Some cytologic characteristics of the hemocytes of Limulus during clotting

The hemolymph of the horseshoe crab, Limulus (Xiphosura) polyphemus, contains a single cell type. The hemocytes are ovoid and contain many refractile granules. One-half to one minute after the onset of clotting the hemocytes swell and a hyaline cytoplasmic ring, essentially devoid of granules, appears about their circumference. During this time the granules disperse and the nucleus becomes visible. Three to five minutes following the initiation of clotting, the cell extends long pseudopodial processes. Phase contrast time-lapse cinematography reveals that the cells are extremely motile during this phase. Concomitant with these changes, many of the granules lose their refractility and one by one disappear from the cytoplasm leaving what appears to be a vacuole. Electron micrographs of native (un-clotted) hemocytes and of clots fixed in glutaraldehyde and post-fixed in osmium reveal that the membrane-bounded granules of native cells are very dense and homogeneous with no evidence of an internal structure. One-half to one minute after clotting, however, they become less dense and 250 Å microtubules spaced at ca. 500 Å intervals appear parallel to the long axis of the granule. Further degradation of the granule ensues and involves (a) change to a spherical shape, and (b) further decrease in density and better resolution of the microtubules. The microtubular component subsequently disintegrates leaving a membrane-bounded granule containing particulate material. Finally, the membrane of the granule fuses with the plasma membrane and the components of the granule are dispersed in the plasma where they presumably contribute to the formation of the gelatinous clot.     

Effects of Electrical Stimulation on Molecular Forms of Butyrylcholinesterase in Denervated Fast and Slow Latissimus Dorsi Muscles of Newly Hatched Chicken

The effects of denervation and direct electrical stimulation upon the activity and the molecular form distribution of butyrylcholinesterase (BuChE) were studied in fast-twitch posterior latissimus dorsi (PLD) and in slow-tonic anterior latissimus dorsi (ALD) muscles of newly hatched chicken. In PLD muscle, denervation performed at day 2 substantially reduced the rate of rapid decrease of BuChE specific activity which takes place during normal development, whereas in the case of ALD muscle little change was observed. Moreover, the asymmetric forms which were dramatically reduced in denervated PLD muscle were virtually absent in denervated ALD muscle at day 14. Denervated PLD and ALD muscles were stimulated from day 4 to day 14 of age. Two patterns of stimulation were applied, either 5-Hz frequency (slow rhythm) or 40-Hz frequency (fast rhythm). Both patterns of stimulation provided the same number of impulses per day (about 61,000). In PLD muscle, electrical stimulation almost totally prevented the postdenervation loss in asymmetric forms and led to a decrease in BuChE specific activity. In ALD muscle, electrical stimulation partially prevented the asymmetric form loss which occurs after denervation. This study emphasizes the role of evoked muscle activity in the regulation of BuChE asymmetric forms in the fast PLD muscle and the differential response of denervated slow and fast muscles to electrical stimulation.     

Extensive changes in cytokeratin expression patterns in pathologically affected human gingiva

The stratified squamous epithelium of the oral gingiva and the hard palate is characterized by a tissue architecture and a cytoskeletal composition similar to, although not identical with, that of the epidermis and fundamentally different from that of the adjacent non-masticatory oral mucosa. Using immunocytochemistry with antibodies specific for individual cytokeratins, in situ hybridization and Northern blots of RNA with riboprobes specific for individual cytokeratin mRNAs, and gel electrophoresis of cytoskeletal proteins of microdissected biopsy tissue samples, we show changes in the pattern of expression of cytokeratins and their corresponding mRNAs in pathologically altered oral gingiva. Besides a frequently, although not consistently, observed increase in the number of cells producing cytokeratins 4 and 13 (which are normally found as abundant components in the sulcular epithelium and the alveolar mucosa but not in the oral gingiva) and a reduction in the number of cells producing cytokeratins 1, 10 and 11, the most extensive change was noted for cytokeratin 19, a frequent cytokeratin in diverse one-layered and complex epithelia. While in normal oral gingiva cytokeratin 19 is restricted to certain, sparsely scattered cells of --or near--the basal cell layer, probably neuroendocrine (Merkel) cells, in altered tissue of inflamed samples it can appear in larger regions of the basal cell layer(s) and, in apparently more advanced stages, also in a variable number of suprabasal cells. Specifically, our in situ hybridization experiments show that this altered suprabasal cytokeratin 19 expression is more extended at the mRNA than at the protein level, indicating that cytokeratin 19 mRNA synthesis may be a relatively early event during the alteration. These changes in cytokeratin expression under an external pathological influence are discussed in relation to other factors known to contribute to the expression of certain cytokeratins and with respect to changes occurring during dysplasia and malignant transformation of oral epithelia.     

Differential associative behaviour of mitotic and meiotic acrocentric chromosomes

Summary A comparative study of the association of mitotic acrocentric chromosomes and acrocentric bivalents at the pachytene stage shows that at least two factors can act in the associative behaviour of these chromosomes: (1) Nor activity and (2) the presence of satellite DNA in the short arms of these chromosomes. These factors do not act with the same intensity in the two cell lines studied. In lymphocytes, Nor activity prevails, whereas satellite DNA plays the main role in the association of acrocentric chromosomes in germ cells at the pachytene stage.     

Nitrification and nitrate uptake: Leaching balance in a declined forest ecosystem in eastern France

Nitrate uptake and leaching were measured during one year in a declined fir forest on the Vosges highlands (eastern France), in order to investigate whether excess nitrification could be responsible for a deleterious acidification of the ecosystem. Nitrate uptake by the vegetation was active mainly from spring to early fall, and then reached about 66 kg N ha^-1. No significant leaching loss occurred during the growth period of the vegetation. Significant nitrate leaching occurred in winter (about 17 kg N ha^-1). During fall and winter the nitrification rate was of the same magnitude as values reported for other ecosystems, and, thus, was not considered to be abnormaly strong. No abnormal temporal discoupling of nitrate production and nitrate uptake occurred in the ecosystem, and forest decline must therefore have some other cause.     

The human genes for the α and γ subunits of the mast cell receptor for immunoglobulin E are located on human chromosome band 1823

The receptor with high affinity for immunoglobulin E (FcERI) is a key molecule in triggering the allergic reaction. It is tetrameric complex of one subunit, one subunit, and two disulfide-linked subunits. This receptor is present exclusively on mast cells and basophils. Molecules identical to the subunit of FcRI also form cell surface complex with other Fc receptors such as mouse FcRIIa in macrophages and most probably with human FcRIII (CD16) in natural killer (NK) cells. Here we show by in situ hybridization that the human genes for the (FCER1A) and subunits (FCER1 G) of FcERI and the gene for FcRIII (FCGR3, CD16) are located on human chromosome band 1823.     

Intraspecific larval competition in two solitary parasitoids, Apoanagyrus (Epidinocarsis) lopezi and Leptomastix dactylopii

Analysis of total aromatic amino acid (free and bound) in some cucumber accessions selected previously for resistance to western flower thrips, Frankliniella occidentalis (Pergande) [Thysanoptera: Thripidae], indicated that low concentrations of these essential nutrients, relative to total leaf protein, were correlated with a reduction in damage by the insect. Further analysis of samples of four important horticultural crops (lettuce, tomato, pepper and cucumber) with unknown levels of resistance to thrips showed a significant genotypic variation in the concentrations of total aromatic amino acids relative to the total leaf protein. Accessions from each crop with low or high concentrations of aromatic amino acids in proteins were exposed to thrips larvae. Regression analysis showed a highly significant positive correlation between aromatic amino acid concentration in leaf protein and thrips damage, regardless of crop species. It is concluded that higher concentrations of aromatic amino acids in plant proteins are important for successful thrips development. These results provide plant breeders with a promising tool for indirect selection without using undesirable insect bioassays.     

Identification of four novel splice site mutations in the ornithine transcarbamylase gene

Ornithine transcarbamylase (OTC) deficiency, the most common inborn error of the urea cycle, shows X-linked inheritance with frequent new mutations. Using polymerase chain reaction (PCR) amplification of the individual exons including adjacent intron sequences followed by direct sequencing of the amplimers we identified four new mutations affecting donor splice sites of introns 2, 5, 6, and 8. The mutation at the first position of intron 2 was a G to A exchange associated with acute neonatal hyperammonemia in a male patient at the age of 5 months. A G to C substitution in intron 5 was detected in a boy who developed 2 days after birth hypotonia, and respiratory distress, followed by severe hyperammonemia and terminal coma. The intron 6 mutation, a G to T substitution, was detected in a girl presenting with first episodes of vomiting and agitation at the age of 2 months. The mutation in intron 8, also a G to T transition, caused fatal hyperammonemia and early death at the age of 15 days in a male patient. We present four donor splice site mutations resulting in severe neonatal or very early onset of the disease in three boys and in one female patient. As the GT dinucleotide of the 5 donor splice site is invariant and required for correct splicing the described mutations may lead to improperly spliced mRNAs and aberrant gene products.