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Biomedical subjects

C A Dangler

Publications and source records attributed to C A Dangler.

At least 37 records · Page 2Linked to original sources

Helicobacter bilis/Helicobacter rodentium co-infection associated with diarrhea in a colony of scid mice.

An outbreak of diarrhea spanning 3 months occurred in a breeding colony of scid/Trp53 knockout mice. Approximately a third of the 150 mice were clinically affected, with signs ranging from mucoid or watery diarrhea to severe hemorrhagic diarrhea with mortality. Helicobacter bilis and the newly recognized urease-negative organism H. rodentium were isolated from microaerobic culture of feces or cecal specimens from affected mice. Dual infection with H. bilis and H. rodentium were confirmed by culture and polymerase chain reaction (PCR) in several animals. Both Helicobacter species rapidly colonized immunocompetent sentinel mice exposed to bedding from cages containing affected mice, but the sentinel remained asymptomatic. Mice with diarrhea had multifocal to segmental proliferative typhlitis, colitis, and proctitis. Several affected mice had multifocal mucosal necrosis with a few focal ulcers in the cecum, colon, and rectum. Mice with diarrhea were treated with antibiotic food wafers (1.5 mg of amoxicillin, 0.69 mg of metronidazole, and 0.185 mg of bismuth/mouse per day) previously shown to eradicate H. hepaticus in immunocompetent mice. Antibiotic treatment resulted in resolution of diarrhea, but not eradication of H. bilis and H. rodentium; mice continued to have positive PCR results after a 2-week treatment regimen, and clinical signs of diarrhea returned in some mice when treatment was suspended. To the authors' knowledge, this is the first report of natural infection with either H. bilis and/or H. rodentium causing acute diarrheal disease and suggests that H. bilis and/or H. rodentium can be an important pathogen for scid mice.

Animals↗

Mice carrying a truncated Apc gene have diminished gastric epithelial proliferation, gastric inflammation, and humoral immunity in response to Helicobacter felis infection.

Helicobacter pylori infection and adenomatous polyposis coli (Apc) gene mutations have been linked to gastric cancer in humans, but possible synergistic interaction(s) between these risk factors have not been examined. Fourteen C57BL/6 wild-type and 14 Apc1638 heterozygous mice were inoculated with Helicobacter felis at 6 weeks of age and compared at various time points with a similar number of uninfected control mice of the same genotype. Both infected and uninfected Apc1638 mice had a limited incidence of atypical proliferation foci in the mucosa of the antrum and pyloric junction at 4.5 and 6 months of age, whereas polyps of the antrum and pylorus were present in all mice, regardless of infection status, at 7.5 months. In contrast, no altered gastric mucosal foci were observed in control or infected C57BL/6 mice at any time point. Interestingly, the infected Apc1638 mice had less epithelial proliferation and inflammation in the body of the stomach, lower anti-H. felis serum IgG antibody responses (although both the wild-type and Apc mutant mice had a Th1-like immune response, based on a predominantly IgG2a immunoglobulin response), and higher bacteria and urease scores than did infected wild-type C57BL/6 mice. In conclusion, the Apc1638 truncating mutation leads to gastric dysplasia and polyposis of the antrum and pyloric junction, but H. felis infection of the Apc mutant mouse does not lead to an increased rate of gastric neoplasia. In addition, our data suggest this Apc mutation may actually lead to decreased immune, inflammatory, and gastric hyperplastic responses to Helicobacter infection, suggesting the possibility of a novel role for this tumor suppressor gene in the immune and local tissue responses to gastric bacterial infection.

Adenomatous Polyposis Coli Protein↗

Exogenous, basal, and flow-induced nitric oxide production and endothelial cell proliferation.

The role of nitric oxide (NO) from endogenous and exogenous sources in regulating large vessel and microvascular endothelial cell proliferation was investigated. Exogenous NO liberated from five different chemical donors inhibited bovine aortic, bovine retinal microvascular, and human umbilical vein endothelial cell proliferation in a dose-dependent manner as determined by 3H-thymidine incorporation. The potency of the donors varied as a function of the donors' half-lives. Donors with half-lives greater than 30 min were more effective than donors with significantly shorter half-lives. Coincubation of endothelial cells with 0.4 mM deoxyadenosine and 0.4 mM deoxyguanosine reduced the percentage of inhibition due to an NO donor. These data are consistent with a ribonucleotide reductase-dependent mechanism of inhibition. Inhibition of basal NO production with four different inhibitors of nitric oxide synthase (NOS) did not modify proliferation. Laminar flow with a wall shear stress of 22 dyn/cm2 inhibited the proliferation of subconfluent bovine aortic endothelial cells. The addition of a NOS inhibitor did not abrogate the flow-induced inhibition of proliferation, suggesting that flow-stimulated release of NO from endothelial cells did not account for flow-induced inhibition of proliferation. Taken together, these data suggest that relatively large concentrations of exogenous NO inhibit endothelial cell proliferation, while endogenous levels of NO are inadequate to inhibit proliferation.

Animals↗

Helicobacter bilis-induced inflammatory bowel disease in scid mice with defined flora.

Helicobacter bilis has been isolated from aged inbred mice with multifocal chronic hepatitis and from scid mice with diarrhea, proliferative typhlitis, and colitis. To determine the pathogenic potential of H. bilis, we inoculated 4-week-old female Tac:Icr:Ha(ICR)-scidfDF mice by intraperitoneal injection of approximately 10(8) CFU of H. bilis in phosphate-buffered saline (PBS) (n = 15) or PBS alone (n = 10) and necropsied them at 7 weeks postinfection. Sham-inoculated mice had no significant gross or histopathological findings. In contrast, all 15 experimentally inoculated mice (confirmed to be H. bilis-colonized by culture and PCR of cecal contents) exhibited varying degrees of inflammatory bowel disease (IBD). Proliferative typhlocolitis was characterized by focal to segmental areas of crypt hyperplasia and a predominantly histiocytic inflammatory cell infiltrate. Labeling indices for 5-bromo-2'-deoxyuridine incorporation were increased approximately 2.5-fold in the ceca and colons of H. bilis-inoculated mice. This is the first study to demonstrate experimentally that infection with H. bilis causes IBD in scid mice with defined flora. This result both confirms a pathogenic role for H. bilis in mice and provides a new model relating a specific microbial agent and IBD.

Animals↗

Inflammatory bowel disease: an immunity-mediated condition triggered by bacterial infection with Helicobacter hepaticus.

Inflammatory bowel disease (IBD) is thought to result from either an abnormal immunological response to enteric flora or a normal immunological response to a specific pathogen. No study to date has combined both factors. The present studies were carried out with an immunologically manipulated mouse model of IBD. Mice homozygous for the severe combined immunodeficiency (scid) mutation develop IBD with adoptive transfer of CD4+ T cells expressing high levels of CD45RB (CD45RB(high) CD4+ T cells). These mice do not develop IBD in germfree conditions, implicating undefined intestinal flora in the pathogenesis of lesions. In controlled duplicate studies, the influence of a single murine pathogen, Helicobacter hepaticus, in combination with the abnormal immunological response on the development of IBD was assessed. The combination of H. hepaticus infection and CD45RB(high) CD4+ T-cell reconstitution resulted in severe disease expression similar to that observed in human IBD. This study demonstrates that IBD develops in mice as a consequence of an abnormal immune response in the presence of a single murine pathogen, H. hepaticus. The interaction of host immunity and a single pathogen in this murine system provides a novel model of human IBD, an immunity-mediated condition triggered by bacterial infection.

Animals↗

Necrotic oophoritis in gilts associated with experimental inoculation of a viral gene-deletion mutant pseudorabies vaccine.

Previous work on the reproductive effects of various herpesviruses has demonstrated adverse effects on reproductive function in several host species. Although herpesviral vaccines are used in several species to ameliorate the clinical effects of infection, pathogenicity for reproductive tissue, associated with diminished reproductive efficiency, has been reported to be retained in a live-attenuated vaccine strain of the herpesvirus, bovine herpesvirus-1. The objective of this study was to determine if a gene-deletion mutant, thymidine kinase negative, pseudorabies virus retained acute pathogenicity for the reproductive tract of swine following intravenous inoculation during estrus. Estrous cycles of nulliparous gilts were synchronized by administration of a gonadotropin and daily exposure to a boar. During estrus, six gilts were inoculated intravenously with twice the recommended intramuscular dose of a commercially available viral gene-deletion mutant pseudorabies vaccine. Six control gilts in estrus were sham inoculated intravenously with vaccine diluent during estrus. All animals were euthanatized 10 days postinoculation, and the ovaries and uterus were collected for histopathology following gross examination. All reproductive tracts were grossly normal. Histologically, four of six treated gilts had a mild to moderate, multifocal, necrotizing oophoritis, with the lesions limited to corpora lutea and the adjacent stroma. Ovaries of control gilts exhibited to necrotizing lesions. Both control and pseudorabies vaccine-inoculated gilts had occasional minimal focal mononuclear infiltrates in the ovaries. These data show that live attenuated viral gene-deletion mutant pseudorabies vaccine administered to swine during estrus can result in acute pathogenicity in ovarian corpora lutea. No endocrinologic data is available in these pigs, so the impact on pregnancy maintenance is unknown.

Animals↗

Helicobacter mustelae-associated gastric adenocarcinoma in ferrets (Mustela putorius furo).

Helicobacter pylori in humans is associated with active, chronic gastritis, peptic ulcer disease, and most recently has been linked epidemiologically to gastric adenocarcinoma. A related organism, Helicobacter mustelae, naturally infects ferrets and also causes a persistent gastritis, a precancerous lesion, and focal glandular atrophy of the proximal antrum. In this report, we document the clinical presentation and histopathologic confirmation of H. mustelae-associated gastric adenocarcinoma in two middle-aged male ferrets. The ferret appears to be well suited to study the pathogenesis of naturally occurring Helicobacter sp.-induced gastric adenocarcinoma.

Adenocarcinoma↗

Processing and proliferative effects of human progastrin in transgenic mice.

Incompletely processed gastrins have been postulated to play a role in growth of the gastrointestinal tract, but few studies have examined the effects of progastrin on mucosal proliferation in vivo. Human gastrin gene expression and progastrin processing were therefore studied in transgenic mice containing a human gastrin (hGAS) minigene, and compared to processing in mice bearing an insulin gastrin (INS-GAS) transgene that overexpresses amidated gastrin. Progastrin processing was studied using region-specific antisera and radioimmunoassays, biosynthetic labeling, immunoprecipitation, and HPLC. Proliferative effects due to overexpression of processed and unprocessed gastrin in INS-GAS and hGAS mice, respectively, were determined using routine histology and BrdU incorporation. The pancreatic islets of INS-GAS mice were able to produce carboxyamidated G-17, resulting in a twofold elevation of serum amidated gastrin, marked thickening of the oxyntic mucosa, and an increased BrdU labeling index (LI) of the gastric body. In contrast, livers of adult hGAS mice expressed abundant human gastrin mRNA and human progastrin but were unable to process this peptide to the mature amidated form, resulting in markedly elevated serum progastrin levels and normal amidated gastrin levels. Nevertheless, there was a marked increase in the BrdU labeling index of the colon in hGAS mice (LI 7.46+/-1.90%), as well as in INS-GAS mice (LI 6.16+/-1.17%), compared to age-matched, wild type control mice (LI 4.01+/-0.98%, P < 0.05). These studies suggest that incompletely processed gastrin precursors may contribute to colonic mucosal proliferation in vivo.

Animals↗

Garlic and associated allyl sulfur components inhibit N-methyl-N-nitrosourea induced rat mammary carcinogenesis.

Our previous studies demonstrated that dietary garlic powder supplementation inhibits N-nitrosamine induced DNA alkylation in liver and mammary tissue. The present studies compared the impact of dietary supplementation with garlic powder or two garlic constituents, water-soluble S-allyl cysteine (SAC) and oil-soluble diallyl disulfide (DADS), on the incidence of mammary tumorigenesis induced by N-methyl-N-nitrosourea (MNU). Female Sprague-Dawley rats were fed semi-purified casein based diets with or without supplements of garlic powder(20g/kg), SAC (57 micromol/kg) or DADS (57 micromol/kg) for 2 weeks prior to treatment with MNU (15 mg/kg body wt). Garlic powder, SAC and DADS supplementation significantly delayed the onset of mammary tumors compared to rats receiving the unsupplemented diet. Tumor incidence 23 weeks after MNU treatment was reduced by 76, 41 and 53% in rats fed garlic, SAC and DADS, respectively, compared to controls (P<0.05). Total tumor number was reduced 81, 35 and 65% by these supplements, respectively (P<0.05). In a separate study the quantity of mammary DNA alkylation occurring 3 h after MNU treatment was reduced in rats fed garlic, SAC or DADS (P<0.05). Specifically, O(6)-methylguanine adducts were reduced by 27, 18 and 23% in rats fed supplemental garlic, SAC and DADS, respectively, compared to controls. N(7)-Methylguanine adducts decreased by 48, 22 and 21% respectively, compared to rats fed the control diet. These studies demonstrate that garlic and associated allyl sulfur components, SAC and DADS, are effective inhibitors of MNU-induced mammary carcinogenesis.

Allyl Compounds↗

Assessing dissimilarity of genes by comparing their RNAse A mismatch cleavage patterns.

We propose a simple algorithm for estimating the number of nucleotide differences between a pair of RNA or DNA sequences through comparison of their RNAse A mismatch cleavage patterns. In the RNAse A mismatch cleavage technique two or more sample sequences are hybridized to the same RNA probe, the hybrids are partially digested with RNAse A, and the digestion products are compared on an electrophoretic gel. Here we provide an algorithm for converting the numbers of unique and matching electrophoretic bands into an estimate of the number of nucleotide differences between the sequences. Computer simulation indicates that the proposed method yields a robust estimate of the genetic distance despite stochastic errors and occasional violation of certain assumptions. Our study suggests that the method performs best when the distance between the sequences is < 15 differences. When the sequences under analysis are likely to have larger distances, we advise to substitute one long riboprobe with a set of shorter nonoverlapping probes. The new algorithm is applied to infer the proximity of several strains of pseudorabies virus.

Algorithms↗

Intimal lipid accretion and elevated serum cholesterol in Marek's disease virus-inoculated chickens.

In our previous studies of the early pathogenesis of the Marek's disease virus (MDV)-associated model of atherosclerosis, the brachiocephalic arteries and ascending aortas of MDV-inoculated chickens failed to develop lipid accretion and intimal/medial proliferation consistent with atherosclerotic lesions, as described in the original reports of this model. The role of cholesterol supplementation in the formation of MDV-associated atherosclerotic lesions was reexamined. At 3 days of age, 40 chicks were inoculated with the CU-2 strain of MDV. Another 40 chicks were sham inoculated. At 15 weeks postinfection, half of the sham- and MDV-inoculated birds received 2% cholesterol supplementation in the diet for the rest of the experimental period. At 30 weeks postinfection, the aortas and brachiocephalic arteries were evaluated. Several observations were different from the original description of the model. None of the chickens among the four experimental groups developed atherosclerotic lesions, regardless of MDV inoculation or cholesterol supplementation. However, intimal thickening and marked oil red O-positive foam cell accumulation were observed in all 11 MDV-inoculated, cholesterol-supplemented chickens. None of the nine MDV-inoculated, unsupplemented chickens manifested arterial lipid accumulation, despite the presence of an intimal cellular infiltrate. Also in contrast to the original model description, both cholesterol-supplemented and unsupplemented MDV-inoculated chickens manifested significant increases in serum cholesterol in comparison with the respective sham-inoculated groups.

Animals↗

Murine cytomegalovirus-associated arteritis.

Unique inflammatory lesions affecting the ascending aorta and pulmonary artery of BALB/c and C57BL/6 mice infected with murine cytomegalovirus (MCMV) were identified in a pilot and two subsequent experiments to characterize the potential effect of MCMV infection on diet-induced atherosclerotic lesions. Suckling BALB/c and C57BL/6 mice were inoculated with MCMV and subsequently fed either a commercial mouse diet or a synthetic atherogenic diet from weaning. The three experiments varied with respect to the age of the mice at the time of MCMV inoculation and the dose of virus given. The conditions of MCMV exposure were progressively modified in the three experiments to increase the prevalence of MCMV-associated inflammatory lesions in the pulmonary artery and aorta. In the final experiment, in which suckling mice were inoculated at 9 days of age, MCMV-associated arteritic lesions had an observed prevalence at 8 weeks post-inoculation of 87.5% (7/8) in BALB/c mice on the normal diet and 100% (8/8) in C57BL/6 mice on the normal diet and in both strains on the atherogenic diet. The inflammatory lesions in both vessels were characterized by mononuclear cell infiltrates containing CD3+, CD4+, and CD8+ lymphocytes. The cellular infiltrates were often more intense on the adventitial surface and infiltrated into the overlying tunica media. The intima was infiltrated by mononuclear cell infiltrates that appeared to contain more macrophages and fewer lymphocytes than did the adventitial infiltrates.(ABSTRACT TRUNCATED AT 250 WORDS)

Age Factors↗

Endothelial MHC class II antigen expression and endarteritis associated with Marek's disease virus infection in chickens.

Experimental Marek's disease virus (MDV) infection in chickens was used to study the early pathogenesis of virus-induced atherosclerosis. Previous investigations using this model have reported the occurrence of atherosclerotic lesions after approximately 7 months postinfection. In this study, a total of 75 susceptible Cornell P-line chickens were inoculated intraperitoneally with the CU-2 strain of MDV at 3 days of age and subsequently perfused for histologic examination. At 2, 4, 8, 13, and 20 weeks postinoculation, the ascending aorta and the brachiocephalic and coronary arteries were evaluated for early changes. Expression of class II major histocompatibility complex (Ia) antigen by the vascular endothelium was demonstrated by indirect immunodetection as early as 2 weeks after virus inoculation. This change was followed by significant thickening of the intimal layer associated with mononuclear cell infiltration. All the arteries examined from the MDV-infected chickens were affected. Preliminary immunohistochemical staining showed the presence of CD3+ CD4+, and CD8+ cells among the infiltrating cells. The results suggest that an immunopathologic mechanism may be involved in the early pathogenesis of MDV-induced atherosclerosis in chickens.

Animals↗

Prevalence of coagulase gene polymorphism in Staphylococcus aureus isolates causing bovine mastitis.

This study was conducted to investigate polymorphism of the coagulase gene of Staphylococcus aureus causing bovine mastitis. One hundred eighty-seven strains of S. aureus were isolated from bovine mastitic milk samples obtained from 187 different Danish dairy farms. The isolates were characterised for restriction fragment length polymorphism (RFLP) of the coagulase gene. A variable region of the coagulase gene was amplified using the polymerase chain reaction (PCR) followed by AluI restriction enzyme digestion. A total of 15 different RFLP patterns were observed. The predominant pattern was found in 35% of the isolates. The ease of analysing coagulase gene polymorphisms among a large number of strains, and the multiple distinct polymorphic patterns generated, supports the use of this technique in epidemiological investigations of bovine mastitis. The predominating variants may have predelection for causing intramammary infections.

Animals↗

Genetic recombination between two strains of Aujeszky's disease virus at reduced multiplicity of infection.

The potential for in vivo genetic recombination has been suggested for Aujeszky's disease virus (ADV) stemming from the use of modified-live vaccines carrying vaccine-specific deletion mutations. This scenario serves as a model for the study of biological factors that affect in vivo herpesviral recombination and, ultimately, the natural evolution of herpesviruses. This report describes experiments evaluating the efficiency of ADV recombination under in vitro conditions of low multiplicity infection which were felt to represent in vivo conditions more closely than previous in vitro measurements of ADV recombination. A series of experiments were performed to measure in vitro recovery of recombinant viral progeny following co-infection of cell monolayers with non-saturating concentrations of two parental strains of ADV. A simple statistical model was generated to estimate the rate of cellular co-infection. After a single replicative cycle infectious viral progeny were recovered and their genotypes determined at two alleles by characterization of two gene loci using a battery of PCR assays. Recovery rates of parental and recombinant progeny genotypes were calculated from the PCR data. The observed frequency of recovery of recombinant viral progeny closely approximated the values projected by the model calculations. The data from our system suggest that at low multiplicity of infection, the rate of genetic recombination is a function of the probability of cellular co-infection.

Genotype↗

Measurement of Aujeszky's disease virus recombination in vitro under conditions of low multiplicity of infection.

The natural development of Aujeszky's disease virus (ADV) vaccine-derived recombinants has been proposed as a conceivable, although unproven, outcome of the practice of using modified-live vaccines on infected herds. Herpesviral recombination has been studied in vitro using high multiplicity of infection (m.o.i.) to maintain synchronous co-infection of target cells and one-step multiplication cycles. However, natural in vivo recombination is unlikely to occur at high m.o.i. with synchronous infection of all available target cells by both parental virus strains. Using a battery of gene-specific polymerase chain reaction assays, an analysis of recombination frequency at comparatively low m.o.i. was performed, which indicates that at low m.o.i. the rate of in vitro genetic recombination can be expressed as a function of the number of virus-cell interactions. This finding indicates that the rate of generation of recombinant ADV genotypes in swine infected by multiple ADV strains would be modulated by factors that affect the distribution of different virus strains to common target cells, thereby limiting or enhancing the likelihood of cellular co-infection. A full description of this study is available (J. gen. Virol., in press).

Animals↗

Recovery of Aujeszky's disease virus recombinants from experimentally co-infected swine.

Tissue homogenates were obtained from swine experimentally co-infected with two vaccine strains of Aujeszky's disease virus (ADV). Viral isolates were derived by serial plaque purification directly from tissue homogenates, without an intervening step of isolation and amplification on cell cultures. Use of limiting dilutions and recovery of virus isolates as individual plaques minimized the likelihood of in vitro recombination serving as a confounding source of recombinant ADV. The tyhmidine kinase and glycoprotein X gene sequences were classified as wildtype or deleted, using a battery of polymerase chain reaction assays. On the basis of pairwise combinations of the allelic forms of the thymidine kinase and glycoprotein X genes, the isolates were characterized as recombinant and parental genotypes. The results substantiate the observation that ADV vaccine strains can form genetic recombinants in vivo after experimentally induced co-infection. A full description of this study is available (Am. J. Vet. Res. 54 (4) 540-545, 1993).

Animals↗