Biomedical subjects

A L Scott

Publications and source records attributed to A L Scott.

At least 55 records · Page 3Linked to original sources

PubMed1995

Expression of ICAM-1 in airway epithelium after acute ozone exposure in the mouse.

We investigated the time course and regional distribution of the expression of intercellular adhesion molecule-1 (ICAM-1) on airway epithelial cells and the polymorphonuclear leukocyte (PMN) inflammatory response in the lung after acute exposure to ozone (O3). C57BL/6J mice were exposed to air or 2 ppm O3 for 3 h and killed immediately or 3, 6, 9, or 21 h after exposure. Expression of ICAM-1 was examined by immunohistochemical staining of frozen sections. PMN influx was evaluated by lavage and by histochemical staining of myeloperoxidase (MPO) and measurement of tissue MPO activity. ICAM-1 expression exhibited regional selectivity and temporal patterns that were unique to each region. Upregulation of ICAM-1 expression on the epithelial cells in the trachea, and to a lesser extent in the lobar and segmental bronchi, was observed 3-9 h after exposure and remained present at 21 h. Enhanced ICAM-1 expression in bronchioles and terminal bronchiole/alveolar duct regions was evident earlier (immediately to 3 h after exposure) but returned to baseline levels by 21 and 9 h, respectively. Maximal ICAM-1 expression and PMN influx in the lung parenchyma were concurrently observed at 3 h, followed by transepithelial migration of PMNs to the airway lumen. These results demonstrate regional variations in airway inflammatory activity and are supportive of the notion that upregulation of ICAM-1 on the airway epithelium may play a role in local regulation of PMN influx to the airways after acute O3 exposure.

Administration, Inhalation↗

PubMed1994

Effect of vitamin A supplementation on immunoglobulin G subclass responses to tetanus toxoid in children.

Previously, we demonstrated that administering vitamin A supplements to children resulted in a significant increase in the immunoglobulin G (IgG) response generated against a vaccine dose of tetanus toxoid (TT) (R. D. Semba et al., J. Nutr. 122:101-107, 1991). However, from these analyses we could not determine whether there was an increase in levels of IgG of the subclass presumed to be important for protection against challenge by the toxin or whether there was simply a general increase in the levels of all the IgG subclasses expressing anti-TT activity. The goal of this study was to determine the profile of the anti-TT IgG subclasses in children receiving vitamin A supplementation or a placebo in order to assess the potential utility of the enhanced anti-TT response. In a randomized, double-masked, placebo-controlled clinical trial, the levels of the different anti-TT IgG subclasses were measured in 139 Indonesian preschool children (3 to 6 years of age) 2 weeks before and 3 weeks after immunization. Baseline anti-TT levels and immunization histories were used to separate those children who were responding to TT for the first time from those who responded in a secondary fashion because of previous exposure to TT. Children who were given vitamin A prior to immunization had significant increases in IgG1 levels regardless of whether they were undergoing primary or memory reactions. In the group of individuals who underwent a secondary response to TT, vitamin A supplementation was also associated with a modest but significant change in the levels of anti-TT IgG3. There were only minor changes in the levels of anti-TT IgG2 and IgG4. Since IgG1 is the subclass associated with a protective response to TT immunization, these results suggest that vitamin A supplementation may be a safe and effective intervention to enhance the relevant humoral response to TT and other vaccine antigens.

Adjuvants, Immunologic↗

PubMed1993

Abnormal T-cell subset proportions in vitamin-A-deficient children.

Although vitamin A deficiency in children seems to increase susceptibility to infection and community trials have shown that vitamin A supplementation can reduce childhood mortality from infectious diseases, the underlying biological mechanisms are largely unknown. We conducted a randomised, double-masked, placebo-controlled clinical trial among children in West Java, Indonesia, to determine whether vitamin A deficiency is associated with abnormalities in T-cell subsets and whether vitamin A supplementation affects T-cell subsets. We studied 55 children aged 3-6 years--30 with xerophthalmia and 25 without. Acutely malnourished children (< 80% of reference weight-for-height) were excluded. CD4/CD8 ratios and the proportions of circulating CD4 naive, CD4 memory, CD8, CD45RA, and CD8, CD45RO T-cell subsets were measured. Children with xerophthalmia had lower CD4/CD8 ratios (p < 0.08), lower proportions of CD4 naive T cells (p < 0.03), and higher proportions of CD8, CD45RO T cells (p < 0.04) than those without xerophthalmia. 26 children were given vitamin A supplementation (60 mg retinol equivalent) and 29 received placebo. 5 weeks later the vitamin A group had higher CD4/CD8 ratios (p < 0.001), higher proportions of CD4 naive T cells (p < 0.01), and lower proportions of CD8, CD45RO T cells (p < 0.05) than the placebo group. Vitamin-A-deficient children have underlying immune abnormalities in T-cell subsets and these abnormalities are reversible with vitamin A supplementation.

Analysis of Variance↗

PubMed1992

Surface-associated antigens of Brugia malayi L2 and L3 parasites during vector-stage development.

Surface and metabolic labeling procedures were used to characterize the composition and the time of expression of Brugia malayi L2 and L3 surface-associated molecules as the larvae develop within the mosquito vector. Larvae were harvested from mosquito tissues at 5 (early L2), 8 (late L2) and 11 (L3) days post-infection and labeled with 125I-Iodo-Gen. The results of one-dimensional analysis showed that there is a progressive increase in the complexity of peptides associated with the surface of developing larvae, culminating in the expression of 7 major labeled components on L3s. Both L2 and L3 parasites have surface-associated components of 42, 35, 33, 19 and 17 kDa. Between days 8 and 11 of development in the insect vector, Brugia malayi undergoes the L2 to L3 molt and acquires additional major immunogenic peptides of 40 and 22 kDa. Two-dimensional analyses of extracts from 125I-labeled L2s and L3s revealed that the major 35-, 33-, 19- and 17-kDa molecules are part of a peptide complex that forms a 'ladder' between 17 and 150 kDa. To gain information on the times during which the major surface-associated molecules are produced by the parasite, larvae were labeled with [35S]methionine either in situ as they developed within the mosquito or during culture after exiting the vector. For in situ labeling, [35S]methionine was introduced into the hemolymph of infected mosquitoes by micro-injection at days 2, 5 and 8 post-infection and the larvae were allowed to develop for an additional 3 days. The results of 1- and 2-dimensional analyses of [35S]methionine-labeled extracts from vector-stage or post-vector-stage larvae indicate that the molecules associated with the surface of B. malayi L3s are synthesized between day 5 and day 11 of development in the insect host. Immediately after the larvae exit the vector, the synthesis of the 40 and 22-kDa peptides is drastically reduced or terminated.

Animals↗

PubMed1992

Depressed immune response to tetanus in children with vitamin A deficiency.

A randomized, double-masked, placebo-controlled clinical trial was conducted with 236 preschool children, age 3-6 y, in Indonesia to assess immune status in mild vitamin A deficiency. The immune response to tetanus immunization was used as a measure of immune competence. Clinically normal children (n = 118) and children with mild xerophthalmia (n = 118) were randomly assigned to receive oral vitamin A (60,000 micrograms retinol equivalent) or placebo treatment for a total of four study groups. Two weeks after treatment, children were immunized with diphtheria-pertussis-tetanus vaccine. The immunoglobulin G (IgG) responses to tetanus at baseline and 3 wk following immunization were measured by ELISA. After adjusting for previous tetanus immunization, clinically normal and xerophthalmic children receiving vitamin A had a significantly greater IgG response to tetanus than clinically normal and xerophthalmic children receiving placebo (P less than 0.05). These results suggest that children with mild vitamin A deficiency have a relative immune depression compared with children who have been supplemented to normal vitamin A levels.

Antibody Formation↗

PubMed1992

Anti-Onchocerca volvulus immunoglobulin subclass response in children from Sierra Leone.

The humoral immune responses of a paediatric population of 224 children, 2-15 years of age, residing in an onchocerciasis endemic area of Sierra Leone, were studied to gain information that may be useful in the serodiagnosis of prepatent and low-level infections with Onchocerca volvulus. The antigen-specific levels of IgG and IgG4 were determined and evaluated in the context of the parasitological status of the child. Over 87% of the microfilaria-negative children had significant levels of antigen-specific antibodies against O. volvulus, suggesting that a large percentage of this paediatric population were harbouring prepatent or low-level infections. Over 60% of the children in the group that had detectable numbers of microfilariae in skin snips had elevated levels of O. volvulus-specific IgG4. In contrast, only 23% of the microfilaria-negative children had detectable IgG4 responses. The results presented here indicate that measures of O. volvulus-specific IgG4 may be a useful indicator of the parasitological status of children during the very early stages of onchocerciasis.

Adolescent↗

PubMed1992

Cardiac angiotensin receptors: effects of selective angiotensin II receptor antagonists, DUP 753 and PD 121981, in rabbit heart.

Angiotensin II (AII) elicits a positive inotropic response in cardiac muscle preparations from several species including humans. The purpose of this study was to characterize the AII binding sites and inotropic responses in rabbit ventricle using the selective AII receptor antagonists/ligands, DuP 753 (AT1) and PD 121981 (AT2). Biphasic displacement of specific 125I-Sar1,Ile8-AII binding was observed with both DuP 753 and PD 121981, suggesting the presence of two AII binding sites. The high affinity site for DuP 753 (29 nM) was a low affinity site for PD 121981 (91 microM), and the high affinity site for PD 121981 (78 nM) was a low affinity site for DuP 753 (81 microM). Of the specific AII binding, 70% was DuP 753 (AT1)-sensitive sites. Positive inotropic responses to AII in isolated papillary muscles from rabbit heart were antagonized competitively by both DuP 753 and PD 121981. The potencies of DuP 753 (pA2 = 7.99) and PD 121981 (pA2 = 4.28) to antagonize AII inotropic responses were similar to their potencies to displace 125I-Sar1,Ile8-AII from AT1 sites. There was no apparent functional consequence of AII interaction with AT2 site. Inotropic responses to isoproterenol were unaffected by DuP 753 and PD 121981. Therefore, there are two binding sites for AII in rabbit ventricle; however, only one site, AT1, participates in the inotropic response to AII. The roles of these receptor subtypes in other cardiac responses to AII have yet to be determined. Also, DuP 753 and PD 121981 are useful tools to study these two AII binding sites in cardiac preparations.

Angiotensin II↗

PubMed1991

Leukotriene D4 increases both the force of contraction and polyphosphoinositide formation in guinea-pig papillary muscle.

Leukotriene D4 (LTD4) increased the force of contraction in guinea-pig papillary muscle. A rapid (less than 1 min), transient (less than 5 min) response to LTD4 (1 microM) reached 19.3 +/- 5.4% of isoproterenol maximum. A single exposure to LTD4 resulted in complete and homologous desensitization which was not influenced by indomethacin. LTD4 (0.1-3.0 microM) increased total inositol phosphates released from [3H]inositol-labeled tissue. ICI 198,615, a selective LT receptor antagonist, blocked both the increase in force of contraction and the increase in inositol phosphates by LTD4, but had no effect on the inotropic response to isoproterenol. These data support the existence of specific functional LTD4 receptors in myocardial tissue of guinea-pigs.

Animals↗

PubMed1991

Interleukin-1 is a mucus secretagogue.

Explant cultures of mouse duodenum were used to show that interleukin-1 (IL-1) causes release of mucus from epithelial goblet cells. Our experiments made use of a newly described enzyme-linked lectin assay (ELLA) which employs enzyme-conjugated soybean agglutinin to detect mucus glycoproteins secreted from explant cultures of mouse duodenum. Supernatants from cultures of lipopolysaccharide-stimulated peritoneal macrophages as well as partially purified rabbit alveolar macrophage-derived IL-1 and human rIL-1 beta all induced mucus release in a rapid and dose-dependent fashion. This observation may be important for investigating a link between the immune response and mucus hypersecretion from inflamed intestinal mucosa.

Animals↗

PubMed1991

Experimental ocular onchocerciasis in cynomolgus monkeys. IV. Chorioretinitis elicited by Onchocerca volvulus microfilariae.

Onchocerciasis is a major cause of blindness worldwide, and much of the blindness is caused by onchocercal chorioretinitis. In an experimental animal model for ocular onchocerciasis, intravitreal injections of 10,000 live Onchocerca volvulus microfilariae isolated from infected humans into the eyes of cynomolgus monkeys (Macaca fascicularis) resulted in patchy, progressive loss of retinal pigment with pigment clumping. Areas of pigment loss were less extensive in animals that had been sensitized with microfilariae. Intravitreal injections of dead O. volvulus microfilariae resulted in mild vitritis with relatively less clinical change noted in the retina and choroid. Histopathologic examination revealed thinning and loss of outer retinal layers with pigment migration into the retina, and inflammation was more pronounced in eyes that received live microfilariae. Clinical changes appeared in eyes receiving live microfilariae before the development of significant antibody or cell-mediated immune responses. O. volvulus microfilariae appear to be more suitable than O. lienalis microfilariae in producing lesions which resemble human onchocerciasis in the primate model.

Animals↗

PubMed1990

Surface-associated antigens of second, third and fourth stage larvae of Dirofilaria immitis.

The surface-associated molecules of the second (L2), third (L3) and fourth (L4) larval stages of Dirofilaria immitis were characterized employing radiolabeling techniques and SDS-PAGE analysis. Major labeled components of 35 kDa and 6 kDa were present in extracts from IODO-GEN-labeled L2 and L3 parasites. The results of lactoperoxidase-catalyzed reactions also demonstrated that L2 and L3 stages of D. immitis have a similar repertoire of surface-associated antigens. However, in contrast to the results obtained with IODO-GEN, lactoperoxidase reactions labeled components with apparent molecular weights of 66, 48, 25, 16.5 and 12 kDa. The similarities in the molecular weights of the L2 and L3 surface-associated components and the results of immunoprecipitation experiments which demonstrated that antibodies produced against the 35 kDa molecule from D. immitis L3s also recognize the 35 kDa component from L2 parasites suggest that synthesis of the molecules found at the surface of mature infective larvae begins as early as day 6 of development in the mosquito, D. immitis L4s emerged from the molting process with a repertoire of surface-associated antigens distinct from those found on L2s and L3s. IODO-GEN labeling of D. immitis L4s showed major surface-associated molecules with apparent molecular weights of 57, 40, 25, 12 and 10 kDa when analyzed under non-reducing conditions. In addition to molecules of 57, 40, 25, 12 and 10 kDa, extracts of D. immitis L4s labeled with lactoperoxidase contained additional major bands at 45, 43 and 8 kDa. Metabolic labeling experiments demonstrated a shift in the amount and complexity of the excretory/secretory products released by D. immitis during L3 to L4 development.

Animals↗

PubMed1990

Onchocerca volvulus: molecular cloning, primary structure, and expression of a microfilarial surface-associated antigen.

Although the filarial nematode parasite Onchocerca volvulus is an important human pathogen in large areas of Africa and Latin America, little is known of the molecular interactions that govern the clinical status of patients with this chronic, debilitating disease. As a step toward defining the parasite molecules important to the immunobiology of host-parasite interactions, we have identified and cloned a major surface-associated antigen expressed by O. volvulus microfilariae. Radiolabeling experiments demonstrated that O. volvulus microfilariae have a limited repertoire of peptides at the surface. Prominent among these labeled peptides is an 18-kDa component. Immunological cross-reactivity between a surface-associated component of Dirofilaria immitis microfilariae and the 18-kDa surface-associated molecule from O. volvulus was exploited in a strategy to clone this potentially important O. volvulus microfilarial antigen. The cross-reacting antibodies were used to immunoscreen O. volvulus cDNA expression libraries. One clone, M2f.e, contained an open reading frame of 495 bp encoding an 18.1-kDa protein (OVMS18). Antibodies produced against the expression product of M2f.e recognized an 18-kDa component in extracts of O. volvulus microfilariae and bound to the surface of intact O. volvulus and D. immitis microfilariae. Southern blot analyses showed that M2f.e-like sequences are present in the genomic DNA of a number of filarial nematode species, but not in DNAs from nonfilarial nematode species.

Amino Acid Sequence↗

PubMed1990

Negative lusitropic effect of DPI 201-106 and E4031. Possible role of prolonging action potential duration.

In open-chest anesthetized dogs, the time constant of isovolumic left ventricular pressure decay increased following the intravenous administration of either E4031, a class III antiarrhythmic agent which acts by K+ channel blockade, or DPI 201-106 (DPI), a cardiotonic agent which acts by delaying Na+ channel inactivation. In addition to prolonging cardiac refractoriness, both E4031 and DPI increased left ventricular +dP/dt but without significantly altering -dP/dt. Consequently, the value of the ratio (+dP/dt)/(-dP/dt) increased. There were no significant changes in heart rate, mean arterial pressure, or left ventricular end diastolic pressure. Since both E4031 and DPI prolonged the action potential duration (APD) and the refractory period, and slowed relaxation in vivo, the possibility of a causal link between these effects was further investigated under in vitro conditions. In isometrically contracting rabbit papillary muscles, E4031 and DPI increased peak developed tension (DT) and its maximal rate of rise (+T). Since the maximal rate of fall of DT (-T) did not increase by the same factor that +T increased, the value of the ratio +T/-T increased. Time to half relaxation increased, whereas time to peak tension was not significantly changed by either E4031 or DPI. These negative lusitropic effects produced by E4031 or DPI were not observed when equivalent increases in contractility were produced by increasing the extracellular Ca2+ concentration. The effective refractory period measured in the papillary muscles increased following superfusion with either of the two drugs, consistent with their known ability to increase APD. A causal link between the prolongation of APD and the negative lusitropic effects of E4031 and DPI is postulated as the possible mechanism.

Action Potentials↗

PubMed1989

Major sperm protein genes from Onchocerca volvulus.

Nematode spermatozoa, unlike their mammalian counterparts, are nonflagellated crawling cells. The pseudopod of these cells contains the major sperm protein (MSP) which comprises more than 15% of the protein in the sperm. MSP is presumed to function as a cytoskeletal element involved in motility. An Ascaris MSP cDNA sequence was used as a probe to identify and isolate Onchocerca volvulus MSP clones from a lambda gt11 genomic library. Two clones, OVGS-1 (765 bp) and OVGS-2 (1765 bp), were characterized by restriction endonuclease mapping and sequence analysis. Both genomic clones contain MSP protein coding regions of 99 and 282 bp separated by an intervening sequence of 153 bp. The genes OVGS-1 and OVGS-2 are 95% similar in nucleotide sequence in the protein coding regions, but only 79% similar in their intron sequences. A number of potential regulatory sequences in the flanking regions and at the exon/intron junctions of the O. volvulus MSP genes are in good agreement with consensus sequences in other eukaryotic cells. The nucleotide sequence of the O. volvulus MSP genes were over 80% similar to the Ascaris MSP cDNA sequence and 79% similar to the Caenorhabditis MSP-3 cDNA. The predicted amino acid sequence of the O. volvulus MSPs were 96% similar to each other, 90-91% similar to Ascaris MSP and 81-82% similar to Caenorhabditis MSP-3. These results offer evidence that the MSP sequences have been highly conserved throughout nematode evolution but are variable in their genomic organization and the presence of introns.

Amino Acid Sequence↗

PubMed1989

Antigen shedding from the surface of the infective stage larvae of Dirofilaria immitis.

A qualitative and quantitative analysis was made of the release of surface-associated molecules from developing Dirofilaria immitis infective-stage larvae (L3). D. immitis L3s were labelled with 125I using an Iodogen catalysed reaction and either maintained in culture or placed in chambers that were implanted into Lewis rats. The larvae released 10-20% of the labelled material each day during the first 4 days of in vitro and in vivo development. The loss of surface-labelled peptides from developing larvae corresponded with an increase in the amount of trichloroacetic acid-precipitable radioactivity found in the culture medium. SDS-PAGE analysis of the labelled material showed that the same 35 and 6 kDa components found in larval extracts were shed into culture medium by the developing parasites. Metabolic labelling studies and experiments in which larvae were labelled after different times in culture indicated that, once released, the surface-associated molecules were not replaced, and that this net loss of surface peptides resulted in a reduction in the antigenic potential of the cuticular surface. Antibodies from both immunized rabbits and naturally infected dogs immunoprecipitated the 35 kDa component. In contrast, the 6 kDa molecule was not recognized by the antibodies in any of the sera tested. Shedding of surface peptides and reducing surface antigenicity may represent mechanisms by which D. immitis infective-stage larvae evade immune attack.

Animals↗

PubMed1989

Major sperm protein and actin genes in free-living and parasitic nematodes.

The DNA from a number of free-living and parasitic nematode species was examined to determine the genomic number and distribution of DNA sequences encoding two evolutionarily conserved proteins; the major sperm protein (MSP) and nematode actin. Ascaris and Caenorhabditis MSP cDNA sequences and Ascaris genomic actin sequences were used to probe Southern blots of Eco RI and Hin d III digested nematode DNA. The number of MSP genes varied widely between the 1 MSP gene in Ascaris and the 60 MSP genes in Caenorhabditis. Filarial nematodes appeared to have 1-4 MSP genes while the plant and insect parasitic species showed from 5-12 MSP-hybridizing restriction fragments. Mammalian intestinal parasites showed between 1 and 13 bands hybridizing with the MSP probes. Blots probed to estimate the number of actin genes showed that, with the exception of Ascaris which contains more than 20 germ line sequences that encode actin, all of the nematodes tested had between 3 and 9 bands that hybridized to the Ascaris genomic actin probe. The possible use of highly conserved sequences such as MSP and actin to differentiate between nematode species in diagnostic and taxonomic studies is discussed.

Actins↗

PubMed1989

Enhanced release of atrial natriuretic factor by endothelin in atria from hypertensive rats.

Intravenous (bolus) administration of endothelin results in a transient fall in blood pressure that is accentuated in spontaneously hypertensive rats (SHR) compared with Wistar-Kyoto normotensive rats (WKY). In attempting to discern possible mechanisms underlying this depressor response, we examined the ability of endothelin to release atrial natriuretic factor (ANF) from isolated, spontaneously contracting atria from SHR and WKY. Isolated right atria were suspended under 3.0 g of resting force in tissue baths with the amount of immunoreactive ANF (irANF) released after exposure to endothelin assessed by radioimmunoassay. Endothelin (10(-8) and 10(-7) M) caused a concentration-dependent increase (1.5-4.5-fold) in the release of irANF, which was significantly greater in atria of SHR compared with WKY. The greater release of irANF in atria of SHR versus WKY was not related to tissue weight or changes in contractile rate or force induced by endothelin. Therefore, endothelin appears to cause a direct release of irANF from rat right atria in vitro. As found for the depressor response in vivo, endothelin is more efficacious in the hypertensive compared with the normotensive atrial preparation. Release of ANF may be important in the hypotensive response to endothelin in vivo.

Animals↗

PubMed1989

Dirofilaria immitis: studies on anti-microfilarial immunity in Lewis rats.

The effect of vaccination on rates of microfilarial clearance using Dirofilaria immitis in male Lewis rats was examined. Animals were immunized with whole, dead microfilariae or a PBS extract of microfilariae in Freund's adjuvant. The immunized animals, as well as untreated and adjuvant controls, were challenged intravenously with 4 x 10(5) viable microfilariae (mf). The duration of microfilaremia was 15.5 days in rats vaccinated with whole mf, 17.7 days in those vaccinated with a PBS extract, 36.3 days for those vaccinated with adjuvant alone, and greater than 70 days for the untreated group. Analysis of the anti-microfilarial IgG response by ELISA and Western blots demonstrated that immunization induced significant amounts of antibody against high molecular weight peptides, particularly a peptide located at 105 kDa. Antibody levels in both groups of immunized animals continued to rise following challenge, reaching peak levels of 78-80 micrograms/ml on the day of microfilarial clearance. Decreasing microfilaremia following challenge was associated with an enhanced recognition of low molecular weight peptides.

Animals↗