Amino acid sequence homologies between rabbit, rat, and human serum retinol-binding proteins

The main transporting protein for vitamin A in rabbit serum, the retinol-binding protein (RBP), was isolated and its amino acid sequence determined. Rabbit RBP was found to be highly homologous to human RBP, whose amino acid sequence was elucidated earlier, and to rat RBP. The rat RBP sequence was obtained by combining information deduced from the nucleotide sequences of two overlapping cDNA clones with the NH2-terminal sequence of the isolated protein determined by automated Edman degradation. The identity between the three proteins is approximately 90%. The high degree of homology between RBP molecules from different species is probably explained by the fact that RBP participates in at least three types of molecular interactions: in the binding of prealbumin, in the interaction with retinol, and in the recognition of a specific cell surface receptor. All these interactions should lead to a conservation of RBP structure. The amino acid differences between rabbit, rat, and human RBP are discussed in light of the recent elucidation of the three-dimensional structure of human RBP. Hybridization of a probe isolated from a rat RBP cDNA clone to restriction enzyme-digested genomic DNA from rat and mouse suggests that RBP is encoded by a single gene.     

Genetic polymorphism of complement component C8

Summary Extensive genetic polymorphism of complement component C8 was demonstrated by isoelectric focusing of serum or plasma samples followed by immunoblotting procedures. Using these methods, we could detect both - (C81) and (C82) chain polymorphisms in the same gel. Two-dimensional (2D) electrophoresis of C8 immunoprecipitates was used to obtain further information of the C8 patterns. Evidence was obtained that the C81 polymorphism resides in the structural gene of the C8 chain. Both C8 systems show autosomal, chiefly codominant inheritance, and the distribution of phenotypes agrees with the Hardy-Weinberg equilibrium. Our findings suggest at least five different alleles in the C81 system; the gene frequencies of the two most common ones, C81 ^*A and C81 ^*B being 0.59 and 0.39, respectively. In C82 we found evidence for at least three codominant alleles, the gene frequencies for the two most common ones, C82 ^*B and C82 ^*A being 0.94 and 0.05, respectively. In addition, family studies disclosed the existence of a null allele, C82 ^* Q0.     

Plasma levels of arginine vasotocin,prolactin, aldosterone and corticosterone during prolonged dehydration in the domestic flowl: effect of dietary NaCl

Three groups of White Plymouth Rock laying hens were adapted to three levels of dietary NaCl: low-NaCl food with tap water (LOW), high-NaCl food (1% NaCl w/w added) with tap water (HT), and high-NaCl food with 0.5% NaCl for drinking (HS). The birds were subjected to water deprivation (dehydration) for 18 days. Blood sampling was done at 2-4 day intervals. Plasma concentrations of arginine vasotocin (AVT), prolactin (PRL), aldosterone (ALDO) and corticosterone (CS) were determined by radioimmunoassay. Plasma osmolality, sodium, chloride, and potassium were also determined. In the normally hydrated hens fully adapted to the diets, there was a stepwise increase from LOW to HS in plasma osmolality (305, 315, 332 mOsm, for LOW, HT and HS, respectively), [Na+] (144, 153, 161 mM) and [Cl-] (109, 119, 127 mM) as well as in [AVT] (6, 14, 18 pg/ml) and [PRL] (16, 24, 34 ng/ml). Regressing [AVT] on osmolality gave a slope of 0.30 pg . ml-1/mOsm and a threshold of 273 mOsm. The slope of [PRL] on osmolality was 0.73 ng . ml-1/mOsm. The correlation coefficient of [AVT] and [PRL] was 0.67. LOW had high [ALDO] (165 pg/ml) which was suppressed to low levels in HT and HS (5-8 pg/ml), while [CS] was the same in all groups (0.9-1.1 ng/ml). Plasma [K+] was decreased in the high-NaCl groups (5.8 mM in LOW, 4.4 and 4.7 mM in HT and HS). Dehydration resulted within 2 days generally in a sharp (5-15%) increase in osmolality, [Na+] and [Cl-], which thereafter increased more slowly during the remaining 16 days in all groups, with the slowest increase in LOW. The levels of osmolality [Na+] and [Cl-] were 5% lower in LOW than in HT and HS, which showed the same levels during the dehydration period. Plasma [AVT] and [PRL] increased 2-4 fold within 2 days of dehydration; [AVT] reached a plateau at 29 pg/ml in all groups, but [PRL] continued to rise in all groups, fastest in LOW, reaching similar levels in all groups after 14-18 days of dehydration, about 85 ng/ml. The correlation coefficient of [AVT] and [PRL] was decreased by half (to 0.32) during dehydration. Plasma [ALDO] increased in all groups with dehydration, 1.7 fold in LOW and 3-6 fold in HT and HS, but the levels reached in HT and HS were only 15-30% of that seen in LOW.(ABSTRACT TRUNCATED AT 400 WORDS)     

A BR 1 gene in Chironomus tentans has a composite structure: a large repetitive core block is separated from a short unrelated 3''-terminal domain by a small intron.

The large Balbiani ring (BR) genes in the dipteran genus Chironomus have been considered to be homogeneous repetitive structures. Analysis of a genomic DNA segment now reveals that a BR 1 gene in C. tentans is a composite gene, consisting of two different types of sequences. A 15-20 kb core block of tandemly arranged repeat units extends close to the 3' end of the BR 1 gene and ends in repetitive structures partly different from the repeat units in the core block. A 55 bp long intron separates the core block, which probably constitutes a single exon, from a non-related 3'-exon, comprising the final 332 bp of the translated part of the gene. According to hydrophobicity and secondary structure predictions, the 3'-exon encoded peptide is distinctly different from the repetitive core block domain and attains a globular structure. The carboxyl-terminal peptide domain is likely to be a general feature of BR encoded proteins and may have important functions in the excretion and polymerisation of the secretory proteins.     

Histocompatibility Gene Organization and Mixed Lymphocyte Reaction

TRANSFORMATION of allogenic lymphocytes in mixed cultures depends chiefly on an incompatibility between the lymphocyte donors at the major histocompatibility locus in man (HL-A), mouse (H-2) and rat (H-l)¹. Although the mouse H-2 locus can be divided into several regions each of which controls one or more antigenic specificities² and two or more subloci control HL-A antigens in man³, it is not known whether all parts of the major histocompatibility locus are equally important in eliciting transformation in mixed lymphocyte cultures. We now show that capacity to elicit lymphocyte transformation is different for different parts of the mouse H-2 locus.     

Studies on phytohemagglutinins XXIII. A receptor which does not conform to the so called sugar specificity of the pea phytohemagglutinin

A receptor glycopeptide for hemagglutinin of the pea (Pisum sativum L.) was isolated from rabbit erythrocytes by the method originally applied for isolation of a receptor glycopeptide from human erythrocytes (Kubánek, J., Entlicher, G. and Kocourek, J. (1973) Biochim. Biophys. Acta 304, 93–102). The receptor isolated from rabbit erythrocytes contains galactose, mannose, N-acetylglucosamine, asparic acid, threonine, serine glutamic acid and glycine in approximate molar ratios of 2:1:3:3:1:1:1:1. It has a minimum molecular weight of about 2000. According to the sequence analysis the following structure has been proposed for the receptor:

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About 90% of the receptor-site activity for the pea hemagglutinin is due to the presence of two galactose residues on non-reducing terminals of the saccharide chain in spite of the fact that the pea haemagglutinin is not inhibited by this sugar.     

The extent and differences in mitotic activity of the root tip ofVicia faba L.

Mitotic activity does not stop for different meristematic cells of the root apex at the same distance from the initials. The differences are connected with the functional heterogeneity of the apical meristem of the root. The arrangement of vascular bundles,i.e. the alternation of independent xylem and phloem groups, is of major importance. In broad bean roots, the protophloem sieve elements stop dividing first. The centre of the stelei. e. late metaxylem elements stop dividing next. Division in the stele gradually ceases centrifugally, while it ceases centripetally in the peripheral part of the root. The cylindrical region with prolonged cell division includes internal layers of the cortex including endodermis, pericycle and adjoining cells of the stele. Proximally apical meristem is reduced to isolated strands of cells adjacent to the protoxylem poles. Pericycle cells stop dividing last at a distance of approx. 9–10 mm from the initials. The number of the division cycles is limited and is specific for individual cell types. Epidermal and cortical cells divide in broad bean roots transversely approximately seven times, cells of late metaxylem approximately five times. Root apical meristem is an asynchronous cell population with a different duration of the mitotic cycle. We determined local variations in the duration of the mitotic cycle in the apical meristem of broad bean root by means of colchicine-induced polyploidy. The cells of the quiescent centre had the longest mitotic cycle after colchicine treatment. The region of the proper root adjacent to the quiescent centre was mixoploid (2n and 4n). Isolated cells with a long cycle occurred also in the cortex and in the central cylinder. Cells with a division cycle of 18h were found in the root cap, in the epidermis, in the cortex and in the central cylinder. Relatively numerous cells with the shortest division cycle, approx. 12 h, occurred farther of the quiescent centre in the epidermis, in the cortex, in the pericycle, and in adjacent layers of the stele through-out the entire meristematic region. The results derived from the analysis of the apical meristem are discussed in connection with the ontogenesis of different types of cells taking part in the primary structure of the root.     

Der Einfluß einiger Hormone auf die heteroplastische Knorpel- und Knochenbildung im Herzmuskel der Ratte

Zusammenfassung Es wird über histopathologische Untersuchungen des chronologischen Verlaufs an der durchSelyes Ventrikel-Ligatur hervorgerufenen Herzspitzennekrose bei Ratten berichtet. In dem der Nekrose verfallenen Myokard setzt alsbald Organisation ein; dann wandelt sich allmählich das Granulationsgewebe in eine zellarme und faserreiche Narbe um. In den subendokardialen Schichten des Narbengewebes entstehen erst Knorpel-, später Knochenherde. Der Prozeß der Knorpelbildung wird nicht von Verkalkung eingeleitet; vielmehr werden zuerst die Fibroblasten zu Knorpelzellen, um die sich dann sekundär Kalziumsalze ablagern; danach spielt sich eine typische endochondrale Ossifikation ab. Schließlich erscheint Knochenmarkgewebe zwischen den Knochenbalken.Triamcinolon hemmt geringgradig die Bindegewebsproliferation, Thyroxin steigert die Knorpel- und Knochenbildung, während Östradiol diese Vorgänge nicht beeinflußt.

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The effect of hormones on the heteroplastic cartilage and bone formation in the cardiac muscle of the rat

Summary A chronologic study was made of the histopathologic changes which occur in the cardiac apex of the rat following the application of Selye's ventricular ligature. Organisation in the necrotic cardiac muscle begins soon after ligature. Later, the granulationtissue is gradually replaced by scar tissue which is poor in cells but rich in fibers. In the subendocardial fibrous tissue, cartilage and bone develop. It is emphasized that cartilage formation is not initiated by calcification. Instead, the fibroblasts are converted to cartilage cells and, later, calcium salts are deposited in the matrix. This is followed by endochondral bone formation. Finally, bone marrow appears in the intertrabecular spaces.Triamcinolone mildly hindered connective-tissue proliferation, thyroxine increased cartilage and bone formation, while estradiol did not influence these processes.

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Die Versuche, die diesem Bericht zugrunde liegen, wurden durch das Ministère de la Santé, Québec, die Quebec Heart Foundation, Montreal, das Medical Research Couneil of Canada (Block Term Grant MT-1829) und das USPHS, Child Welfare (Grant HDO 2612-02) unterstützt. Die Autoren danken an dieser Stelle der Firma Lederle Laboratories Div., Pearl River, N.Y., USA für das bei diesen Versuchen verwendete Triamcinolon (Aristocort^®) und der Firma Schering Corporation Ltd., Pointe Ciaire, Quebec, für die Bereitstellung von Estradiol.