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Biomedical subjects

D A Anderson

Publications and source records attributed to D A Anderson.

At least 73 records · Page 4Linked to original sources

Effect of riboflavin-binding protein deficiency on riboflavin metabolism in the laying hen.

Normal chicken eggs contain substantial amounts of riboflavin, all of which is bound to a specific, high-affinity, riboflavin-binding protein (RfBP). Two hens, genetically unable to produce RfBP and thus unable to deposit sufficient riboflavin in their eggs, were compared to two normal hens with respect to the biological half-life of [14C]riboflavin, the tissue distribution of 14C-labeled flavins, and the relative contributions of tissue and dietary riboflavin to flavins deposited in the egg. The biological half-life of [14C]riboflavin was slightly but insignificantly less in the RfBP-deficient hens (11.5 +/- 1.7 days vs 15.1 +/- 3.3 days). The 14C-labeled flavin content of a variety of tissues 3 weeks after the intraperitoneal injection of 5 microCi of riboflavin was also very similar among the four hens. In contrast, the 14C-labeled flavin content of egg yolk, egg albumen, and blood plasma from RfBP-deficient birds was less than 10% of normal. For all hens, the specific radioactivity of flavins in yolk and albumen was similar to that in liver but less than that in heart. We conclude that riboflavin deposited in egg had equilibrated with the large hepatic flavin pool and was not derived preferentially from unlabeled dietary riboflavin. Other than the inability to deposit riboflavin in their eggs, hens of the mutant strain have normal riboflavin metabolism.

Administration, Oral↗

Glomerulotubular function in patients undergoing moderate surgical stress.

Glomerular function and renal tubular function assessed by measurements of creatinine clearance rates (CCR), lithium clearance rates (CLi, and N-acetyl-beta-D-glucosaminidase (NAG) enzymuria were measured immediately prior to and within 24 h of operation (average operative time 150 min) in 16 patients undergoing a moderate surgical stress. Although serum creatinine concentrations and CCR were similar pre- and postsurgery at 105 +/- 21 and 108 +/- 21 mumol/L and 108 +/- 67 and 110 +/- 59 mL/min/100 kg body weight (mean +/- SD), respectively, both CLi and NAG were significantly increased following the surgical insult. CLi was increased from 19.7 +/- 6.1 to 31.7 +/- 16.7 mL/min/100 kg (p less than 0.01) and NAG from 71 +/- 58 to 164 +/- 10 U/mmol urinary creatinine (p less than 0.001). In addition, the absolute distal tubular reabsorption of sodium and water increased from 18.3 +/- 5.9 and 15.8 +/- 9.7 to 30.6 +/- 16.4 and 27.6 +/- 12.2 mL/min/100 kg body weight (both p less than 0.05), respectively. These results indicate early postoperative renal tubular dysfunction following a moderate surgical stress, undetected by conventional screening.

Aged↗

Characterization of hepatitis A virus capsid proteins with antisera raised to recombinant antigens.

The capsid proteins of hepatitis A virus (HAV) were expressed as fusion proteins of beta-galactosidase in E. coli using the expression vector lambda gt11. Four fusion proteins were stably expressed and used to immunize rabbits to obtain mono-specific antisera. The antisera were unable to neutralize viral infectivity or react with HAV by radioimmunoassay. Three of the antisera were able to recognize HAV antigens in infected BS-C-1 cells by immunofluorescence and denatured capsid proteins by immunoblot analysis. The antisera were used to investigate the migration of the capsid proteins in gels by immunoblot analysis using standard SDS-PAGE conditions and in gels containing urea. The migration of VP1 and VP3 correlated with their molecular weights predicted from the nucleotide sequence and was consistent in either the presence or absence of urea. However, VP2 migrated with an apparent molecular weight significantly higher than the predicted value and, in gels containing urea, migrated as a doublet. It is proposed that the upper band of this doublet represents VP0, the proteolytic precursor of VP2 and VP4. The relative molecular mass (Mr) of VP4 was estimated to be less than 1 kDa, which is substantially lower than the 2.5 kDa predicted from the nucleotide sequence.

Animals↗

Morphogenesis of hepatitis A virus: isolation and characterization of subviral particles.

The morphogenesis of hepatitis A virus (HAV) in BS-C-1 cells was examined by immunoblotting with antisera to capsid proteins and labeling of virus-specific proteins with L-[35S]methionine. Antiserum to VP2 detected two virus-specific proteins with apparent molecular masses of 30.6 and 30 kDa, representing VP0 and VP2, while antiserum to VP1 detected proteins with molecular masses of 33 and 40 kDa, representing VP1 and a virus-specific protein which we designated PX, respectively. Sedimentation of cell lysates revealed the presence of virions, procapsids, and pentamers, but particles analogous to the protomers of other picornaviruses were not detected. Although provirions and virions were not found as discrete species in our gradient system, it was evident that the rate of sedimentation was proportional to the relative amounts of VP0 and VP2 in particles, with slower-sedimenting particles (provirions) containing predominantly VP0 rather than VP2. Procapsids contained VP0 in addition to VP1 and VP3. Pentamers also contained VP0, but PX was present rather than VP1. These results suggest that PX is a precursor to VP1 and is most likely 1D2A. Primary cleavage of the viral polyprotein also occurs at the 2A-2B junction in cardioviruses and aphthoviruses, but assembly of pentamers containing 1D2A has not been reported for those viruses. The absence of detectable levels of protomers suggests a high efficiency of pentamer formation, which may be related to the high efficiency of viral RNA encapsidation for HAV (D.A. Anderson, B.C. Ross, and S.A. Locarnini, J. Virol. 62:4201-4206, 1988). The results of this study reveal further unusual aspects of the HAV replicative cycle which distinguish it from other picornaviruses and may contribute to its restricted replication in cell culture.

Capsid↗

Protection against enzootic pneumonia of pigs: intraperitoneal inoculation with live LKR strain of Mycoplasma hyopneumoniae.

Pigs obtained from a mycoplasma-free piggery were randomised into 4 groups of 9. Groups 1 and 2 were injected by the intraperitoneal route with liquid culture of the LKR strain of Mycoplasma hyopneumoniae. Group 1 was injected once and group 2 twice. Group 3 was made up of pigs inoculated by the intranasal route with the virulent Beaufort strain of M. hyopneumoniae; they served as the source of infection for the challenge. Group 4 were uninfected, uninjected controls. Six weeks after the last injection, groups from 1 to 4 were placed in contact. Seven of the pigs in the 1-dose group and 6 in the 2-dose group were free of lesions at necropsy 6 weeks after challenge. Of the two pigs with lesions in the 1-dose group one had only a small lesion but the other had extensive lesions; it had not shown an antibody response after injection of culture. The lesions in the 3 pigs in the 2-dose group were all small. All 9 control pigs had lesions which varied from medium to large in size. The difference in the incidence of pneumonia between the injected and control groups was significant (P less than 0.05) and the proportion of severely affected pigs in the vaccinated groups was significantly lower (P greater than 0.01). There was no difference between those given one dose of vaccine and those receiving 2 doses.

Administration, Intranasal↗

Effects of altered dietary iron intake in Mycobacterium paratuberculosis-infected dairy cattle: sequential observations on growth, iron and copper metabolism and development of paratuberculosis.

Twenty calves were orally inoculated with Mycobacterium paratuberculosis at six weeks old. At six months old, 10 of these, plus four uninfected controls were maintained on limited dietary copper and supplemented iron intake for a further 27 months. During this time all these animals, together with a further four untreated controls, were bred before being killed and examined for evidence of paratuberculosis. Despite significant reduction in weight gain, attributable to both iron supplementation and infection, no significant difference was found in the numbers of iron-supplemented and unsupplemented animals that developed clinical signs nor in the extent and severity of intestinal lesions between groups. Accumulation of iron in paratuberculosis lesions was not affected by iron supplementation but was positively correlated with the frequency of shedding of M paratuberculosis in faeces (P less than 0.05). Dietary iron supplementation alone resulted in serum hyperferraemia, hepatic siderosis and slight hypocuprosis, whereas, in infected animals, this resulted in marked hypocuprosis and anaemia within groups (P less than 0.05). Infection alone resulted in serum hypoferraemia and intestinal and hepatic siderosis which was positively correlated with the severity of infection within groups (P less than 0.05). Susceptibility to paratuberculosis may result from failure ultimately to limit monokine-mediated iron sequestration in intestinal tissue.

Animals↗

Persistence of infection in mice inoculated intranasally with Cryptococcus neoformans.

Cryptococcus neoformans was instilled intranasally into mice which were periodically sacrificed to determine the course of infection. Cryptococci persisted within the nasal passages throughout the 90 day study. Extranasal dissemination began 14-28 days after instillation and was still demonstrable 90 days post-exposure. Ten percent mortality was observed in mice receiving 10(6) cryptococci, while no mortality was observed in mice exposed to 10(3) or 10(4) cryptococci. Our research suggests that nasal colonization with C. neoformans can precede pulmonary and systemic cryptococcosis by weeks or months.

Animals↗

Restricted replication of hepatitis A virus in cell culture: encapsidation of viral RNA depletes the pool of RNA available for replication.

The replication of hepatitis A virus (HAV) in BS-C-1 cells was examined under single-cycle growth conditions by using strand-specific probes for detection of viral RNA species. No measurable lag phase was demonstrated between accumulation of positive-strand HAV RNA and production of infectious virions, indicating that replication of virion RNA is rate limiting for the production of infectious virus. Intracellular viral RNA was further analyzed by using 2 M LiCl to fractionate the insoluble nonvirion 35S RNA and replicative intermediates (RI) from the soluble virions and double-stranded replicative forms, in conjunction with sucrose density gradient ultracentrifugation to separate the different forms of viral RNA. Throughout the productive phase of HAV infection, 95 to 97% of positive-strand HAV RNA was soluble in 2 M LiCl and was shown to be contained in mature virions. Of the LiCl-insoluble HAV RNA, more than 99% was positive-stranded 35S RNA, whereas 0.4% was negative stranded and had the sedimentation and partial RNase resistance characteristics of RI. The pattern of RNA accumulation in HAV-infected cells is thus very different from that seen in poliovirus-infected cells, where large pools of RI and mRNA are produced before RNA is sequestered into mature virions. The results of this study suggest that encapsidation of positive-strand HAV RNA inhibits transcription at all times during the growth cycle, thereby reducing the pool of replicating RNA and the final yield of infectious HAV.

Capsid↗

Memories of labor and birth: reliability of post partum questionnaire reports.

The stability of reports of feelings during vaginal births was assessed in three studies. In Study I, post partum reports from 475 primiparous women who participated in a childbirth preparation program were compared to identify differences between reports completed from the day of the birth to more than six weeks after the birth. In Study II, the post partum reports of 46 primiparous and 95 multiparous women completed 1, 3, or 6 mo. after giving birth were compared. In Study III, test-retest reliabilities were calculated for post partum reports completed by 22 primiparous and 16 multiparous women during the 5th and 7th post partum weeks. In these studies, reports of pain during labor were stable.

Female↗

Prediction of intravenous theophylline dosage based on a single, nonsteady-state concentration: a clinical study of childhood status asthmaticus.

A pharmacokinetic model was applied to achieve therapeutic serum theophylline concentrations rapidly in 25 children with status asthmaticus. A sustained release theophylline preparation had been taken within 36 hours by 12 children; within 14 hours, seven had taken an immediate release preparation; for six children, no theophylline was taken before hospital admission. Single serum theophylline concentrations were determined at nonsteady-state conditions within 13.5 hours of admission (median 6.75 hours). An iterative program was applied to predict the steady-state theophylline concentration as well as necessary adjustments in dosage. Measured steady-state concentrations were then compared with the predicted values. The median measured steady-state concentration was 15 mg/L, and the median predicted steady-state level was 13 mg/L. The least squares regression line was: Measured = 0.738 predicted + 4.77; r = .721, P less than .01. No patient experienced symptoms of toxicity. This technique affords the possibility of accurate prediction of steady-state theophylline concentrations and dosing requirements with a minimum number of serum concentration determinations in children with status asthmaticus.

Asthma↗

Cytopathology, plaque assay, and heat inactivation of hepatitis A virus strain HM175.

Hepatitis A virus (HAV) strain HM175 derived after repeated subculture of persistently infected B-SC-1 cells caused a specific cytopathic effect upon acute infection of B-SC-1 cells. The virus formed visible plaques on B-SC-1 cell monolayers after 9 to 14 days of incubation at 34 degrees C, and virus can therefore be titrated by plaque assay. Virus could be re-isolated from plaques of infected cells, which allows the clonal isolation of HAV variants. The stability of a plaque-purified variant of HAV at elevated temperatures exceeded that of poliovirus type 1, but this variant is less stable than previously reported strains of HAV.

Animals↗

Cellular changes associated with persistent hepatitis A infection in vitro.

The rate of division, morphology and ultrastructure of BSC-1 cells, persistently infected with hepatitis A virus (HAV), were compared with uninfected cells for 60 days after splitting of the cells. Both control and infected cells showed a biphasic growth pattern marked firstly by increasing cell density and high mitotic rate (exponential phase) and then high constant cell density and little mitosis (stationary phase). Immunoperoxidase studies showed that hepatitis A antigen (HAAg) appeared as cytoplasmic granules approximately one third of the way through the exponential phase in infected cells. The percentage of cells with HAAg rose until the early stationary phase when virtually all cells contained antigen. Radioimmunoassay demonstrated an increase in HAAg per cell in the stationary phase. Radioimmunofocus assay and immune electron microscopy confirmed the presence of HAV in infected cells in the stationary phase. Thin sectioning electron microscopy showed cytoplasmic annulate lamellae in infected cells of both phases but not in control cells.

Animals↗

Early serological responses to Mycoplasma hyopneumoniae infection.

A complement fixation (CF) test for Mycoplasma hyopneumoniae antibody, using the complement dilution method, will detect infection in inoculated pigs at an early stage. The interval between inoculation and the first positive CF response--defined as 4.6 u of complement fixed--was determined for four separate methods of inoculation and for pigs with artificially induced pneumonia. Sera from two groups were tested by the enzyme-linked immunosorbent assay (ELISA). By CF, the i.v. group showed positive titers at 9.8 +/- 0.4 (mean +/- SE) days. All pigs in this group had positive titers, most of them at the maximum level of 31 u fixed. The i.p. group first showed positive titers at 13.3 +/- 1.0; 17 of 70 had no CF response. The s.c. group first showed titers at 21.1 +/- 2.6; 7 of 24 did not respond. Intranasal inoculation first produced a CF response at 14.3 +/- 0.9, and all pigs had positive titers. Pigs in contact with those with induced pneumonia first showed titers at a mean of 27.2 days (SE +/- 1.3), and all pigs exhibited a CF response. There was no significant difference between the CF test and ELISA results.

Administration, Intranasal↗

Studies into immunisation of cattle against interdigital necrobacillosis.

Calves were immunised with fractions of Fusobacterium necrophorum incorporated in mineral oil adjuvant and then each foot was experimentally exposed to interdigital necrobacillosis (foot abscess) by subcutaneous injection of homologous organisms through the interdigital skin. The number of cells from an 18 h liquid culture that might be expected to cause 50% of the feet of control calves to develop marked swellings following subcutaneous injection was shown to be approximately 2.2 X 10(8) cells. Immunity was shown to be associated with antigens that were located in the supernatant of the culture, and which may be identical with or closely associated with the exotoxins.

Abscess↗

Indirect immunofluorescence assay for the detection of hepatitis A virus-specific serum immunoglobulins.

Hepatitis A virus-specific BSC-1 cells were used for the detection of serum immunoglobulins to hepatitis A virus by indirect immunofluorescence. Of 150 serum samples tested, specific immunoglobulin M was detected only in patients with serologically confirmed acute hepatitis A, while specific immunoglobulin G was detected in patients with acute or past clinical hepatitis A as well as many patients with no known history of hepatitis.

Antibodies, Viral↗

Restrictive events in the replication of hepatitis A virus in vitro.

Hepatitis A virus was purified from the feces of 2 patients with unrelated, naturally acquired infections and was inoculated into FRhK-4 cells. Analysis of protein synthesis by double-label coelectrophoresis and subtraction allowed the resolution of virus-specific proteins synthesized during infection. In FRhK-4 cells the two strains of virus studied produced markedly different profiles of virus-specified proteins, with an accumulation of high-molecular-weight proteins for strain HM790 relative to strain HM175, suggesting a level of restriction in the processing of the viral polyprotein.

Antigens, Surface↗