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Biomedical subjects

R Ling

Publications and source records attributed to R Ling.

At least 19 recordsLinked to original sources
PubMed1996

Selection of mutations in the hepatitis B virus polymerase during therapy of transplant recipients with lamivudine.

We describe mutations in the hepatitis B virus (HBV) polymerase gene in viruses which reactivated in two patients during therapy with -2'-deoxy-3'-thiacytidine, or lamivudine (3TC), and following orthotopic liver transplantation for chronic hepatitis B. Virus resistance to 3TC is associated with mutations which lead to amino acid substitutions in the highly conserved tyr-met-asp-asp (YMDD) motif, part of the active site of the polymerase, and which parallel those seen in resistant human immunodeficiency virus (HIV). Substitutions of valine and isoleucine for methionine were found in the two cases. The significance of single secondary mutations, which differ between viruses from the two patients, remains to be determined. Thus, viral resistance to lamivudine of hepatitis B virus mimics that of HIV and can occur in the setting of immunosuppression after liver transplantations.

Adult↗
PubMed1996

Sequence of the nucleocapsid protein gene of subgroup A and B avian pneumoviruses.

The nucleocapsid protein (N) gene of two subgroup A and one subgroup B strains of avian pneumovirus has been cloned and sequenced. The gene of all three isolates comprised 1197 nucleotides (nt), which formed a single major open reading frame, potentially encoding a protein of 391 amino acid residues. The N gene of the two subgroup A isolates differed by only 1 nt but differed by 282 (24%) nt and 35 (11%) amino acids from the B isolate. The predicted protein was identical in length to that of human, bovine and ovine respiratory syncytial viruses, the amino acid identity being approximately 41% overall but with some regions of identity > 90%.

Amino Acid Sequence↗
PubMed1995

Sequence and in vitro expression of the phosphoprotein gene of avian pneumovirus.

The phosphoprotein (P) gene of two subgroup A strains of avian pneumovirus comprised 855 nucleotides containing only one substantial open reading frame encoding a protein of 278 amino acids, with a predicted M(r) of 30,323. In vitro translation of P mRNA in a wheat germ system resulted in the synthesis of two polypeptides of M(r) 35,000. Comparison of the deduced P protein sequence with that of the known mammalian pneumoviruses revealed overall amino acid identities ranging from 31 to 34.5%, suggesting a distant relationship. However, there was a much higher identity (63.2-68.4%) in a region of 57 residues, which included a heptad repeat sequence.

Amino Acid Sequence↗
PubMed1995

Case report: malignant fibrous histiocytoma following radiation therapy of fibrous dysplasia.

Malignant fibrous histiocytoma commonly occurs spontaneously. In some cases it follows previous therapeutic or incidental irradiation, or miscellaneous pre-existing osseous conditions. Recently, it has been associated with total hip arthroplasty. We report a case of malignant fibrous histiocytoma following radiation therapy of fibrous dysplasia and review the relevant literature.

Bone Neoplasms↗
PubMed1994

Use of single-cysteine mutants to probe the location of the disulfide bond in LamB protein from Escherichia coli.

The two cysteine residues of the LamB protein of Escherichia coli outer membrane have been shown to form an intrasubunit disulfide whose location differs greatly in the two current topology models of the LamB protein. This study probes the location of the disulfide by examining conditions for intersubunit disulfide formation in single-cysteine mutants of LamB protein. Formation of an intersubunit bond in the purified mutant proteins, which resulted in a disulfide-linked dimer, only occurred after heat treatment, suggesting the disulfide is not exposed on the surface in the native protein.

Bacterial Outer Membrane Proteins↗
PubMed1992

Effect of cup geometry and the presence of cement on acetabular component fixation.

Two series of implanted conical, polyethylene Ring cups were studied clinically and radiographically. In one series the cups were uncemented while in the second cement was used. The results using cemented conical cups were then compared with results using a cemented cup of hemispherical design to study the effect of cup geometry. At 7-8 years a total of 3.8% of the cemented conical series and 2.9% of the uncemented were revised for aseptic loosening. At 9 years survivorship was identical. Migration of the sockets occurred in approximately 25% of both series and was directly related to cup cover of less than 80% (P < 0.001). The series of cemented hemispherical cups were reviewed after an identical period. Although the numbers revised for loosening were comparable in this and the cemented conical group, radiological migration was statistically greater in the latter (P < 0.001). Alternative methods of socket preparation and cement technique were thought to be the most likely explanation for the observed differences.

Acetabulum↗
PubMed1992

Sequence analysis of the 22K, SH and G genes of turkey rhinotracheitis virus and their intergenic regions reveals a gene order different from that of other pneumoviruses.

The nucleotide and deduced amino acid sequences of three genes of turkey rhinotracheitis virus (TRTV) together with the nucleotide sequences of the relevant intergenic regions were determined. The deduced amino acid sequence of one of the genes shows significant identity (42%) to that of the 22K protein of human respiratory syncytial virus (RSV). The TRTV 22K gene, like that of RSV, has a second open reading frame, although the amino acid sequence deduced from this reading frame does not show any similarity to the equivalent predicted RSV protein. The other two genes and their deduced amino acid sequences do not show any sequence similarity to the genes of other pneumoviruses. However, the hydrophobicity profiles of the predicted proteins do show similarities to those of the small hydrophobic (SH) and attachment protein (G) genes of RSV. The TRTV G gene is 1193 nucleotides in length and encodes a protein of 391 amino acids (M(r) 42984), which is rather larger than the RSV G protein (predicted M(r) 36000). The TRTV SH gene is 589 nucleotides in length, encoding a protein of 174 amino acids (M(r) 18797), which is considerably larger than the size of the RSV SH protein (M(r) 7500). The sequences of the intergenic regions derived from clones of polycistronic mRNAs and polymerase chain reaction products obtained with primers from different genes reveal the order on the virus genome to be 3' F-22K-SH-G 5'. This differs from the gene order of paramyxoviruses and morbilliviruses, which lack a 22K gene (and in some cases a SH gene), and the pneumoviruses RSV and pneumonia virus of mice, which have the F and 22K genes located after the G gene.

Amino Acid Sequence↗
PubMed1992

Method to measure acoustic impedance and reflection coefficient.

A frequency-domain based system for measuring acoustic impedance and reflection coefficient is described. The calibration procedure uses a least-mean-squares approximation to the Thevenin parameters describing the source and receiver characteristics in which the data measured on closed, cylindrical tubes are matched to a viscothermal tube model. The system is intended for use in acoustical measurement in human ear canals, in which the cross-sectional area of the ear canal at the point of insertion is imprecisely known. This area is acoustically estimated from the impedance data, and the reflection coefficient is calculated in terms of this area and the impedance data. Measurements on a variety of closed tubes show the method is accurate over the frequency range investigated (less than 10.7 kHz). The time-domain reflection function is evaluated by transforming the reflection coefficient from the frequency domain, but the finite bandwidth of the measured data limits the accuracy of time-domain response measurements. The method is well suited for frequency-domain measurements in human ear canals.

Acoustic Impedance Tests↗
PubMed1991

Role of a disulfide bond in the thermal stability of the LamB protein trimer in Escherichia coli outer membrane.

In order to understand the unusual heat resistance of LamB protein (the outer membrane component of the maltose transport system in Escherichia coli and its receptor for bacteriophage lambda), we investigated the role of its 2 cysteinyl residues. Our studies show that Cys22 and Cys38 form an intrasubunit disulfide bond which contributes to the heat stability of the LamB protein trimer. Physical evidence for the disulfide was obtained by using site-directed mutagenesis to convert Asn36 to Met, which allowed cyanogen bromide cleavage between the 2 cysteines. Upon reduction one of the N36M fragments migrated as two pieces, resolved by two-dimensional polyacrylamide gel electrophoresis. Other mutagenized LamB proteins, in which 1 or both Cys residues were converted to Ser, exhibited a sharp loss of thermal stability. In contrast to wild-type LamB protein trimer, which does not dissociate to monomers even after 60 min at 100 degrees C, only 10-15% of the mutant LamB proteins remain trimeric after boiling 10 min. The disulfide bond in LamB protein is not required for its transport function, since both mutagenized LamB protein and N-ethylmaleimide-labeled LamB protein exhibit normal uptake of sugars in proteoliposomes. Finally, the disulfide bond must not be between subunits of the LamB trimer since reversible dissociation of trimer is achieved by low pH or denaturants in the absence of reducing agent.

Amino Acid Sequence↗
PubMed1989

A comparison of the effects of various stimulatory agents on t-PA secretion by normal and malignant cell lines.

Succinyl con A and acetyl con A both stimulated epithelial cells to produce similar yields of tissue plasminogen activator (t-PA) to those previously obtained with native con A. However, unlike con A, the derivatized lectins did not adversely affect cell morphology and viability, and cells treated with succinyl con A could secrete t-PA for a prolonged period. Con A and the two derivatives produced similar morphological effects in Bowes melanoma cells, but t-PA production was not increased. Elevated cyclic nucleotide concentrations did not affect t-PA production from epithelial cells, but calcium ionophore treatment generated t-PA yields similar to those obtained with lectins. Azacytidine, which enhanced t-PA production from epithelial cells, did not increase yields from Bowes melanoma cells, and also sodium butyrate, reported to increase t-PA yields from human endothelial cells, had no effect on either cell line.

Azacitidine↗
PubMed1989

Polypeptides of pneumonia virus of mice. I. Immunological cross-reactions and post-translational modifications.

A murine polyclonal antiserum and monoclonal antibodies have been employed to identify pneumonia virus of mice (PVM) polypeptides in infected cells and to study post-translational modifications. Immunoprecipitation experiments using a murine polyclonal antiserum and a monoclonal antibody directed against a 39K protein have established an antigenic relationship between two PVM proteins and the N and P proteins of human respiratory syncytial virus. Although 20 virus-specific polypeptides have been identified in lysates of infected cells, evidence is presented that some of these are related and that the number of unique polypeptides probably does not exceed 11 or 12. The phosphoproteins of PVM have a pattern of mobilities more like that of the recently described pneumovirus causing rhinotracheitis in turkeys than that of human or bovine respiratory syncytial virus.

Animals↗
PubMed1989

Polypeptides of pneumonia virus of mice. II. Characterization of the glycoproteins.

The kinetics of synthesis and the nature of the oligosaccharides of the glycoproteins of pneumonia virus of mice (PVM) were studied. Tryptic peptide mapping showed that the two major glycosylated polypeptides G1 and G2 were different forms of the same protein. G2 was derived from G1 which in turn appeared to be derived from an unidentified precursor. The G1/G2 protein of PVM is probably a haemagglutinin since a monoclonal antibody directed against it has a high haemagglutination inhibition titre. On the basis of experiments with inhibitors and glycosidases it was deduced that G1 and G2 have both N-linked and O-linked oligosaccharides. The putative fusion protein-equivalent of PVM was shown to possess N-linked oligosaccharides. In the presence of tunicamycin a high mobility form (F1t) appeared to be derived from a precursor (F0t) with the same mobility as the fully glycosylated protein. If by analogy with other paramyxoviruses this represents a cleavage event, the difference in mobility of the precursor and product suggests that the putative F2 product is smaller than the corresponding F2 protein of other paramyxoviruses. However, no F2 candidate protein was detected and evidence for an F1,2 dimer was inconclusive. The glycoproteins of PVM resemble those of respiratory syncytial virus in terms of their pattern of glycosylation, but differ in their processing.

Animals↗
PubMed1988

Turkey rhinotracheitis virus: in vivo and in vitro polypeptide synthesis.

Turkey rhinotracheitis (TRT) virus is a paramyxovirus associated with recent outbreaks of acute respiratory disease in turkeys. On morphological criteria it resembles the pneumoviruses more than the other members of the paramyxovirus family. In this communication we report the identification of five virus-induced polypeptides in TRT virus-infected BS-C-1 cells. The Mr of these polypeptides were estimated as 38K, 35K, 30K, 19K and 15K from their electrophoretic mobility in SDS-polyacrylamide 6 to 15% gradient gels. The virus specificity of four (the 38K, 35K, 30K and 19K Mr polypeptides) of these five was confirmed by their predominance as products of in vitro translation of mRNA from TRT virus-infected BS-C-1 cells. An additional virus-specific polypeptide with an estimated Mr of 22K was revealed by in vitro translation and may be either a non-structural polypeptide or a minor structural protein. All six polypeptides were immunoprecipitated by a murine antiserum. Three other polypeptides (129K, 57K and 45K) were also immunoprecipitated. The 57K, 45K and 15K Mr polypeptides were glycosylated, and the 57K glycopolypeptide may be a disulphide-bonded dimer of the 45K and 15K glycopolypeptides. Another major glycopolypeptide of 83K Mr was observed but its virus-specificity could not be confirmed. Taken together our results tentatively identify eight or nine TRT virus-specified polypeptides and are consistent with the classification of TRT virus as a new member of the genus Pneumovirus.

Animals↗
PubMed1983

Production of a fibrinolytic enzyme from cultures of guinea pig keratocytes grown on microcarriers.

Microcarrier technology was investigated as a means of producing a fibrinolytic enzyme, for the treatment of occlusive thrombotic vascular disorders, in sufficient quantity to carry out an evaluation of its clinical potential and the economics of large scale production. Preliminary results showed that the cells grew well and produced significant quantities of the enzyme at high (10 g/1) microcarrier concentrations.

Animals↗