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Biomedical subjects

M K Jones

Publications and source records attributed to M K Jones.

At least 109 records · Page 6Linked to original sources
PubMed1996

Immunolocalization of the dengue virus nonstructural glycoprotein NS1 suggests a role in viral RNA replication.

Although many workers have investigated the maturation and processing of the flavivirus nonstructural glycoprotein NS1 in infected cells, these studies have provided little insight into a possible function for NS1. In this study we investigated the subcellular localization of NS1 both by immunofluorescence and cryo-immuno electron microscopy of infected Vero and C6/36 cells. NS1 was found to be tightly associated with intracellular membranes, in particular with vesicle packets and large cytoplasmic vacuoles. Surprisingly, NS1 did not associate with mature virus particles, a finding that is inconsistent with the postulated role for this protein in virion assembly and/or maturation. However, dual-labeling experiments did reveal in both the confocal immunofluorescence and cryo-immuno EM studies the colocalization of NS1 with the viral dsRNA replicative form. Furthermore, localization of the dsRNA to the vesicle packets and cytoplasmic vacuoles seen in infected Vero and C6/36 cells, respectively, suggests that these structures may comprise the flavivirus replication complex. These findings provide further insights into the maturation pathway of NS1 and suggest a possible role for this protein in viral RNA replication.

Animals↗
PubMed1996

Danger expectancies and insight in spider phobia.

Differences between phobic and normal subject perceptions of danger were examined. Nineteen spider-fearful subjects and a matched set of controls gave danger ratings before and during a spider-avoidance test. When detached from the phobic stimulus, spider-fearful subjects: (1) gave higher estimates of the probability of being bitten than did controls, (2) gave higher estimates of the injuries that would result from being bitten, and (3) believed their high levels of anticipated anxiety were more reasonable and appropriate to the demands of the situation than did controls. These findings are inconsistent with both the traditional account and Beck and Emery's (1985) more recent view that, when detached from the phobic situation, patients can accurately evaluate the danger of potential phobic encounters. The present results question the view that phobic individuals have complete insight into the inappropriateness of their own distress.

Adaptation, Psychological↗
PubMed1996

Improved membrane preservation of flavivirus-infected cells with cryosectioning.

Ultra-cryomicrotomy and electron microscopy were used to investigate membranous structures in dengue virus-infected mammalian and insect cells. The cryo-sectioned samples displayed ultrastructure comparable to their resin-embedded counterparts with all previously identified virus-induced structures being observed. Structures not previously identified were also found. In particular, membrane-bound packets of vesicles, 100-200 nm in diameter were seen distributed throughout areas of virus-induced membrane proliferation. These packets were clearly distinct from virion arrays. Small smooth membrane vesicles, previously found to contain thread-like enclosures (M.L. Ng, J. Gen. Virol. 68 (1987) 577-582), were frequently observed to contain dense staining material, however the exact nature of this material remains unclear. Virus-induced modification of golgi-like and/or ER membranes was also observed and may represent early events in the generation of the smooth membrane vesicles seen during infection. We suggest that cryosectioning is the method of choice to investigate membrane rearrangement induced by this family of viruses and that a diamond knife and modified staining techniques, as utilised in this report, be employed to enhance morphology and section preservation.

Animals↗
PubMed1996

The ultrastructural localization of Echinococcus granulosus antigen 5.

Murine monoclonal and polyclonal antisera, raised against the 38 kDa subunit of Echinococcus granulosus antigen 5, were used to investigate the tissue distribution of the antigen in hydatid cysts. Immunoreactivity was visualized by indirect immunofluorescence on whole protoscoleces, and ultrastructural immunocytochemistry utilizing colloidal gold-based labelling procedures on unsectioned and cryosectioned brood capsules and protoscoleces. In protoscoleces, the 38 kDa subunit of antigen 5 was localized at the interface of parenchymal cells and associated extracellular matrices, as well as along the interface of the tegumentary syncytium in the somal region and its basal matrix. Cytoplasmic labelling of parenchymal cells was rare; when observed, it was associated with vesicles and membranes in cytoplasmic extensions of parenchymal cells. In brood capsules, the antigen was associated with the external face of the plasma of degenerating parenchymal cells. The 38 kDa subunit occurs along the extracellular face of cell membranes, suggesting that antigen 5 is either a component of the membranes or associated extracellular matrices.

Animals↗
PubMed1996

Characterization of the cathepsin-like cysteine proteinases of Schistosoma mansoni.

Adult Schistosoma mansoni parasites synthesize and secrete both cathepsin L and cathepsin B cysteine proteinases. These cysteine proteinase activities, believed to be involved in hemoglobin digestion by adult schistosomes, were characterized by using specific fluorogenic peptide substrates and zymography. Both cathepsin L- and B-like activities with pH optima of 5.2 and 6.2, respectively, predominated in soluble extracts of worms, and both these activities were secreted by adult worms into the culture medium. The specific activity of cathepsin L was about double that of cathepsin B when each was assayed at its pH optimum, and moreover, the specific activities of cathepsins L and B in extracts of female schistosomes were 50 to 100% higher than in extracts of male schistosomes. Analysis of the primary structure of two cloned S. mansoni cathepsins L, here termed cathepsin L1 and cathepsin L2, revealed that they are only 44% similar and that cathepsin L2 showed more identity (52%) with human cathepsin L than with schistosome cathepsin L1. Moreover, differences in their active site, propeptide region, and potential for glycosylation suggest separate functions for schistosome cathepsin L1 and cathepsin L2.

Amino Acid Sequence↗
PubMed1995

The partial mu opiate agonist buprenorphine protects a sub-population of thalamic reticular neurons following cardiac arrest in rats.

Extensive loss of neurons occurs from the middle region of the thalamic reticular nucleus (RT) in rats resuscitated after 10 min of cardiac arrest. Administration of the partial mu opiate agonist buprenorphine 45 min after resuscitation produces a small but significant increase in spared neurons along the medial and lateral margins of the middle RT, regions where mu receptors have been localized. Systemic administration of mu agonists may augment endogenous opiate mechanisms that contribute to the relative ischemic resistance of subpopulations of RT neurons.

Animals↗
PubMed1995

Ultrastructure of the scolex and tentacles of the metacestode of Polypocephalus species (Cestoda: Lecanicephalidae) from the blue-swimmer crab Portunus pelagicus.

The ultrastructure and histochemistry of the scolex of a Polypocephalus species is described. This cestode is distinguishable from all known Polypocephalus spp. in that it has approximately 400 tentacles and inhabits the musculature of the blue swimmer crab, Portunus pelagicus. The metacestode has a large scolex, divided into an anterior par apicalis, and a posterior pars basalis. An elongate cercomer lies posteriorly to the scolex. The pars basalis lacks accessory suckers. The tegument of this region is armed with numerous spinous microtiches. The pars apicalis consists of approximately 400 unarmed tentacles, a large cavity ( = scolex cavity) containing some glandular tissue, and lying posteriorly to this cavity, a large gland ( = scolex gland). The tegument of the tentacles and pars apicalis is typically syncytial, and contains abundant electron-opaque secretory vesicles. These vesicles are produced by glandular tissue of the scolex gland and scolex cavity. Histochemistry revealed the presence of proteins and neutral mucosubstances in tentacles, scolex gland and scolex cavity, suggesting the presence of glycoproteins. It is argued that the secretion produced by the tentacles has an adhesive role. When applied to the large interface between the tentacles and the mucosa of the hosts, this secretion may provide the adult parasite with a strong holdfast to prevent dislodgement.

Animals↗
PubMed1995

Identification and characterization of myophilin, a muscle-specific antigen of Echinococcus granulosus.

A muscle-specific gene of Echinococcus granulosus has been identified and characterized. A lambda gt11 clone (10P1), containing an incomplete copy of the gene, was originally isolated from a larval E. granulosus cDNA library by serum antibodies from dogs infected with the parasite. The full-length cDNA sequence was obtained by PCR amplification of cDNA from an adult E. granulosus lambda gt22A library. Southern blot analysis indicated the presence of the gene as a single copy in the genome of E. granulosus and also detected homologous genes in genomic DNA of E. multilocularis and Taenia saginata. The 21.2-kDa protein deduced from the complete cDNA sequence contains two regions of 12 amino acids with similarity to the EF-hand motif of calcium binding proteins. Antibodies raised against the purified 10P1-GST fusion protein detected a 22-kDa antigen in the E. granulosus developmental stages examined. Immunoelectron microscopy localized the native protein in the muscle of the parasite. The amino-acid sequence of the E. granulosus protein shows significant homology to the muscle proteins mp20 of Drosophila melanogaster, chicken SM22 alpha and mammalian calponin, and also to the neuronal protein NP25 of rats. A conserved carboxy-terminal motif of 17 amino acids is present in all the homologous proteins and is proposed to be the characteristic feature of a novel protein family. The term myophilin is proposed for the E. granulosus protein due to its localization and homology to other muscle proteins.

Amino Acid Sequence↗
PubMed1995

Cutaneous scleroderma in association with carcinoid syndrome.

A case of scleroderma in a woman with carcinoid syndrome is described and the similarities between our case and those in the literature are reviewed. The carcinoid tumours were all of midgut origin and liver metastases were present. All subsequently developed fibrotic heart disease and none had clinical features or autoantibodies suggestive of systemic sclerosis. The association between carcinoid syndrome and particular features of scleroderma is likely to be more than fortuitous.

Carcinoid Tumor↗
PubMed1995

Antimicrobial mode of action of secretions from the metapleural gland of Myrmecia gulosa (Australian bull ant).

Secretions from exocrine metapleural glands of Myrmecia gulosa (Australian bull ant) exhibit broad-spectrum antimicrobial activity. Treatment of the yeast Candida albicans with metapleural secretion resulted in the rapid and total leakage of K+ ions from cells within 10 min. Ultrastructural analysis of the bacteria Bacillus cereus, Escherichia coli, and Pseudomonas aeruginosa, and cells and protoplasts of Candida albicans demonstrated gross damage of the cell membrane and aggregation of the cytoplasmic matrix of treated cells. Degradation of membrane-bound organelles was also observed in Candida albicans. The antimicrobially active components of metapleural secretions were nonpolar and interacted with the phospholipid bilayer, causing damage to the structural integrity of liposomes and the release of carboxyfluorescein. The data suggest that the antimicrobial agents in metapleural secretion act primarily by disrupting the structure and function of the phospholipid bilayer of the cytoplasmic membrane.

Animals↗
PubMed1995

Circumvallate placenta: sonographic diagnosis.

We prospectively identified 12 patients who had a shelf at the margin of the placenta consistent with the diagnosis of circumvallate placenta. Ten patients were followed to delivery and three placentas were sent for pathologic analysis. Circumvallate placenta was diagnosed by gross inspection in all three but was confirmed by microscopic analysis in only two. One of these patients had placental abruption and cesarean delivery for fetal distress at 27 weeks. Circumvallate placenta is another cause of uterine band, sheet, or shelf. The finding can be associated with a normal obstetric outcome but may predispose to placental complications.

Abruptio Placentae↗
PubMed1995

Identification and cholesterol quantification of low density lipoprotein subclasses in young adults by VAP-II methodology.

Low density lipoprotein (LDL) particles are heterogeneous in size, density, and chemical composition; small, dense LDL may be more atherogenic than large, buoyant LDL. We have developed a rapid microscale method called LDL VAP-II (Vertical Auto Profile-II) for quantification of cholesterol in LDL subclasses. The method is based upon a short (1 h) single vertical spin density-gradient ultracentrifugation and on-line VAP-II analyzer. LDL VAP-II is rapid and reproducible. Using this method five LDL subclasses, designated as LDL-1 (most buoyant) through LDL-5 (most dense), have been identified in a population consisting of 195 medical students (ages, 22-29 years). The Rf (relative position of the major LDL peak in the density gradient; the higher the Rf value, the lower the peak density) was significantly positively correlated with cholesterol levels of high density lipoprotein (HDL) (r = 0.594), HDL3 (0.350) and HDL2 (0.625), and significantly negatively correlated with triglycerides (TG) (-0.355) and cholesterol levels of very low density lipoprotein (VLDL) (-0.386) and intermediate density lipoprotein (IDL) (-0.432). These results are consistent with those obtained by other investigators. The Rf value was significantly correlated with peak particle diameter as determined by non-denaturing gradient gel electrophoresis (r = 0.859). In a forward stepwise multivariate analysis comparing Rf with sex, VLDL, LDL, Lp[a], IDL, HDL3, HDL2, and triglyceride, only HDL2 remained in the model.

Adult↗
PubMed1994

Transcatheter tubal sterilization in rabbits. Technique and results.

RATIONALE AND OBJECTIVES: A potential method of nonsurgical tubal sterilization was tested in rabbits. METHODS: Metal coils were designed which could be placed into the uterotubal junction using transvaginal fluoroscopic fallopian tube catheterization. These metal coils were successfully placed unilaterally in the uterotubal junction of 32 rabbits. The contralateral fallopian tube and uterus were used as a control. The rabbits were bred, and the presence of pregnancies was confirmed by palpation. RESULTS: In 21 rabbits (66%), the coil stayed in place. Sixteen rabbits had multiple gestations on the side without the coil and no gestations on the side with the coil. Three rabbits had gestations on both sides, even though the coil was in place, and two rabbits never conceived. In 11 rabbits (34%) the coil was dislodged as early as 5 days and as late as 18 weeks after the procedure. Five of these 11 rabbits had bilateral embryos, 4 had embryos only on the side contralateral to where the coil had been, and 2 never conceived. CONCLUSIONS: The metal coil does prevent conception if it stays in place at the uterotubal junction. However, the coil failed to prevent pregnancy in 3 of 19 rabbits, and was dislodged in 11 rabbits, giving an overall failure rate for contraception of 44%.

Animals↗
PubMed1994

Localization of contraceptive implant capsules for removal.

Implanted contraceptive capsules occasionally cannot be removed by means of palpation with local dissection, and imaging guidance is necessary. The authors report obtaining a single antero-posterior radiograph of the upper arm followed by sonography to localize the capsule and to allow precise marking of the skin over the capsules. The method can be performed quickly and is inexpensive and effective, allowing the localization and removal procedures to be scheduled independently.

Adult↗
PubMed1994

apoB-100 has a pentapartite structure composed of three amphipathic alpha-helical domains alternating with two amphipathic beta-strand domains. Detection by the computer program LOCATE.

Due to the great length of apolipoprotein (apo) B-100, the localization of lipid-associating domains in this protein has been difficult. To address this question, we developed a computer program called Locate that searches amino acid sequences to identify potential amphipathic alpha-helixes and beta-strands by using sets of rules for helix and strand termination. A series of model chimeric protein test datasets were created by tandem linking of amino acid sequences of multiple proteins containing four different secondary structural motifs: motif A (exchangeable plasma apolipoproteins); motif G (globular alpha-helical proteins); motif C (coiled-coil alpha-helical proteins); and motif B (beta pleated-sheet proteins). These four test datasets, as well as randomly scrambled sequences of each dataset, were analyzed by Locate using increasingly stringent parameters. Using intermediately stringent parameters under which significant numbers of amphipathic helixes were found only in the unscrambled motif A, two dense clusters of putative lipid-associating amphipathic helixes were located precisely in the middle and at the C-terminal end of apoB-100 (a sparse cluster of class G* helixes is located at the N-terminus). The dense clusters are located between residues 2103 through 2560 and 4061 through 4338 and have densities of 2.4 and 2.2 amphipathic helixes per 100 residues, respectively; under these conditions, motif A has a density of 1.4 amphipathic helixes per 100 residues. These two domains correspond closely to the two major apoB-100 lipid-associated domains at residues 2100 through 2700 and 4100 through 4500 using the principle of releasability of tryptic peptides from trypsin-treated intact low-density lipoprotein. The classes of amphipathic helixes identified within these two putative lipid-associating domains are considerably more diverse than those found in the exchangeable plasma apolipoproteins. Interestingly, apoB-48 terminates at the N-terminal edge of the middle cluster. By using a similar strategy for analysis of amphipathic beta-strands, we discovered that the two gap regions between the three amphipathic helix clusters are highly enriched in putative amphipathic beta-strands, while the three amphipathic helical domains are essentially devoid of this putative lipid-associating motif. We propose, therefore, that apoB-100 has a pentapartite structure, NH2-alpha 1-beta 1-alpha 2-beta 2-alpha 3-COOH, with alpha 1 representing a globular domain.

Algorithms↗
PubMed1994

An improved method of needle localized biopsy of nonpalpable lesions of the breast.

We have created a modified technique of localization biopsy of the breast that more easily identifies the position of the wire tip after placement and before biopsy. After wire placement, metallic skin markers are placed on the nipple and wire entry site. The patient is then positioned on a roentgenography table in the same position to be used during biopsy. A supine roentgenogram of the breast is obtained. At the time of the biopsy, this roentgenogram is aligned with the corresponding markers on the skin of the patient and the position of the wire tip is noted. A curvilinear incision along Langer's lines can then be made at that site. This procedure was used in 53 consecutive biopsies of nonpalpable mammary lesions. In all instances, the biopsy incisions were independent of the wire insertion sites and in each instance, the lesion was excised in a single specimen. Compared with other techniques, our method is simple, inexpensive, requires no special equipment and results in only a minimal increase in radiation dose to the breast (50 millirad). The decrease in the amount of tissue dissection without decreasing accuracy afforded by our technique can improve compliance with the American Cancer Society's recommended surgical guidelines for breast preservation therapy.

Biopsy, Needle↗