), a rod (38.2%) or self-paired (1 cell). Thirty-four (38.2%) of the univalents were paired or associated with the sex bivalent. Twenty-five (21.2%) of the 118 pachytene cells analysed demonstrated different types of trivalent pairing: the 17(7) chromosome paired with chromosome pair 7 in 24 cells and with pair 17 in one cell. Moreover, in 4 cells, chromosome 17(7) was paired with both pair 7 and pair 17, forming a pentavalent. Trivalent association with XY was observed in 4 cells. Nineteen bivalents plus a univalent (8 cells), and eighteen bivalents plus a trivalent (11 cells), were found during conventional meiotic investigation of diakinesis-metaphase I. Though the boar demonstrated a normal testicular histology, a qualitatively deviant semen picture was evident.

Animals↗
PubMed1979

Synergistic protection against experimental cholera by immunization with cholera toxoid and vaccine.

Rabbits were immunized with two parenteral injections of Wellcome toxoid PX389A, Wyeth toxoid 20101, or Merck bivalent vaccine. Other groups of rabbits were immunized with combinations of the Merck vaccine and each of the two toxoids. Antitoxin responses were monitored in each group of rabbits before livecell challenge of each animal by the ligated intestinal loop assay. Inaba and Ogawa strains of Vibrio cholerae were used for challenge experiments. Basically, the data indicate that the toxoids were equivalent in antigenic potency and antitoxin responses were unaffected by combination of the toxoids with the whole-cell vaccine. The 50 microgram doses of each toxoid as well as the 4 X 10(9) cells of the bivalent vaccine provided the same magnitude of protection against live-cell challenge with either Inaba or Ogawa vibrios. Immunization with either toxoid in combination with the bivalent vaccine resulted in a synergistic protective response against live-cell challenge of intestinal loops with V. cholerae. Synergistic protection was observed when toxoid and vaccine were administered together by the oral and parenteral routes. Maximum protection was obtained when rabbits were immunized with the combined toxoid-whole-cell vaccine administered by both oral and parenteral routes.

Animals↗
PubMed1977

Clinical evaluation of commercial conjugates for direct immunofluorescence of herpes simplex virus.

Commercially prepared antibody conjugates that are now available allow extensive application of direct immunofluorescence for diagnosing herpetic infections. Five of these conjugates--Microbiological Associates bivalent herpes antiserum (MBA-bivalent), MBA anti-herpes types 1 (MBA-1) and 2 (MBA-2), and Flow Laboratories anti-herpes types 1 (Flow-1) and 2 (Flow-2)--were compared for antibody titer, diagnostic accuracy, and nonspecific background staining on known infected tissue. Four of these were tested simultaneously on 75 clinical specimens. No attempt was made to differentiate between herpes simplex virus type 1 or type 2 infections. All antisera were comparable in titer and exhibited fluorescence with both herpes simplex virus types 1 and 2 antigens. The most significant difference was excessive nonspecific staining consistently seen with the MBA-2 conjugate. Overall, Flow-2 and MBA-bivalent antisera were preferable.

Antibodies, Viral↗
PubMed2006

Affinity chromatography of GroEL chaperonin based on denatured proteins: role of electrostatic interactions in regulation of GroEL affinity for protein substrates.

The chaperonin GroEL of the heat shock protein family from Escherichia coli cells can bind various polypeptides lacking rigid tertiary structure and thus prevent their nonspecific association and provide for acquisition of native conformation. In the present work we studied the interaction of GroEL with six denatured proteins (alpha-lactalbumin, ribonuclease A, egg lysozyme in the presence of dithiothreitol, pepsin, beta-casein, and apocytochrome c) possessing negative or positive total charge at neutral pH values and different in hydrophobicity (affinity for a hydrophobic probe ANS). To prevent the influence of nonspecific association of non-native proteins on their interaction with GroEL and make easier the recording of the complexing, the proteins were covalently attached to BrCN-activated Sepharose. At low ionic strength (lower than 60 mM), tight binding of the negatively charged denatured proteins with GroEL (which is also negatively charged) needed relatively low concentrations (approximately 10 mM) of bivalent cations Mg2+ or Ca2+. At the high ionic strength (approximately 600 mM), a tight complex was produced also in the absence of bivalent cations. In contrast, positively charged denatured proteins tightly interacted with GroEL irrespectively of the presence of bivalent cations and ionic strength of the solution (from 20 to 600 mM). These features of GroEL interaction with positively and negatively charged denatured proteins were confirmed by polarized fluorescence (fluorescence anisotropy). The findings suggest that the affinity of GroEL for denatured proteins can be determined by the balance of hydrophobic and electrostatic interactions.

Calcium↗
PubMed1981

Immune responses of breeding chickens to trivalent oil emulsion vaccines: responses to Newcastle disease and infectious bursal disease.

Bivalent Newcastle disease (ND)/infectious bursal disease (IBD) and trivalent ND/IBD/infectious bronchitis (IB) inactivated oil emulsion vaccines were prepared in the laboratory and evaluated under field conditions. Broiler breeder parent chickens previously vaccinated with live vaccines were inoculated with commercial monovalent ND and experimental bivalent or trivalent oil emulsion vaccines. The commercial vaccine induced a higher initial ND haemagglutination inhibition (HI) response than the experimental vaccines but, by 34 weeks after vaccination, the mean ND HI levels were not significantly different in any of the three flocks. All three vaccines provided sufficient ND immunity to protect against the clinical disease and egg production losses. The IBD responses of both flocks vaccinated with oil emulsion vaccine were similar to each other and only slightly lower than those flocks vaccinated with monovalent IBD oil emulsion vaccine in earlier experiments. Six weeks after vaccination, sufficient immunity was transferred to protect all the progeny against IBD challenge up to 33 days of age and some of them up to 45 days of age. Thirty-four weeks after vaccination of the parents with oil emulsion vaccine, the progeny were totally immune up to 27 days of age and some of them were immune until 37 days. Application of oil emulsion vaccines in bivalent or trivalent form did not impair the responses of the chickens to the monovalent components.

Animals↗
PubMed2002

FISH-mapping of rDNAs and Arabidopsis BACs on pachytene complements of selected Brassicas.

To improve resolution of physical mapping on Brassica chromosomes, we have chosen the pachytene stage of meiosis where incompletely condensed bivalents are much longer than their counterparts at mitotic metaphase. Mapping with 5S and 45S rDNA sequences demonstrated the advantage of pachytene chromosomes in efficient physical mapping and confirmed the presence of a novel 5S rDNA locus in Brassica oleracea, initially identified by genetic mapping using restriction fragment length polymorphism (RFLP). Fluorescence in situ hybridization (FISH) analysis visualized the presence of the third 5S rDNA locus on the long arm of chromosome C2 and confirmed the earlier reports of two 45S rDNA loci in the B. oleracea genome. FISH mapping of low-copy sequences from the Arabidopsis thaliana bacterial artificial chromosome (BAC) clones on the B. oleracea chromosomes confirmed the expectation of efficient and precise physical mapping of meiotic bivalents based on data available from A. thaliana and indicated conserved organization of these two BAC sequences on two B. oleracea chromosomes. Based on the heterologous in situ hybridization with BACs and their mapping applied to long pachytene bivalents, a new approach in comparative analysis of Brassica and A. thaliana genomes is discussed.

Arabidopsis↗
PubMed2004

M chromosome of the wild silkworm, Bombyx mandarina (n = 27), corresponds to two chromosomes in the domesticated silkworm, Bombyx mori (n = 28).

Chromosomes of Bombyx mori (n = 28) and of Bombyx mandarina (n = 27) were studied cytogenetically to resolve the origin of the large M chromosome in the Japaneses type of B. mandarina. In the F1 progeny from the reciprocal cross between B. mandarina and B. mori, the mitotic chromosome number was 2n = 55, and a chromosome configuration of 26 bivalents plus 1 trivalent was observed at metaphase I of germ cells. The trivalent chromosome consisted of the M chromosome from B. mandarina and two chromosomes from B. mori. When males of B. mori were mated to the F1 females, nuclei with two types of chromosome number (2n = 55 and 2n = 56) and two sets of chromosome pairs (26 bivalents plus 1 trivalent versus 28 bivalents) were observed in the metaphase I stage. Linkage analysis showed that the 14th chromosome of B. mori was involved in these two types of chromosome segregation. This result indicates that the M chromosome in B. mandarina arose from a fusion between a chromosome corresponding to the 14th linkage group and another, yet unidentified linkage group.

Animals↗
PubMed1976

Improved resolution of the meiotic chromsomes in both sexes of Euxoa species and their hybrids (Lepidoptera: noctuidae).

An air drying and Giemsa staining technique produces consistently good quality cytological preparations when applied to different species of Euxoa (Lepidoptera: Noctuidae) and, in particular, it allows an imporved resolution of the meiotic chromosomes in both sexes. All species so far investigated have a basic haploid chromosome complement of n (male) = 31(XX): n(female) = (XY). A single chiasma per individual bivalent is clearly visible in the male sex. Some chiasmata are formed in interstitial positions, but, by metaphase I, they have all undergone complete terminalization while the bivalents orient axially on the first division spindle. Direct evidence for lack of chiasma formation in the 31 hormorphic bivalents in the female sex is reported for the first time for species of Noctuidae. Evidence in favor of an XY system in the female sex is discussed. Preliminary studies of the meiotic divisions in hybrid material reveal the presence of cryptic structural differences between certain taxa, and will help to elucidate taxonomic problems within this complex genus.

Animals↗
PubMed1988

Quadrivalent formation in a tetraploid chicken oocyte.

Synaptonemal complex analysis of an exceptional tetraploid oocyte from a diploid chicken heterozygous for the MN t (Z;1) rearrangement was performed by electron microscopy of a spread preparation. Ten separate quadrivalents (26% of the chromosomal axes) were analyzed, as well as 50 autosomal bivalents. All the axes less than 2.5 microns in length formed bivalents (38) only, while axes in the 2.5-4.2 micron range formed 5 quadrivalents and 12 bivalents. The longer, separate axes formed quadrivalents only. Partner switches in excess of one were documented. The two identical W chromosomes paired only at the ends of their short arms. Quadrivalent formation may require a threshold length (2.5 microns), at least in this species. The tip of the short arm of the W chromosome may be a pairing initiation point, and it corresponds to the region associated with a localized recombination nodule previously described in diploid oocytes.

Animals↗
PubMed1994

Synaptonemal complex analysis of domestic sheep (Ovis aries) with Robertsonian translocations. I. Pachytene karyotype substaging of normal sheep.

Meiotic chromosome pairing behaviour in three normal sheep was surveyed by synaptonemal complex (SC) analysis at the electron microscope level. The SC karyotype of normal rams is composed of a sex pair, three metacentric or submetacentric pairs, and 23 acrocentric autosomal bivalents, with the total autosomal SC length ranging from 309 to 315 mircrons. Five nucleoli are terminally located on the three metacentric, and one large and one small acrocentric autosomal bivalents. XY morphology was used to divide pachytene into five substages. Although pairing abnormalities (mostly SC interactions) were recorded in an average of 16% of the spermatocytes, bivalent-XY association occurred in only 4.7% of the cells.

Animals↗
PubMed1999

Recombination nodule mapping and chiasma distribution in spermatocytes of the pigeon, Columba livia.

Pigeon spermatocytes were processed with a drying-down technique and their synaptonemal complex (SC) complements were analyzed by electron microscopy. The synaptonemal complex karyotype of the macrobivalents shows an excellent correspondence with the mitotic karyotype. The number and distribution of recombination nodules (RNs) were scored in complete nuclei stained with phosphotungstic acid. The average number of RNs per nucleus is 64.7. The number of nodules per bivalent shows a clear linear relationship with SC length in the 10 longest synaptonemal complexes, while the microbivalents usually bear a single RN. The location of RNs has a non-random distribution along the largest synaptonemal complexes, with lower frequencies near kinetochores and higher frequencies toward the telomeres. The ZZ bivalent is the fourth in size and shows free recombination, having on average 3.8 RNs. The mean number of nodules per cell and the mean number of nodules in the largest bivalents show very good agreement with the corresponding number of chiasmata scored in metaphase-I spermatocytes. It is concluded that the recombination nodules provide a good check for reciprocal exchanges in this and other species of birds. Additionally, a new morphology for the recombination nodules is presented, consisting of groups of electron-dense particles measuring 43 nm in diameter.

Animals↗
PubMed2003

Recombination nodules in coleopteran species: Palembus dermestoides (Tenebrionidae) and Epicauta atomaria (Meloidae).

This work describes the first report about the occurrence of recombination nodules (RNs) in spread pachytene cells of two species of Coleoptera: Palembus dermestoides (Tenebrionidae) and Epicauta atomaria (Meloidae). The RNs were observed in preparations contrasted with phosphotungstic acid. Considering RN morphology and its occurrence in pachytene bivalents (one per autosome bivalent) these structures were interpreted to be late RNs. P. dermestoides and E. atomaria have 2n = 20 chromosomes including an Xy(p) sex determination system. In spite of most frequently subtelocentric morphology observed in the autosomes of both species, the occurrence of RNs is limited only to the synaptonemal complex (SC) structure of the long arms. These findings are in agreement with those obtained using light microscopy analysis in which only one chiasma or terminalization event is observed per autosomal bivalent in early or late metaphase I cells. The RNs have the same average width of the SC of each analyzed species, a circular shape, strong electron density, and are observed mainly between the lateral elements of the SC. The RNs of P. dermestoides and E. atomaria have approximately the same average size (width), 180 +/- 20 nm and 160 +/- 80 nm, respectively. The absence of RNs in the short arms and its occurrence in the long arms are discussed considering the short arm pericentromeric and pro-centric heterochromatin.

Animals↗