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Biomedical subjects

L D Smith

Publications and source records attributed to L D Smith.

At least 109 records · Page 6Linked to original sources
PubMed1981

Subcellular compartmentalization of maize storage proteins in Xenopus oocytes injected with zein messenger RNAs.

Maize storage proteins synthesized in oocytes were compartmentalized in membrane vesicles because they were resistant to hydrolysis by protease, unless detergent was present. The site of storage protein deposition within the oocyte was determined by subcellular fractionation. Optimal separation of oocyte membranes and organelles was obtained when EDTA and high concentrations of NaCl were included in the homogenization and gradient buffers. Under these conditions, fractions in sucrose gradients containing a heterogeneous mixture of smooth membranes (presumably endoplasmic reticulum, Golgi apparatus, and plasma membrane, density = 1.10-1.12 g/cm3), mitochondria (densities = 1.14 and 1.16 g/cm3), yolk platelets (density = 1.21 g/cm3), and a dense matrix material (density = 1.22 g/cm3) could be separated. Some zein proteins were recovered in the mixed membrane fraction, but the majority occurred in vesicles sedimenting with yolk platelets and granular material at a density of approximately 1.22 g/cm3. When metrizamide was included in the gradient to increase the density, little of the dense matrix material was isolated, and vesicles containing zein proteins were separated from other oocyte components. These vesicles were similar to protein bodies in maize endosperm because they were of identical density and contained the same group of polypeptides.

Animals↗
PubMed1981

Calmodulin triggers the resumption of meiosis in amphibian oocytes.

The calcium-binding protein, calmodulin, has been purified from Xenopus laevis oocytes. This 18,500-dalton protein, pl 4.3, has two high-affinity calcium-binding sites per mole protein having a dissociation constant of 2.8 x 10(-6) M. Full-grown Xenopus oocytes, arrested in late G2 of the meiotic cell cycle, resumed meiosis when microinjected with 60-80 ng (3-4 pmol) of calmodulin in the form of a calcium-calmodulin complex. The timing of the meiotic events in these recipient oocytes was the same as that normally induced by progesterone. Xenopus ovarian calmodulin stimulated bovine brain phosphodiesterase (PDE) 3- to 10-fold in a calcium-dependent manner, but it had no apparent effect on ovarian PDE activity. A calcium-calmodulin-dependent protein kinase has been isolated from Xenopus oocytes using a calmodulin-Sepharose 4B affinity column. The possible role for this kinase in regulating the G2-M transition in oocytes has been discussed.

Animals↗
PubMed1980

Progesterone induces a rapid increase in [Ca2+]in of Xenopus laevis oocytes.

Progesterone causes a rapid increase in the intracellular free calcium level in fully grown amphibian oocytes. When albino Xenopus laevis oocytes were microinjected with the Ca-specific photoprotein aequorin, the calcium-induced luminescence from this protein increased at 40-60 sec after the addition of progesterone and returned to the control level within 5-6 min. No further change in the aequorin glow could be detected through the remainder of the maturation period. This transient increase in cytoplasmic free calcium may be involved in controlling the resumption of oocyte maturation.

Aequorin↗
PubMed1979

Synthesis and processing of maize storage proteins in Xenopus laevis oocytes.

Xenopus oocytes injected with zein mRNAs efficiently synthesized maize storage proteins for prolonged periods. Under optimal conditions, zein was synthesized at 3 ng/hr and represented approximately 10% of the total protein synthesized in the oocyte. The mRNA from the normal maize inbred line directed synthesis of all the major zein components; however, products of mRNA from the opaque-2 mutant did not contain the largest zein component. Zein proteins synthesized in the oocyte were 2000 daltons smaller than proteins synthesized by cell-free translation of mRNAs in the wheat germ and reticulocyte systems. This result, which suggested that the oocyte processed prezein polypeptides into native zein proteins, was confirmed by amino-terminal sequence analysis of zein proteins from the oocytes. Cyanogen bromide cleavage of translation products from oocytes and the wheat germ system confirmed the existence of several proteins within each of the major zein components. However, we were unable to detect the presence of signal sequences on zein peptides by this technique.

Journal Article↗
PubMed1979

Effect of hepatocarcinogens on the adenine purine nucleotide cycle during the initiation phase of carcinogenesis.

Activities of the adenine purine nucleotide cycle enzymes, i.e., adenylosuccinate (SAMP) synthetase, SAMP lyase, and adenosine 5'-monophosphate deaminse, were determined in hepatic tissue of rats fed and/or given injections of 3'-methyl-4-dimethylaminoazobenzene, 4'-methyl-4-dimethylaminoazobenzene, thioacetamide, ethionine, or 2-acetylaminofluorene. SAMP lyase activity showed an early increase in all regimens containing hepatocarcinogens. Adenosine 5'-monophosphate deaminse showed increases with 3'-methyl-4-dimethylaminoazobenzene and thioacetamide but not with ethionine or 2-acetylaminofluorene. SAMP synthetase either was nonresponsive or else showed inhibition to the carcinogens. Increase in SAMP lyase activity was noted as early as 48 to 72 hr following i.p. injections of these carcinogens. The response of SAMP lyase was not duplicated by analogs of carcinogens such as 4'-methyl-4-dimethylaminoazobenzene or methionine. These data imply interaction of active carcinogens with SAMP lyase and to some extent adenosine 5'-monophosphate deaminse or to some mechanism responsible for their synthesis and/or release. This interaction may be a significant component of the initiation phase of carcinogenesis.

2-Acetylaminofluorene↗
PubMed1978

The cyclic behavior of a cytoplasmic factor controlling nuclear membrane breakdown.

The activity of a cytoplasmic factor (MPF), capable of inducing nuclear membrane breakdown (germinal vesicle breakdown) when injected into amphibian oocytes, has been studied during the course of early cleavage in amphibian embryos. Mature egg cytoplasm was found to contain high levels of this activity, but this was quickly lost after fertilization or artificial activation. MPF activity later reappeared in the egg cytoplasm and started to cycle with time. The peak of embryonic MPF activity during each cycle coincided with the time the embryonic nuclei were entering the G2-M transition, i.e., mitosis. However, in colchicine-arrested embryos, this activity remained at an elevated level and no longer oscillated. The timing of the appearance and disappearance of this activity appeared to be under the control of the cytoplasm because such behavior was still observed in enucleated eggs. Continued protein synthesis in the embryo was required for the reappearance, but not for the disappearance, of this activity. MPF, previously thought to be restricted to oocyte maturation, may play a more general role in controlling nuclear membrane breakdown during mitosis as well as meiosis.

Animals↗
PubMed1978

Comparison of high pressure liquid chromatographic and chemical methods for vitamin D3 concentrates. II. Collaborative study.

A collaborative study was carried out which compared the official chemical method, 43.B14-43.B24, the official rat bioassay, 43.165, and the high pressure liquid chromatographic method for vitamin D3 resin, vitamin D3 resin in oil, and dry concentrate. A total of 340 samples were distributed to 17 collaborators for analysis. Five laboratories performed both the chemical and HPLC methods on 5 sets of blind duplicates. A 2-way analysis of variance comparing both methods for each sample showed a significant (P less than 0.01) difference between methods only for Sample 5. When the 2 methods were compared over all the samples, no significant (P less than 0.05) difference was found. Except for Sample 5, there were no differences in the repeatability of the methods. Per cent recoveries on Sample 3, which contained exactly 0.200 X 10(6) IU/g, showed 98.2% for the chemical method and 100.6% for the HPLC method for the 5 laboratories that performed both methods. The assay results of the HPLC and chemical methods are in good agreement with those found by the biological assay on Samples 1-4, but not for Sample 5. Evidence indicates that Sample 5 degraded partially to isotachysterol, and while the HPLC method yielded a reasonable value on this material, the chemical method erroneously showed full potency. An amendment is included for the collaboratively studied HPLC method which detects and eliminates 5,6-trans-vitamin D3, a possible interferant.

Animals↗
PubMed1978

The occurrence of Clostridium botulinum and Clostridium tetani in the soil of the United States.

Soil samples taken every fifty miles on four east-west transects across the United States were examined for C. botulinum and C. tetani, organisms that could inhibit the growth of C. botulinum type A, and for various soil properties. Type A strains were found mostly in the western part of the United States, in neutral to alkaline soil. Type B strains were more uniformly distributed, with a majority of them occurring east of the Mississippi River; none, however, were found in samples taken in the southermost transect. They were associated with soil of high organic content. Type C strains were found only in acid soil of the Gulf Coast, and type D strains in alkaline soil of some western states. Type E strains were mostly associated with damp to wet soil. Organisms inhibitory to type A strains were found in 4 of 21 samples of soil in which type A strains were demonstrated and in 7 of 20 samples in which they were not. Trypsin activation of culture fluids was necessary for the demonstration of most strains of types B, C, D, and E. C. tetani was demonstrated in 30 per cent of the soil samples. Its occurrence was not correlated with any particular soil type or climatic area.

Clostridium botulinum↗