Suppression of overt diabetes in NOD mice by anti-thymocyte serum or anti-Thy 1, 2 antibody

Effects of anti-thymocyte serum (ATS) and anti-Thy 1, 2 monoclonal antibody on the spontaneously occurring diabetes in NOD mice were examined. Spontaneous diabetes in female mice was markedly suppressed by intravenous injection of rabbit anti-mouse thymocyte serum diluted to 1:4 on three consecutive days during the time period from 70 to 100 days after birth; the cumulative incidence of overt diabetes upto 195 days of age was greatly reduced and the onset of diabetes was delayed. Similar effect was observed with anti-Thy 1, 2 antibody treatment. These findings suggest that T lymphocytes play a role in the production of spontaneous diabetes in this mouse strain.     

Glycinin A3B4 mRNA. Cloning and sequencing of double-stranded cDNA complementary to a soybean storage protein

The cDNA clones encoding the precursor form of glycinin A3B4 subunit have been identified from a library of soybean cotyledonary cDNA clones in the plasmid pBR322 by a combination of differential colony hybridizations, and then by immunoprecipitation of hybrid-selected translation product with A3-mono-specific antiserum. A recombinant plasmid, designated pGA3B41425, from one of six clones covering codons for the NH2-terminal region of the subunit was sequenced, and the amino acid sequence was inferred from the nucleotide sequence, which showed that the mRNA codes for a precursor protein of 516 amino acids. Analysis of this cDNA also showed that it contained 1786 nucleotides of mRNA sequence with a 5'-terminal nontranslated region of 46 nucleotides, a signal peptide region corresponding to 24 amino acids, an A3 acidic subunit region corresponding to 320 amino acids followed by a B4 basic subunit region corresponding to 172 amino acids, and a 3'-terminal nontranslated region of 192 nucleotides, which contained two characteristic AAUAAA sequences that ended 110 nucleotides and 26 nucleotides from a 3'-terminal poly(A) segment, respectively. Our results confirm that glycinin is synthesized as precursor polypeptides which undergo post-translational processing to form the nonrandom polypeptide pairs via disulfide bonds. The inferred amino acid sequence of the mature basic subunit, B4, was compared to that of the basic subunit of pea legumin, Leg Beta, which contained 185 amino acids. Using an alignment that permitted a maximum homology of amino acids, it was found that overall 42% of the amino acid positions are identical in both proteins. These results led us to conclude that both storage proteins have a common ancestor.     

Purification and physicochemical characterization of murine T cell replacing factor (TRF)

Murine T cell replacing factor (TRF) was purified from a cellfree supernatant of a T cell hybridoma (B151K12) that constitutively produces TRF. Two assay systems for TRF activity were employed: 1) induction of anti-DNP IgG PFC responses in cultures of splenic B cells from DNP-KLH-primed BALB/c mice, and 2) induction of IgM PFC in chronic B cell leukemic cells (BCL1). The purification scheme consisted of ammonium sulfate precipitation, DEAE-cellulose chromatography, Blue-Sepharose chromatography, hydroxylapatite chromatography, gel permeation with fast protein liquid chromatography (FPLC), and disc polyacrylamide gel electrophoresis. Overall, TRF was purified approximately 34,000-fold with a maximum 3.8% recovery of activity, and the specific activity of the purified TRF was approximately 9.6 X 10(4) U/mg. The TRF that is active in these systems is distinct from the other lymphokines such as IL 1, IL 2, BCGFI (now known as BSFp1), and gamma-interferon. The TRF is extremely hydrophobic, with an apparent m.w. of 50,000 to 60,000 on gel permeation chromatography and 18,000 on SDS-PAGE under reducing conditions. Highly purified B151-TRF abrogated the activity by treatment with trypsin but not with RNase. Moreover, it bound to lima bean agglutinin-Sepharose specific for N-acetylgalactosamine residues, indicating that B151-TRF is a glycosylated glycoprotein containing N-acetylgalactosamine residues. The role of N-acetylgalactosamine residues on TRF activity was additionally substantiated by the fact that the addition of appropriate amounts of N-acetylgalactosamine in the assay systems for TRF preferentially induced a profound suppression for TRF-mediated PFC responses.     

Incorporation of sialoglycoprotein containing lacto-series oligosaccharides into chicken asialoerythrocyte membranes and restoration of receptor activity toward hemagglutinating virus of Japan (Sendai virus)

Bovine erythrocyte sialoglycoprotein (GP-2) (1) containing lactoseries oligosaccharide chains, which showed highly specific inhibition of hemagglutination by HVJ (Hemagglutinating virus of Japan, Sendai virus), was incorporated into neuraminidase-treated chicken erythrocytes which had lost their biological responsiveness to the virus. The GP-2-incorporated erythrocytes were agglutinated and lyzed again by the virus. Incorporation of 1,900 molecules of GP-2 per asialoerythrocyte restored fairly well the susceptibility of the cells to HVJ-mediated agglutination and hemolysis. Treatment of the erythrocytes with neuraminidase again resulted in the complete abolishment of the response to HVJ. The above observations are consistent with the view that exogenous sialoglycoprotein, GP-2, can be functionally integrated into the surface membrane of asialoerythrocytes and serve as the receptor for HVJ during the initial adsorption-fusion phase of the virus infection of the target cells.     

Coordinate repression of arginine aminopeptidase and three enzymes of the arginine deiminase pathway in Streptococcus mitis

Streptococcus mitis contains two arginine aminopeptidases (I and II) as an arginine-supplying system and the arginine deiminase pathway as an arginine-utilizing system. The levels of arginine aminopeptidase I and three enzymes of the arginine deiminase pathway were suppressed by glucose in an apparently coordinate manner. Enzyme II appeared to be constitutive.     

Suppressive effect of elcatonin, an eel calcitonin analogue, on excessive urinary hydroxyproline excretion in polyostotic fibrous dysplasia (McCune-Albright's syndrome)

A 12-year-old Japanese girl with polyostotic fibrous dysplasia and endocrine concomitants, was treated with elcatonin, a synthetic eel calcitonin analogue, 10 MRC unit/twice a week given by intramuscular injection. Significant decreases in 24 hr urinary content of hydroxyproline and other amino acids from bone collagen were observed during the course of treatment over 5 months. This biochemical result suggests that the synthetic eel calcitonin analogue exhibits the therapeutic effect in patients with polyostotic fibrous dysplasia by inhibiting bone resorption.     

Uptake of liposome-encapsulating plasmid DNA by plant protoplasts and molecular fate of foreign DNA

Summary Conditions faborable for the uptake of artificial lipid vesicles (liposomes) encapsulating plasmid DNA byDaucus carota protoplasts were investigated. Incubation period necessary for the maximum uptake of liposome-DNA was in the neighborhood of 10 minutes under the circumstances where liposomes (approximately 1.28 moles lecithin/ml) were mixed with 5 × 10⁶ protoplasts/ml. Results obtained by Southern hybridization indicated that the recombinant DNA vector, pBR325 was found in protoplasts after 20 hours incubation in the open circular, linear and some complexed forms. There were some variations in DNA-uptake among protoplasts from different plant species. The gradual disappearance of plasmid molecules was confirmed after 1 week culture, suggesting instability of pBR325 in prolonged cell culture.     

Formation of Recombinants Between Nontransmissible Drug-Resistance Determinants and Transfer Factors

Noninfectious drug-resistance determinants acquired conjugal transmissibility by the formation of recombinants with transfer factors, suggesting the origin of R factors.