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Biomedical subjects

D Todd

Publications and source records attributed to D Todd.

At least 55 records · Page 3Linked to original sources
PubMed2000

Biochemical characterization of salmon pancreas disease virus.

Salmon pancreas disease virus (SPDV) has been shown to cause severe economic losses in farmed Atlantic salmon (Salmo salar) and has been reported to occur in Europe, Scandinavia and the United States. This paper describes the biochemical characterization of SPDV in terms of its RNA and protein composition. SPDV was purified by precipitation from infected Chinook salmon embryo (CHSE-214) cell-culture supernatant and sucrose density-gradient centrifugation. Fractions containing virus were identified by an immunodot blot assay using an SPDV-specific MAb. Two major proteins with molecular masses of approximately 55 and 50 kDa, putatively identified as the E1 and E2 alphavirus glycoproteins respectively, were detected when purified virus preparations were analysed by PAGE. Radiolabelling experiments indicated that SPDV infection of CHSE-214 cells did not shut-off host-cell protein synthesis, making attempts to identify virus-specific proteins unsuccessful. However, radioimmunoprecipitation assay (RIPA) experiments showed that two SPDV-specific MAbs reacted with a protein in the 50-55 kDa range. Northern blot hybridization with cloned cDNA probes indicated that infected cells contained RNA species of approximately 11.4 and 4 kb, which correspond to the genomic and subgenomic RNAs specified by SPDV. The results described are consistent with SPDV being characterized as an alphavirus.

Alphavirus↗
PubMed1999

A new isolation method for rat intraepithelial lymphocytes.

Intraepithelial lymphocytes (IELs) play critical roles in gut immunity. In mice, gammadelta T cells are a large component of the IEL population. In the rat, gammadelta IELs are reportedly much less common, but technical issues suggest that previous analyses should be interpreted cautiously. The study of IELs in rats has been impeded by isolation procedures that are lengthy and complex, leading to small cell yields. For this reason, it is possible that rat IELs analyzed in previous studies have not been representative of the entire IEL compartment. We report a new method for the isolation of rat IELs that is based on the selective removal of intestinal epithelial cells under conditions that leave the basement membrane undisturbed. The method is rapid and requires neither enzymatic digestion, nor surgical removal of Peyer's patches, nor vigorous mechanical manipulation of the intestine. The yield of rat IELs using this method is 5- to 10-fold greater than that reported for other methods. Morphological and phenotypic analyses demonstrated that the purified cell population is comprised of IELs and is not contaminated with lamina propria or Peyer's patch lymphocytes. Phenotypic analysis revealed five major subsets of IELs based on differential cell surface expression of CD4, CD8, and alphabeta T cell receptor (TcR). Among the alphabetaTcR- cells was a population of gammadelta T cells present at levels not previously detected. The isolation of IEL sub-populations using this methodology should facilitate studies of the function of these cells in gut immunity.

ADP Ribose Transferases↗
PubMed1999

Salmon pancreas disease virus, an alphavirus infecting farmed Atlantic salmon, Salmo salar L.

A 5.2-kb region at the 3' terminus of the salmon pancreas disease virus (SPDV) RNA genome has been cloned and sequenced. The nucleotide and predicted amino acid sequences show that SPDV shares considerable organizational and sequence identity to members of the genus alphavirus within the family Togaviridae. The SPDV structural proteins encoded by the 5.2-kb region contain a number of unique features when compared to other sequenced alphaviruses. Based on cleavage site homologies, the predicted sizes of the SPDV envelope glycoproteins E2 (438 aa) and E1 (461 aa) are larger than those of other alphaviruses, while the predicted size of the alphavirus 6K protein is 3.2 K (32 aa) in SPDV. The E2 and E1 proteins each carry one putative N-linked glycosylation site, with the site in E1 being found at a unique position. From amino acid sequence comparisons of the SPDV structural region with sequenced alphaviruses overall homology is uniform, ranging from 32 to 33%. While nucleotide sequence analysis of the 26S RNA junction region shows that SPDV is similar to other alphaviruses, analysis of the 3'-nontranslated region reveals that SPDV shows divergence in this region.

3' Untranslated Regions↗
PubMed1999

Antigenicity and pathogenicity characteristics of molecularly cloned chicken anaemia virus isolates obtained after multiple cell culture passages.

The Cux-1 isolate of chicken anaemia virus (CAV), which had received 310 (P310) cell culture passages, was substantially less pathogenic than virus that had been passaged 13 times (P13). Molecularly cloned virus isolates, selected from the P310 and P13 virus populations using recombinant DNA cloning and transfection procedures, reacted differently with 4 CAV-specific monoclonal antibodies (MAbs), which had been raised to low-passage Cux-1 virus. In contrast to the strong immunofluorescence (IF) reactivities exhibited by all P13 cloned isolates tested, 80% and 57% of the P310 cloned isolates reacted weakly with MAbs 2A9 and 4H4, which are directed against conformational epitopes on the capsid protein, VP1. Sequence analysis of the VP1 coding regions possessed by ten P310 and two P13 cloned isolates showed that 6 amino acid changes within VP1 had been selected by multiple-cell culture passage. One of these at position 89 in VP1 appeared to be crucial for determining reactivity with MAb 2A9. Of nine P310 cloned isolates evaluated, 8 were substantially attenuated compared to the low-passage Cux-1 virus pool. It is concluded that the individual virus variants comprising the P310 virus pool differ with regards to their antigenicity and pathogenicity.

Amino Acid Sequence↗
PubMed1999

Development of a blocking enzyme-linked immunosorbent assay for the serological diagnosis of chicken anaemia virus.

The development and evaluation of an enzyme-linked immunosorbent assay (ELISA) for the detection of serum antibody to chicken anaemia virus (CAV) are described. This test depends on the abilities of CAV-specific antibodies present in convalescent chicken serum to block the reaction between virus antigen, adsorbed to the ELISA plate. and a CAV-specific mouse monoclonal antibody (MAb), 2A9, that has been conjugated to horseradish peroxidase. The 2A9 MAb has been shown to react with 10 geographically different field isolates of CAV, a finding which indicates that the test will find worldwide application. In comparative experiments involving 525 serum samples from specific pathogen free and commercial breeder flocks, there was 98.5% agreement between the results obtained with the blocking ELISA and those obtained with an indirect ELISA developed previously in this laboratory. The blocking ELISA was found to have advantages in terms of speed and cost compared with the indirect ELISA format.

Adsorption↗
PubMed1999

Treatment of de novo acute myeloid leukaemia in Hong Kong: a twenty-year experience (1975 to 1996).

BACKGROUND: Small patient numbers and short follow-up are common in some acute myeloid leukaemia (AML) studies and data on secondary malignancies after treatment of AML are rare. AIMS: To determine the prognostic factors and long-term treatment results. METHODS: A retrospective study of patients with de novo AML under the age of 60 over a 20-year period in which two induction therapy regimens: 7:3 (1975-1983) and 7:3:7 (1984-1996) and three consolidation chemotherapy regimens: 5:2 (1975-1983), 5:2:5 (1984-1990) and Ara-C/mitoxantrone (1991-1996) were used. Disease-free (DFS), overall survivals (OS) and prognostic factors were analysed. RESULTS: Two-hundred and two of 276 (73%) patients attained complete remission (CR). The CR rates of 7:3 and 7:3:7 regimens were 70.5% and 74.5% respectively (p=0.92). The median DFS was 12 months and the projected DFS at 10- and 20-years were 23% and 21% respectively. For patients consolidated with 5:2, 5:2:5 and Ara-C/mitoxantrone, the median DFS was 15 m, 12 m and 11 m respectively and the projected ten-year DFS were 27%, 21% and 18% respectively (p=0.2). Ninety per cent of relapses occurred within two years from remission but there were two late relapses at 109 m and 120 m respectively. Young age and FAB M3 subtype were favourable prognostic factors to OS (p=0.04) and DFS (p=0.006) respectively. There was no secondary solid tumour in the long-term survivors. CONCLUSION: Our experience confirmed the efficacy of standard-dose Ara-C/daunorubicin and the prognostic value of age and FAB subtype. Median and projected DFS were similar to western studies.

Acute Disease↗
PubMed1999

The inducible expression of dominant-negative epidermal growth factor receptor-CD533 results in radiosensitization of human mammary carcinoma cells.

Ionizing radiation activates the epidermal growth factor receptor (EGFR) and downstream signaling involving the cytoprotective mitogen-activated protein kinase (MAPK) pathway. In our effort to investigate the role of EGFR in cellular responses to radiation, we generated mammary carcinoma cell clones, MCF-TR5-EGFR-CD533 and MDA-TR15-EGFR-CD533, that inducibly express EGFR-CD533, a truncated EGFR mutant lacking mitogenic and transformation activity. EGFR-CD533 expression inhibits radiation- and EGF-induced EGFR autophosphorylation and MAPK activation and, therefore, functions as a dominant-negative mutant without blocking the expression of EGFR or erbB-2, another member of the erbB receptor Tyr kinase family. Expression of EGFR-CD533 only minimally inhibited cell growth and did not alter radiosensitivity to single radiation exposures. However, repeated 2 Gy radiation exposures of cells, under conditions of EGFR-CD533 expression, essentially abolished their ability for subsequent cell growth. These results identify the inhibition of EGFR function through genetic manipulation as a potential therapeutic maneuver. The concept of such an intervention would be the radiosensitization of cells by counteracting a radiation-induced cytoprotective proliferation response.

Breast Neoplasms↗
PubMed1999

Functional magnetic stimulation: a new modality for enhancing systemic fibrinolysis.

OBJECTIVE: To investigate whether functional magnetic stimulation (FMS) of the leg muscles could enhance systemic fibrinolysis. DESIGN: A within-subject analysis of systemic fibrinolysis before and after 60 minutes of FMS. SETTING: The Functional Magnetic Stimulation and Special Coagulation Laboratories in the Spinal Cord Injury Service at a Veterans Administration health care facility. PARTICIPANTS: Twenty healthy volunteers were recruited. The exclusion criteria were: (1) personal or family history of venous or arterial thrombosis, (2) personal or family history of cardiac arrythmias, (3) current use of medication, and (4) the presence of a cardiac pacemaker or other metallic implants. MAIN OUTCOME MEASURES: Whole blood clot lysis time (WBCLT) and tissue plasminogen activator (t-PA) antigen determined at baseline and 10 and 60 minutes after FMS. RESULTS: A significant decrease was observed in the mean WBCLT after FMS. The WBCLT decreased from 17+/-1.3 h before FMS to 12+/-1.0 h and 11+/-0.8 h at 10 and 60 minutes post-FMS, respectively. The mean t-PA antigen levels did not show a significant change (p = .6701) from pre-FMS (6.7+/-0.91 ng/mL) to 10 min post-FMS (6.8+/-0.91 ng/mL) and 60 min post-FMS (7.0+/-1.02 ng/mL). Several patterns of fibrinolytic response to FMS were observed. They differed in relation to the degree of enhancement, the period at which maximum enhancement occurred, and the corresponding t-PA antigen levels observed. CONCLUSIONS: The FMS-induced contractions of the leg muscles enhanced systemic fibrinolysis ex vivo. The improvement in fibrinolysis occurred immediately after FMS and was also observed at 60 minutes post-FMS. FMS appears to produce a sustained enhancement of systemic fibrinolysis that may prove useful in deep-vein thrombosis prophylaxis.

Adult↗
PubMed1999

Retinopathy of prematurity in infants < 32 weeks' gestation at birth in New South Wales in 1993 and 1994.

OBJECTIVE: To study the incidence and severity of retinopathy of prematurity (ROP) in infants < 32 weeks' gestation. DESIGN: Review of the records maintained in the New South Wales Neonatal Intensive Care Unit Study (NICUS) database on infants admitted to the neonatal intensive care units (NICU) in NSW from 1 January 1993 to 31 December 1994. RESULTS: In the more premature infants, 23-26 weeks' gestation, 65% developed ROP (102 of 157 examined for ROP). Forty-four infants (28%) developed severe ROP (Stage >/= 3 ROP), 19 infants (12.1%) required cryo/laser therapy and one infant (0.6%) in this group had a retinal detachment. One hundred and fifty-seven of 159 surviving infants (98.7%) were examined for ROP. In the infants 27-28 weeks' gestation, 38.3% developed ROP (103 of 269 examined for ROP). Fifteen infants (5.6%) developed severe ROP, seven infants (2. 6%) required cryo/laser therapy for threshold ROP and three infants (1.1%) in this group had a retinal detachment. Two hundred and sixty-nine of 299 surviving infants (90%) were examined for ROP. In the infants 29-31 weeks' gestation, 10.8% developed ROP (48 of 443 examined for ROP). Six infants (1.4%) developed severe ROP, one infant (0.2%) required cryo/laser therapy for threshold ROP and no infant in this group had a retinal detachment. However, only 443 of 681 surviving infants (65.1%) in this group were examined for ROP. Of the four infants with detached retinas, one was a 25 week gestation infant weighing 840 g, two were 27 weeks' gestation weighing 960 and 980 g and one infant was a 28 week gestation infant weighing 620 g. No infant developed Stage 5 ROP. CONCLUSION: In the more mature infants 29-31 weeks' gestation, the rate of ROP is low, although severe ROP still occurs. However, only 65.1% of these infants were examined for ROP and we should be diligent in screening for ROP in the sicker infants in this group. The incidence of severe ROP as well as the rate of cryo/laser therapy in premature infants 23-26 weeks' in NSW has not changed since the increases seen in the early 1990s. Retinal detachment also occurs in the infants 27-28 weeks' gestation and it is important that all these infants are screened for ROP.

Gestational Age↗
PubMed1999

Halnan Lecture 1998. Medicine: quo vadis.

Medicine has made rapid progress but has it lost its direction? Medicine is but one of the factors on which the health of a community depends; others include sanitation, housing, education, and life-style. Developments in medical education and practice must take into account the needs of society, changes in demography, patient expectations, costs, and the importance of preventive medicine. While the humanities, communication skills, and medical economics are important, these should not be taught at the expense of a sound core curriculum and clinical skills, which may be declining. Can human kindness and understanding be taught? Role models may help, but are their numbers sufficient? Postgraduate organisations have been successful in vocational training, but 'professionalism' must be further nurtured. This concerns doctors' responsibilities and relationships with patients, colleagues, the profession, and society. Specialists and subspecialists are needed but so are the 'generalists'. The education and training of doctors must be done in the context of the overall training of health care personnel. The Hong Kong Academy of Medicine and its Colleges must have a significant voice in making decisions and policies that affect health and medical matters. A patient should be a satisfied patient and not a statistic or a client. Where is medicine heading? Let us pause, think, and then act to lead it straight ahead.

Journal Article↗
PubMed1998

Hemosiderosis with diabetes mellitus in untransfused Hemoglobin H disease.

A 37-year-old untransfused, non-drinking man with Hemoglobin H-CS disease presented with insulin-dependent diabetes mellitus, markedly elevated serum ferritin level, and marked iron deposition in hepatocytes. He did not carry either of the two common mutations of the HLA-H gene for hereditary hemochromatosis, namely, Cys282Tyr and His68Asp, nor did he have the associated HLA marker (HLA-A3, B7 nor B-14) for the disease. Patient with HbH disease should be monitored for iron overload.

Adult↗
PubMed1998

Characterization of novel circovirus DNAs associated with wasting syndromes in pigs.

Porcine circovirus (PCV) was initially recognized as a contaminant of continuous pig kidney cell lines and was not thought to be pathogenic. Antibodies reactive to the cell culture isolate of PCV (PCV PK-15) are prevalent in the swine population worldwide. Recently, PCV PK-15-like antigen and nucleic acid were demonstrated in lesions associated with wasting syndromes in pigs in North America and Europe. Monoclonal antibodies raised to circoviruses isolated from pigs with wasting syndromes highlighted differences between these circoviruses and the PCV PK-15 cell culture isolate. This has led to speculation that a new pathogenic PCV may have emerged in the swine populations of several countries. We report the cloning and characterization of novel circovirus DNAs purified from virus isolates made from tissues of North American and European pigs with wasting syndromes. These North American and European circoviruses form a closely related group at the nucleotide sequence level (> 96% intra-group nucleotide sequence identity) but exhibit < 80% nucleotide sequence identity with the PCV PK-15 cell culture isolate. This report provides evidence for a new type of possibly pathogenic PCV. We propose that these new circoviruses should be referred to as PCV2 as opposed to the original PK-15 cell culture isolate, which should be referred to as PCV1.

Amino Acid Sequence↗
PubMed1998

Isolation of circovirus from lesions of pigs with postweaning multisystemic wasting syndrome.

Postweaning multisystemic wasting syndrome (PMWS), an apparently new disease, has been recognized in swine herds in western Canada. Young pigs with this disease have progressive weight loss, tachypnea, dyspnea, and jaundice, accompanied by interstitial pneumonia, lymphadenopathy, hepatitis, and nephritis. We examined more than 400 pigs from more than 70 herds in Alberta, Saskatchewan, and Manitoba with cases of PMWS. A small virus was isolated from a range of tissues from 8 of 8 affected pigs examined. The agent was identified as a circovirus-like virus using electron microscopy, immunohistochemical staining with porcine and rabbit immune serum, and in situ hybridization. Immunohistochemical examination of tissues from more than 100 affected pigs has revealed widespread viral antigen, often contained in circovirus-like inclusion bodies, in lesions from numerous organs. Although Koch's postulates remain to be fulfilled, these results demonstrate a high degree of association between the presence of the circovirus-like virus and PMWS in affected swine.

Alberta↗
PubMed1997

Close phylogenetic relationship between egg drop syndrome virus, bovine adenovirus serotype 7, and ovine adenovirus strain 287.

A cloned egg drop syndrome (EDS) virus genomic DNA fragment containing the protease gene has been identified and the complete nucleotide sequence of the protease and partial nucleotide sequence of the hexon genes has been determined. Phylogenetic analysis of the protease gene has revealed EDS virus to be genetically more closely related to bovine adenovirus type 7 (BAV-7) and ovine adenovirus isolate 287 (OAV287) than either of these two viruses are to other members of the genus Mastadenovirus or EDS virus is to an other member of the Aviadenovirus genus. The three viruses share further similarities in that they have a high percentage AT content in their genome and are characterized by having more compact genomes than other adenoviruses. The protease gene from all three viruses contained the active site residues (H55-D72-C122 triad) and C104 (providing a disulfide bond to cofactor pVIc). However, P137, found in all other members of the Mastadenovirus genus, and thought to be involved in trafficking, was missing from the protease of the EDS virus, BAV-7, and OAV287. These results suggest that EDS virus should be classified along with BAV-7 and OAV287 in a separate taxon.

Adenoviridae↗
PubMed1997

CD56+ NK lymphomas: clinicopathological features and prognosis.

The surface molecule CD56 marks a category of malignant lymphoma of putative natural killer (NK) cell origin. We conducted a retrospective analysis of 24 cases of CD56+ NK lymphoma/leukaemia to define the clinicopathologic and prognostic features of this specific group of lymphomas. 56 cases of nasal lymphomas and 204 cases with an initial diagnosis of peripheral T-cell lymphoma were retrospectively analysed. To specifically examine lymphomas of putative NK origin, only those that were negative for surface expression of CD3 but positive for CD56 were analysed. 24 cases were identified. The initial predominant sites of involvement were nasal (n = 18), palate (n = 1), nodal (n = 1) and multi-organ (n = 4). Clinically, in patients with disease localized to one anatomical site (n = 20), most had symptoms confined to the nose, with a high percentage in early stage (I: 91%; IV: 9%). The marrow was not involved in any of these cases. However, patients with multi-organ involvement at presentation (n = 4) behaved differently. All presented acutely with pancytopenia, hepatosplenomegaly, and marrow infiltration with haemophagocytosis. A leukaemic phase was observed in one case. Anthracycline containing combination chemotherapy resulted in complete remission in 75% of patients with localized disease, but only in 25% with multi-organ involvement. The median survival of patients with localized disease was 12 months, compared with 2 months in the multi-organ group (P = 0.06); the disease-free survival was significantly better in the former (P < 0.01). The overall median survival of all patients was still poor at 11 months. We conclude that CD56+ NK lymphomas could be divided into two main patterns of disease presentations: localized (predominantly nasal), and multi-organ involvement. Each has different clinicopathologic and prognostic features. Conventional chemotherapy appeared ineffective for the majority of patients, and innovative treatment modalities are needed to improve outcome.

Adolescent↗
PubMed1997

Molecular defects in Hb H hydrops fetalis.

The molecular defect in two unique cases of Hb H hydrops fetalis has been characterized. Both cases are due to co-inheritance of a 'non-deletion' defect affecting the alpha2 gene: at codon 30 delta GAG, Glu) and codon 59 (G --> A, Gly --> Asp) respectively, and a zeta-alpha thalassaemia (thal) 1 or alpha thal 1 genotype. These two non-deletion defects, unlike previously described cases, resulted in severe anaemia of the fetuses and emphasize the importance of performing prenatal diagnosis for these families.

Gene Deletion↗