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H Craig

Publications and source records attributed to H Craig.

15 recordsLinked to original sources

A preliminary proteomic survey of the in vitro excretory/secretory products of fourth-stage larval and adult Teladorsagia circumcincta.

The nature of the proteins which comprise the in vitro excretory/secretory products (ES) of the fourth-stage larva (L4) and adult Teladorsagia circumcincta are largely undefined, despite the fact that this nematode induces profound changes, in part related to parasite ES, in the cellular architecture of the glands lining the abomasal surface of infected sheep and goats. In this study, the protein components of L4 and adult ES were fractionated using 1D gel electrophoresis and the major protein bands, detected by Coomassie blue staining, excised from the gel and subjected to tryptic digest and subsequent mass spectrometric analysis. The resultant peptide mass fingerprints were used to identify 15 L4 and 13 adult ES proteins. Several proteins, such as globin and some metabolic enzymes, were present in both ES. L4 ES alone contained thioredoxin peroxidase, an enzyme that can detoxify free radicals resulting from host inflammatory responses to the parasite, a cysteine proteinase which may aid penetration of the gastric mucosa and 2 different galectins which may influence cell differentiation and morphogenesis. Adult ES contained a nucleoside diphosphate kinase homologue, an enzyme which has been linked to cellular changes and can affect liquid secretion and goblet cell degranulation.

Animals↗

Nef-induced CD4 downregulation: a diacidic sequence in human immunodeficiency virus type 1 Nef does not function as a protein sorting motif through direct binding to beta-COP.

The Nef protein from the human immunodeficiency virus (HIV) induces CD4 cell surface downregulation by interfering with the endocytic machinery. It has been recently proposed that binding of HIV type 1 Nef to the beta subunit of COPI coatomers participated in the Nef-induced CD4 downregulation through recognition of a novel diacidic motif found in the C-terminal disordered loop of Nef (V. Piguet, F. Gu, M. Foti, N. Demaurex, J. Gruenberg, J. L. Carpentier, and D. Trono, Cell 97:63-73, 1999). We have mutated the glutamate residues which formed this motif in order to document this observation. Surprisingly, mutation of the diacidic sequence of Nef did not significantly affect its ability (i) to interact with beta-COP, (ii) to downregulate CD4 cell surface expression, and (iii) to address an integral resident membrane protein containing Nef as the cytoplasmic domain to the endocytic pathway. Our results indicate that these acidic residues are not involved in the connection of Nef with the endocytic machinery through binding to beta-COP. Additional studies are thus required to characterize the residues of Nef involved in the binding to beta-COP and to evaluate the contribution of this interaction to the Nef-induced perturbations of membrane trafficking.

Adaptor Protein Complex gamma Subunits↗

Virological importance of the protease-cleavage site in human immunodeficiency virus type 1 Nef is independent of both intravirion processing and CD4 down-regulation.

The HIV-1 Nef protein is present within the virion and is processed there by the viral protease. Mutational analysis indicated that residues 54-60 in HIV-1 Nef were required for intravirion cleavage. When viruses were produced using T cell lines or primary lymphoblasts, these residues were also required for optimal viral infectivity. However, substitution of native Nef residues with those of a functional Gag cleavage site demonstrated that intravirion cleavage was insufficient for the virological function of this domain. Furthermore, the importance of certain cleavage site residues to infectivity was conditional on the producer cell type. In particular, a mutant containing a deletion of residues 54-57 was phenotypically nef defective when produced using T cells (CEM, A2.01, or primary lymphoblasts) but was minimally impaired when produced from 293 or HeLa cells. This mutant was cleavage resistant, indicating that proteolytic processing of Nef was dispensable for infectivity enhancement when virions were assembled in certain non-T cells. Residues 54-61 of the cleavage site, including 54-57, were also required for Nef-mediated down-regulation of CD4. However, the surface expression of CD4 on HeLa cells in amounts comparable to that on the surface of primary T lymphoblasts did not create a producer cell environment in which residues 54-57 acquired greater virological importance. Furthermore, these residues were required for optimal infectivity even during virion assembly in T cells (A2. 01) that expressed a CD4 molecule that is unable to respond to Nef. These data suggested that in producer T cells, certain cleavage site residues (54-57) contribute to a Nef-mediated virological effect that is unlikely to be linked causally to CD4 down-regulation. Conversely, in the context of 293 cells as viral producers, the Delta54-57 mutant separated genetically down-regulation of CD4 (for which it was defective) from enhancement of infectivity (for which it was functional). Together, these data indicate that the virological function of the cleavage site domain is both independent of intravirion proteolytic processing of Nef and independent of CD4 down-regulation.

Amino Acid Sequence↗

Nonequilibrium air clathrate hydrates in Antarctic ice: a paleopiezomdter for polar ice caps.

"Craigite," the mixed-air clathrate hydrate found in polar ice caps below the depth of air-bubble stability, is a clathrate mixed crystal of approximate composition (N2O2).6H2O. Recent observations on the Byrd Station Antarctic core show that the air hydrate is present at a depth of 727 m, well above the predicted depth for the onset of hydrate stability. We propose that the air hydrate occurs some 100 m above the equilibrium phase boundary at Byrd Station because of "piezometry"--i.e., that the anomalous depth of hydrate occurrence is a relic of a previous greater equilibrium depth along the flow trajectory, followed by vertical advection of ice through the local phase-boundary depth. Flowline trajectories in the ice based on numerical models show that the required vertical displacement does indeed occur just upstream of Byrd Station. Air-hydrate piezometry can thus be used as a general parameter to study the details of ice flow in polar ice caps and the metastable persistence of the clathrate phase in regions of upwelling blue ice.

Journal Article↗

Type 1 plasminogen activator inhibitor gene expression following partial hepatectomy.

A murine model of partial hepatectomy (PH) was employed to investigate type 1 plasminogen activator inhibitor (PAI-1) gene expression in regenerating liver. Mice were anesthetized, and a portion of the left lobe of the liver was ligated and resected distal to the ligature, and at various times thereafter, total liver RNA was prepared and analyzed by Northern blotting. PH caused a transient increase in PAI-1 messenger (m)RNA that was apparent within 1 to 2 hours after surgery, was maximal at 8 hours (ninefold increase over sham-operated controls), and then slowly declined. Analysis of discrete liver segments demonstrated much greater induction of PAI-1 mRNA in the region adjacent to PH than in more distal regions. Further analysis of the adjacent tissue by in situ hybridization revealed that PAI-1 mRNA was induced primarily in hepatocytes in the transition zone created by the occluding hemostatic ligature between viable and necrotic tissue. Expression of PAI-1 mRNA could also be detected in this transition zone in capsular mesothelial cells, subcapsular hepatocytes, and venous endothelial cells bordering the area. A much weaker signal was evident in hepatocytes dispersed throughout the remaining intact lobes of PH mice, and no signal was detected in the livers of sham-operated mice. These observations suggest that PAI-1 may be of importance in local tissue remodeling events accompanying liver regeneration.

Animals↗

Oxygen supersaturation in ice-covered Antarctic lakes: biological versus physical contributions.

Freezing in ice-covered lakes causes dissolved gases to become supersaturated while at the same time removing gases trapped in the ablating ice cover. Analysis of N2, O2, and Ar in bubbles from Lake Hoare ice shows that, while O2 is approximately 2.4 times supersaturated in the water below the ice, only 11% of the O2 input to this lake is due to biological activity: 89% of the O2 is derived from meltwater inflow. Trapped bubbles in a subliming ice cover provide a natural "fluxmeter" for gas exchange: in Lake Hoare as much as 70% of the total gas loss may occur by advection through the ice cover, including approximately 75% of the N2, approximately 59% of the O2, and approximately 57% of the Ar losses. The remaining gas fractions are removed by respiration at the lower boundary (O2) and by molecular exchange with the atmosphere in the peripheral summer moat around the ice.

Antarctic Regions↗

Cosmogenic He in terrestrial rocks: The summit lavas of Maui.

We have identified terrestrial cosmic rayproduced (3)He in three lava flows on the crest of Haleakala Volcano on Maui, 3 km above sea level, and approximately 0.5 million years old. Although these lavas, like all oceanic basalts, contain primordial (3)He from the mantle, the "cosmogenic" component ((3)He(C)) can be identified unambiguously because it is extractable only by high-temperature vacuum fusion. In contrast, a large fraction of the mantle helium resides in fluid inclusions and can be extracted by vacuum crushing, leaving a residual component with (3)He/(4)He ratios as high as 75x those in the atmosphere, which can be liberated by melting the crushed grains. Cosmogenic (3)He is present in both olivines and clinopyroxenes at 0.8-1.2 x 10(-12) ml(STP)/g and constitutes 75% +/- 5% of the total (3)He present. The observed (3)He(C) levels require a cosmic ray exposure age of only some 64,000 years, much less than the actual age of the lavas, if there is no erosion. Using a model that includes effects of uplift or submergence as well as erosion, we calculate an apparent "erosion rate" of the order of 8.5 m/10(6) years for the western rim of the summit crater, as an example of the application of measurements of cosmogenic rare gases to terrestrial geological problems.

Journal Article↗

The ultrastructure of the neonatal pig colon.

The neonatal pig colon has several unique structural and developmental features. At birth it has a variable population of epithelial cells which in their arrangement on villus-like protrusions and in their capability for protein uptake into large preformed supranuclear vacuoles closely resemble neonatal ileal cells. Such villus-like protrusions and vacuolated cells are not present in the 2-day-old piglet. On the first day after birth absorptive epithelial cells which lack supranuclear vacuoles transiently accumulate a large number of lipid droplets, each separated from the cytoplasm only by a proteolipid interface. None of the much smaller lipid droplets bounded by a unit membrane of the smooth endoplasmic reticulum and characteristic of normal small intestinal fat uptake were ever seen in these cells. Very few of the large lipid drops remain on the second day after birth. This initial capacity of the colon for protein and lipid uptake never reappears. The pattern of colonic amino acid transport also changes markedly in the first four days of independent life and this may be correlated with the observation that the absorptive cells at birth have microvilli which are twice the length of those on similar cells at and after two days old. These morphological results are discussed in terms of implied functional changes in the neonatal period.

Amino Acids↗

"Sunbursts" and "christiesomes": cellular fragments in normal cow and goat milk.

Goats' milk includes numerous cell fragments ("christiesomes") which originate from the mammary secretory cells, contain well preserved endoplasmic reticulum, mitochondria and lipid droplets, and are responsible for the considerable triglyceride synthesising capacity of fresh goat milk. Cows' milk shows a few such particles only after repeated oxytocin-aided milkings. Cows' milk does contain quite different particles which have a dense content with a few small vesicles and numerous microvillus-like protrusions on one side ("sunbursts"). These have not been found in goats milk. Cytoplasmic particles similar to sunbursts have been found on the surface of the mammary secretory epithelium. It is suggested that they are residues of dead cells.

Animals↗

Abnormal distribution of pulmonary blood flow in aortic valve disease. Relation between pulmonary function and chest radiograph.

Wasted ventilatory volume (V(D)) and its ratio to tidal volume (V(D)/V(T)) were measured at rest and during exertion in 17 patients with aortic valve disease. We considered V(D)/V(T) to indicate abnormal ventilation: perfusion relations if it did not decrease on exertion, or if the exercising value was greater than 40 per cent. Plain chest radiographs were independently examined for evidence of diversion of pulmonary blood to the upper lobes. There was significant agreement (p<0.05) between radiographic and pulmonary function estimations of abnormality. This suggests that the raised pulmonary venous pressure associated with left ventricular failure creates an abnormal pattern of blood flow through the lung, which is responsible for causing inadequate perfusion with respect to ventilation.

Aortic Valve Insufficiency↗

Oxygen-18 variations in sulfate ions in sea water and saline lakes.

A new method measures the oxygen-isotopic composition of dissolved sulfate ions and barium sulfate. Sulfate ions in the oceans are enriched in oxygen-18 by 9.5 per mille relative to mean ocean water and show only minor variations, probably because the exchange rate with water is slow enough to prevent local equilibrium with surface waters. Sulfate ions in saline lakes and brines have oxygen-18 enrichment of from 7 to 23 per mille relative to mean ocean water;the value of the highest enrichment observed is about the same as that of atmospheric oxygen.

Carbon Dioxide↗