Large-scale purification of hepatitis B surface antigen using affinity chromatography

Summary Large-scale purification of hepatitis B surface antigen, applicable to the preparation of potential vaccines for prevention of hepatitis B, is described. The method involves the following steps: precipitation of serum with polyethylene glycol 6000, affinity chromatography on concanavalin A-Sepharose and on -aminononyl-Sepharose, and isopycnic centrifugation.This study was supported by grant 09011 from the National Heart, Lung and Blood Institute. We thank Dr. B. Hollinger for testing HB_sAg samples by a complement fixation test.     

Cytotoxicity of mouse peritoneal cells after alloimmunization]

CBA Mice were immunized by two intraperitoneal injections of 30 X 10(6) DBA/2 or C57BL/6 spleen cells at days--12 and--2. Peritoneal cell population was obtained at day zero by washing the peritoneal cavity of Mice. Adherent cells were then separated using a 2 hrs. incubation in "Falcon" plates followed by washing. This macrophage-rich peritoneal cell population was found nonspecifically cytotoxic against 51Cr labeled tumoral target cells: P815 X DBA/2 mastocytoma cells, EL4 X C57BL/L lymphoma cells and spontaneous lymphoma AKR cells (same H--2k as CBA). This adherent peritoneal cell cytoxicity was demonstrated after 24 hrs. incubation with the target cells. It was found in nonspecific combination as well as when using target cells syngeneic to the donor. These findings suggest that adherent peritoneal cell cytotoxicity could be at least partly due to macrophages and result from factor (s) released by sensitized lymphocytes in vivo in the same way as has been previously demonstrated in vitro.     

Spatial conformation of human serotransferrin glycans]

The construction of molecular models for the human serotransferrin glycans shows that they present one compact section linked to the protein and constituted by the pentasaccharide alpha-Man-(1 leads to 3)-[alpha-Man-(1 leads to 6)]-beta-Man-(1 leads to 4)-beta-GlcNAc-(1 leads to 4)-beta-GlcNAc-(1 leads to)-Asn to which are attached two "antennae" consisting of the trisaccharide alpha-NANA-(2 leads to 6)-beta-Gal-(1 leads to 4)-beta-GlcNAc. The trisaccharide sequence beta-Man-(1 leads to 4)-beta-GlcNAc-(1 leads to 4)-beta-GlcNAc adopts a flat and rigid conformation, stabilised by hydrogen bonds. In contrast, the sequence alpha-NANA-(1 leads to 6)-beta-Gal-(1 leads to 4)-beta-GlcNAc-(1 leads to 2)-alpha-Man takes up a helical configuration. The two "antennae" can be disposed on the pentasaccharide core to give two possible configurations, one Y-shaped and the other T-shaped. In both cases, the general conformation of the glycans is perfectly compatible with their postulated role as a recognition signal.     

Effects of thyrotrophic-releasing hormone (TRH) on thermoregulation in the rat

At ambient temperatures (Ta) of both 8 and 22 degrees C, intraventricular administration of TRH (10-80 microgram) produced a dose-dependent hypothermia in rats. The hypothermia was due to both decreased metabolic heat production and cutaneous vasodilatation. In contrast, at 30 degrees C Ta, TRH increased metabolic heat production (due to behavioral excitation) and led to hyperthermia.     

Nucleolus organizer region location and ‘ring’ chromosomes in the bharal

Summary Silver-staining has been used to identify the nucleolus organizer regions (NORs) in the bharal. These show homology with sheep, goat, cattle and aoudad. The association of the NORs on both telomeres of chromosome 3 results in a ring chromosome.     

H-Y Evolution

Summary The female: male sex ratio in litters born to C57 black mice immunized with male spleen preparations is 21 and males were stillborn. In additionDrosophila busckii males are H-Y antigen positive. It is argued that H-Y is the primary testicular determinant and that this role is evolutionarily ancient.     

Presence of natural killer cells in Peyer's patches of the mouse

Summary Natural cell-mediated cytotoxicity of normal murine Peyer's patch cells against Ehrlich ascites carcinoma was found in a short-term⁵¹Cr release assay. Peyer's patch and lymph node cells showed natural cytotoxicity at approximately the same level.Acknowledgments. This work was supported by grant No. 10.5 from the Polish Academy of Sciences.     

Giant collagen fibres in the gonopodium of the mosquitofishHeteradria formosa,Agassiz, 1853 (Pisces,Poeciliidae)

Summary The gonopodium of the mosquitofishHeteradria formosa was studied by transmission electron microscopy. In cross-section the gonopodium shows the following structure from the outside inwards: mutilayered epidermis, basal lamina and central supporting tissue, in which vessels and nerves are embedded. At the top of the gonopodium giant collagen fibres are found, which measure up to 150 m in length and 6 m in diameter. These fibres reinforce the gonopodium.This research was supported by the Deutsche Forschungsgemeinschaft. Address for reprints: Forschungsgruppe Dermatologie der Universität Heidelberg, Im Neuenheimer Feld 324, D-6900 Heidelberg.     

Epithelial-mesenchymal interaction in differentiation of duodenal epithelium of fetal rats in organ culture

Summary Epithelial-mesenchymal interaction in the differentiation of duodenal epithelium of fetal rats was investigated by recombination experiments in vitro. The proportion of goblet cells in duodenal epithelium was significantly greater on recombination of developing duodenal epithelium with mesenchyme of the glandular stomach than on recombination with that of the duodenum. Mesenchyme of the glandular stomach or forestomach was better than duodenal mesenchyme in supporting morphogenesis of duodenal epithelium. Treatment of tissues with N-methyl-N-nitro-N-nitrosoguanidine (MNNG) did not affect these tissue interactions.This work was supported by grants-in-aid for cancer research from the Ministry of Education, Science and Culture, Japan.Acknowledgment. The authors wish to express their gratitude to Prof. T. Mizuno of the University of Tokyo for valuable suggestions.     

Inheritance and expression ofEsterase-1 allozymes inPeromyscus leucopus

Summary We analyzed inheritance of 5Es-1 alleles inP. leucopus and found them to be co-dominant and segregating from a single autosomal locus, thereby verifying assumptions of Mendelian inheritance imfield data. We also described an allele that is silent in hemolysate, but is active in liver extract.Acknowledgment. Laboratory analysis was partially supported by a grant-in-aid (No. 0171-02-240-76) to Kaufman from the Research Foundation of State University of New York.