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PubMedDate not supplied

[Binding of reversible spin-labeled inhibitors with an butyrylcholinesterase active center].

The interaction of spin-labeled metacyn, procaine, carbolin and bivalent cations (Ca2+, Co2+, Ni2+) with butyrylcholinesterase (BChE) was studied by ESR and enzyme kinetic methods. The effect of pH, ionic strength and organic solvent was analysed. Spin-labeled metacyn binds at the anionic site of BChE active centre. This complex is stabilized both with coulombic and hydrophobic interactions, ionizing group of active centre with pK 6-7 also affects the binding. Spin-labeled procaine appeared to be enzyme competitive inhibitor (Ki = 4 X 10(-5) M) and is located, most probably, at the same site. Activating effect of Ca2+ ions on BChE was confirmed. Simultaneous application of spin labels and paramagnetic ions demonstrates that cations Co2+ and Ni2+ bind with BChE in the close vicinity of spin-labeled inhibitor site. Paramagnetic cations are located more closely to the cationic part of the inhibitor molecule than to the hydrophobic one, and can be displaced by surplus of Ca2+ ions. The experimental data testify the model of anionic centre which consists of bivalent metal ions and aminoalcyl cationic group subsites and is located in a hydrophobic pocket of the enzyme surface.

Binding Sites↗
PubMed1986

Valency of CD3 binding and internalization of the CD3 cell-surface complex control T cell responses to second signals: distinction between effects on protein kinase C, cytoplasmic free calcium, and proliferation.

We have studied the relationship of valency of CD3 stimulation and modulation of the CD3 receptor complex with biochemical and proliferative responses of T cells. Anti-CD3 Fab, as well as F(ab')2 and whole antibody caused rapid modulation of the CD3 antigen, whereas anti-CD3 conjugated to Sepharose did not. In the absence of monocytes, T cells stimulated with anti-CD3 Fab, F(ab')2, or F(ab')2-Sepharose showed differences in their ability to respond to second signals given by PMA, IL 1, IL 2, or antibodies to Tp67 and Tp44. None of the anti-CD3 signals alone caused resting T cells to produce IL 2, and only the Sepharose-bound anti-CD3 F(ab')2 caused T cells to express high levels of functional IL 2 receptors. Anti-CD3 F(ab')2-Sepharose-stimulated T cells produced IL 2 and proliferated in response to each of the second signals. Because anti-CD3-Sepharose did not cause modulation of the CD3 antigen, the ability of the Sepharose-bound antibody to induce T cells to express IL 2 receptors and to respond to individual second signals may be related to lack of modulation rather than valency of binding. Anti-CD3 Fab-stimulated T cells responded to PMA but required combinations of other second signals. T cells stimulated with unmodified anti-CD3 antibody or F(ab')2 fragments responded to PMA but did not respond to any other second signals alone or in combination. Stimulations that resulted in modulation (i.e., anti-CD3 whole antibody, anti-CD3 F(ab')2, or anti-CD3 Fab fragments) caused an increase in cytoplasmic calcium levels in resting T cells but blocked proliferation of T cells in response to mitogenic lectins or CD2 stimulation. Anti-CD3 F(ab')2 on Sepharose, however, did not block T cell proliferation. Whole bivalent anti-CD3 antibody or F(ab')2 fragments, but not monovalent Fab fragments, caused a rapid translation of protein kinase C activity from cytosol to membrane in the Jurkat T cell line. Because all of these modulate the receptor, these data indicate that the functional difference between monovalent and bivalent binding to CD3 is related to antibody valency and not to antigenic modulation. The use of Fab anti-CD3 stimulation that requires combinations of second signals for proliferation allowed an analysis of the functional relationships between IL 1, anti-Tp67, and anti-Tp44.

Animals↗
PubMed1986

Ferritin versus ferrous sulphate preparations: a controlled study in sideropaenic patients.

A double-blind study was carried out in 54 patients requiring iron therapy to compare the effects of orally administered bivalent and trivalent iron over a period of 12 weeks. Patients were allocated at random to receive ferrous sulphate (100 mg/day), ferritin (80 mg/day as Fe3+) or a combination of ferritin (80 mg Fe3+/day), folinic acid and vitamin B12 coenzyme. Haematological measurements made before and during treatment showed that, whilst there were significant improvements in all of the parameters studied with both the bivalent and trivalent forms of iron, the results were more progressive and consistent with the two ferritin preparations, indicating that ferritin is clinically effective as a physiological iron donor in sideropaenic conditions.

Adolescent↗
PubMed1988

On the nature of IgG dimers. II. Idiotype--anti-idiotype complexes of polyclonal and monoclonal origin: size distribution patterns and molecular geometries.

Electron micrographs of a fraction containing dimers isolated from pooled human polyclonal immunoglobulin G (IgG) suggest essentially a cyclic geometry compatible with bivalently associated monomers. It is obvious that such a structure can be produced by idiotype (Id)--anti-idiotype (anti-Id) interactions where the latter are able to neutralize certain combining site related Id functions. Accordingly, antibody (ab) activities against tetanus toxoid (tt) and rubella antigen (ag) were found to be almost exclusively confined to the monomeric molecules in preparations composed of monomers and dimers only. Moreover, electron micrographs of complexes prepared from a murine monoclonal Id as well as anti-Id reveal the presence of ring complexes, especially of cyclic tetramers. Gel filtration patterns of mixtures containing equimolar concentrations (concns) of such abs (1.6 x 10(-6) M) show, correspondingly for 9 different Id--anti-Id pairs and therefore probably representing a more common feature, mainly the formation of even-numbered complexes, especially tetramers. That is basically in accordance to an equilibrium model developed by Archer and Krakauer but not from a quantitative point of view because non-ideality terms had not been originally included. Despite taking strain energies determined by Schumaker et al. for cyclic complexes of polyclonal rabbit abs and a bivalent hapten into account for computation of size distribution patterns, the predominant formation of dimers was, nevertheless, again predicted by the modified theory in contrast to the experimental results. Fundamental conformity could only be achieved by further decreasing one of the statistical factors, namely the ring closing factor, which theoretically influences the generation of cyclic dimers. Therefore, referring to the experimental results of Schumaker et al., we postulate a strain energy well above 700 cal/mol for cyclic dimers produced by interacting Ids and anti-Ids. In general, the findings confirm predictions based on the interpretation of recent kinetic studies.

Animals↗
PubMed1986

Activation of the first component of human complement, C1, by monoclonal antibodies directed against different domains of subcomponent C1q.

Two monoclonal antibodies directed against C1q, and their (Fab)2 and Fab fragments, were used to study the mechanism of C1 activation. Monoclonal antibody 2A10, an IgG2a, was digested by pepsin to yield fully immunoreactive (Fab')2. Monoclonal antibody 1H11, an IgG1, was digested by papain to yield fully immunoreactive, bivalent (Fab)2. Previously 1H11 had been shown to bind to the C1q "heads," whereas 2A10 bound to stalks. Activation of C1 was followed by the cleavage of 125I-C1s in the presence of C1 inhibitor (C1-Inh) at 37 degrees C. Spontaneous activation was minimal at inhibitor concentrations above 0.4 micron (1.3 X physiologic inhibitor concentration); all results were corrected for the spontaneous activation background. Heat-aggregated IgG activated completely in this system and was taken as 100% activation. Monoclonal antibody 2A10 caused precipitation of C1 and slow activation; neither the (Fab')2 nor the Fab' derived from 2A10-caused activation. Probably, aggregates of intact 2A10 and C1 were serving as immune complexes to activate other molecules of C1. In contrast, both 1H11 and its (Fab)2 activated completely and stoichiometrically; that is, maximal activation was achieved at a ratio of one C1q head to one antibody combining site. The monovalent Fab derived from 1H11 bound well to C1q, but no activation of C1 was observed. Thus, bivalent binding of this head-binding monoclonal is required for C1 activation, but not the presence of the antibody Fc portion. Neither 1H11 nor its (Fab)2 fragments caused C1 precipitation; however, the 1H11 did form complexes composed of two C1q cross-linked by multiple 1H11, which were visualized by electron microscopy. The presence of these dimeric complexes correlated well with activation. A model for C1 activation is proposed in which two C1q subcomponents are held together by multiple (Fab)2 bridging C1q heads. The model is roughly analogous to touching opposing pairs of fingers and thumb tips, the two hands representing the two C1q, forming a cage. C1-Inh, which probably binds to C1r through the open end of the C1 cone, is too long asymmetric to be included within the cage. Thus, according to this model, the dimers of C1 are released from the inhibitory action of C1-Inh, and activation proceeds spontaneously and rapidly at 37 degrees C.

Animals↗
PubMed1986

Effect of simultaneous administration of live measles vaccine on the "take rate" of live mumps vaccine.

During the course of clinical studies to develop a new bivalent measles-mumps vaccine, it was established that simultaneous administration of live measles and mumps vaccines at doses contained in the monovalent vaccines resulted in a reduced seroconversion rate against mumps but not against measles. This one-way interference was resolved by increasing the dose of the mumps component in the bivalent measles-mumps and the trivalent measles-mumps-rubella vaccines above that in the monovalent mumps vaccine. On the other hand, it was not necessary to adjust the doses of the measles and rubella components in the combined vaccines. This observation requires that not only the titres of vaccine viruses must be carefully adjusted in combined live mumps vaccines but also that the heat-stability of each component in such vaccines be carefully determined to ensure that interference is not a problem during the complete shelf-life of the product. The relative doses at release of combined mumps vaccines using the Urabe Am 9 strain and the heat-stability characteristics of these products will be presented and discussed.

Antibodies, Viral↗
PubMed1987

Bioavailability of bi- and trivalent oral iron preparations. Investigations of iron absorption by postabsorption serum iron concentrations curves.

Since 1977 the bioavailability of 14 bi- and trivalent oral iron preparations has been investigated in five separate orientated clinical studies by using postabsorption serum iron concentration curves. The range of relative bioavailability was 46 to 100% in the first group of preparations, 31 to 47% in the second group and 6 to 29% in the third group. The first group contained mainly bivalent quick release preparations, the second group slow or sustained release preparations and the third mainly trivalent iron preparations. The postabsorption serum iron concentration curves which show a good congruence with exact 59Fe whole body retention tests again confirmed the nearly 50-year-old rule that bivalent iron is up to 16 times better absorbed than trivalent iron. There is no doubt that the oral iron preparations of good bioavailability are able to cure an iron deficiency more rapidly than iron preparations with a low bioavailability. This therefore has a clear influence on the overall expense of iron therapy. Only those preparations from the first group can be recommended for oral iron therapy. The preparations in the second group are less suitable and those in the third group should be excluded from iron therapy in all countries.

Administration, Oral↗
PubMed1987

Acid alpha naphthyl acetate esterase reacting lymphocytes in the peripheral blood of chickens challenged with Marek's disease after vaccination with three different vaccines.

The average percentage of acid alpha naphthyl acetate esterase reacting lymphocytes (APARL) was enumerated in the peripheral blood of chickens challenged with Marek's disease after vaccination with either turkey herpesvirus (HVT), inactivated Marek's disease virus (IMDV) or a mixture of the two (bivalent vaccine). A gradual increase in APARL value was noticed in the vaccinated chickens from day 7 to 70 after challenge with a virulent Marek's disease virus. The increase was consistent and significantly higher in bivalent (HVT plus IMDV) than in HVT-vaccinated chickens while the slight increase noticed in IMDV vaccinated-challenged birds was inconsistent.

Animals↗
PubMed1985

[Patterns in the occurrence and the submicroscopic organization of elongated metaphase chromosomes isolated from the cell].

Chinese hamster metaphase chromosomes were investigated under different conditions of isolation. Light microscopic study demonstrated different forms of stretched chromosomes, from those in which merely a small region is stretched to rope-like structures 20-25 microns long with diameter of about 0.4 micron. The ratio between the number of stretched and of compact chromosomes is dependent on the concentration of bivalent cations, on the pH and temperature of the isolation buffer. A study of the submicroscopic organization of stretched chromosomes revealed lengthwise fibrils that disappeared after the treatment with 0.6 NaCl and staphylococcal nuclease. Distinct aggregates were seen, whose array is maintaining the stretched chromosome structure. It is suggested that stretched chromosomes appear due to the existing in vivo lability of bonds between the main chromosome components involved in organization of chromatin fiber packing. It is proposed that the structure obtains rigidity in the course of isolation with bivalent cations.

Animals↗
PubMed1973

Trials of aqueous killed influenza vaccine in Canada, 1968-69.

The appearance of the pandemic A/Hong Kong/1/68 (H3N2) influenzavirus strain provided an opportunity for a clinical field trial of influenza vaccines in Canada during the winter of 1968-69. As by November 1968 there were reports of influenza B activity and as supplies of A2/HK/68 vaccines were limited, it was decided to make a series of strictly randomized double-blind trials comparing A2/HK/68 vaccines not only with B/Mass/66 vaccines but also with a bivalent vaccine that was already in production and contained B/Mass/66 and A2/Mtl/68, the latter a strain isolated in Canada during January 1968. In 4 trials, a total of 13 729 military personnel and 4 795 primary schoolchildren were vaccinated. Reported vaccine reactions were less than 0.1% with zonally-purified vaccines and 2.6% with the "standard" aqueous killed bivalent vaccine. Three children had serious reactions. Surveillance detected an outbreak of influenza in the first two trials on the military. The 3 vaccines containing A2 strains gave similar clinical protection conservatively estimated at 42-55% but probably about 80%. The effectiveness of the A2/Mtl/68 vaccine, which was in production before the Hong Kong variant had been isolated, was unexpected. In the absence of a vaccine specific to a new pandemic strain, it should not be assumed that a vaccine made from another recent strain could not be useful.

Canada↗
PubMed1969

An evaluation of influenza immunization: influence of route of administration and vaccine strain.

A field study was undertaken in Tampa, Fla., to assess the efficacy of subcutaneous and aerosol methods of administering vaccine, and to compare the protection afforded by bivalent (A2 and B) influenza virus vaccine and by A2/Hong Kong/68 virus vaccine. Further objectives of the study included a comparison of the effectiveness of single-dose and 2-dose immunization. Approximately 2100 volunteers received, in a double-blind manner, both an injection and an aerosol administration on 2 occasions 3 weeks apart. The results showed that aerosol administration gave a lower over-all protection rate, although the booster dose seemed to have a marked effect. The protection afforded by A2/Hong Kong/68 virus vaccine was considerably greater than that afforded by the bivalent vaccine, particularly when administration was subcutaneous. Results are also given on the occurrence of side-effects and on the correlation between cigarette smoking and the occurrence of influenza-like illness.

Aerosols↗
PubMed1979

[Cytology of egg maturation in the silkworm (Bombyx mori L.) in artificial parthenogenesis].

The orientation of bivalentss in the first meiotic metaphase was studied on the squash preparations and sections of eggs in Bombyx mori L., Antheraea pernyi Guèr and Carpocapsa pomonella L. The reduction fissure of bivalents separating the homologous chromosomes lies in the plane of spindle equator; the equation fissure separating the sister chromatids is parallel to the long spindle axis. Complicated rearrangements are observed in the nuclei of the silkworm eggs stimulated to ameiotic parthenogenesis by the high temperature dissociation of bivalents, full destruction of spindle, formation of a new spindle and metaphase plate. The latter includes univalent chromosomes involved in the equation division. As a result, two genetically identical pronuclei with the somatic set of chromosomes are formed.

Animals↗
PubMed1982

E-rosette forming cells and humoral antibody titres in humans after vaccination with three different inactivated influenza virus vaccines A/USSR/92/77 (H1N1).

The number of E-rosette forming cells and the serum haemagglutination inhibition (HI) antibody titres were examined in 37 volunteers immediately before and 14, 28, 35 and 63 days after immunization with three inactivated influenza virus vaccines A/USSR/92/77 (H1N1)--NIB 6 and in 11 non-vaccinated controls. From the former, 10 volunteers were immunized with 1000 haemagglutinin (HA) IU per dose, 11 volunteers with the NIB 6 adsorbate vaccine (340 HA IU/dose) and 16 volunteers with a bivalent vaccine composed of 180 HA IU/dose NIB 6 and 180 HA IU/dose of influenza virus A/Bangkok X-73 (H3N2). The percentage of E-rosette forming cells was decreased in all vaccinated volunteers 14 days after vaccination; later on the values reached normal level of non-vaccinated controls or of subjects before vaccination. The number of E-rosette forming cells was in correlation with the applied virus vaccine dose, i.e. for the 1000 HA IU/dose: 29.95 +/- 11.74%, p less than 0.001 and for the 340 HA IU/dose: 47.75 +/- 11.15%, p less than 0.005; however, after administration of 180 HA IU/dose of NIB 6 in the bivalent vaccine, the value 58.65 +/- 11.5% was not significantly decreased in comparison to non-vaccinated donors. The serum HI antibody titres reached the highest level 14 days after vaccination and remained constant during the next 6 weeks. There was a correlation between decreased E-rosette values and increased serum antibody titres (p less than 0.05). The current study indicates that the number of E-rosette forming cells may serve as a further laboratory criterion for controlling the effect of inactivated influenza virus vaccines on the immune system of man.

Adolescent↗
PubMedDate not supplied

Metabolism of styrene oxide in the rat and guinea pig.

The metabolism of styrene oxide has been studied in the rat and guinea pig, with emphasis upon bivalent sulfur metabolites. Methylthio analogs of phenylethylene glycol, with the methylthio group in both possible positions, were found as urinary metabolites in both species. These compounds were present in more than trace amounts. The excretion of 2-hydroxy-1-methylthio-1-phenylethane amounted to about 7% of the administered dose in the guinea pig, and about 2% in the rat, in o-24 hr urine samples. The positional isomer 1-hydroxy-2-methylthio-1-phenylethane was excreted in lesser amounts in both species. Acidic urinary metabolites derived from glutathione conjugates are species dependent. In this study, the only products observed in the rat were the mercapturic acids expected as a result of reaction of the oxide with glutathione. In the guinea pig, the major bivalent sulfur acids were the corresponding mercaptoacetic acids. Other related metabolites included a mercaptolactic and a mercaptopyruvic acid, together with one of the mercapturic acids. These metabolites result from partial acetylation or acetylation/deacetylation of cysteine or cysteinylglycine adducts. The hitherto unobserved dihydrodiol formed via an arene oxide was found as a minor metabolite for both styrene and styrene oxide.

Animals↗
PubMed1976

Insulin--anti-insulin complexes.

When guinea pig antibodies (ab) bind insulin (ag), they can make complexes of different sizes. We propose the following model: In ab excess: (see article) Intermediate: (see article) In ag excess: (see article). An insulin molecule acts as a bivalent antigen, although more than two different antigenic determinants may be present. In vivo the large C II type disappears more rapidly from the blood than does the C I. The C II binds to complement factor C1q, whilst C I and C III do not. In sera from insulin treated patients we found C I and C III. The lack of lattice formation, due to the bivalency, may explain the difficulty in obtaining precipitation. The different complexes may influence calculations of antibody concentrations and affinity constants of the binding sites. The in vivo effects and possible clinical effects of antibodies to insulin may depend on the type of complex formed. Possibly, prevailing C II formation tends to cause large insulin requirements, although C II may seldomly be detected in the blood, because of rappid trapping. The immune complexes could affect the progression of angiopathy a) by interfering with insulin metabolism and control of diabetes, and b) by complement activation (mainly C II) and trapping in the vascular bed.

Animals↗
PubMed1984

The binding of monoclonal antibodies to cell surface molecules. Quantitative analysis of the reactions and cross-reactions of an antibody (MB40.3) with four HLA-B molecules.

The MB40.3 monoclonal antibody binds to four distinct HLA-B molecules; B7, B40, B40*, and B27. With Fab' fragments only the interaction with B7 and B40 was detected and the affinity for both was the same (1-2 X 10(8) M-1) suggesting the epitope is shared by the two molecules. Unlike many antibodies for which low affinity is due to a high-dissociation constant, that of MB40.3 results from a very low-association rate constant, coupled with a low-dissociation constant. In consequence, the affinity and avidity of Fab', F(ab')2, and IgG for B7 and B40 were found to be of a similar magnitude, soluble B7 was a more efficient competitor for antibody than cell surface B7 and in practice antibody bivalency was of little importance. The forward rate constant could be increased by removing Fc from the antibody or by removing sialic acid from the cells by treatment with neuraminidase. The neuraminidase treatment also produced an increase in the number of detectable cell surface HLA-A,B molecules. The affinity of MB40.3 for B40* and B27 was estimated to be less than 4 X 10(6) as no binding with Fab' was detected due to a high-dissociation rate. For these two HLA-B molecules bivalent attachment was critical, and it increased the strength of interaction with cell surface B40* and B27 to a point where the avidities were comparable to those obtained with B7 and B40, with B40* interacting more strongly than B27. The epitopes recognized by MB40.3 on B40* and B27 were thus shown to be structurally different from each other and from those on B7 and B40. The properties of this antibody contrast with those of other anti-HLA-A,B we have studied (Ways, J.P., and Parham, P. (1983) Biochem. J. 216, 423-432).

Antibodies, Monoclonal↗
PubMed1978

[Nature and role of accessory chromosomes in silver-gray foxes. IV. The behavior of accessory chromosomes in meiosis].

Comparison of chromosome number at somatic and spermatogonial mitoses has demonstrated the increase in the number of additional chromosomes in cells of germinal tissue. This may evidence a mechanism of B-chromosomes accumulation in foxes. B-chromosomes may lag as univalents, may form bivalent associations, or occasionally form trivalents at the stage of diakinesis-metaphase I, and they may associate with macrobivalents (A-chromosome bivalents). The analysis of metaphase II has shown that the distribution of B-chromosomes in the second metaphase is random resulting in gametes with various numbers of B-chromosomes.

Animals↗
PubMed1984

The synaptonemal complex, recombination nodules and chiasmata in human spermatocytes.

Reconstruction from serial sections of 120 human spermatocyte nuclei, ranging in stage from leptotene to anaphase I, has led to the following conclusions. The reconstructed nuclei form a continuous sequence of developmental stages. With few exceptions, all of the bivalents are identifiable by length, centromere index and morphological markers. Crossing over is accompanied by a transformation of the recombination nodule (RN) into a fusiform bar lying across the synaptonemal complex (SC). The evolution of the crossover distribution at late pachytene is regulated at two levels both involving the SC. Each bivalent displays a pattern of SC regions - domains - with high affinity for RNs in the middle and low affinity at the ends. The occurrence of crossing over in a domain reduces the probability of attachment of new RNs and causes RNs not yet involved in crossing over to be released from that domain. Positive interference between crossovers is a consequence of these phenomena. At diplotene the degradation of the SC is initiated in regions with low affinity for RNs and small segments of the SC are retained at the crossover sites until late diakinesis. The site of crossing over is only identifiable by an RN for a certain period of time during pachytene. The distribution of total crossovers in a nucleus is similar but not identical to the distribution of chiasmata as seen in the light microscope. The number of crossovers has been estimated to be 70 and exceeds the number of chiasmata determined by light microscopy by a factor of 1.4.

Animals↗