Purification and Characterization of the Egg Jelly Macromolecules, Sialoglycoprotein and Fucose Sulfate Glycoconjugate, of the Sea Urchin Hemicentrotus Pulcherrimus

A sialoglycoprotein and a fucose sulfate glycoconjugate (FSG) were purified from egg jelly of the sea urchin Hemicentrotus pulcherrimus . Sialoglycoprotein which consisted of sialic acid (90%, w/w) and protein (10%, w/w) did not cause induction of the acrosome reaction and sperm isoagglutination. FSG which contained one mol sulfate/mol fucose possessed 2.0 times protein to fucose by weight. The proteins in intact FSG were separated to two major (258 kDa and 237 kDa) and one minor (120 kDa) proteins by SDS-polyacrylamide gel electrophoresis (SDS-PAGE) in the presence of 2-mercaptoethanol (2-ME) while the proteins could not be separated by HPLC in the presence of 0.1% SDS or SDS-PAGE without 2-ME. However, after carboxymethylation of FSG, two major (260 kDa and 240 kDa) proteins and two minor (140 kDa and 135 kDa) proteins were separated from the fucose sulfate moiety by HPLC in the presence of 0.1% SDS or SDS-PAGE without 2-ME. When FSG was first carboxymethylated with non-radioactive iodoacetic acid and then reduced with 2-ME and finally carboxymethylated with ¹⁴C-iodoacetic acid, the most of radioactivity was detected in 140 kDa and 135 kDa proteins. Carboxymethylted-FSG was less potent than intact FSG in induction of the acrosome reaction. Fucoidan, a fucose sulfate polymer, did not induce the acrosome reaction.     

Effects of pantolactone and butyrolaectone on the pleiotropic phenotypes of lon mutants and on thermal induction of the SOS phenomena in a tif mutant of Escherichia coli K12

Hydrophobic and charge-charge interactions of Salmonella typhimirium and Serratia marcescens were determined and related to their content of fimbriae and lipopolysaccharide (LPS). The cell surface structures were characterized with hydrophobic interaction chromatography (HIC), electrostatic interaction chromatography (ESIC) and particle electrophoresis measurements. The degree of interaction at the air-water interface was tested using a monolayered lipid film applied to an aqueous surface. The cell surface hydrophobicity of S. typhimurium in the presence of fimbriae was less in smooth than in rought bacteria. Examination of a series of rough mutants of S. typhimurium indicates that reduction of the O-side chain and core oligosaccharides was correlated with increased cell hydrophobicity. The enrichment factors at the air-water interface were significantly higher for fimbriated than for non-fimbriated S. typhimurium cells. Fimbriated S. marcescens cells were less hydrophobic and adhered to a lesser degree at the air-water surface than non-fimbriated counterparts. Electrophoretic measurements and adsorption to ion exchangers gives different information about the surface charge of bacteria. The latter technique gives the interaction between localized charged surfaces.Abbreviations HIC hydrophobic interaction chromatography - ESIC electrostatic interaction chromatography - LPS lipopolysaccharide - PBS phosphate buffered saline solution     

Antioxidative effect of α-tocopherol incorporation into lecithin liposomes on ascorbic acid-Fe2+-induced lipid peroxidation

The antioxidative effect of α-tocopherol incorporated into lecithin liposomes was studied. Lipid peroxidation of liposome membranes, assayed as malondialdehyde production, was catalyzed by ascorbic acid and Fe²⁺. The peroxidation reaction, which did not involve the formation of singlet oxygen, superoxide, hydrogen peroxide, or a hydroxyl radical, was inhibited by α-tocopherol and a model compound of α-tocopherol, 2,2,5,7,8-pentamethyl-6-hydroxy-chroman (TMC), but not by phytol, α-tocopherylquinone, or α-tocopheryl acetate. One mole of α-tocopherol completely prevented peroxidation of about 100 moles of polyunsaturated fatty acid. Decrease in membrane fluidity by lipid peroxidation, estimated as increase of fluorescence polarization of 1,6-diphenyl-1,3,5-hexatriene (DPH) embedded in the membrane, was also inhibited by α-tocopherol and TMC, reflecting their antioxidant functions. Cholesterol did not act as an antioxidant, even when incorporated in large amount into the liposome membranes, but it increased the antioxidative efficiency of α-tocopherol. When a mixture of liposomes with and without α-tocopherol was incubated with Fe²⁺ and ascorbic acid, α-tocopherol did not protect the liposomes not containing α-tocopherol from peroxidation. However, preincubation of the mixture, or addition of Triton X-100 allowed the α-tocopherol to prevent peroxidation of the liposomes not containing α-tocopherol. In contrast, in similar experiments, liposomes containing TMC prevented peroxidation of those without TMC without preincubation. Tocopherol in an amount so small as to exhibit only a slight antioxidative effect was oxidized when incorporated in egg lecithin liposomes, but it mostly remained unoxidized when incorporated in dipalmitoyllecithin liposomes, indicating that oxygen activated by ascorbic acid-Fe²⁺ does not oxidize α-tocopherol directly. Thus, decomposition of α-tocopherol may be caused by its interaction with peroxy and/or alkoxyl radicals generated in the process of lipid peroxidation catalyzed by Fe²⁺ and ascorbic acid.     

Oligouridylates as a template for nonenzymatic synthesis of oligoadenylates

Summary Chemical polymerization of adenosine-5-phosphorimidazolide was conducted in the presence of oligouridylate templates. Oligo U with chain length more than eight served as a template and facilitated oligoadenylate formation. No template activity was observed when oligo U up to a hexamer was used. These results correlate with thermal transition temperatures of oligo U-pA complexes.     

Highly Sensitive Assay for Dopamine-β-Hydroxylase Activity in Human Cerebrospinal Fluid by High Performance Liquid Chromatography-Electrochemical Detection: Properties of the Enzyme

Abstract: This paper describes a new, sensitive assay for dopamine-β-hydroxylase (DBH) activity in human cerebrospinal fluid (CSF), serum and brain tissues by high performance liquid chromatography (HPLC) with electrochemical detection (ED). Dopamine (DA) was used as a substrate and was incubated under optimal conditions. Norepinephrine (NE) formed enzymatically from DA was isolated by a double-column procedure, the first column of Dowex-50-H⁺ and the second column of aluminum oxide. NE was adsorbed on the second aluminum oxide column and then eluted with 0.5 M-hydrochloric acid and assayed by HPLC-ED. Epinephrine (EN) was added to each incubation mixture as an internal standard, and this assay was therefore highly reproducible. The peak height in HPLC was linear from 500 fmol to 100 pmol of NE and EN. The lower limit of detection for NE formed enzymatically was about 30 pmol, which indicated that the sensitivity of this procedure was comparable to that of radioassay procedures. We applied the method to measurement of the activity of and examination of some of the characteristics of DBH in human CSF. DBH activity in CSF of Parkinsonian patients was lower than that of control patients. The properties of DBH in human CSF were similar to those in serum and adrenal medulla.     

Cartilage-derived factor (CDF) : II. Somatomedin-like action on cultured chondrocytes

Previously, we showed that fetal bovine cartilage contains a polypeptide that stimulates the incorporation of [³⁵S]sulfate into proteoglycans synthesized by rat and rabbit costal chondrocytes in culture. In this paper, we report that the cartilage-derived factor (CDF) increases not only [³⁵S]sulfate incorporation but also [³H]thymidine incorporation into rabbit chondrocytes in monolayer culture. The dose-response curve of CDF stimulation of DNA synthesis was similar in profile to that of CDF stimulation of proteoglycan synthesis. In addition, CDF markedly enhanced [³H]uridine incorporation into rabbit chondrocytes and significantly enhanced [³H]serine incorporation into total protein. These findings indicate that fetal bovine cartilage contains a factor that shows somatomedin-like activity in monolayer cultures of rabbit chondrocytes.     

Seroimmunological and parasitological surveys of leucocytozoon caulleryi infection in chickens in several asian countries

Epizootiological surveys of Leucocytozoon caulleryi infection in chickens in Japan, Taiwan, Philippines, Singapore, Malaysia and Thailand were undertaken by means of the immunodiffusion test. The rate of infection of L. caulleryi confirmed by the examination of parasites in the peripheral blood of chickens coincided with that of positive antibody response in the immunodiffusion test. Antibodies against L. caulleryi were found in chickens in all the countries surveyed in the present investigation. The prevalence of L. caulleryi infection in chickens was confirmed by the immunodiffusion test. Several chickens in each country showed the presence of serum antigens of L. caulleryi at the times of serum sample collection. These results seemed to indicate that the immunodiffusion test is a method efficient enough to be applicable to the epizootiological surveys and diagnosis of L. caulleryi infection in chickens in the field. As a result, the antibodies or soluble antigens in the sera of chickens infected with L. caulleryi present, respectively, in each country may have the same immunological characters.     

Dental pulp collagenase: initial demonstration and characterization.

A collagenase, active against native helical collagen, was initially found in the explant medium of bovine dental pulp. In contrast to the collagenases from other oral tissues, all the pulp enzyme released was in a latent form which was activated by trypsin treatment, 4-aminophenylmercuric acetate, and some chaotropic agents. The activated enzyme was inhibited by low concentrations of EDTA and calf serum. The molecular weight of activated enzyme was tentatively estimated at 45,000 daltons by gel filtration. The enzyme attacked undenatured collagen in solution at 20°C producing characteristic products $\text{α}{}^{\mathrm{A}}\text{(}\text{3}\text{4}\text{)}$ and $\text{α}{}^{\mathrm{B}}\text{(}\text{1}\text{4}\text{)}$.     

Incidence of major chromosome aberrations in 12,319 newborn infants in Tokyo

Summary In order to ascertain the frequency of chromosome aberrations among newborn infants in Japan, a chromosome survey of a large number of newborn infants is in progress. A consecutive series of 12,319 newborn babies, 6382 male and 5937 female, have been screened for clinical manifestations of autosomal aberrations and for sex chromatin and sex chromosome aberrations. Chromosome studies were carried out on 694 infants with suspected chromosome aberrations. The clinically abnormal infants were screened by conventional staining, and banding techniques have been used in the part of the study performed since 1974. Of the clincally abnormal infants, 25 had abnormal karyotypes, including two males with a 47,XXY complement, one female with a 45,X complement, three male infants with a 47,XYY complement, two with trisomy 13 syndrome, three with trisomy 18 (including one case of mosaicism), eleven with Down's syndrome (including one case of mosaicism), one with B5p partial trisomy, one with cri-du-chat syndrome, and one with Y/D translocation. The overall results are comparable to those of previous population cytogenetic studies only in the autosomal trisomies and sex chromosome abnormalities and in that the observed frequencies were comparable to those found in studies in Caucasians.To whom offprint requests should be sent