Studies on the interactions between glycosylated β-peptides and the lectin Vicia villosa by saturation transfer difference NMR spectroscopy

Saturation transfer difference (STD) NMR spectroscopy was used to study the interaction of the lectin Vicia villosa (VVLB₄) with α-d-GalNAc glycosylated β³-peptides. The data were compared to those obtained with the monosaccharides d-Gal, d-GalNAc, and d-Glc as well as with those obtained with the Tn antigen α-glycopeptide (d-GalNAc-α-O-Ser/Thr), molecule naturally recognized by V. villosa. Evidence that the lectin also recognizes glycosylated β³-peptides and has close contact with both the sugar and amino acid moieties was obtained.     

Purification of complement components by hydrophobic affinity chromatography on phenyl—Sepharose: Purification of human C5

Human complement components C5 and C3 were purified with 41% and 20% yields, respectively, by euglobulin precipitation, DEAE—Sephacel ion-exchange chromatography and gel filtration. Phenyl—Sepharose chromatography allowed the complete separation of C3 and C5. C3 bound loosely on the resin whereas C5 bound firmly and was eluted with 50% glycerin solution. Gel filtration on Sephacryl S-300 allowed the depletion of C4bp and H that contaminated C5 preparations. Homogeneity of C5 and C3 preparations was demonstrated by SDS—PAGE and immunochemical analysis. C5 and C3 consisted of two chains (α, 110000; β, 75000) linked by disulfide bridges.     

Estimation of protein digestibility—III. Studies on the digestive enzymes from the pyloric ceca of rainbow trout and salmon

Digestive proteinases were isolated and partially purified from the pyloric ceca of trout and salmon. Their stability and some catalytic properties were compared with those of a three-enzyme system that is used for determination of in vitro protein digestibility. In contrast to the three-enzyme system, pyloric ceca trypsin and total proteinase activity were least stable at pH values below 5.0 and most stable under alkaline conditions up to pH 10.0. Thermal inactivation (50%) occurred in 60 min at 55°C for trypsin activity of trout and salmon ceca proteinases and at 40°C for the three-enzyme system at the pH (8.0) of the in vitro assay. Thermal inactivation (50%) of total proteinase activity occurred in 60 min at about 55, 50 and 35°C for chinook, trout and three-enzyme preparations, respectively. SDS-PAGE zymograms of the ceca enzymes showed the presence of several proteolytic activity bands. Two of the bands corresponded in molecular weight to trypsin and chymotrypsin. Ceca proteinases differ from the three-enzyme system in their response to inhibitors; in particular, the ceca proteinases are much more sensitive to soybean trypsin inhibitor than the procine trypsin used in the three-enzyme system when assayed for trypsin, but less sensitive when assayed for total proteinase. The distinctive properties of ceca enzymes help explain why they are more appropriate than the three-enzyme system, and other enzyme cocktails for in vitro protein digestibility assay of saunonid feed components.     

Quality evaluation of Semen persicae (seed of Prunus persica L. Batsch) by gas chromatography–mass spectrometry

Seeds of Prunus persica L. Batsch were collected at various times from different regions of China. The fatty acid composition of the seeds was analysed using gas chromatography-mass spectrometry (GC-MS). The aim was to evaluate whether genetic and environmental factors influenced the chemical profile of fatty acids in Semen persicae. Saturated and unsaturated fatty acids were present at 4.8-8.7% and 90.7-94.8% of total fatty acids, respectively. Oleic and linoleic acids were dominant in all samples and ranged from 59.3 to 81.4% and from 11.6 to 31.0%, respectively. All samples had high levels of unsaturated fatty acids. Hierarchical clustering analysis and principal components analysis were performed to differentiate and classify the samples based on the contents of the characteristic fatty acid constituents. This study provides evidence that metabolites may reflect genetic and environmental similarities, such as management practices used in cultivars (irrigation, fertilization and sanitary treatments), and evidence of the variability of genetic make up and local environmental conditions in samples grown in the wild.     

Quantification of diadenosine polyphosphates in blood plasma using a tandem boronate affinity–ion exchange chromatography system

Endogenous diadenosine polyphosphates (Ap_nAs) have been associated with a variety of biological effects but quantifying their concentration in blood is difficult. We report on the development of a tandem affinity–ion exchange high-performance liquid chromatography (HPLC) system that employs boronate affinity upstream of ion exchange chromatography for automated rapid (45-min) resolution and extraction of Ap_nAs from human plasma. This system obviates previous requirements for multiple column separations and handling steps, so it is ideally set up for time- and cost-efficient screening of blood samples for Ap_nA pharmacokinetic and biodistribution studies.     

Studies on the Constituents of Typha L.—IV. A Preliminary Study of Chemosystematics of Typha L.

From pollen grains of Typha davidiana, T. latifolia, T. angustata the same eight flavonoids have been isolated. They are identified as naringenin I, isorhamnetin II, quercetin III, isorhamnetin-3-O-(2^G-α-L-rhamnopyranosyl)-rutioside IV, quercetin-3-O-(2^G-α-L-rhamnopyranosyl)-rutinosida, V, isorhamnetio-3-O-rutinoside VI, isorhamnetino-3-O-neohesperidoside VII, kampferol-3-O-neohesperidoside VIII. Flavonoids of pollen grains of five species of Typha, including the above three species, were analysed by TLC with the result showing that the constituents in the pollen grains of the five species are very similar. The chemical comparison among Typha and Sparganium and 16 possibly related families shows that Typha is different from Pandanaceae or Pandanales and is similar to Restionaceae, Flagellariaceae, Juncaceae and Cyperaceae in some respects. Typha and Sparganium are very similar in many respects, and they could be treated in the same family, Typhaceae, which merit the rank of order, Typhales.     

14. Studies on Poliomyelitis Virus—Concentration and Purification of the Virus

ABSTRACT

For the purpose of determining the immunogenic potency of polio virus, relatively large amounts of concentrated virus material were prepared which had titres of the order of 10¹⁰ T.C.I.D.jo per ml. These were obtained by pervaporating large quantities of tissue culture fluid containing approximately 10⁶⁵ T.C.I.D.JQ per ml.     

Studies on the C-24 configurations of Δ7-sterols in theseeds of Cucurbita maxima

Uncertainties surrounding the structures of the Δ⁷-sterols in the seeds of Cucurbita maxima have been resolved. Seven components were found by TLC, GLC, HPLC, mass spectrometry and ¹H NMR. They were 24β-ethyl-5α-cholesta-7,22,25(27)-trien-3β-ol, 24β-ethyl-5α-cholesta-7,25(27)-dien-3gb-ol, avenasterol, spinasterol, 24-dihydrospinasterol, 24ζ-methyllathosterol and 25(27)-dehydrofungisterol. The ¹H NMR spectra indicated that the sterols with an ethyl substituent at C-24 occurred in the absence of their C-24 epimers. This seems to be the first instance of the detection of 25(27)-dehydrofungisterol in a higher plant.     

Studies on the adenosine 3′,5′-monophosphate-dependent protein kinase of Blastocladiella emersonii

Using a combination of Chromatographic and sucrose density gradient techniques under carefully controlled conditions of pH and protease inhibitors, we demonstrate that there is only one form of adenosine 3′,5′-monophosphate-dependent protein kinase in the cytosol fraction of the Blastocladiella emersonii zoospore. If any of these conditions are omitted during extract preparation, one obtains what are apparently multiple forms of the enzyme, which are in reality artifacts due to extensive endogenous proteolytic activity. This endogenous protease is stimulated by alkaline pH and inhibited by antipain. The zoospore protein kinase is similar to type II protein kinase from mammalian cells in several aspects including Chromatographic behavior on DEAE-cellulose column, conditions for subunit dissociation and reassociation, as well as the molecular weight value of the regulatory subunit.     

The foraging behavior of honeybees on hairy vetch. III. — Differences in the vetch

Summary The time required for each of 50 honeybees to forage from a hairy vetch blossom, and the time required for each bee that tripped blossoms to trip a different blossom was measured three times each in two fields. There were highly significant differences in the speed with which bees tripped and foraged from blossoms in the two fields. It appeared that the blossoms in one field were easier to trip than the blossoms in the other. The bees spent less time foraging from the blossoms in the field in which tripping was most rapid, probably because there was less nectar in the blossoms in that field than in the other field. The bees that did not trip blossoms foraged from blossoms more rapidly than the bees that did trip the blossoms. After all known sources of variation had been removed there was a highly significant correlation between the times required for bees to trip and to forage from blossoms; this is interpreted as indicating differences in the efficiency of individual bees. In one of the fields many of the bees foraged through punctures in the blossoms made by carpenter bees. An attempt was made to measure the relationship of the amount of sugar collected by bees per unit time to the attractiveness of competing plants.Published as Technical Contribution No. 4636 from theTexas Agricultural Experiment Station.     

Nomenclatural notes on Aster (Asteraceae)—III. The status of a. Sandwicensis

Plants ofAster sandwicensis (A. Gray in H. Mann) Hieron. belong inA. subulatus Michaux but are recognized as taxonomically distinct from those of var.subulatus. A new combination is needed:A. subulatus var.sandwicensis (A. Gray in H. Mann) A. G. Jones, comb. nov.Chamisso s.n. (G-DC!) is chosen as the lectotype.     

The detoxification of α-tomatine by Fusarium oxysporum f. sp. lycopersici

Fusarium oxysporum f. sp. lycopersici detoxifies α-tomatine by producing an inducible extra-cellular enzyme which cleaves the glycoalkaloid into the tetrasaccharide lycotetraose and tomatidine.     

Isolation of heart specific isophosphorylase by affinity chromatography on 5′-AMP sepharose from pig heart

• 1.1. 5′-AMP Sepharose was used for adsorption and separation of the isophosphorylases from pig heart.
• 2.2. The heart specific isophosphorylase was selectively eluted by glucose-6-phosphate from the 5′-AMP Sepharose.
• 3.3. This preparation was homogeneous, the homogeneity was tested by SDS-gel electrophoresis and immunotitration using skeletal muscle anti-phosphorylase.