Influence of age,weaning, season and body weight on the levels of progesterone,oestradiol-17β and luteinizing hormone in growing buffalo heifers (Bubalus bubalis)

The influence of age, weaning, season of the year and body weight on the peripheral levels of progesterone, oestradiol-17β and luteinizing hormone (LH) were studied during neonatal, perinatal and peripubertal periods in buffalo heifers. The buffalo heifers exhibited oestrus only after 30 months of age and had higher levels of LH and oestradiol-17β and a lower level of progesterone on the day of oestrus. The progesterone concentration was affected significantly (P < 0.01) by different seasons, by weaning (P < 0.05) and varied between pubertal and neonatal periods (P < 0.01), whereas the oestradiol-17β level was affected significantly (P < 0.01) by weaning and varied at different seasons and with body weight. However, the LH concentration was greater during the neonatal period than the pre- and peripubertal periods and changed significantly (P < 0.01) between groups of ages and body weights. The results suggest that increases in the levels of oestradiol-17β and progesterone after 30 months of age are probably indicative of the onset of puberty in buffalo heifers. However, a further increase in oestradiol-17β, LH, and a decrease in progesterone are essential for oestrus and cyclicity to be exhibited in buffalo heifers.     

Purification and immunological properties of an NAD(H) dependent glutamate dehydrogenase from soybean cells (Glycine max L.)

Studies were carried out on glutamate dehydrogenase (GDH, EC 1.4.1.2) isolated from the SB1 and SB3 soybean (Glyciene max L. cv. Mandarin) cell cultures. The NAD(H) dependent enzyme from SB1 and SB3 cells was purified to homogeneity, and that from the SB3 cells studied in detail. It was shown to be activated by calcium. The molecular weight of the native enzyme was found to be 263 000 ± 12 000. The molecular weight of the subunits was shown to be 41 000 ± 2000, which indicates that the enzyme has a hexameric structure. Anti-GDH antibodies were produced in rabbits, to GDH purified to homogeneity from both cell cultures. Each antibody preparation reacted with the purified enzyme produced from either cell culture. Antibodies to GDH from SB3 cells were utilized to study the apparent induction of GDH, which occurs when these cells are grown in a medium with ammonium ions as the sole nitrogen source. The increase in GDH activity was shown to be due to de-novo protein synthesis. The anti-SB3-GDH antibody preparation was also tested for cross reactivity with crude GDH preparations from a number of plant sources, and purified GDH from a number of other organisms. The antibody was shown to cross react with a number of the GDH preparations.     

Dissociation of fibrinogen and fibronectin binding from transglutaminase-mediated cross-linking at the hepatocyte surface

The interaction of fibrinogen and fibronectin with hepatocytes has been dissociated into distinct binding and cross-linking steps. Binding and cross-linking of 125I-labeled ligands were both decreased by transglutaminase inhibitors, but not by heparin or hirudin. Transglutaminase activity was manifest by Ca2+-dependent incorporation of [14C]putrescine into cells. Preferential cross-linking of fibrinogen A alpha over gamma chains, and lack of inhibition by heparin or hirudin indicates the involvement of tissue transglutaminase, and not Factor XIIIa. Hepatic transglutaminase activity, as well as binding and cross-linking of fibrinogen and fibronectin, were maximally supported by Ca2+, partially supported by Mn2+ and Sr2+, and markedly decreased by Mg2+ and Ba2+. In contrast, Co2+ supported binding but not cross-linking or transglutaminase activity, indicating that binding and cross-linking are dissociable events. This conclusion was corroborated by the finding that fibrinogen fragments D95 and D78 both inhibited Ca2+-dependent fibrinogen binding without being cross-linked themselves. Ligand binding in the presence of either cation was localized to the cell surface as evidenced by its trypsin sensitivity. Thus, fibrinogen and fibronectin binding to hepatocytes is independent of transglutaminase activity, whereas cross-linking of these adhesive macromolecules requires an enzymatically active cellular transglutaminase. In addition, fibrinogen binding appears to be mediated by molecular determinants present in fragment D78.     

Structural and functional relationships of human ferritin H and L chains deduced from cDNA clones

We have isolated essentially full-length cDNA clones for human ferritin H and L chains from a human liver cDNA library. This allows the first comparison of H and L nucleotide and amino acid sequences from the same species as well as ferritin L cDNA sequences from different species. We conclude that human H and L ferritins are related proteins which diverged about the time of evolution of birds and mammals. We also deduce the secondary structure of the H and L subunits and compare this with the known structure of horse spleen ferritin. We find that residues involved in subunit interaction in shell assembly are highly conserved in H and L sequences. However, we find several interesting differences in H subunits at the amino acid residues involved in iron transport and deposition. These substitutions could account for known differences in the uptake, storage, and release of iron from isoferritins of different subunit composition.     

Population biology of Avena

Summary Surveys for polymorphisms in natural populations of A. barbata sampled in California grasslands had provided evidence for widespread monomorphism and rather localized polymorphic areas in the north coastal and San Francisco regions, based on a set of morphological and isoenzymatic marker loci. Since this species, like many other annuals, was introduced from the Mediterranean region during the Spanish mission period, a comparative study of the Canadian-Welsh collections of Avena species from the Mediterranean region was undertaken using various plant characters and starch gel electrophoresis to analyze variants for esterase, phosphatase and peroxidase systems. A total of 96 samples including 73 of A. barbata and 23 of A. hirtula were studied and the results were scored to compute the polymorphism indices. In both species, only 10 to 15 percent sites showed any significant degree of polymorphism of which a majority seemed to originate from localized regions in Italy and Turkey; a part of this observed lack of within-sample variation might be the result of small sample size. In general, the patterns of variation in A. barbata from the California surveys and the present analyses seemed to be very similar and raised some interesting questions on (a) the colonizing history of introduced materials (b) the factors underlying such marked patterns of geographical variation, and (c) the current evolutionary changes occurring in these two broad, disjunct areas of species distribution.

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Zusammenfassung Untersuchungen der Polymorphismen in natürlichen Populationen von A. barbata im kalifornischen Weideland hatten einerseits zum Nachweis eines weit verbreiteten Monomorphismus und andererseits streng lokalisierter polymorpher Bereiche in der nördlichen Küsten- und San Francisco-Region geführt, wobei eine Anzahl morphologischer und isoenzymatischer Markerloci zugrunde gelegt wurde. Da diese Art, wie viele andere Annuelle auch, während der spanischen Missionsperiode aus der Mittelmeerregion eingeführt wurde, wurde eine vergleichende Untersuchung der Canadian-Welsh-Sammlungen von Avena-Arten aus der Mittelmeerregion anhand verschiedener Merkmale der Pflanzen und der Stärkegelelektrophorese-Untersuchung auf Esterase-, Phosphatase- und Peroxydase-Systeme durchgeführt. Es wurde eine Gesamtheit von 96 Stichproben, bestehend aus 73 A. barbata und 23 A. hirtula, untersucht und die Ergebnisse zur Berechnung von Polymorphismus-Indices verwendet. In beiden Arten zeigten nur 10 bis 15% der Herkünfte einen signifikanten Polymorphismusgrad. Von ihnen scheint die Mehrzahl von lokalisierten Regionen in Italien und Griechenland abzustammen. Ein Teil des beobachteten Fehlens einer Variation innerhalb der Stichproben könnte eine Folge des geringen Stichprobenumfangs sein. Im allgemeinen scheinen die Variationsmuster der kalifornischen Untersuchungen und die der vorliegenden Analysen von A. barbata sehr ähnlich zu sein. Das führt zu einigen interessanten Fragen nach a) der Besiedelungsgeschichte des eingeführten Materials, b) den Faktoren, die derart auffallenden Mustern der geographischen Variation unterliegen und c) den laufenden evolutionären Änderungen, die in diesen beiden großen, voneinander getrennten Gebieten der Artverteilung auftreten.

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This work was supported in part by a grant from the National Science Foundation (GB 8627).