Distribution of sulphate and iduronic acid residues in heparin and heparan sulphate

1. A method was developed for determination of the uronic acid composition of heparin-like glycosaminoglycans. Polymers or oligosaccharides are degraded to monosaccharides by a combination of acid hydrolysis and deamination with HNO₂. The resulting uronic acid monosaccharides (accounting for about 70% of the uronic acid contents of the starting materials) are isolated and converted into the corresponding aldono-1,4-lactones, which are separated by g.l.c. The calculated ratios of glucuronic acid/iduronic acid are reproducible within 5%. 2. Samples of heparin from pig intestinal mucosa (molar ratio of sulphate/disaccharide unit, 2.40) and heparan sulphate from human aorta (sulphate/disaccharide ratio, 0.46) were subjected to uronic acid analysis. l-Iduronic acid constituted 77% and 19% respectively of the total uronic acid contents. 3. The correlation between the contents of sulphate and iduronic acid indicated by this finding also applied to the fractionated deamination products of the two polymers. The sulphated fragments varied in size from disaccharide to octasaccharide (or larger) and showed sulphate/disaccharide molar ratios in the range of 0.05–2.0. The proportion of iduronic acid increased with increasing ester sulphate contents of the oligosaccharides. 4. Previous studies on the biosynthesis of heparin in a cell-free system have shown that l-iduronic acid residues are formed by C-5 epimerization of d-glucuronic acid units at the polymer level; the process requires concomitant sulphation of the polymer. The results obtained in the present structural study conform to these findings, and suggest further that similar mechanisms may operate in the biosynthesis of heparan sulphate. The epimerization reaction appears to be linked to the sulphation of hydroxyl groups but does not seem to require sulphation of the target uronic acid residues. The significance of sulphamino groups in relation to the formation of iduronic acid is unknown.     

An investigation of bidirectional translocation in the Phloem

Patterns of (14) CO(2) , assimilate movement in Vicia jaba plants having 7 nodes were studied. Bidirectional translocation occurred throughout most of the stem length when tracer was applied to leaves of various ages. To determine whether this bidirectional translocation occurs within single sieve tubes, a O.1 % solution of the fluorescent dye K-fluorescein was applied to a lightly scraped area on the stem in the middle of a young internode. After one hour the dye was present short distances above and below the treated area. Free-hand sections of the internode showed the dye to be localized in the traces of the larger leaves below tbe treated area and in the traces of the younger leaves above the treated area. The dye was never present in the same bundle both above and below the treated area, indicating that each bundle and sieve tube translocated the dye in only one direction. These results were confirmed using Phaseolus vulgaris, Vinca rosea, and Pelargonium hortum. A similar study in which petioles of young Ecballium elaterium leaves were treated showed that usually the phloem of one bundle translocated the dye in only one direction but in some cases the external phloem of the bicollateral bundles carried the dye toward the stem while the internal phloem carried the dye toward the blade. When longer time intervals were used in all these experiments, the dye sometimes appeared in the same phloem areas both above and below the treated area. This is explained by a lateral transfer of tracer within the phloem, either through secondary phloem or through bundle anastomoses at the nodes.     

Deoxyribonucleic Acid Homology and Relative Genome Size in Mycoplasma

The deoxyribonucleic acid homologies of Mycoplasma laidlawii type A and type B, M. pulmonis (#47 and #63), and M. hominis were determined by membrane methodology. The homology data revealed a difference in genome size between M. laidlawii type A and type B. This difference also held with stringent conditions of annealing (high temperature). Little or negligible homology was shown to exist between the M. laidlawii strains type A and type B and M. pulmonis strains 47 and 63 and M. hominis. M. hominis showed less than 10% homology to the M. pulmonis and M. laidlawii strains. Neither of the M. laidlawii strains showed more than 2% annealing to the M. pulmonis strains. Reaction rate studies are suggested as a means of demonstrating the phylogenetic relationship between the Mycoplasma and other microorganisms.     

An unstable locus in soybeans

Summary Investigation of a variegated condition in the soybean variety Lincoln indicates instability at the Y locus. Leaf sectors of chlorophyll-less yellow tissue occur in distinct heritable patterns; some leaves have small flecks of yellow tissue (late occurring mutations) and others possess large areas or whole leaflets (early occurring mutations).There is evidence that this allele, Y ₁₈ ^m , mutates to the wild type, Y, which is green and stable and to the recessive, y, which is yellow and lethal in the seedling condition. (With an increase in the amount of yellow tissue there is an increase in the frequency of lethals.) However, changes from one type to the other are observed, and patterns of variegation representing different states of the instability are described. These depend upon the time and frequency of mutation events.Evidence is presented to support the hypothesis that this instability is controlled by a factor that resides at the locus. Such a factor governs the timing of the mutation events and is related to similar elements in maize, which are part of specific mutable systems. Control of variegation of the Y ₁₈ ^m locus is compared with the models proposed for the cases of instability in maize.

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Zusammenfassung Die Untersuchung eines variegaten Zustandes bei der Sojabohnensorte Lincoln führte zum Nachweis einer Instabilität des Y-Locus. Blattsektoren mit chlorophyllfreiem gelbem Gewebe traten in bestimmten erblichen Mustern auf. Einige Blätter wiesen kleine Flecken gelben Gewebes auf (spät eingetretene Mutationen), während andere große Flächen oder vollständig gelbe Blättchen besaßen (früh eingetretene Mutationen).Es gibt Beweise dafür, daß das entsprechende Allel Y ₁₈ ^m sowohl zum stabilen Wildtypallel Y, mit grünem Phänotyp, als auch zum rezessiv gelben y, das im Sämlingsstadium letal wirkt, mutiert. (Eine Zunahme der Menge gelben Gewebes ist mit einer Zunahme der Letalfrequenz verbunden.) Umwandlungen eines Typs zu einem anderen werden beobachtet und Variegationsmuster beschrieben, die unterschiedliche Stadien der Instabilität verkörpern. Diese hängen von dem Zeitpunkt und der Frequenz der Mutationsereignisse ab.Es werden Beweise vorgelegt, die die Hypothese stützen, daß diese Instabilität durch einen Faktor kontrolliert wird, der sich am Locus befindet. Ein Faktor dieser Art kontrolliert das zeitliche Auftreten der Mutationsereignisse. Er ist mit ähnlichen Elementen des Maises verwandt, die Teile eines spezifisch mutablen Systems sind. Die Kontrolle der Variegation durch den Y ₁₈ ^m -Locus wird mit den Modellen verglichen, die für die Fälle der Instabilität beim Mais vorgeschlagen wurden.

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Joint contribution from the Iowa Agricultural and Home Economics Experiment Station, Ames, Iowa (Projects 1335 und 1179) as Journal Paper No. 5635.

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Former Agronomist, Crops Research Division, ARS, USDA, and Professor Iowa State University; now Research Director, Peterson Seed Company.     

Amino Acid incorporation in developing fucus embryos

The incorporation of ¹⁴C-leucine and ¹⁴C-amino acid mixture into protein in unfertilized eggs and developing embryos of the brown alga Fucus vesiculosus L. was studied. Bacterial contamination was initially a problem, but it was found that the addition of 40 μg/ml chloramphenicol to the incubation medium would inhibit bacterial protein synthesis without affecting early development of the Fucus embryos. The kinetics of uptake and incorporation of ¹⁴C-leucine into the trichloroacetic acid-soluble and -insoluble fractions indicated that the exogenous precursor did not equilibrate with the main soluble leucine pool before incorporation into protein. Uptake and incorporation of leucine by embryos 90 to 175 minutes old were proportional to exogenous leucine concentration over the range 5 × 10⁻⁶ m to 5 × 10⁻³ m. Unfertilized eggs will incorporate ¹⁴C-leucine into protein. The rate of this incorporation increases dramatically in newly fertilized eggs with a maximum rate at 3.5 hours, a period of cell wall formation and increasing metabolic rates. Thereafter, the rate of incorporation declines until approximately 15 to 17 hours when it increases again concurrently with the onset of rhizoid initiation and cell division.     

Phenylketonuria—Experience at One Center in the First Year of Screening in California

One year''s experience with phenylketonuria during the calendar year 1966, the first year for compulsory newborn screening in California, was reviewed. The over-all prevalence rate from reported cases in California during this period was one case per 19,500 persons tested. Fifty-seven persons suspected of having pku were evaluated, and 25 of them were determined to be phenylketonuric. Eleven of the 25 were infants in whom the abnormality was detected through the newborn screening program or because it was detected in a sibling through a screening program. All the newborn phenylketonuric patients were developing normally at the time of last report (although the follow-up periods were short).In nine of the other children, pku was detected because they were retarded. Five retarded children who were diagnosed as phenylketonuric at another clinic were given dietary assistance.Five additional infants had elevated serum phenylalanines but did not have the classic biochemical findings of pku and are being evaluated further. Nine infants with positive screening tests exhibited biochemical and clinical findings consistent with transient tyrosinemia. Eighteen other children were evaluated and found to have no metabolic abnormality.The newborn screening program for pku is of decided benefit in early identification of a group of infants who have a high rate of potentially serious metabolic disease. Early identification permits treatment soon enough to prevent mental retardation. Newly identified patients should be evaluated in a medical setting capable of careful pediatric, biochemical and nutritional surveillance.