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Biomedical subjects

N Allison

Publications and source records attributed to N Allison.

At least 19 recordsLinked to original sources

The transduction of rat submandibular glands by an adenoviral vector carrying the human growth hormone gene is associated with limited and reversible changes at the infusion site.

Adenoviral vectors have been shown to efficiently deliver exogenous genes to salivary glands and have therefore been investigated as tools for the treatment of human disease. The purpose of this study was to evaluate the response of F344 rats to intraductal infusion of the right submandibular salivary gland with an adenoviral vector encoding the gene for human growth hormone (AdCMVhGH). Co-administration of hydroxychloroquine (HCQ) was used to redirect the secretion of human growth hormone (hGH) from saliva into serum. This paper documents the findings of the pathology evaluation of this National Toxicology Program study. The right submandibular salivary gland (infusion site) was the primary target organ, with microscopic lesions characteristic of a mild to moderate insult observed at 3 days post infusion in vector exposed animals. These lesions were characterized by variable degrees of acute glandular inflammation, degeneration and necrosis, with more severe lesions in the higher dose groups. Rats at 28 days post infusion had milder inflammation, degeneration and necrosis compared to day 3 rats, with variable degrees of regeneration. In conclusion, the effects on the salivary glands are reversible as indicated by the milder inflammation and degeneration in the day 28 rats concomitant with mild to moderate regeneration. Therefore, the vector appears relatively innocuous with limited tissue toxicity. [The supplemental data referenced in this paper is not printed in this issue of Toxicologic Pathology. It is available as a downloadable file in the online edition of Toxicologic Pathology, 34(4). In order to access the full article online, you must have either an individual subscription or a member subscription accessed through www.toxpath.org.].

Adenoviridae↗

Fatal herpes simplex infection in a pygmy African hedgehog (Atelerix albiventris).

An adult pygmy African hedgehog developed acute posterior paresis attributed to a prolapsed intervertebral disc diagnosed by C-T scan. Corticosteroid therapy resulted in prompt resolution of the ataxia, but 2 weeks later the animal became anorexic and died. Macroscopically, the liver was stippled with punctate off-white foci which were confirmed microscopically to be foci of necrosis. Numerous hepatocytes contained intranuclear inclusions and syncytial cell formation was also present. A herpes virus was isolated and identified by fluorescent antibody and polymerase chain reaction studies as herpesvirus simplex type 1. To our knowledge, this is the first report of herpes infection in the African hedgehog and the first time herpes simplex has been identified as a cause of disease in insectivores.

Animals↗

Macracanthorhynchus ingens infection in a dog.

A 4-month-old sexually intact female Siberian Husky was examined because of diarrhea and 4 flesh-colored annulated helminths that were recovered from the dog's feces. Infection with Macracanthorhynchus ingens was diagnosed on the basis of morphologic features of the adult parasites and ova. Spindle-shaped eggs (mean length, 91 microm; mean width; 54 microm) were obtained from the body cavity of a gravid female specimen. The dog was treated empirically with epsiprantel (5.5 mg/kg [2.5 mg/lb] of body weight) and ivermectin (250 to 500 microg/kg [114 to 227 microg/lb]), and the diarrhea resolved. Infection with this parasite has been reported in raccoons from the same geographic area. Macracanthorhynchus ingens is typically a parasite of raccoons, wolves, badgers, foxes, skunks, mink, and moles; transmission from wildlife to dogs may occur via ingestion of infected intermediate hosts (millipedes).

Acanthocephala↗

Cesium and strontium ion exchange on the framework titanium silicate M2Ti2O3SiO4.nH2O (M = H, Na).

The ion exchange properties of the titanium silicate, M2Ti2O3SiO4.nH2O (M = H, Na), toward stable and radioactive 137Cs+ and 89Sr2+, have been examined. By studying the cesium and strontium uptake in the presence of NaNO3, CaCl2, NaOH, and HNO3 (in the range of 0.01-6 M) the sodium titanium silicate was found to be an efficient Cs+ ion exchanger in acid, neutral, and alkaline media and an efficient Sr2+ ion exchanger in neutral and alkaline media, which makes it promising for treatment of contaminated environmental media and biological systems.

Cesium↗

Transferrin-binding protein B isolated from Neisseria meningitidis discriminates between apo and diferric human transferrin.

Neisseria meningitidis utilization of human serum transferrin (hTF)-bound iron is an important pathogenicity determinant. The efficiency of this system would clearly be increased through preferential binding of diferric hTF over the iron-free form. To characterize this process, functionally active meningococcal transferrin-binding protein A (TbpA) and TbpB have been purified from N. meningitidis using a novel purification procedure. The association of isolated Tbps and Tbps in the presence of hTF was investigated by gel filtration. Co-purified TbpA+B formed a complex of molecular mass 300 kDa which bound 1-2 molecules of hTF. Purified TbpA formed a complex of 200 kDa, indicating association as a dimer, whereas TbpB aggregated to form multimers of variable sizes. On recombining TbpA and TbpB, a stable complex of equivalent size to co-purified TbpA+B was formed. This complex may be composed of a single TbpA dimer and 1 molecule of TbpB. The technique of surface plasmon resonance (SPR) was used to demonstrate clearly that TbpB of either high (85 kDa) or low (68 kDa) molecular-mass preferentially bound diferric hTF in comparison with iron-free hTF. This selectivity was not observed with TbpA, but was found at low levels with co-purified TbpA+B. Individual TbpA and TbpB, recombined in a 1:1 molecular ratio, showed iron-mediated discriminatory binding at an intermediate level. SPR was also used to show that TbpA and TbpB bound to distinct regions of hTF, and that prior saturation with TbpB reduced subsequent TbpA binding. The results demonstrated that hTF bound more TbpA than TbpB, with an approximate ratio of 2:1. We have demonstrated that in vitro, TbpA+B exists as a receptor complex composed of a TbpA dimer and one molecule of TbpB, and that TbpB selectively binds diferric hTF. We propose that, in vivo, TbpA and TbpB also exist as a receptor complex, with TbpB selectively binding diferric hTF, bringing it close to TbpA, the transmembrane component, where the ferric iron can be transported to the periplasm.

Apoproteins↗

Accuracy of an automated blood pressure device in stable inpatients: optimum vs routine use.

BACKGROUND: Despite widespread use of the automated blood pressure (BP) device (IVAC model 4200, IVAC Corporation, San Diego, Calif), there is little formal validation in the literature on its accuracy. OBJECTIVE: To assess the accuracy of the IVAC 4200 device, both under standardized conditions and as routinely used by ward staff, compared with the true indirect BP measured by mercury manometer (MM). METHODS: One hundred forty-five stable inpatients were randomly selected for BP measurements by 3 randomly ordered protocols: (1) MM performed by certified investigators, (2) IVAC 4200 BP performed by trained investigators (research automated [RA]), and (3) IVAC 4200 BP performed by ward personnel (ward automated [WA]). RESULTS: For RA compared with MM ("true" indirect BP), 59% of systolic and 54% of diastolic readings were within 5 mm Hg and 83% of systolic and 86% of diastolic were within 10 mm Hg for a British Hypertension Society grade C for both. For WA compared with MM, 40% of systolic and 50% of diastolic readings were within 5 mm Hg and 70% of systolic and 80% of diastolic readings were within 10 mm Hg for British Hypertension Society grades D and C, respectively. The presence of arrhythmias and/or low K5 values (fifth phase of Korotkoff sounds <30 mm Hg) significantly increased the inaccuracy for diastolic values. Inappropriate cuff selection significantly increased inaccuracy of systolic BP (WA vs MM). CONCLUSIONS: The IVAC 4200 yields substandard estimates of systolic and diastolic BP even under standardized, thus optimum conditions. The presence of arrhythmias or low K5 values and the selection of inappropriate cuff size by the ward staff also contributed to inaccuracy.

Adult↗

Metabolic characterisation of a novel vanillylmandelate-degrading bacterium.

A newly isolated gram-negative bacterium, possibly Brevundimonas diminuta, utilised D,L-vanillylmandelate (D,L-VMA) as a sole carbon and energy source. The organism converted D,L-VMA to vanillylglyoxylate using a soluble NAD-dependent dehydrogenase specific for D-VMA and a dye-linked, membrane-associated L-VMA dehydrogenase. Vanillylglyoxylate was further metabolised by decarboxylation, dehydrogenation and demethylation to protocatechuate. A 4,5-dioxygenase cleaved protocatechuate to 2-hydroxy-4-carboxymuconic semialdehyde. Partially purified d-VMA dehydrogenase exhibited optimal activity at 30 degrees C and pH 9.5 and had an apparent Km for D-VMA of 470 microM. Although induced by several substituted mandelates, the enzyme had a narrow substrate specificity range with virtually no activity towards D-mandelate. Such properties render the enzyme of potential use in both diagnostic and biosynthetic applications.

Bacteria↗

Degradation of homovanillate by a strain of Variovorax paradoxus via ring hydroxylation.

A newly isolated strain of Variovorax paradoxus could grow on homovanillate and several monohydroxylated phenylacetic acids. During growth on homovanillate, the organism formed separate NAD(P)H-dependent hydroxylases with activity towards 4-hydroxyphenylacetic acid and homovanillate. Homovanillate hydroxylase catalysed a typical monooxygenase reaction and had little activity towards 4-hydroxyphenylacetic acid GC-MS and TLC analysis suggested that homovanillate was 1-hydroxylated to yield a dihydroxymonomethoxyphenylacetic acid which served as a substrate for homogentisate 1,2-dioxygenase. Methanol, but not formaldehyde, was released either during ring-cleavage or subsequent metabolism of the ring-cleavage product.

Alcaligenes↗

Trypanosoma cruzi infection in Walker hounds from Virginia.

Trypanosomiasis has been reported in dogs from Texas, Oklahoma, Louisiana, and South Carolina. We describe the first isolation and characterization of Trypanosoma cruzi from a Walker Hound pup in Virginia that also had postvaccinal distemper. The mother of the pup and 7 of its 8 siblings also were found to be infected with T cruzi, suggesting that the parasite had been transmitted transplacentally or through lactation. Parasitologic, serologic, histologic, and molecular methods were used to establish the diagnosis of T cruzi infection in these dogs. In a serologic survey of 12 dogs (including the sire of the pups) from the area in which the index case occurred, none were found to have antibodies to T cruzi. However, 2 of a further 52 dogs from different areas (to the index case), but in the same county, were seropositive to T cruzi. These findings indicate that canine trypanosomiasis is present in an area of the United States not previously known to be enzootic.

Animals↗

Rapid enzymatic method for the measurement of mannitol in urine.

A previously described method for mannitol in urine has been modified and improved. End product inhibition by fructose in the mannitol dehydrogenase method for mannitol has been minimized; the assay is linear over a sample mannitol concentration range of 0-12 mmol/L; no significant interference from other sugars or sugar alcohols could be demonstrated. The method is precise (within-batch CV less than 1%), rapid and shows excellent recovery of mannitol in spiked samples. Comparison with gas liquid chromatography shows excellent correlation (r = 0.994) between the two methods.

Chromatography, Gas↗

Enteric pythiosis in a horse.

Enteric pythiosis was diagnosed in a 2-year-old Quarter Horse gelding. The horse had signs of colic, which appeared to be alleviated by administration of mineral oil and analgesics, but only temporarily. Intestinal distention developed after initial examination. At surgery, a thick stenotic area was observed in the middle portion of the jejunum. The thick intestine and associated mesentery contained multiple firm nodules of gritty caseous material. Histopathologic findings included sclerosing eosinophilic granulomatous enteritis and peritonitis. Although the lesion resembled a response to migrating parasites and lacked hyphae in initial sections, the gross appearance of the lesion prompted the acquisition of additional sections to detect the etiologic agent.

Animals↗

A rapid, quantitative microplate assay for NAD-linked D-mannitol dehydrogenase.

A 96-well microtitre plate assay for NAD-linked D-mannitol dehydrogenase based on the reduction of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) by reduced NAD is described. The assay allows rapid measurement of D-mannitol dehydrogenase in crude bacterial extracts derived by sonic disruption, in acetone permeabilized cells and in column eluates during enzyme purification. The absorbance of reaction mixtures in a microtitre plate is measured at 620 nm over a 3-4 min period using a programmable microplate reader. The rate of increase in absorbance is directly proportional to the amount of enzyme present and there is excellent correlation between activities derived using the microplate assay with those determined using conventional spectrophotometric methods.

Lactobacillus↗

Reductive acetylation of tandemly repeated lipoyl domains in the pyruvate dehydrogenase multienzyme complex of Escherichia coli is random order.

In vitro deletion and site-directed mutagenesis of the aceF gene of Escherichia coli was used to generate dihydrolipoamide acetyltransferase (E2p) polypeptide chains containing various permutations and combinations of functional and non-functional lipoyl domains. A lipoyl domain was rendered non-functional by converting the lipoylatable lysine residue to glutamine. Two- and three-lipoyl domain E2p chains, with lipoyl-lysine (Lys244) substituted by glutamine in the innermost lipoyl domains (designated +/- and +/+/-, respectively), and similar chains with lipoyl-lysine (Lys143) substituted by glutamine in the outer lipoyl domains (designated -/+ and -/-/+), were constructed. In all instances, pyruvate dehydrogenase complexes were assembled in vivo around E2p cores composed of the modified peptide chains. All the complexes were essentially fully active in catalysis, although the complex containing the -/-/+ version of the E2p polypeptide chain showed a 50% reduction in specific catalytic activity. Similarly, active-site coupling in the complexes containing the +/-, +/+/- and -/+ constructions of the E2p chains was not significantly different from that achieved by the wild-type complex. However, active-site coupling in the complex containing the -/-/+ version of the E2p chain was slightly impaired, consistent with the reduced overall complex activity. These results indicate that during oxidative decarboxylation there is no mandatory order of reductive acetylation of repeated lipoyl domains within E2p polypeptide chains, and strongly suggest that the three tandemly repeated lipoyl domains in the wild-type E2p chain function independently in the pyruvate dehydrogenase complex.

Acetylation↗

Eumycotic mycetoma caused by Pseudallescheria boydii in a dog.

Abdominal eumycotic mycetoma caused by Pseudallescheria boydii was diagnosed in a 3-year-old male Siberian Husky. The dog was examined because of weight loss and signs of depression. Initially, pyrexia was the only clinical finding. Antibiotic and corticosteroid treatment was ineffective. Two weeks later, the dog's appetite had decreased, it had vomited a few times, and the caudal portion of the abdomen was sensitive to palpation. Hematologic and serum biochemical abnormalities consisted of anemia, leukocytosis, hypoglycemia, hypoalbuminemia, hyperglobulinemia, and high alkaline phosphatase activity. One week later, the dog's condition continued to worsen, and testicular swelling was observed. The dog was castrated. Microscopic examination of specimens obtained at surgery revealed pyogranulomatous periorchitis with mycetoma granules. Ketoconazole treatment was initiated and continued until the dog died one month later. Necropsy revealed multifocal duodenal ulcers, with transmural pyogranulomatous enteritis, pancreatitis, and peritonitis. This case is unique because the etiologic agent apparently entered via the intestinal tract rather than by contamination of an external wound.

Animals↗