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Biomedical subjects

C A Hopkins

Publications and source records attributed to C A Hopkins.

At least 19 recordsLinked to original sources
PubMed2001

Production of biologically active equine interleukin 12 through expression of p35, p40 and single chain IL-12 in mammalian and baculovirus expression systems.

Interleukin-12 (IL-12) is a key cytokine in the development of cell-mediated immune responses. Bioactive IL-12 is a heterodimeric cytokine composed of disulphide linked p35 and p40 subunits. The aim of this study was to verify biologically activity of the products expressed from equine interleukin-12 (IL-12) p35 and p40 cDNAs and to establish whether equine IL-12 could be expressed as a p35/p40 fusion polypeptide, as has been reported for IL-12a of several mammalian species. We report production of equine IL-12 through expression of p35 and p40 subunits in mammalian and insect cells and of a p35:p40 fusion polypeptide in mammalian cells. Conditioned medium recovered from cultures transiently transfected with constructs encoding equine p35 and p40 subunits or single chain IL-12 enhanced IFN-gamma production in cells derived from equine lymph nodes. Preincubation of IFN-gamma inducing preparations with anti-p40 monoclonal antibody resulted in a significant decrease in IFN-gamma induction capacity. Medium recovered from p35 and p40-expressing baculovirus infected cultures enhanced target cell IFN-gamma production and proliferation. Experimental studies in mice and other animals have revealed a therapeutic benefit of IL-12 in cancer, inflammatory and infectious disease and an adjuvant effect in prophylactic regimes. Production of a bioactive species-specific IL-12 is a first step towards an investigation of its potential application in equine species.

Amino Acid Sequence↗
PubMed1991

Inhibition of growth of a tapeworm Hymenolepis diminuta in its normal host (rat).

The biomass of 8-day-old worms of Hymenolepis diminuta in secondary infections, administered to rats 3-10 days after chemotherapeutically expelling a primary infection, was 70-90% less, and the worms were more posteriorly distributed, than in naive controls. The strong depressive effect on growth waned rapidly over 2-5 weeks, but even in rats not challenged until 17 months later, worm growth was weakly depressed by 30%. The extent to which growth was depressed in a secondary infection was independent of the number of worms in the challenge but increased with number of worms in the immunizing infection up to four to eight worms. Further increase up to 64 worms had little effect. This suggests, as it is known that the biomass of worms in a rat reaches a maximum with infections of between five and 10 worms, that the change in the intestine is proportional to biomass, not number, of worms. It is argued that partially suppressed immuno-inflammatory changes in the intestine, which will affect secondary worms so strongly, will also have depressed growth and fecundity effects on the primary worms, that a dynamic equilibrium is reached between the strength of the intestinal response and the biomass of the tapeworm, and that it is reaching this equilibrium, not a 'crowding effect', which limits H. diminuta to a level compatible with the survival of the rat.

Animals↗
PubMed1988

Purified capsular polysaccharide-induced immunity to Staphylococcus aureus infection.

In this study, we determined that immunization with capsular polysaccharide from Staphylococcus aureus could protect mice against nonlethal infections induced by encapsulated staphylococci. We immunized mice with either formalin-killed bacteria or purified capsular polysaccharide (PCP) and challenged them with one of three related S. aureus strains that varied in capsule size. Quantitative cultures of blood and kidney from the animals were performed to evaluate protection. Immunization with whole bacteria protected mice against infection with the homologous strain. Mice immunized with PCP were protected when challenged intravenously with either a highly encapsulated S. aureus strain or a microencapsulated mutant but not with an unencapsulated mutant. Protection correlated with capsular antibody levels in the immunized animals. Immunity to staphylococcal infection could be passively transferred to naive animals by using immune serum. These experiments suggest that the S. aureus capsular polysaccharide merits further study as a potential vaccine candidate for preventing staphylococcal infection.

Animals↗
PubMed1987

Virulence studies, in mice, of transposon-induced mutants of Staphylococcus aureus differing in capsule size.

We used three related strains of Staphylococcus aureus to determine whether capsule size influenced bacterial virulence. Strain SA1 mucoid elaborated a large capsule demonstrable by transmission electron microscopy (TEM). Nonmucoid isolates were derived from strain SA1 mucoid by Tn551 insertional mutagenesis. By TEM, strain JL24 produced a "microcapsule," whereas strain JL25 was unencapsulated. Strain SA1 mucoid had a 50% lethal dose for mice greater than 3,000-fold lower than that of strains JL24 and JL25. Quantitative cultures of blood and kidney from animals challenged intravenously revealed that strain SA1 mucoid was cleared less readily from the bloodstream and kidneys than the nonmucoid mutants. In an in vitro assay, only strain SA1 mucoid demonstrated antibody-dependent, complement-mediated opsonophagocytosis by human leukocytes. Strains JL24 and JL25 were opsonized for phagocytosis by complement alone. Thus a highly encapsulated strain of S. aureus was more virulent in mice than two related nonmucoid strains. The microencapsulated mutant was not more virulent than the unencapsulated mutant.

Abscess↗
PubMed1987

Chemical characterization and immunogenicity of capsular polysaccharide isolated from mucoid Staphylococcus aureus.

In this study we report the isolation and purification of the capsular polysaccharide elaborated by Staphylococcus aureus SA1 mucoid. The capsule was isolated from bacterial extracts and culture supernatants by a series of ethanol precipitations and enzyme digestions, followed by ion-exchange chromatography. Teichoic acid contamination was eliminated by oxidation with sodium metaperiodate, and the final product eluted in the void volume of a Sephacryl S-300 column. The purified capsular polysaccharide was analyzed by gas-liquid chromatography-mass spectroscopy, 13C and 1H nuclear magnetic resonance, amino acid analysis, immunelectrophoresis, and numerous biochemical assays. The major constituents of the capsule were 2-acetamido-2-deoxy-alpha-galacturonic acid (4-O linked), 2-acetamido-2-deoxy-alpha-fucose (3-O linked), and taurine. The polysaccharide also contained O-acetyl groups which were removed by mild alkaline hydrolysis. Serologically and biochemically, the capsule from strain SA1 mucoid appeared very similar to that produced by strain M. Purified capsular polysaccharide was immunogenic in both rabbits and mice. The optimal immunizing dose in mice was 0.1 microgram of purified capsular polysaccharide administered intraperitoneally. SA1 mucoid resisted opsonophagocytic killing by human leukocytes and complement. However, antibodies raised to the purified capsular polysaccharide neutralized the antiphagocytic effect of the capsule.

Animals↗
PubMed1986

Hymenolepis diminuta: ultrastructural abnormalities in worms from C57 mice.

Ultrastructural studies, including stereological analyses of micrographs, have been made of five-worm primary infections of Hymenolepis diminuta from C57 mice to determine whether the immune destrobilation/rejection process was accompanied by significant changes in the fine structure of the scolex tegument. Destrobilation/rejection of worms occurred from Days 9-12 after infection. For the first 5 days after infection, the scolex tegument showed no detectable differences in ultrastructure compared with that of "control" worms from either Wistar rats or immunosuppressed C57 mice. By Day 6, large lipid deposits were observed in the tegument and associated musculature of worms from untreated C57 mice. Further, worms recovered from Days 6-8 after infection also showed increased activity of the Golgi apparatus, GER, and mitochondria of the tegument, resulting in increased numbers of discoidal secretory bodies. Concomitant with destrobilation/rejection from Day 9 was a drop in the number of secretory bodies, an increase in autophagic activity throughout the tegument, and a blistering of the tegument surface plasma membrane. The possible functional significance of the results is discussed in relation to host immunity.

Animals↗
PubMed1986

Pulmonary infection with opportunist mycobacteria on Merseyside 1974-1983.

During a 10 year period 49 patients were recorded as having pulmonary infection caused by opportunist mycobacteria. Six different species were identified of which M. kansasii (65%) and M. avium-intracellulare (20%) were the most common. Cough and sputum (82%) or haemoptysis (26%) were frequent symptoms on presentation and over two thirds of patients had pre-existing respiratory disease. Chest radiographs showed predominantly apical disease with the right apex (44%) being twice as commonly affected as the left (22%). In 30% the radiograph showed bilateral disease. Clinicians had an individual approach to treatment with no uniform pattern of drug prescribing. A majority of patients (59%) received rifampicin for at least 9 months and those patients with M. kansasii infection responded well with no bacteriological relapses in 20 patients followed for a mean period of 3.9 years. M. avium-intracellulare, M. malmoense and M. xenopi were less responsive to treatment and in four patients receiving chemotherapy death was attributed to mycobacterial infection.

Adult↗
PubMed1982

Immunological memory in mice to adult Hymenolepis diminuta (Cestoda).

Following a primary infection with Hymenolepsis diminuta mice were found to be strongly resistant to a challenge infection over a period of eight months. Challenge was by three cysticercoids administered orally, or by inserting into the duodenum one 7-day-old strobilate worm recovered from a donor mouse. Autopsy of immunized mice 6, 8, and 9 days after challenge showed that challenge worms established but that, with few exceptions, only shunted or destrobilated worms remained. Autopsy of recently immunized mice, at daily intervals following challenge, confirmed that transplanted worms establish as well in immunized as in naive mice, and showed that worms grow normally for the first 48 hr but between 48 to 120 hr most worms destrobilate or are lost. In naive mice, transplanted worms survive and grow, approximately doubling their weight daily for at least 6 days (144 hr). The results provide insight to the problem of developing a useful vaccine and the location of memory cells.

Animals↗
PubMed1981

Raillietina cesticillus: rejection by bursa-deficient chickens.

Chickens in which antibody-forming capacity was abrogated by bursectomy and irradiation developed protective immunity against Raillietina cesticillus as effective as controls which had specific antiworm antibodies in their sera and immunoglobulin positive cells in splenic and intestinal tissues. It is concluded that antibodies are not essential for the rejection of the tapeworm by chickens and may even retard rejection.

Animals↗
PubMed1980

Immunologically mediated rejection of Hymenolepis diminuta by its normal host, the rat.

The adult tapeworm, Hymenolepis diminuta, living in the lumen of the intestine of its normal rat host was shown to sensitize the intestine. The growth of worms in a secondary infection, given 8, 23, or 41 days after the primary infection had been removed by anthelmintic treatment, was 0.5, 3, and 25%, respectively, of that in the control worms. Thus, there is a strong initial resistance to a challenge infection, but this resistance weakens fifty-fold between days 8 and 41 postinfection. Even primary infections of five worms, which would have survived indefinitely if not expelled by anthelmintics, depressed the growth by over 80% of secondary infections given 8 days later. Rats given a light primary infection manifested their sensitivity only by depressing the growth of challenge worms, but in strongly sensitized rats, worm growth was stunted severely (or the worms destrobilated), partial worm expulsion occurred, and the remaining worms migrated to a more anterior position in the intestine. We suggest that the rapid waning of memory indicates that the protective mechanism evoked in the intestine has as its primary biological function the limitation of the number of worms that can accumulate in the intestine in concurrent infections, not the prevention of reinfection after the loss of worms from previous infections.

Animals↗
PubMed1979

Hymenolepis diminuta: the role of the tail in determining the position of the worm in the intestine of the rat.

One-worm infections of Hymenolepis diminuta in rats had their strobila severed surgically, in the neck region, on day 14 of an infection. The scolex and remaining strobila survived but were recovered from a more posterior region of the intestine where small worms are attached during development. The movement to the new region was usually not complete in 24 h, but was complete by 72 h, and probably by 48 h. The operation, involving laparotomy and an incision in the duodenal wall which avoided severing the strobila, had no effect on the position of the worm but did depress the growth of the worm during the ensuing 24 h. It is suggested that (1) the preferred site for H. diminuta is 30-50% down the small intestine, (2) the worm monitors information about its position from all over its strobila and (3) as the worm grows, its position is determined by balancing the input of adverse information from its tail and head ends. The slowness with which surgically shortened worms return to the preferred site may be due either to delay in the worm "realising" it has no tail, or to the location stimuli in the intestine being disturbed for 24 h by the operation.

Animals↗
PubMed1978

Effects of ultrafiltration of diffusion and convection in two newer coils.

Clearances of sodium and B12 were measured with increasing ultrafiltration in two newer coils with new types of membrane support. In all other coil studies previously reported, clearance increases with ultrafiltration did not equal predictions from contributions of convection implying simultaneous decreases in diffusive solute transport with increasing transmembrane pressure. Such deterioration was attributed to masking of membrane surface area by the membrane support, widening of blood channels, and/or channeling of dialysate flow. In these newer coils, clearance increases were very near to predicted enhancement by convection while diffusion was stable in one type and decreased only modestly in the other. Coil volumes assessed with the kerosene technique increased with higher transmembrane pressure similar to findings in previous coils studies. Thus, although coil blood path dimensions are altered with increasing pressure as in older coils, diffusive clearances remain stable and increases in total clearances equal that predicted from convective solute transport. These results suggest that membrane masking is probably the major mechanism for decreases in diffusion with ultrafiltration in other coils and that the problem has been minimized with improved membrane supports.

Chemical Phenomena↗
PubMed1978

Effects of ultrafiltration on solute clearances in cuprophan and cellulose hollow fiber dialyzers: in vitro and clinical studies.

Hollow fiber dialyzers are currently available with either regenerated cellulose or thinner walled cuprophan fibers. In vitro and clinical studies were undertaken to compare a cellulose with a cuprophan dialyzer in regard to (1) UF's, (2) small and large solute clearances at high and low TMP's, (3) SC's, and (4) effects of ultrafiltration on diffusive and convective components of clearances. The results show that per square meter of surface area, the cuprophan dialyzer has higher rates of UF per TMP and greater clearances, particularly of larger solutes; also with cuprophan the SC for vitamin B12 is higher. Thus all findings suggest greater permeability of cuprophan fibers. In both dialyzers, total clearances increased with UF by amounts compatible with predicted increases in convective transport; diffusive transport remained stable as TMP was increased.

Cellulose↗
PubMed1978

Comparison of polymer, glucose, and hydrostatic pressure induced ultrafiltration in a hollow fiber dialyzer: effects on convective solute transport.

Ultrafiltration induced by (1) poly(sodium acrylate), (2) glucose, and (3) hydrostatic pressure was studied in a hollow fiber dialyzer. Poly(sodium acrylate) added to dialysate induced large amounts of ultrafiltration without crossing the dialyzer membrane. Sodium ions of the acrylate polymer were osmotically active but were held in dialysate by the impermeant anionic polymer. The hydrostatic pressure equivalent of osmotic pressure induced primarily by the sodium ions approximated that predicted for a completely impermeant molecule. The apparent (net) sieving coefficients for vitamin B12 observed during polymer and hydrostatic ultrafiltration studies were both significantly higher than that observed during glucose ultrafiltration but did not differ from each other. These studies suggest that sodium salts of polyanions can provide an osmotic driving force to yield large amounts of ultrafiltration in dialysis systems and yet not cross the membrane. The studies also suggest that relatively less efficient convective transport with glucose as compared to hydrostatic pressure is neither a membrane phenomenon nor a characteristic of all osmotic pressure induced ultrafiltration. Relatively low effective solute sieving appears to be associated with osmotic induced ultrafiltration with a permeant solute. Under such conditions it is proposed that molecular interaction within the membrane impairs convective transport.

Acrylic Resins↗