Quantitative immunogold ultracryomicrotome stidies of the distribution of periimplantation proteins in the sheep

Summary Several mammalian uterine and conceptus proteins are produced at specific stages of implantation. Ovine trophoblast protein-1 (OTP-1) is only synthesised of pregnancy (dpc). This immunogold ultracryosection study shows that, during this period, OTP-1 immunoreactivity is only found in the Golgi body of the trophectodermal cells. A second protein, of 14 kD molecular weight (14K protein), has a more varied distribution being found in membrane-bounded crystals in uterine epithelium and trophectodermal cells, and distributed throughout the cytosol and nucleoplasm of the uterine epithelium. There are only trace amounts of the 14 K protein in the fetomaternal syncytium which replaces the uterine epithelium during implantation, and no crystals are found in the trophectoderm after cotyledonary villus formation is initiated at 24–25 dpc. The crystals containing 14 K protein persist throughout pregnancy in the intercotyledonary areas. The narrow time window of OTP-1 occurrence reinforces the suggestion that this represents an important developmental signal, whereas the distribution of the 14 K protein indicates a more general nutritive function.     

The relative importance of local phase and local amplitude in patchwise image reconstruction

Natural images were subjected to patchwise Fourier analysis, and the local amplitude and phase spectra were swapped between different images. When the patches were large relative to the image size, the appearance of the reconstructed image was similar to that of the image from which the phase information had been derived, in agreement with previous reports of phase-dominance in the global Fourier Transform. However, when the patch size was made sufficiently small, the appearance of reconstructed images was dominated by amplitude rather than phase. This was not simply due to the DC component of the amplitude spectrum. Prior low-pass filtering of the images enhanced the dominance of amplitude information in the patchwise transform. We conclude that patchwise-reconstructed images contain two quite distinct kinds of information for the human observer. The first is the positional information (local sign) of the patches themselves; the second is the textural information within patches, which is dominated by amplitude rather than phase. The reason why the global Fourier Transform is dominated by phase is that in the absence of any other information about local sign, phase is necessary to reconstruct localised features such as edges.     

The use of field-inversion gel electrophoresis for deletion detection in Duchenne muscular dystrophy.

Deletion is a common cause of Duchenne muscular dystrophy (DMD). Field-inversion gel electrophoresis, in conjunction with Southern blot hybridization, was used to detect large SfiI DNA fragments in the DMD locus. Two unrelated boys with DMD were found to have abnormal sized DNA fragments resulting from deletions. Some of the female relatives of these patients were also shown by this method to have deletions in the DMD locus.     

Low genetic variability in a Utah cherry-infesting population of the apple maggot, Rhagoletis pomonella

The apple maggot fly, Rhagoletis pomonella (Walsh), has only recently been found in Utah infesting sour cherry, Prunus cerasus L. An electrophoretic comparison of flies from Utah cherries with flies from Illinois hawthorns, Crataegus mollis (T. & G.) Scheele (a native host within the native range of the fly), show a marked reduction of genetic variability in the Utah sample. This result is indicative of a genetic bottleneck associated with the establishment of the apple maggot population in Utah cherries.

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Résumé R. pomonella (Walsh), est originaire de Crataegus dans l'Amérique du N.E. Il attaque de nombreux autres fruits, y compris les pommes et les cerises aigres (Prunus cerasus). La mouche a été récemment signalée en Utah, à la fois sur cerises et sur Crataegus douglasii. Nous avons comparé les niveaux de variabilité génétique d'une population de l'Utah contaminant les cerises et d'une population de l'Illinois contaminant C. mollis (la population de l'Illinois est représentative des niveaux de variabilité génétique dans l'aire d'origine de la mouche).La variabilité génétique à 17 loci a été évaluée par électrophorèse sur gel d'amidon. 10 de ces loci sont polymorphes dans la population d'Illinois, mais seulement 4 dans la population de l'Utah. Les fréquences alléliques de ces 4 loci de R. pomonella diffèrent significativement en Utah et en Illinois. La population de l'Utah présente nettement moins d'allèles par locus, un plus faible pourcentage de loci polymorphes et une hétérozygotie moyenne plus faible que la population de l'Illinois. Tous ces résultats sont conformes aux conséquences prévisibles d'un goulot d'étranglement.Deux explications existent pour cette perte de variabilité, toutes les deux liées à la combinaison de la faible taille de la population et de la dérive génétique ultérieure. Pour la première, la colonisation du cerisier par les mouches venant de Crataegus peut avoir provoqué un goulot d'étranglement génétique. Au contraire, la réduction de la variabilité peut avoir été la conséquence de la colonisation de l'Utah par R. pomonella. Nous retenons cette dernière comme la cause la plus vraisemblable de la variabilité génétique de la population de R. pomonella contaminant les cerises de l'Utah.

     

Azotobacter vinelandii ferredoxin I: cloning, sequencing, and mutant analysis

The structure of Azotobacter vinelandii ferredoxin I (AvFdI) has been extensively characterized by a variety of techniques. Although its physiological function is unknown, it has long been implicated as being involved in electron donation to nitrogenase. Here we report that the AvFdI gene (fdxA) has been cloned from an EcoRI digest lambda library using a synthetic oligonucleotide probe and that its sequence has been determined. The amino acid sequence deduced from the DNA sequence is identical to the previously published protein sequence. Analysis of the promoter region indicates that AvFdI is not a nif specific gene product. A mutant of A. vinelandii has been constructed which is identical to the wild-type, at the DNA level, except that the fdxA gene has been interrupted by insertion of a kanamycin cartridge. This mutant, called LM100, does not synthesize AvFdI but does synthesize the Fe and MoFe proteins of nitrogenase and grows at wild-type rates under N2-fixing conditions. This demonstrates that AvFdI is not required for N2 fixation by A. vinelandii. There is a small acidic protein, which is present in wild-type A. vinelandii, whose level is dramatically increased in LM100. The nature of this protein is under further investigation.     

The role of sodium restriction in the management of hypertension

More than 50 studies have investigated the effect of altered sodium intake on blood pressure. A regression line drawn through the change in blood pressure and change in sodium intake indicates that blood pressure alters about 10 mmHg (1 mmHg = 133.322 Pa) for every 100 mmol/day alteration in sodium intake, a change similar to that observed in between-population "studies." The studies that have failed to show a change in blood pressure have usually been in people with a blood pressure less than 130/90 mmHg. Normotensive people appear to tolerate a higher intake of sodium before blood pressure rises, but if increased sufficiently, blood pressure rises in most people. Sodium restriction reduces blood pressure in people with severe hypertension, moderate hypertension and mild hypertension. It may be the cause of blood pressure increase associated with age and the reason for the higher prevalence of hypertension and vascular disease in Western communities. Sodium restriction should be used to treat people with elevated blood pressure.     

Detection of the intracellular ras p21 oncogene product by flow cytometry

Rapid identification of the expression of oncogene products in specific cell types could potentially be useful in the diagnosis and treatment of human malignancy. We have now observed that through the use of lysolecithin permeabilization and fluorescence-activated flow cytometry, cells expressing high levels of the v-Ha-ras oncogene product, p21, can readily be distinguished from the nontransformed parent cells in a rapid and quantitative manner.     

Description of Dolichodorus cobbi n. sp. (Nematoda: Dolichodoridae) with Description of Dolichodorus cobbi n. sp. (Nematoda: Dolichodoridae) with Morphometrics and Lectotype Designation of D. heterocephalus Cobb, 1914

Dolichodorus cobbi n. sp. is described and illustrated from soil around roots of "woods" on the Stanley Farm, Scott County, Arkansas. This species is distinguishable from others of the genus by its short tail projection. It is most closely related to D. marylandicus but differs in having a short tail projection, longer stylet, greater body length, larger c value, and also in position of excretory pore. A limited number of Cobb''s original specimens and drawings of D. heterocephalus were examined. A lectotype and paralectotype were designated to establish the taxonomic base for the genus. Cobb''s original modified labeled drawing of D. heterocephalus is also included. Morphometric data on D. heterocephalus from the type locality and several other populations are given. Some variations in female tail shape, body length, and stylet length were noted.     

Binding specificity of the periplasmic oligopeptide-binding protein from Escherichia coli.

The structural properties required for the binding of peptide substrates to the Escherichia coli periplasmic protein involved in oligopeptide transport were surveyed by measuring the ability of different peptides to compete for binding in an equilibrium dialysis assay with the tripeptide Ala-Phe-[3H]Gly. The protein specifically bound oligopeptides and failed to bind amino acids or dipeptides. Acetylation of the peptide amino terminus of (Ala)3 severely impaired binding, whereas esterification of the carboxyl terminus significantly reduced but did not completely eliminate binding. Peptides composed of L-amino acids competed more effectively than did peptides containing D-residues or glycine. Experiments with a series of alanyl peptide homologs demonstrated a decrease in competitive ability with increasing chain length beyond tripeptide. Competition studies with tripeptide homologs indicated that a wide variety of amino acyl side chains were tolerated by the periplasmic protein, but side-chain composition did affect binding. Fluorescence emission data suggested that this periplasmic protein possesses more than one substrate-binding site capable of distinguishing peptides on the basis of amino acyl side chains.