A new DNA profiling system for cell line identification for use in cell banks in Japan

Summary Using the polymorphic DNA probes, ChdTC-15, ChdTC-114, pYNH24, and λTM-18, a DNA profiling system was developed that verified identities of individual cultured cell lines collected in the Japanese cell banks, JCRB, RCB, and IFO. These highly polymorphic DNA probes include both VNTR (Variable Number of Tandem Repeats) sequences and substantial lengths of unique regions. In the mixed probe system, several distinct bands from four to eight can be used for cell line identification. These bands were widely spread in a range of molecular sizes, and were stable and reproducible under stringent conditions of Southern blot hybridization. Because the DNA profile was specific for each individual human cell line, it is useful not only to authenticate many existing cultured cell lines but also to monitor their identity during propagation in a laboratory, and to confirm newly established lines as unique.     

The late-replicating X chromosome in digynous mouse triploid embryos

Using BrdU-labeling and acridine orange staining, the behavior of X-chromosome replication was studied in 28 XXX and 19 XXY digynous mouse triploids. In some of these the paternal and maternal X chromosome could by cytologically distinguished. Such embryos were obtained by mating chromosomally normal females with males carrying Cattanach's X chromosome which contains an autosomal insertion that substantially increases the length of this chromosome. In the XXX triploids there were two distinct cell lines, one with two late-replicating X chromosomes, and the other with only one late-replicating X. The XXY triploids were also composed of two cell populations, one with a single late-replicating X and the other with no late replicating X chromosome. Assuming that the late-replicating X is genetically inactive, in both XXX and XXY triploids, cells from the embryonic region tended to have only one active X chromosome, whereas those from the extra-embryonic membranes tended to have two active X chromosomes. The single active X chromosome was either paternal or maternal in origin, but two active X chromosomes were overwhelmingly maternal in origin, suggesting paternal X-inactivation in extra-embryonic tissues.     

Diterpene carboxylic acids and a heliangolide from Helianthus angustifolius

Two diterpene carboxylic acids, one a new kaurenoid derivative and one the previously characterized labdane, ()-cis-ozic acid, as well as a     

Synthesis of biologically active cyclic peptides

1. The extent of racemization and the coupling yield in peptide synthesis were studied under high dilution conditions. The azide method yielded the best results. 2. Five linear penta-peptide precursors related to gramicidin S were subjected to cyclization in order to study how the difference in the sequence influences the yield and the ratio of cyclic dimer to monomer. The azide with the sequence of -L -Pro-L -Val-L -Orn(Z)-L -Leu-D -Phe- afforded diZ-gramicidin S in a high yield of 63%. 3. Alternaria mali toxin III, a cyclotetradepsipeptide phytotoxin, was synthesized. The activated linear tetradepsipeptide containing a D -Dap(Z) (N³-Z-D -2,3-diaminopropionic acid) residue at the N-terminus afforded the cyclic precursor (53%). The Dap residue in the precursor was converted into a ΔAla residue by Hofmann degradation to give the desired product.     

Adjuvant actions of polyclonal lymphocyte activators: III. Two distinct types of T-initiating adjuvant action demonstrated under different experimental conditions

We demonstrated that each of various polyclonal lymphocyte activators (PLA) exhibits two types of adjuvant action to initiate the carrier-specific helper T-cell response to otherwise nonimmunogenic antigen. Type 1 action was characterized as that to initiate the T-cell response to subcutaneous injection of soluble bovine γ-globulin (BGG), and type 2 as that to initiate the response to intravenous injection of aggregated BGG. Each of various PLA showed these two types of adjuvant action in a dissociated fashion. The capsular polysaccharide of Klebsiella pneumoniae (CPS-K) showed both types of action to the highest degrees. Lipopolysaccharide of Escherichia coli exhibited type 2 action as markedly as CPS-K, but failed to show type 1 action. Concanavalin A showed definite type 1 action, but not type 2 action. Polyadenylic-uridylic acid showed definite type 2 action, but not type 1 action. Type 1 and type 2 actions of dextran sulfate were minimal. A hypothetical view is presented to consider that type 1 adjuvant action is directed to two mutually independent sites whereas type 2 action is directed to one site.     

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     

Isolation and some properties of copper-binding proteins found in a copper-resistant strain of yeast

Copper-binding proteins were extracted from a copper-resistantstrain of Saccharomyces cerevisiae which was obtained by repeatedsubculturing in a copper-containing medium. They were separatedinto three types through purification steps such as salt fractionation,gel filtration and preparative polyacrylamide gel electrophoresis.They resembled each other in amino acid composition. Acidicamino acids, lysine, serine, glycine and half-cystine constituteda large part of the protein, with a small amount of hydrophobicamino acids. Aromatic amino acids and methionine were almostabsent. The molecular weight of the components was estimatedto be about 10,000 by Sephadex gel filtration and electrophoresison polyacrylamide gel (slope method). Absorption spectra ofthe components exhibited a broad band at 275 nm, but none inthe visible region, thus resembling that of copper-thionein.Moreover, the absorption band at 275 nm changed markedly onaddition of Ag⁺, Hg2⁺, CN^– or H₂O₂, which are well knownas thiol reagents. These components were abo produced in theparent cells, if they could grow in a copper-containing medium.Based the results of experiments using various culture conditionsand some other yeast species, a possible role of the componentsis discussed. (Received July 13, 1976; )     

Inhibitory effects of steroidal allenes on growth,development, and steroid metabolism of the silkworm,Bombyx mori

Steroidal allenes, stigmasta-5,24(28),28-trien-3β-ol (allene-I) and cholesta-5,23,24-trien-3β-ol (allene-II), were tested for their inhibitory effects on growth, development, and steroid metabolism in the silkworm, Bombyx mori. The allenic analogue (I) of stigmasta-5,24(28)-dien-3β-ol (2) was found to be a specific inhibitor for the conversion of stigmast-5-en-3β-ol (1) to stigmasta-5, 24(28)-dien-3β-ol (2) and/or stigmasta-5,24(28)-dien-3β-ol (2) to 24,28-epoxy-stigmast-5-en-3β-ol (3) This inhibitor held the larvae in the second instar for more than 20 days without developing to the third instar, when administered alone or with the dietary sterols of stigmast-5-en-3β-ol (1) or stigmasta-5,24(28)-dien-3β-ol (2). The second allene (II) with a similar structure to cholesta-5,24-dien-3β-ol (4) was also found to be an inhibitor for insect growth and development, but it appeared not to be acting via inhibition of sterol dealkylation.     

Changes in serum lipoproteins during metamorphosis of the bullfrog,Rana catesbeiana

Serum lipoproteins of the bullfrog, Rana catesbeiana, were studied during metamorphosis. Adult bullfrog has essentially one lipoprotein, designated β-lipoprotein. This β-lipoprotein migrates during electrophoresis to β-globulin region and it has a low hydrated density such that it exhibits floatation in a solvent of density 1.063. On the other hand, tadpole serum has one more lipoprotein, designated as α-lipoprotein, in addition to the β-lipoprotein. The α-lipoprotein migrates to the α-globulin region in zone electrophoresis and corresponds to the so called high density lipoprotein judging from ultracentrifugal behavior. Serum α-lipoprotein disappears and β-lipoprotein content decreases during metamorphosis.     

Oxidative Degradation of Squalene by Arthrobacter Species

An organism isolated from soil and identified as Arthrobacter sp. was studied for its squalene degradation. The degradation product from squalene, which accumulated in the culture broth, was isolated and identified as trans-geranylacetone by mass spectrometry, gas chromatography, infrared spectrometry, and nuclear magnetic resonance spectrometry. Addition of a high concentration of K₂HPO₄ to the culture medium resulted in accumulation of fairly large amounts of carboxylic acids in addition to geranylacetone. These carboxylic acids were identified as isovaleric, β,β′-dimethylacrylic, geranic, and (+)-(R)-citronellic acids. Among these acids, α,β-saturated carboxylic acids were found to be predominant in quantity.