Differential Responsiveness in Zonal Growth of Cocklebur Seed Axes to CO2, C2H4, O2 and Respiration Inhibitors

The axial growth of de-coated cocklebur (Xanthium pennsylvanicumWallr.) seeds, whose axes were divided into 4 zones, was examinedin relation to the temperature-dependent shift of the effectof C₂H₄ on germination. At 23?C, where both C₂H₄ and CO₂ stimulatedgermination, CO₂ promoted the axial growth at the radicle tipzone, whereas C₂H₄ promoted growth in the proximal portion ofthe axis. At 33?C, C₂H₄ inhibited germination, and stronglysuppressed the growth at the radicle tip, whereas the effectof CO₂ did not change. The inhibition of growth at the radicletip zone was alleviated by O₂ enrichment, which also reversedthe inhibition of germination. It is thus apparent that thetemperature-dependent shift of the action of C₂H₄ is associatedwith a temperature-dependent responsiveness of the radicle tipzone to C₂H₄. Growth of the radicle tip zone was sensitive toNaN₃, whereas the proximal portion was sensitive to benzohydroxamicacid, an inhibitor of alternative respiration, suggesting thatthere may be an increase in the operation of the alternativerespiration path along a gradient of axial tissue from the tiptowards the cotyledonary side. The effects of CO₂ and C₂H₄ arediscussed in relation to the different respiratory activitiesin each axial zone of cocklebur seeds. (Received May 9, 1986; Accepted November 6, 1986)     

Enzymic synthesis,chemical characterisation and Sda activity of GalNAcß1-4[NeuAcα2-3]Galß1-4GlcNAc and GalNAcß1-4[NeuAcα2-3]Galß1-4Glc

The tetrasaccharides GalNAcß1-4[NeuAc2-3]Galß1-4Glc and GalNAcß1-4[NeuAc2-3]Galß1-4GlcNAc were synthesised by enzymic transfer of GalNAc from UDP-GalNAc to 3-sialyllactose (NeuAc2-3Galß1-4Glc) and 3-sialyl-N-acetyllactosamine (NeuAc2-3Galß1-4GlcNAc). The structures of the products were established by methylation and¹H-500 MHz NMR spectroscopy. In Sd^a serological tests the product formed with 3-sialyl-N-acetyllactosamine was highly active whereas that formed with 3-sialyllactose had only weak activity.     

Ouabain-insensitive salt and water movements in duck red cells. II. Norepinephrine stimulation of sodium plus potassium cotransport

Catecholamines induce net salt and water movements in duck red cells incubated in isotonic solutions. The rate of this response is approximately three times greater than a comparable effect observed in 400 mosmol hypertonic solutions in the absence of hormone (W.F. Schmidt and T. J. McManus. 1977 a.J. Gen. Physiol. 70:59-79. Otherwise, these two systems share a great many similarities. In both cases, net water and salt movements have a marked dependence on external cation concentrations, are sensitive to furosemide and insensitive to ouabain, and allow the substitution of rubidium for external potassium. In the presence of ouabain, but the absence of external potassium (or rubidium), a furosemide-sensitive net extrusion of sodium against a large electrochemical gradient can be demonstrated. When norepinephrine-treated cells are incubated with ouabain and sufficient external sodium, the furosemide-sensitive, unidirectional influxes of both sodium and rubidium are half- maximally saturated at similar rubidium concentrations; with saturating external rubidium, the same fluxes are half-maximal at comparable levels of external sodium. In the absence of sodium, a catecholamine-stimulated, furosemide-sensitive influx of rubidium persists. In the absence of rubidium, a similar but smaller component of sodium influx can be seen. We interpret these results in terms of a cotransport model for sodium plus potassium which is activated by hypertonicity or norepinephrine. When either ion is absent from the incubation medium, the system promotes an exchange-diffusion type of movement of the co-ion into the cells. In the absence of external potassium, net movement of potassium out of the cell leads to a coupled extrusion of sodium against its electrochemical gradient.     

Direct atomization atomic absorption spectrometry for intact biological samples: application to trace metal analysis in silkworm eggs and their hatched worms.

Highly sensitive carbon rod atomic absorption for trace elements was observed by direct atomization of samples with no pretreatment for intact silkworm eggs, their hatched worms, or National Bureau of Standards biological samples (orchard leaves and bovine liver). Crucial temperatures for drying, ashing, and atomization processes were found to obtain accurate results. Results were consistent with those of activation analyses and of ordinary atomic absorption with wet ashing. The precision and dynamic range of this method were evaluated. The advantages and disadvantages of the present approach were critically discussed.     

Scanning electron-microscopy of starch granules, with or without amylase attack

Scanning electron-microscopy (SEM) revealed that, for starch granules relatively susceptible to amylase, numerous pin holes could be observed on the surfaces of granules attacked by amylase. We also observed that the pores penetrated into the inner layers of granules during the enzyme action and some of the granules exhibited a terraced or step-shaped apperance in their inner portions. These internal characteristics are most probably indicative of layered internal structures of the granules. The other characteristic observations by SEM were striated structures on the surfaces of starch granules of banana, lily, and lotus attacked by pancreatin.     

In-depth qualitative and quantitative analysis of composite glycosylation profiles and other micro-heterogeneity on intact monoclonal antibodies by high-resolution native mass spectrometry using a modified Orbitrap

Here, we describe a fast, easy-to-use, and sensitive method to profile in-depth structural micro-heterogeneity, including intricate N-glycosylation profiles, of monoclonal antibodies at the native intact protein level by means of mass spectrometry using a recently introduced modified Orbitrap Exactive Plus mass spectrometer. We demonstrate the versatility of our method to probe structural micro-heterogeneity by describing the analysis of three types of molecules: (1) a non-covalently bound IgG4 hinge deleted full-antibody in equilibrium with its half-antibody, (2) IgG4 mutants exhibiting highly complex glycosylation profiles, and (3) antibody-drug conjugates. Using the modified instrument, we obtain baseline separation and accurate mass determination of all different proteoforms that may be induced, for example, by glycosylation, drug loading and partial peptide backbone-truncation. We show that our method can handle highly complex glycosylation profiles, identifying more than 20 different glycoforms per monoclonal antibody preparation and more than 30 proteoforms on a single highly purified antibody. In analyzing antibody-drug conjugates, our method also easily identifies and quantifies more than 15 structurally different proteoforms that may result from the collective differences in drug loading and glycosylation. The method presented here will aid in the comprehensive analytical and functional characterization of protein micro-heterogeneity, which is crucial for successful development and manufacturing of therapeutic antibodies     

Isolation and Characterization of Globulin from Seeds of Amaranthus hypochondriacus L.

Seeds of grain amaranths contain a high amount (about 60% of total nitrogen) of albumin and globulin and a trace amount of prolamin. From salt-soluble extracts of A. hypochondriacus seeds, a globulin (440,000 in apparent molecular weight and ) was purified by Sepharose 6B gel and DEAE-cellulose column chromatographies. The protein comprised at least four kinds of subunits whose molecular weights were 36,000, 32,000, 20,000 and 18,000, respectively. The amino acid composition of the globulin was almost similar to those of soybean and oat globulins.     

Biosynthesis of Cercosporin

¹³C NMR spectra were measured for 19 pyrethroids and their related compounds including allethrin, tetramethrin, resmethrin, furamethrin, phenothrin and permethrin. Complete assignment of chemical shifts was accomplished by relative spectral pattern, single-frequency off-resonance decoupling, benzene substituent effects, proton selective decoupling and use of shift reagents. The use of shift reagent was found to be especially efficient for assignment of ¹³C resonances. In the case of allethrin, the splittings of some resonance peaks were observed originating from diastereomerism.     

Structures and Syntheses of Two Phenolic Amides from Piper nigrum L.

Two phenolic amides were isolated from the fruits of white pepper (Piper nigrum L.) and identified to be N-trans-feruloyl tyramine (2a) and N-5-(4-hydroxyphenyl)-2E, 4E-pentadienoyl piperidine (6a) on the basis of chemical and specrtal evidence. Both compounds were synthesized.