Antigenic determinants at the carboxy terminus of chicken egg white riboflavin carrier protein (RCP): Epitope mapping and antibody-mediated pregnancy curtailment in rodents

The epitopic core sequences recognized by three monoclonal antibodies raised to chicken riboflavin carrier protein (RCP) were mapped to the C-terminal tail-end of the protein using the pepscan method A 21-residue synthetic peptide corresponding to residues 200-219 of the protein and comprising the regions corresponding to the antibodies was synthesized. Administration of polyclonal antibodies specific to this peptide led to termination of early pregnancy in mice. Also, active immunization of rats with the peptide-purified protein derivative conjugate inhibited establishment of pregnancy. These results demonstrate the functional importance of the C-terminal 200-219 region of chicken RCP. Copyright (C) 1996 Elsevier Science Ltd.

Simultaneous purification of biotin-binding proteins-I and -II from chicken egg yolk and their characterization

Chicken egg yolk biotin-binding protein-I (BBP-I) has been purified to homogeneity along with the tetrameric BBP-II by a common protocol. The purification includes delipidation of egg yolk by butanol extraction, DEAE-Sephacel chromatography, treatment with guanidinium chloride and biotin-aminohexyl-Sepharose affinity chromatography. The identity of purified BBP-I was ascertained by its physicochemical properties as well as by its immunological cross-reactivity and precursor-product relationship with BBP-II.

Improved parametric models and DPCM techniques for EGG data compiression

This paper presents a novel algorithm for compression of single lead Electrocardiogram (ECG) signals. The method is based on Pole-Zero modelling of the Discrete Cosine Transformed (DCT) signal. An extension is proposed to the well known Steiglitz-Hcbride algorithm, to model the higher frequency components of the input signal more accurately. This is achieved by weighting the error function minimized by the algorithm to estimate the model parameters. The data compression achieved by the parametric model is further enhanced by Differential Pulse Code Modulation (DPCM) of the model parameters. The method accomplishes a compression ratio in the range of 1:20 to 1:40, which far exceeds those achieved by most of the current methods.

Rhythmic egg-laying behaviour in virgin females of fruit flies Drosophila melanogaster

Fruit fly Drosophila melanogaster females display rhythmic egg-laying under 12: 12 h light/dark (LD) cycles which persists with near 24 h periodicity under constant darkness (DD). We have shown previously that persistence of this rhythm does not require the neurons expressing pigment dispersing factor (PDF), thought to be the canonical circadian pacemakers, and proposed that it could be controlled by peripheral clocks or regulated/triggered by the act of mating. We assayed egg-laying behaviour of wild-type Canton S (CS) females under LD, DD and constant light (LL) conditions in three different physiological states; as virgins, as females allowed to mate with males for 1 day and as females allowed to mate for the entire duration of the assay. Here, we report the presence of a circadian rhythm in egg-laying in virgin D. melanogaster females. We also found that egg-laying behaviour of 70 and 90% females from all the three male presence/absence protocols follows circadian rhythmicity under DD and LL, with periods ranging between 18 and 30 h. The egg-laying rhythm of all virgin females synchronized to LD cycles with a peak occurring soon after lights-off. The rhythm in virgins was remarkably robust with maximum number of eggs deposited immediately after lights-off in contrast to mated females which show higher egg-laying during the day. These results suggest that the egg-laying rhythm of D. melanogaster is endogenously driven and is neither regulated nor triggered by the act of mating; instead, the presence of males results in reduction in entrainment to LD cycles.

Structural Transformations in Protein Crystals caused by controlled Dehydration

Recent experiments in this laboratory on structural transformations caused by controlled dehydration of protein crystals have been reviewed. X-ray diffraction patterns of the following crystals have been examined under varying conditions of environmental humidity in the relative humidity range of 100-75%: a new crystal form of bovine pancreatic ribonuclease A grown from acetone solution in tris buffer (I), the well-known monoclinic form of the protein grown from aqueous ethanol (II), the same form grown from a solution of 2-methyl pentan-2,4-diol in phosphate buffer (III), tetragonal (IV), orthorhombic (V), monoclinic (VI) and triclinic (VII) hen egg white lysozyme, porcine 2 Zn insulin (VIII), porcine 4 Zn insulin (IX) and the crystals of concanavalin A(X). I, II, IV, V and VI undergo one or more transformations as evidenced by discontinuous changes in the unit cell dimensions, the diffraction pattern and the solvent content. Such water-mediated transformations do not appear to occur in the remaining crystals in the relative humidity range explored. The relative humidity at which the transformation occurs is reduced when 2-methyl pentan-2,4-diol is present in the mother liquor. The transformations are affected by the crystal structure but not by the amount of solvent in the crystals. The X-ray investigations reviewed here and other related investigations emphasize the probable importance of water-mediated transformations in exploring hydration of proteins and conformational transitions in them.

Evolution of sex ratios in social hymenoptera: consequences of finite brood size

Evolutionarily stable sex ratios are determined for social hymenoptera under local mate competition (LMC) and when the brood size is finite. LMC is modelled by the parameter d. Of the reproductive progeny from a single foundress nest, a fraction d disperses (outbreeding), while (1-d) mate amongst themselves (sibmating). When the brood size is finite, d is taken to be the probability of an offspring dispersing, and similarly, r, the proportion of male offspring, the probability of a haploid egg being laid. Under the joint influence of these two stochastic processes, there is a nonzero probability that some females remain unmated in the nest. As a result, the optimal proportion of males (corresponding to the evolutionarily stable strategy, ESS) is higher than that obtained when the brood size is infinite. When the queen controls the sex ration, the ESS becomes more female biased under increased inbreeding (lower d), However, the ESS under worker control shows an unexpected pattern, including an increase in the proportion of males with increased inbreeding. This effect is traced to the complex interaction between inbreeding and local mate competition.

Estrogen Induction Of Biotin-Binding Protein In Immature Chicks - Kinetics, Hormonal Specificity And Modulation

Employing a specific radioimmunoassay for quantification, the kinetics of estrogen-induced elevation in the plasma concentration of biotin-binding protein (BBP) in immature male chicks was investigated. A single injection of the steroid hormone enhanced the plasma BBP content several-fold at 6 h, reaching peak levels around 48 h and declining thereafter. A 2-fold amplification of the response was evident during secondary stimulation with the hormone. The magnitude of the response was hormonal dose-dependent while the initial lag phase and the time of peak protein accumulation were unaltered within the hormonal doses tested. The circulatory half-life of the specific protein in normal and estrogenized birds was 10 h. Hyperthyroidism markedly decreased the hormonal response while the opposite effect was seen during hypothyroidism. The antiestrogens E- and Z-clomiphene citrate effectively blocked the protein induction whereas progesterone, either alone or in combination with estrogen, was ineffective in modulating the induction. Cycloheximide administration drastically inhibited the inductive response. The above observations clearly suggest that the genes corresponding to the two isofunctional proteins of chicken egg, viz. BBP and avidin, are differentially regulated.

Behavior of the indian social wasp ropalidia-cyathiformis on a nest of separate combs (hymenoptera, vespidae)

Observations were made on a nest of Ropalidia cyathiformis consisting of three combs. The number of eggs, larvae, pupae and adults were monitored at about 3-day intervals for a 2-month period. The behaviour of the adults was observed with special reference to the proportion of time spent on each of the three combs, the proportion of time spent away from the nest site and the frequencies of dominance interactions and egg laying. The adults moved freely between the three combs suggesting that all of them and all the three combs belonged to one nest. However, most of the adults preferred combs 2 and 3 over comb 1. Of the 10 animals chosen for a detailed analysis of behaviour, seven spent varying periods of time away from the nest site and oRen brought back food or building material. Five of the 10 animals laid at least one egg each but two adults monopolized most of the egg-laying. The animals showed a variety of dominance interactions on the basis of which they have been arranged in a dominance hierarchy. The dominant individuals laid most of the eggs and spent little or no time foraging, while the subordinate individuals spent more time foraging and laid few eggs or none. It is argued that R. cyathiformis is different from R. marginata, the only other Indian social wasp whose behaviour has been studied, in being at a more primitive stage of social organization.

Vitamin A economy of the developing chick embryo and of the freshly hatched chick

. The changes in the net amounts of retinol, retinyl esters and retinal in both the developing chick embryo and the newly hatched chick were investigated. The embryo requires about 68nmol of the vitamin for its growth, whereas the baby chick requires about 108nmol during the first 7 days after hatching. 2. Retinal was present in the egg in fairly high concentrations at the beginning of the incubation but it virtually disappeared from the extra-embryonic tissue after day 17 of incubation. It was not found in the liver of the embryo or of the newly hatched chick up until day 7.

Oestrogen induction of riboflavin-binding protein in immature chicks. Nature of the secretory protein.

The riboflavin-binding protein isolated from sera of oestrogen-treated male chicks as well as that synthesized and secreted in vitro by the chicken liver have the same molecular size as that of the egg-yolk protein. Functionally the serum and yolk proteins are similar. This is in contrast with the hormone-induced synthesis, secretion and deposition of phosvitin and lipovitellin in the ovary.

Additional binding sites in lysozyme. X-ray analysis of lysozyme complexes with bromophenol red and bromophenol blue

The binding sites in hen egg-white lysozyme for neutral bromophenol red (BPR) and ionized bromophenol blue (BPB) have been characterized at 2 Å resolution. In either case, the dye-bound enzyme is active against the polysaccharide, but not against the cell wall. Both binding sites are outside, but close to, the hexasaccharide binding cleft in the enzyme. The binding site of BPR made up of Arg5, Lys33, Phe34, Asn37, Phe38, Ala122, Trp123 and possibly Arg125, is dose to subsite F while that of BPB made up of Tyr20, Arg21, Asn93, Lys96, Lys97 and Ser100, is close to subsites A and B. The binding sites of the neutral dye and the ionized dye are thus spatially far apart. The peptide component of the bacterial cell wall probably interacts with these cells during enzyme action. Such interactions are perhaps necessary for appropriately positioning the enzyme molecule on the bacterial cell wall.

Social evolution - Has nature ever rewound the tape?

Social insects such as ants, bees, wasps and termites exhibit extreme forms of altruism where some individuals remain sterile and assist other individuals in reproduction. Hamilton's inclusive fitness theory provides a powerful framework for investigating the evolution of such altruism. Using the paper wasp Ropalidia marginata, we have quantified and delineated the role of ecological, physiological, genetic and demographic factors in social evolution. An interesting feature of the models we have developed is their symmetry so that either altruism or selfishness can evolve, depending on the numerical values of various parameters. This suggests that selfish/solitary behaviour must occasionally re-emerge even from the eusocial state, It is useful to contemplate expected intermediate states during such potential reversals. We can perhaps envisage three successive steps in such a hypothetical process: i) workers revolt against the hegemony of the queen and challenge her status as the sole reproductive, ii) workers stop producing queens and one or more of them function as egg layers (functional queen/s) capable of producing both haploid as well as diploid offspring and iii) social evolution reverses completely so that a eusocial species becomes solitary, at least facultatively. It appears that the third step, namely transition from eusociality to the solitary state, is rare and has been restricted to transitions from the primitively eusocial state only. The absence of transitions from the highly eusocial state to the solitary state may be attributed to a number of 'preventing mechanisms' such as (a) queen control of workers (b) loss of spermathecae and ability to mate (c) morphological specialization (d) caste polyethism and (e) homeostasis, which must each make the transition difficult and, taken together, perhaps very difficult. However, the discovery of a transition from the highly eusocial to the solitary state can hardly he ruled out, given that little or no effort has gone into its detection. In this paper I discuss social evolution and its possible reversal and cite potential examples of stages in the transition from the social to the solitary.

Oestrogen-induced synthesis of thiamin-binding protein in immature chicks. Kinetics of induction, hormonal specificity and modulation

A specific radioimmunoassay procedure was developed to monitor the plasma concentrations of thiamin-binding protein, a minor yolk constituent of the chicken egg. By using this sensitive assay, the kinetics of oestrogen-induced elaboration of this specific protein in immature chicks was investigated. After a single injection of the steroid hormone, with an initial lag period of 4–5h the thiamin-binding protein rapidly accumulated in the plasma, attaining peak concentrations around 75h and declining thereafter. A 4-fold amplification of the response was noticed during the secondary stimulation, and this increased to 9-fold during the tertiary stimulation with the steroid hormone. The magnitude of the response was dependent on the hormone dose, and the initial latent period and the duration of the ascending phase of induction were unchanged for the hormonal doses tested during both the primary and secondary stimulations. The circulatory half-life of the protein was 6h as calculated from the measurement of the rate of disappearance of the exogenously administered 125I-labelled protein. Simultaneous administration of progesterone, dihydrotestosterone or corticosterone did not alter the pattern of induction. On the other hand, hyperthyroidism markedly decreased the oestrogenic response, whereas propylthiouracil-induced hypothyroidism had the opposite effect. The anti-oestrogen E- and Z-clomiphene citrates, administered 30min before oestrogen, effectively blocked the hormonal induction. α-Amanitin and cycloheximide administered along with or shortly after the sex steroid severely curtailed the protein elaboration. A comparison of the kinetics of induction of thiamin- and riboflavin-binding proteins by oestrogen revealed that, beneath an apparent similarity, a clear-cut difference exists between the two vitamin-binding proteins, particularly with regard to hormonal dose-dependent sensitivity of induction and the half-life in circulation. The steroid-mediated elaboration of the two yolk proteins thus appears to be not strictly co-ordinated, despite several common regulatory features underlying their induction.