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Biomedical subjects

P Roy

Publications and source records attributed to P Roy.

At least 109 records · Page 6Linked to original sources
PubMed1998

New generation of African horse sickness virus vaccines based on structural and molecular studies of the virus particles.

African horse sickness virus (AHSV) is a member of the genus Orbivirus, which also includes bluetongue virus (BTV) and epizootic haemorrhagic disease (EHDV) virus. These orbiviruses have similar morphological and biochemical properties, with distinctive pathobiological properties and host ranges. Sequencing studies of the capsid proteins have revealed evolutionary relationships between these viruses. Biochemical studies of the viruses together with the expression of individual proteins and protein complexes have resulted in the development of new generation vaccines. Baculovirus expressed AHSV VP2 provides protection against death caused by AHSV challenge. Similarly, BTV VP2 alone elicits protective neutralising antibodies against BTV in sheep, which is enhanced in the presence of VP5. Recent developments in biotechnology (multiple gene expression baculovirus systems) have made it possible to synthesise orbivirus particles that biochemically and immunologically mimic authentic virions but lack the genetic material. Particle doses as low as 10 micrograms elicit responses that are sufficient to protect sheep 15 months post vaccination, against virulent virus challenge. Moreover, knowledge of the three dimensional structure of these particles enables us to engineer them to deliver multiple foreign peptide components representing other viral epitopes (e.g. foot and mouth disease virus and influenza virus) in order to elicit protective immunity.

African Horse Sickness Virus↗
PubMed1998

Induction of T cell response by bluetongue virus core-like particles expressing a T cell epitope of the M1 protein of influenza A virus.

A CD4+ T cell epitope of the influenza virus matrix protein corresponding to the C terminus (QAYQKRMGVQMQRFK) was inserted into the VP7 gene of bluetongue virus (BTV). The chimeric protein was expressed by a dual recombinant Autographa californica polyhedrosis virus (AcNPV), which encodes the two inner capsid proteins VP3 and VP7 of BTV. When Spodoptera fRugiperda cells (Sf9 cells) were infected with this recombinant BTV, core-like particles (CLPs) were formed as demonstrated by electron microscopy. To study the immunogenicity of a foreign epitope deprived of its natural flanking sequences in vitro, purified CLPs expressing the T cell epitope were used to stimulate two different MHC class II-restricted CD4+ human T cell clones. One of these T cell clones, ALF 3.7 was specific for the inserted epitope, whereas the other T cell clone ALF 4.4 recognized shorter derivates of the given epitope. CLPs with the inserted epitope were presented as efficiently as purified influenza virus matrix protein to the clone ALF 3.7, whereas clone ALF 4.4 showed no proliferative response.

Baculoviridae↗
PubMed1998

Rapid onset and dissipation of left atrial spontaneous echo contrast during percutaneous balloon mitral valvotomy.

BACKGROUND: Thromboembolism after percutaneous balloon mitral valvotomy (PBMV) has been attributed to dislodement of preexisting thrombus during transseptal puncture and instrumentation of the left atrium. The occurrence of thromboembolic events after PBMV in the absence of demonstrable left atrial thrombus before PBMV suggests that thrombus might form during the procedure. Spontaneous echo contrast (SEC) is a swirling pattern of blood echogenicity that is a marker of blood stasis in the left atrium. Exacerbation of left atrial SEC during PBMV may be indicative of an increased thromboembolic risk. METHODS: Transesophageal echocardiography was performed during PBMV in 20 patients with mitral stenosis. Grades of severity of left atrial SEC [0 (nil) to 4+ (severe)] were allocated before and after each balloon inflation. RESULTS: Before PBMV, SEC was present in 17 patients. New SEC or increased severity of SEC was observed during 49 of 56 balloon inflations. SEC was unchanged after six deflations, decreased after 14 deflations, and disappeared after 36 deflations. The mean times to onset and dissipation of SEC after balloon inflation and deflation were 3.1+/-1.5 and 3.9+/-1.6 seconds, respectively. After successful PBMV, SEC was unchanged in three patients, decreased in one, and resolved in 13. CONCLUSIONS: SEC is a dynamic and acutely reversible phenomenon that is highly sensitive to changes in left atrial hemodynamic conditions. Left atrial blood stasis induced by balloon inflation may promote thrombogenesis during PBMV.

Adult↗
PubMed1998

The complete sequence of four major structural proteins of African horse sickness virus serotype 6: evolutionary relationships within and between the orbiviruses.

The amino acid sequences of four major capsid proteins of African horse sickness virus (serotype 6, AHSV-6) have been determined from analyses of cDNA clones representing the L2, L3, M6 and S7 RNA segments. The AHSV-6 L3 RNA segment has an open reading frame of 2715 base pairs and encodes the inner capsid protein VP3 which comprises 905 amino acids. The VP3 layer forms the subcore of the virion and is surrounded by the VP7 protein which is encoded by the S7 gene. The AHSV-6 S7 gene was found to be 1047 nucleotides in length with a coding capacity for the VP7 protein of 349 amino acids. These core proteins are encapsulated by the outer capsid proteins VP5 and VP2 which are encoded by the M6 and L2 genes respectively. The M6 gene of AHSV-6 was determined to be 1564 nucleotides in length and encoded a protein product of 504 amino acids while the L2 gene comprised 3203 nucleotides which encoded a predicted protein product of 1051 amino acids. Comparison of these four sequences with the core protein sequences of other serotypes of African horse sickness virus, Bluetongue virus which infects sheep, and Epizootic haemorrhagic disease virus of deer, demonstrated that despite the pathobiological properties and host range of these distinct orbiviruses, extreme conservation is evident within the capsid genes. Sequence analyses also suggested that the similarity levels between serogroups depict the structure and function of the individual capsid proteins. The data indicated that the evolution of the capsid genes of gnat transmitted orbiviruses is strongly influenced by functional and structural constraints.

African Horse Sickness Virus↗
PubMed1998

[Local control of the primary tumor. Necessary step in control of cancer].

There is clear evidence for a cancer cure. The hypothesis behind a treatment with curative intent is the ability to eradicate all the cancer cells of a tumour. Out of three cancer deaths, one is related to local failure. In some cancers like breast carcinoma, death is mainly in relation with the development of distant metastases. Even in such a tumour, an improvement in local control can translate as better survival. Radiation therapy, often in association with surgery, is playing a major role in tumour local control. Such a local control is mandatory if cure is at aim. One of the main goals of clinical research is to find a good compromise between local control of the disease and a non mutilating surgical approach.

Antineoplastic Agents↗
PubMed1998

Comparison of Newcastle disease vaccines by serology using serum, tears and feather pulp samples.

Seroconversion of 3 lentogenic commercial Newcastle disease (ND) vaccines and experimental V4 vaccines was compared based on the haemagglutination inhibition (HI) test against ND. It was found that for primary vaccination all the vaccines produced similar response but for vaccinations V4 and LaSota were better than RDVF. Eight-five samples each of serum, tears and feather pulp were collected from respective birds and antibody assessment was done against ND by HI test. The geometric mean HI titres (GMT) of serum samples were highest followed by tears and feather pulp samples before vaccination and 3 weeks after vaccination by oculonasal route and the difference was statistically significant (p < 0.01). Three weeks after booster vaccination by oculonasal route, however, the GMT of serum samples were highest followed by feather pulp and tear samples. The ease of collection of feather pulp samples and their role in ND serology is discussed.

Animals↗
PubMed1998

Velogenic Newcastle disease virus in captive wild birds.

Newcastle disease virus (NDV) was isolated from the faeces of seven different species of clinically healthy captive wild birds. All seven NDV isolates were characterized as velogenic based on the mean death time in embryonated hens' eggs and the intracerebral pathogenicity index in day-old chicks. Three of the isolates were placed in group C1 based on the reactions with monoclonal antibodies. The role of captive wild birds in the epidemiology of Newcastle disease is briefly discussed.

Animals↗
PubMed1998

Prognostic comparison of the pathological classifications of gastric cancer: a population-based study.

AIMS: There is controversy over the value of the pathological classifications of gastric carcinomas in the prediction of patient survival. This study was designed to assess the prognostic value of four widely used pathological classifications, in addition to classical prognostic factors. METHODS AND RESULTS: Records from the population-based registry of digestive tract tumours in the department of Côte d'Or (France) have been analysed. All available histopathological slides of gastric cancer resected between 1976 and 1985 were reviewed and classified according to World Health Organization (WHO), Laurén, Ming and Goseki pathological coding systems. A relative survival analysis was performed using a relative survival model with proportional hazard applied to net mortality by interval. WHO, Laurén or Goseki classifications were not found to be independent prognostic factors. In addition to advanced age group, depth of parietal involvement, nodal involvement, presence of metastases, tumour site and gross appearance of the tumour, the Ming's infiltrative type was associated with a lower survival. CONCLUSION: This study suggests an independent prognostic value of the Ming subtypes with respect to survival in patients resected for gastric carcinoma.

Adult↗
PubMed1998

Cisplatin-DNA adducts and protein-bound platinum in blood of testicular cancer patients.

DNA adducts formed by cisplatin [cis-diamminedichloroplatinum(II)] were measured in blood samples from 48 testicular cancer patients treated in four centers in Europe during four to six cycles with cisplatin infusions on five successive days (total samples, 112). Total protein-bound platinum (Pt) in blood was also measured (total samples, 84). The mean on the main DNA adduct, cis-Pt(NH3)2d(pGpG) (Pt-GG), was 0.75 fmol/microg DNA [standard deviation (SD) = 0.66] on the first day of the first cycle, increased after the infusion at day 5 of the cycle (mean 1.74 fmol/microg DNA, SD = 0.90) and decreased on the following day (mean 1.09 fmol/microg DNA, SD = 0.62). In subsequent cycles, there was a tendency to an increase in the mean Pt-GG levels. The values of protein-bound Pt in blood showed little reduction between day 5 and 6 of each cycle, and a stable increase during the course of the therapy. Strong correlations were seen between day 1, 5 and 6 of the first cycle for both Pt-GG and protein-bound Pt in blood. A strong correlation (r = 0.62, p < 0.001, 69 pairs) was found between the levels of Pt-GG and protein-bound Pt. Only two patients relapsed during the follow-up; therefore, the analysis of the association between Pt-GG levels and response to therapy was not informative. The results of this study suggest that DNA adducts formed by cisplatin at the beginning of chemotherapy are predictive of values found during later days and cycles, and that the value of protein-bound Pt in blood is predictive of the value of DNA adducts.

Adult↗
PubMed1998

Bluetongue virus core protein VP4 has nucleoside triphosphate phosphohydrolase activity.

The intact virion of bluetongue virus comprises ten segments of dsRNA enclosed in two concentric protein capsids. The core, which is transcriptionally active, includes three minor proteins (VP1, VP4 and VP6) which are considered to be the candidates for the core-associated enzymes that transcribe and modify full-length mRNA copies for each of the ten genome segments. Using purified recombinant VP4 protein and core-like particles containing VP4, in this report it is demonstrated that VP4 has nucleoside triphosphatase (NTPase) activity. VP4 is a nonspecific NTPase that hydrolyses four types of ribonucleoside triphosphate (NTP) to the corresponding nucleoside diphosphate. The substrate preference was GTP>ATP>UTP>CTP. NTP hydrolysis by VP4 was maximal when the Mg2+ or Ca2+ ion concentrations were 4 mM or 6 mM, respectively. The presence of single-stranded polynucleotides poly(A), poly(U) and poly(C) had little effect on the NTPase activity. Although the enzyme exhibited a broad temperature optimum around 40 degrees C, the pH optimum was sharp, between pH 7.5 and 8. The Km and Vmax of ATP hydrolysis were calculated to be 0.25+/-0.05 microM ATP and 55+/-4 pmol ATP hydrolysed min(-1) microg(-1), respectively. The Km was affected by the addition of poly(A) to only a small extent in contrast to the Vmax, which was increased by at least twofold.

Acid Anhydride Hydrolases↗
PubMed1998

A leucine zipper-like domain is essential for dimerization and encapsidation of bluetongue virus nucleocapsid protein VP4.

The bluetongue virus (BTV) minor protein VP4, with molecular mass of 76 kDa, is one of the seven structural proteins and is located within the inner capsid of the virion. The protein has a putative leucine zipper near the carboxy terminus of the protein. In this study, we have investigated the functional activity of this putative leucine zipper by a number of approaches. The putative leucine zipper region (amino acids [aa] 523 to 551) was expressed initially as a fusion protein by using the pMAL vector of Escherichia coli, which expresses a maltose binding monomeric protein. The expressed fusion protein was purified by affinity chromatography, and its size was determined by gel filtration chromatography. Proteins of two sizes, 51 and 110 kDa, were recovered, one equivalent to the monomeric form and the other equivalent to the dimeric form of the fusion protein. To prove that the VP4-derived sequence was responsible for dimerization of this protein, a mutated fusion protein was created in which a VP4 leucine residue (at aa 537) within the zipper was replaced by a proline residue. Analyses of the mutated protein demonstrated that the single mutation indeed prevented dimerisation of the protein. The dimeric nature of VP4 was further confirmed by using purified full-length BTV-10 VP4 recovered from recombinant baculovirus-expressing BTV-10 VP4-infected insect cells. Using chemical cross-linking and gel filtration chromatography, we documented that the native VP4 indeed exists as a dimer in solution. Subsequently, Leu537 was replaced by either a proline or an alanine residue and the full-length mutated VP4 was expressed in the baculovirus system. By sucrose density gradient centrifugation and gel filtration chromatography, these mutant forms of VP4 were shown to lack the ability to form dimers. The biological significance of the dimeric forms of VP4 was examined by using a functional assay system, in which the encapsidation activity of VP4 into core-like particles (CLPs) was studied (H. LeBlois, T. French, P. P. C. Mertens, J. N. Burroughs, and P. Roy, Virology 189:757-761, 1992). We demonstrated conclusively that dimerization of VP4 was essential for encapsidation by CLPs.

ATP-Binding Cassette Transporters↗
PubMed1998

Membrane organization of bluetongue virus nonstructural glycoprotein NS3.

The smallest RNA segment (S10) of bluetongue virus (an orbivirus, family Reoviridae) encodes two closely related nonstructural proteins, the 229-amino-acid (aa) NS3 and the 216-aa NS3A. The proteins are found in glycosylated and nonglycosylated forms in infected cells (X. Wu, H. Iwata, S.-Y. Chen, R. W. Compans and P. Roy J. Virol. 66:7104-7112, 1992). The NS3/NS3A proteins have two hydrophobic domains (aa 118 to 141 and 162 to 182) and two potential asparagine-linked glycosylation sites (aa 63 and 150), one of which is located between the hydrophobic domains. To determine whether these features were used in the mature protein forms, we generated a series of mutants of the S10 gene and expressed them by using the vaccinia virus T7 polymerase transient-expression system. Our data indicate that both hydrophobic domains of NS3 span the cell membrane and that only the site at aa 150 is responsible for N-linked glycosylation of the NS3 proteins.

Amino Acid Sequence↗
PubMed1998

Lower extremity: nonstepping digital angiography with photostimulable imaging plates versus conventional angiography.

PURPOSE: To compare nonstepping digital subtraction angiography (DSA) (ie, storage phosphor radiography adapted to a stationary imaging plate changer) with conventional screen-film angiography in the evaluation of the lower extremities. MATERIALS AND METHODS: Fifty-one patients with peripheral vascular disease underwent both nonstepping DSA and screen-film angiography. The angiographic and radiologic techniques of both systems were kept identical for each patient. Three radiologists independently rated the overall quality of each angiogram. In their evaluations for each of 12 arterial segments on all 102 angiograms, they also rated the degree of opacification, the diameter reduction of the most severe stenosis, and their level of confidence. RESULTS: Mean overall quality scores and levels of confidence were better for nonstepping DSA than for screen-film angiography (P < .001). Full opacification was reported in 95.6% and 89.2% of all 1,836 segments with nonstepping DSA and screen-film angiography, respectively (P < .0001). The difference between the mean stenosis grades obtained with screen-film angiography and nonstepping DSA was not statistically significant. Intertechnique agreements were good (kappa = 0.77, 0.81, and 0.81), whereas interobserver agreements were influenced by the observer's experience with the imaging techniques. CONCLUSION: Nonstepping DSA images of the lower extremity were of better diagnostic quality than were screen-film angiograms. The development of dedicated nonstepping DSA equipment is warranted.

Adult↗
PubMed1998

Thyroid hormone stimulates progesterone release from human luteal cells by generating a proteinaceous factor.

Blood samples collected from 29 women (aged between 19 and 35 years) during the luteal phase of the menstrual cycle (between days 18 and 23 of the cycle) showed that deficiency in thyroid hormone level is related to a decrease in progesterone (P4) secretion. To observe the effect of thyroid hormone on human ovarian luteal cells, 3,5,3'-triiodothyronine (T3; 125 ng/ml) was added to luteal cells in vitro. T3 significantly stimulated progesterone release (P < 0.01) from luteal cells and this could be blocked by cycloheximide, indicating a protein mediator for the T3 effect. The T3 stimulatory effect was inhibited by anti-T3 antibody suggesting specificity of T3 action. Addition of T3 caused a more than threefold increase in cellular protein synthesis which was inhibited by cycloheximide. Preparation of partially purified thyroid hormone-induced factor (TIF) (from peak II of Sephadex G 100 chromatography of T3-incubated cells), and its addition to luteal cell incubations caused a significant increase in P4 release (P < 0.05). Incubation with trypsin or treatment with heat destroyed the stimulatory effect of TIF on P4 release, indicating the proteinaceous nature of TIF. Purified thyroid hormone-induced protein. (TIP) from rat granulosa cells and fish ovarian follicles greatly stimulated P4 release from human luteal cells. These results suggest that T3 stimulation of P4 release from human luteal cells is not direct, but is mediated through a putative protein factor, which appears to be a protein conserved through evolution as far as its biological activity is concerned.

Adult↗