Strongyloides stercoralis hyperinfection in the acquired immunodeficiency syndrome.
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Acquired Immunodeficiency Syndrome↗
Biomedical subjects
J Pasternak
Publications and source records attributed to J Pasternak.
The T-cell accessory molecule CD4 recognizes a monomorphic determinant on isolated Ia.
The membrane protein CD4 is commonly found on mature T cells specific for antigen in association with class II major histocompatibility complex (MHC; Ia) proteins. This correlation has led to the suggestion that CD4 binds to a monomorphic region of the Ia molecule on the antigen-presenting cell (APC) and functions either by enhancing interaction between the T cell and the APC, or conversely, by transducing negative signals to the T cell. To address this hypothesis, we have made use of sublines from an unusual T hybrid that is class I MHC restricted but also CD4+. By incorporating purified MHC proteins into a planar membrane system, we show that different Ia molecules can greatly enhance the ability of a CD4+ but not a CD4- variant of this class I-restricted T hybrid to respond to isolated class I molecules. T-cell responses can be strongly augmented by the concurrent expression of CD4 on the T cell and any of four different Ia proteins on planar membranes, thus supporting the idea that CD4 binds to a monomorphic region of the Ia molecule and increases the avidity with which the T cell can interact with its target.
Animals↗
Ligand-induced desensitization of B-cell membrane immunoglobulin-mediated Ca2+ mobilization and protein kinase C translocation.
Binding of ligand to B-cell membrane immunoglobulin (mIg) can lead to activation of a number of distinct biologic responses, including altered expression of genes encoding c-fos, c-myc, and Ia, as well as proliferation and immunologic tolerance. Tolerance could reflect a functional uncoupling of receptors from systems that generate intracellular second messengers (i.e., receptor desensitization). To better understand the molecular basis of immune regulation, we examined the ability of mIg to function as a signal transducer after the cell's initial contact with mIg-binding ligand. The results show that ligand binding to as little as 2-10% of mIgM or mIgD renders the cell unresponsive to ligand binding to the reciprocal isotype as judged by Ca2+ mobilization and protein kinase C translocation responses. This heterologous receptor desensitization lasts longer than 24 hr and does not reflect loss of receptor from the cell surface. Studies with the calcium ionophore ionomycin, 1,2-dioctanoyl-sn-glycerol, and the protein kinase inhibitor staurosporine indicate that both protein kinase C-dependent and protein kinase C-independent (staurosporine-insensitive) mechanisms mediate heterologous desensitization after mIg crosslinking.
Animals↗
Experimentally induced myopia does not affect post-hatching development of the chick lens.
Myopia, as characterized by a large refractive error (e.g. -10.7 +/- 0.4 D), was induced in post-hatch chicks by a 14 day application of a goggle that was designed to blur the retinal image. In comparison to untreated eyes, the treated eye showed significant changes in wet eye weight and both axial and equatorial lengths. However, the lenses of myopic and non-myopic eyes were not significantly different in focal characteristics, light transmittance or total soluble protein content. Thus the lens neither contributes to, nor compensates for the large refractive error observed in experimentally induced myopia.
Animals↗
Ascaris lumbricoides: characterization of the collagenous components of the adult cuticle.
The proteins of the cuticle of adult Ascaris lumbricoides suum were characterized with respect to heterogeneity, glycosylation, and susceptibility to collagenase. Pepsin digestion of intact cuticles was used to determine the extent of stable triple-helical structures of the cuticular components. With sodium dodecyl sulfate-poly-acrylamide gel electrophoresis, it was shown that treatment of purified cuticles with beta-mercaptoethanol released three components (99, 90, and 68 kDa) which comprise 95% of the total solubilized material. The remaining fraction consists of at least four components (16, 28, 154, and 173 kDa). Periodic acid-Schiff staining showed that the only glycoprotein was the 173-kDa component. All cuticular components, except the 173-kDa protein, were degraded by bacterial collagenase. Pepsin digestion of intact cuticles for 24 hr at 4 C produced, after reduction, a 95-kDa fragment; by 96 hr, four fragments (95, 90, 83, and 77 kDa) were evident. When the 96-hr pepsin digest was treated with fresh pepsin, the 77-kDa fragment became the major constituent. With agarose gel electrophoresis, analysis of non-reduced, pepsin-released material revealed intact aggregates that were greater than 2 X 10(3) kDa. The enzyme digestion studies indicate that, with the exception of the 173-kDa component, each cuticular protein contains collagenous domains and that, within the cuticle, the longest contiguous collagen chain in a triple-helical conformation has a uniform molecular size of 77 kDa.
Animals↗
Isolation and characterization of the cuticle from the free-living nematode Panagrellus silusiae.
The cuticle (0.15 to 0.5 microns thick) of the microscopic free-living nematode Panagrellus silusiae was isolated intact by incubating worms with 1% sodium dodecyl sulfate at 37 degrees C overnight. After shearing and further treatment with detergent, electron microscopy revealed that the cuticular pieces were free of contaminating material and retained their characteristic in situ ultrastructure. From amino acid determinations, the cuticle is collagen-like with high levels of glycine (approximately equal to 31 residue %), proline (approximately equal to 20 residue %) and alanine (approximately equal to 21 residue %) although the hydroxyproline (2.6 residue %) content is low. Half-cystine (approximately equal to 1 residue %) is present in purified cuticles. Treatment with 8 M guanidine hydrochloride-2% beta-mercaptoethanol can solubilize more than 85% of the cuticular preparation. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the solubilized cuticles from juvenile, adult and old dead worms revealed, at least, 18 discrete components. Estimated molecular weights ranged from about 26 000 (peak 1) to 250 000 (peak 18).
Amino Acids↗
The presence of two major protein components in the bovine photoreceptor disc membrane.
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Adenosine Triphosphate↗
Induction of gel-phase lipid in plasma membrane of chick intestinal cells after coccidial infection.
When chickens are infected with the coccidial parasite Eimeria necatrix, the plasma membrane of intestinal cells harbouring second-generation schizonts becomes refractory to mechanical shearing, hypotonic shock and ultrasonication. Plasma membrane from these infected cells was isolated to high purity as judged by enriched levels of ouabain-sensitive (Na+ + K+)-stimulated Mg2-dependent ATPase activity and sialic acid content, the lack of detectable cytochrome oxidase and glucose-6-phosphatase activities and electron microscopic analysis of the final preparation. Wide-angle X-ray diffraction patterns recorded from the isolated membranes revealed that during the later stages of parasite maturation the host cell plasma membrane acquires increasing proportions of gel-phase lipid. By contrast, purified membrane from isolated parasites is in a liquid-crystalline state. The transition temperature of host cell plasmalemma at 100 h postinfection is 61 degrees C, about 20 degrees C above physiological temperature. By contrast, liposomes of plasma membranes from infected cells undergo a thermal transition at about 28 degrees C. The accumulation of gel-phase lipid in the host cell plasma membrane is not attributable either to an increase in the constituent ratio of saturated to unsaturated fatty acids or to a significant change in the cholesterol to phospholipid ratio. During the late stages of infection, the cells become stainable with trypan blue which suggests that the acquisition of crystalline phase lipid disrupts the permeability of the host cell plasmalemma.
Animals↗
Characterization of the genome of the free-living nematode Panagrellus silusiae: absence of short period interspersion.
The complexity of the DNA of the free-living nematode Panagrellus silusiae has been examined. Reassociation kinetics of pressure-sheared fragments (approximately 290 nucleotides) in 0.18 M Na+ at 60 degrees C showed the presence of foldback, repetitive, and unique DNA sequence elements. The three classes comprise 9.3%, 26.1%, and 61.3% of the total DNA, respectively. The mean length of the foldback duplex DNA after digestion with S1 nuclease is about 185 nucleotides. There are about 1.8 x10(4) inverted repeats per genome. Sequence arrangement was deduced from (1) renaturation kinetic profiles of long and short fragments on hydroxylapatite; (2) the pattern of renaturation of tracer DNA, labeled in vitro with 125I, of various sizes after incubation with excess short fragments; and (3) thermal denaturation behavior of DNA that had been reassociated to various C0t values. It was found that DNA fragments of the repetitive fraction that are, at least, 2000 nucleotides in length are virtually free of unique sequences. Moreover, it is estimated that the repeated segments in this species could extend for 10,000 nucleotide pairs. Thus, Panagrellus DNA lacks the pattern of extensive short period interspersion that is typified by the DNA of Xenopus.
Animals↗
Isolation and translation in vitro of poly(A)+RNA from the free-living nematode Panagrellus silusiae.
Polysomal RNA was isolated from the free-living nematode Panagrellus silusiae. Passage of this RNA through a cellulose column resulted in the fractionation of the input RNA into poly(A)-RNA (ca. 97.5% of the total) and poly(A)+ RNA (ca. 2.5% of the total). RNase digestion, followed by polyacrylamide gel electrophoresis, revealed that the poly(A)+ RNA contained poly(A) tracts that ranged from 75 to 104 nucleotides in length with a mean value of about 90 residues. There was no evidence of poly(A) sequences in the poly(A)- RNA fraction. Poly(A)+ RNA gave a 25- to 50-fold stimulation (over background) of amino acid incorporation in the wheat germ cell-free protein-synthesizing system. At least 26 proteins were evident after electrophoresis in cylindrical sodium dodecyl sulfate-polyacrylamide gels. Poly(A)-RNA was capable of stimulating protein synthesis in vitro with about five discrete proteins being produced. In summary, the properties of mRNA from a simple organism such as P. silusiae are very similar to those of more complex eukaryotes.
Animals↗
In vitro translation of nematode cuticular collagens.
Phenanthroline treatment of growing cultures of the free-living nematode Panagrellus silusiae was used to lower the degree of hydroxylation of nascent collagen chains at the polysomal level. Under these conditions, the bound pentasome-hexasome fraction provided substrate for prolyl hydroxylase. When this polysomal fraction was subsequently tested in a cell-free wheat germ system, collagenase-susceptible translation products were observed after sodium dodecyl sulfate-acrylamide gel electrophoresis. The electrophoretic mobilities of each of these four major collagen products were similar to four collagens that are isolated from intact cuticles. In addition, purified polysomal RNA that adhered to unmodified cellulose directed the synthesis of four pepsin-resistant polypeptides that had molecular weights that coincided with four pepsin-resistant collagens that can be purified from the cuticle of this species. Thus, the polysomal site of the messenger RNAs for the cuticular collagens of P. silusiae was located. Although precursor forms of the cuticular collagens were not produced in the cell-free system, the question whether additional amino acid segments occur on the primary translational products of the cuticular collagens in vivo remains open.
Cell-Free System↗
Rapid deterioration in subacute sclerosing panencephalitis after measles immunisation.
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Child↗
Isolation of chick intestinal cells infected with second-generation schizonts of Eimeria necatrix.
After infection with Eimeria necatrix, chick intestinal cells undergo a profound increase in size. Herein, we describe methods for isolating and purifying infected cells containing second-genration schizonts at various times post-infection. Our purification procedures depends upon three fundamental properties of infected cells: namely (1) large size, (2) resistance to shearing and (3) decreased susceptibility to osmotic shock in hypotonic solutions. The availability of simple techniques for isolating infected whole cells should prove valuable in elucidating various intracellular aspects of infection during schizogony.
Animals↗
Isolation of host-cell nuclei from chick intestinal cells infected with second-generation schizonts of Eimeria necatrix.
A procedure is described for the preparation of purified nuclei of chick intestinal cells infected with Eimeria necatrix. During second-generation schizogony the host-cell membrane becomes resistant to homogenization, hypotonic shock and ultrasonication. However, brief treatment with a low concentration of trypsin (0.001%) will disrupt the cell membrane. Thereafter the nuclei can be selectively harvested from contaminating nuclear types and schizonts by a series of centrifugations. Electron microscopy and Feulgen-DNA microspectrophotometry showed that the nuclear preparations retained their integrity with little, if any, loss of DNA. Chemical determinations confirmed that host-cell nuclear DNA synthesis was stimulated by coccidial infection and further established that quantitatively there was a coincident increase in the amount of host nuclear RNA during schizogony. It is now feasible to examine directly the effects of coccidial infection upon the host's nuclear apparatus.
Animals↗
Relationship between nuclear morphology and the phases of the cell cycle during cercarial development of the digenetic trematode Trichobilharzia ocellata.
During the proliferative phase of cercarial development in the digenetic trematode Trichobilharzia ocellata, nuclei varied in size, appearance of the chromatin, and intensity of Feulgen staining. On the basis of interphase nuclear morphology six nuclear classes were identified. Data from microspectrophotometric determinations and 3H-TdR labeling experiments were used to correlate each kind of interphase nucleus with a phase of the cell cycle. Marked variability in nuclear area developed between cells by time they reached late G1. Increases in nuclear area could not be correlated with the onset of DNA synthesis. Throughout all stages of development of T. ocellata cercariae, proliferating cells divide mitotically; meiotic divisions were never seen. Thus, the mode of reproduction appears to be asexual. No evidence was found to support previous suggestions of diploid parthenogenesis.
Animals↗
Cell cycle analysis in developing cercariae of Trichobilharzia ocellata (Trematoda: Schistosomatidae).
Cellular proliferation has been analyzed during cercarial development of the digenetic trematode Trichobilharzia ocellata. Prior to the tail-bud stage (about 1,000 cells), cells were actively involved in cellular proliferation. The mean cell cycle was 15.2 hr. The time for mitosis was 1.6 hr; for G1, 5.6 hr; for G2, 3.2 hr, and for the S phase, 4.8 hr. Beginning with the tail-bud stage, an increasing proportion of cells accumulate in the G1 phase. Cytological evidence of changes in the amount of cytoplasm per cell revealed that these noncycling cells were differentiating. During organ development and differentiation, the proportion of proliferating cells decreases and by the 2,000-celled stage proliferation ceases. Our results do not support a germinal lineage theory of cercarial development since none of the observed nuclear types could be unequivocally identified as belonging to the germ line.
Animals↗
Cytophotometric study of nuclear proteins during gametogenesis in Ascaris lumbricoides.
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Animals↗
Relative ribosomal RNA cistron multiplicity in oocytes and postembryonic stages of the eutelic nematode Panagrellus silusiae.
The number of ribosomal RNA cistrons has been measured in the total DNA extracted from L2 juvenile and adult stages of the free-living nematode Panagrellus silusiae. Saturation hybridization studies with homologous rRNA indicate that both stages have about 275 ribosomal genes per haploid equivalent. Using homologous 125I-labelled rRNA for in situ hybridization, the mean number of silver grains per DNA content for oocyte, hypodermis and gut nuclei was similar. The mean DNA contents of maturing oocyte, hypodermis and gut nuclei are about 20C, 2C, and 10C respectively. We conclude that rDNA amplication alone is insufficient to account for the variation in DNA content of oocytes and that postembryonic development in this eutelic organism occurs without a significant differential increase in the number of ribosomal cistrons per worm.
Animals↗