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.     

The behavioral development of the mutant "staggerer" mouse]

The behavioural study, in particular rearing environmental conditions, of the mutant mouse staggerer has shown that such animals may live more than 90 days. (he behavioural diagnosis of this mutation has been possible from the second week of life, using specific tests. A typical "bat posture" permits one to recognize the mutant from the normal Mouse. Locomotory and feeding behaviours also present late and various qualitatige particularities.     

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.     

Degradation of ribosomal RNA from Escherichia coli by ribonuclease U2 and in vitro transcription of "RNA-fragments" into complementary DNA]

Under well defined conditions ribosomal RNAs purified from Escherichia coli can be degraded by ribonuclease U2 giving rise to RNA fragments of 60--70 nucleotides. In vitro, these fragments are efficiently transcribed into a complementary DNA by DNA polymerase RNA dependent, partially purified from extracts of E. coli. In vivo, "RNA-fragments-U2" inhibit the development of plant tumors.     

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.     

New lectin receptors in carcinoembryonic antigen (CEA).

The glycoprotein CEA (carcinoembryonic antigen) carries carbohydrate groups, which react with the plant lectins from Agaricus bisporus, Arachis hypogaea (peanut), with Tridacnin from invertebrate clams and with the anti-A lectins from snails. Accordingly, it has cryptantigenic structures, which correspond to the T or T-like antigen, the Tridacnin receptor and to the so called A-like antigen.     

Demonstration of generators of the locomotor and respiration rhythms in the cervico-thoracic spinal cord of rabbits

In curarized Rabbits whose cervico-thoracic cord has been isolated through C2 and Th12 transections, a pharmacological activation (nialamide-DOPA) disclosed distinct rhythmic efferent activities, locomotor-like bursts in forelimb muscle nerves and "respiratory" discharges in the phrenic nerves: they originate respectively from a cervico-thoracic locomotion generator and from a "respiration" generator; these spinal generators appear to be interconnected with each other.     

Hypnotic effect of serotonin administered in the vago-aortic afferent pathway

A minute amount of serotonin injected in the nodose ganglion circulation area develops abrupt myosis and general electrocortical synchronization activity in "encéphale isolé" Cat preparation. This hypnogenic effect of serotonin can still be reproduced after transection of vago-aortic nerves caudally to the nodose ganglia. The same injections become ineffective after rostral transection of the same pathway. These results suggest that serotonin may trigger some signs of sleep through peripheric nervous elements in which are probably localized in the nodose ganglia.     

Conversion of mammalian cyclic GMP-dependent protein kinase into modulator-dependent protein kinase (type II) in vitro

The spontaneous conversion of mammalian cyclic GMP-dependent protein kinase (G-PK) into modulator-dependent protein kinase (type II) (M-PKII) in the absence of cGMP or histone was observed in vitro. The findings, together with similarity in substrate protein specificity, suggest that M-PKII is the catalytic subunit of mammalian G-PK.     

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.