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Biomedical subjects

I H Cheng

Publications and source records attributed to I H Cheng.

11 recordsLinked to original sources

Sequence analysis of Pasteurella multocida major outer membrane protein (OmpH) and application of synthetic peptides in vaccination of chickens against homologous strain challenge.

Pasteurella multocida major outer membrane protein (OmpH) has been previously characterized as a porin. The native OmpH from strain X-73 (serotype 1) but not recombinant protein from Escherichia coli induced homologous protection in chickens. In this study OmpH sequences from 15 P. multocida serotypes as well as the CU vaccine strain were compared by sequence alignment and revealed high homology, with major variations confined to two discrete regions which were correspondingly predicted as two largest external loops. Secondary structures of OmpHs were predicted by sequence alignment of OmpHs with well defined porins and analyses of amphiphilicity, hydrophobic moment and antigenic index plots. Several synthetic peptides derived from predicted loop 2 and loop 5 of X-73 OmpH were synthesized as vaccine candidates. Vaccination studies in chickens showed that the cyclic synthetic peptide (Cyclic-L2) mimicking the predicted loop 2 induced 70% protection in chickens against strain X-73 challenge. This is the first report that a synthetic peptide mimicking the conformational epitopes of a native protein provide practical protection in target animal against bacterial infection.

Amino Acid Sequence↗

Cloning and characterization of the major outer membrane protein gene (ompH) of Pasteurella multocida X-73.

The major outer membrane protein (OmpH) of Pasteurella multocida X-73 was purified by selective extraction with detergents, followed by size exclusion chromatography. The planar lipid bilayer assay showed that OmpH has pore-forming function. The average single channel conductance in 1.0 M KCl was 0.62 nS. The gene (ompH) encoding OmpH has been isolated and sequenced by construction of a genomic library and PCR techniques. The coding region of this gene is 1,059 bp long. The predicted primary protein is composed of 353 amino acids, with a 20-amino-acid signal peptide. The mature protein is composed of 333 amino acids with a molecular mass of 36.665 kDa. The ompH gene encoding mature protein has been expressed in Escherichia coli by using a regulatable expression system. The ompH gene was distributed among 15 P. multocida serotypes and strain CU. Protection studies showed that OmpH was able to induce homologous protection in chickens. These findings demonstrate that OmpH is a protective outer membrane porin of strain X-73 and is conserved among P. multocida somatic serotypes.

Amino Acid Sequence↗

Use of clinical findings in the diagnosis of urinary tract infection in women.

To develop a decision rule for predicting urinary culture results in patients suspected of having urinary tract infection, we used discriminant analysis to identify the optimum combination of clinical findings. Thirty variables identified in a pilot study were recorded from 248 patients in a second study. Five findings were independent predictors of positive urinary culture: history of urinary tract infection, back pain, microscopic pyuria, hematuria, and bacteriuria. An additive decision rule that assigned one point for each of the five variables was tested in a third group of 258 patients. These scores stratified patients into subsets with increasing likelihood of positive culture. Higher scores identified patients who can confidently be treated without documentation of bacteriuria. If the rule applies successfully to other populations, cost savings could result from identification of patients who do not require quantitative urinary culture to demonstrate significant bacteriuria.

Adolescent↗

Omaha childhood blood lead and environmental lead: a linear total exposure model.

The majority of experimental and population studies of blood lead (PbB) and environmental lead, including the Omaha study, have utilized the Goldsmith-Hexter log-log or power function model. Comparison was made of the log-log model and a linear model of total exposure to describe the Omaha Study of 1074 PbBs from children ages 1-18 years as related to air (PbA), soil (PbS), and housedust (PbHD) lead. The data fit of the linear model was statistically equivalent to the power model and the predicted curves were biologically more plausible. The linear model avoids the mathematical limitations of the power model which predicts PbB zero at PbA zero. From the Omaha data, this model, ln PbB = ln (beta 0 + B1 PbA + B2 PbS + beta 3 PbHD) predicts that PbB increases 1.92 micrograms/dl as PbA increases 1.0 microgram/m3. Since PbS and PbHD increase with PbA, however, the increases in total exposure predict a PbB increase of 4-5 micrograms/dl as PbA increases 1.0 microgram/m3.

Adolescent↗

In vivo antigen expression by Pasteurella multocida.

Pasteurella multocida was purified from the blood of turkeys affected with acute fowl cholera, and membrane preparations from those bacteria were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and visualized on immunoblots. Antigens were detected in the membranes of these in vivo-propagated bacteria that were not detected in membrane preparations of the same P. multocida strain grown in vitro. The unique antigens were detected in the detergent-insoluble phase and were enriched to various degrees by different detergents.

Animals↗

Alternative injection sites for a Pasteurella multocida bacterin.

Seven different injection sites for a Pasteurella multocida bacterin were evaluated by measuring the immune response and the local tissue reaction. Injection into the ventral surface of the tail or subcutaneously along the dorsal midline of the neck were the most suitable procedures. Ease of application was judged subjectively, and the tail site was found to be easier to inject accurately than the subcutaneous neck site. The tail injection site was found to be the best overall when immune response, tissue reaction, and ease of application were all considered.

Animals↗

The effect of oxytetracycline on the severity of airsacculitis in chickens infected with Mycoplasma gallisepticum.

Four groups of mycoplasma-free commercial broilers were challenged with the R strain of Mycoplasma gallisepticum (MG) at 14 days of age. Groups received feed containing either no medication, or 500 ppm or 1000 ppm oxytetracycline (OTC) beginning at age 13 days, or 1000 ppm OTC beginning at age 15 days. All broilers were vaccinated with a live mild Massachusetts infectious bronchitis vaccine at 17 days of age. Air sac lesions were scored at age 24 days. In two almost identical experiments, all OTC treatment groups had significantly lower mean air sac lesion scores than the unmedicated challenged controls. Groups that were fed 1000 ppm OTC in feed had significantly lower mean air sac lesion scores than groups that were fed 500 ppm OTC in feed. There was no significant difference in mean air sac lesion scores between the groups fed 1000 ppm OTC in feed beginning at 13 days of age and those fed 1000 ppm OTC in feed beginning at 15 days of age.

Air Sacs↗

Pasteurella multocida infection in Japanese quail (Coturnix coturnix japonica).

Three flocks of Japanese quail, approximately 75,000 birds each, experienced acute high mortality beginning at 24 to 28 days of age. Gross lesions were absent or were composed of either multifocal small pale areas on livers and spleens or lungs slightly darker in color than normal. Histopathology revealed multifocal splenic and hepatic necrosis and interstitial pneumonia. Pasteurella multocida, serotype 3,4, was isolated from affected tissues. The quail were successfully treated with chlortetracycline, and the organism was apparently eliminated from the premises by thorough cleaning, disinfection, and insect and rodent control. Experimental studies showed Japanese quail to be highly susceptible to disease caused by the P. multocida isolated from the affected flocks.

Animals↗

Lesions resulting from attempted Shwartzman reaction in turkey poults inoculated with Pasteurella multocida lipopolysaccharide.

Five-week-old turkey poults were given two consecutive intravenous injections (24 hours apart) of highly purified Pasteurella multocida lipopolysaccharide (LPS) in an effort to induce a generalized Shwartzman reaction. There were no gross lesions, and microscopic lesions were limited to focal hepatic necrosis with heterophil infiltration. Hepatic lesions did not differ qualitatively from lesions in turkeys given a single dose of lipopolysaccharide. Margination of heterophils in the pulmonary vasculature was observed in turkeys 4 hours after a single injection of LPS, but it was not present in turkeys given the consecutive injections of LPS. To induce a dermal Shwartzman reaction, turkeys were given intradermal injections of LPS followed by an intravenous injection of LPS 24 hours later. Although no grossly visible hemorrhagic dermal necrosis occurred, microscopic lesions, including heterophil infiltration, vasculitis, thrombosis, and necrosis, were present. Thrombosis and vasculitis were observed only in turkeys given the intravenous and intradermal LPS, whereas the other inflammatory changes were observed in turkeys given the intradermal injection of LPS and intravenous water. Prominent lymphocytic perivascular cuffing at the site of dermal injection was present in all turkeys given intradermal LPS.

Animals↗

Infectious laryngotracheitis virus in commercial hens: a serological study based on enzyme-linked immunosorbent assay.

Serum samples collected from 23 flocks of commercial hens from three different companies were tested by enzyme-linked immunosorbent assay (ELISA) for antibodies against infectious laryngotracheitis (ILT) virus, and data were analyzed statistically. Geometric mean titers (GMTs) were compared from hens that were unvaccinated, once-vaccinated, or twice-vaccinated, from single-age farms or multiple-age farms, from molted or unmolted flocks, and from different companies. There were significant differences among the groups compared by vaccination, between the single-age and multiple-age groups, and between the molted and unmolted groups. The GMT of unvaccinated flocks and the GMT of molted flocks that had been vaccinated once as pullets with a chick-tissue-culture-origin (CTCO) live vaccine could not be differentiated. The ELISA is useful to detect and quantitate ILT vaccine-induced antibody, but it does not reliably identify older flocks that were vaccinated at a young age with CTCO vaccine.

Aging↗

Cross-protection studies with Pasteurella multocida bacterins prepared from bacteria propagated in iron-depleted medium.

Strains X-73 (serotype 1) and P-1059 (serotype 3) of Pasteurella multocida, avian origin, expressed additional membrane proteins (MPs) when grown in brain-heart infusion (BHI) broth containing the iron chelator dipyridyl and when grown in BHI broth treated with the iron chelator Chelex 100. These additional MPs were not detected when both strains were grown in BHI broth. Chickens and turkeys were vaccinated twice with inactivated oil-emulsion vaccines containing bacterial cells expressing these MPs or with vaccines containing bacterial cells grown in BHI broth. Two weeks after the final vaccination, all birds were challenged to determine whether bacterins made from P. multocida that had been propagated in conditions of iron deprivation would induce heterologous serotype immunity. The bacterins produced in medium low in iron did not consistently induce significant protection against heterologous challenge.

Animals↗