Biomedical subjects

M Chen

Publications and source records attributed to M Chen.

At least 541 records · Page 30Linked to original sources

PubMed1996

Use of intrinsic and extrinsic helper epitopes for in vivo induction of anti-hepatitis C virus cytotoxic T lymphocytes (CTL) with CTL epitope peptide vaccines.

The induction of virus-specific cytotoxic T lymphocytes (CTL) is an important part of vaccine strategy. CTL induction in vivo by two hepatitis C virus (HCV) peptides containing CTL epitopes, one from the NS5 region (P17) and one from the core (C7), was compared. P17 required covalent attachment of a helper peptide (PCLUS3 containing a cluster of epitopes from the human immunodeficiency virus envelope protein), whereas C7 did not. However, the minimal decapeptide of C7, C7A10, alone did not induce CTL. The helper cells induced by PCLUS3-17 or by C7 were shown to be CD4+ and to produce interleukin-2 (IL-2). Thus, help can be supplied by a natural helper epitope intrinsic to the CTL peptide, as in C7, or by attaching a helper epitope from another protein, as in the case of P17. The cluster peptides may be useful promiscuous helper peptides for a variety of CTL epitopes from diverse pathogens.

Amino Acid Sequence↗

PubMed1996

Immunogenicity and antigenicity of the ATPase/helicase domain of the hepatitis C virus non-structural 3 protein.

The immunogenicity and antigenicity of an enzymatically functional (ATPase/helicase) recombinant protein encompassing residues 1207-1612 of the hepatitis C virus (HCV) non-structural 3 (NS3) protein was characterized using B10 congenic mice. Previous studies have indicated a high frequency of NS3-specific antibodies in HCV-infected humans. Similarly, all six immunized murine haplotypes were antibody responders to the NS3 ATPase/helicase domain, with the H-2k and H-2s haplotypes as high responders. As also observed in HCV-infected humans, the murine NS3 antibodies were predominantly directed to conformational determinants. Irrespective of the murine haplotype, IgG1 predominated in the primary anti-NS3 response, whereas IgG1 and IgG2b predominated in the secondary response. The antibody responder hierarchy was reiterated at the T cell level, with the H-2k and the H-2s haplotypes as the best responders. In both the H-2d and H-2k haplotypes ATPase/helicase-primed T cells secreted interleukin 2 and interferon gamma, corroborating observations from HCV-infected humans. In the H-2d, H-2k and H-2s haplotypes the fine specificity of the T cell recognition of the ATPase/helicase domain was further characterized. Multiple, although generally weak, T cell recognition sites were found for all three haplotypes. The large size of the NS3 protein together with the presence of multiple class II binding motifs explain the high prevalence of NS3 antibodies in immunized mice and predict a similar explanation for the observed high frequency of NS3-specific antibodies in HCV-infected humans.

Adenosine Triphosphatases↗

PubMed1996

Effect of neonatal axoplasmic transport attenuation in the infraorbital nerve on vibrissae-related patterns in the rat's brainstem, thalamus and cortex.

This study evaluated the effects of neonatal attenuation of axoplasmic transport in the infraorbital nerve (ION) on the organization of vibrissae-related patterns in the rat's CNS. Application of colchicine- or vinblastine- impregnated implants to the ION from birth until postnatal day (P)6 to P10 resulted in a 92.4% reduction in the number of trigeminal (V) ganglion cells labelled by application of horseradish peroxidase to the vibrissa pad and a 44.8% decrease in the number of Nissl-stained ganglion cells in the ophthalamic-maxillary portion of the V ganglion. These implants also decreased the number of myelinated fibres in the ION. In normal rats killed on P6-10, there was an average of 10273 +/- 1259 myelinated axons in the nerve. In the animals with colchicine- or vinblastine-treated implants, this value was 3891 +/- 1965. The highest axon count in an experimental animal was 9859. In all animals, axoplasmic transport attenuation resulted in the disappearance of normal vibrissae-related cytochrome oxidase patterns in the brainstem, thalamus and primary somatosensory cortex. Axoplasmic transport attenuation did not result in the disappearance of vibrissae-related ordering of V primary afferent terminal arbors, as demonstrated by anterograde labelling with neurobiotin. These results suggest that some factor conveyed from the periphery of the V ganglion and perhaps on to the brainstem is necessary for the maintenance of vibrissae-related patterns in the thalamus and cortex.

Afferent Pathways↗

PubMed1996

Effects of beta-adrenoceptor antagonists on Ca(2+)-overload induced by lysophosphatidylcholine in rat isolated cardiomyocytes.

1. The effects of beta-adrenoceptor antagonists including (-)- and (+)-propranolol, (-)- and (+)-penbutolol, timolol, pindolol, atenolol, acebutolol and practolol on the Ca(2+)-overload induced by lysophosphatidylcholine (LPC) were examined in isolated cardiomyocytes of the rat. 2. Fura-2 was used for measurement of the intracellular calcium concentration ([Ca2+]i). LPC (15 microM) produced a rapid increase in [Ca2+]i from 72 +/- 5 to 3042 +/- 431 nM which coincided with a decrease in the percentage of rod-shaped cells from 69 +/- 2 to 5 +/- 2%. 3. Preincubation with (-)-propranolol (20 microM), (+)-propranolol (50 microM), or (-)- or (+)-penbutolol (20 microM), the lipophilicity of which is higher than other beta-adrenoceptor antagonists, significantly inhibited both the increase in [Ca2+]i and the cell-shape change induced by 15 microM LPC. The inhibitory effects of the four drugs on the LPC-induced increase in [Ca2+]i and cell-shape change were concentration-dependent. The IC50S of (-)-propranolol, (+)-propranolol, (-)- and (+)-penbutolol for the increase in [Ca2+]i were 1.28, 10.50, 0.67 and 0.76 microM, respectively. 4. Pretreatment with pindolol, timolol, acebutolol, practolol, atenolol or lignocaine did not inhibit the increase in [Ca2+]i and the morphological change induced by LPC. 5. LPC markedly increased the release of creatine phosphokinase from 9 +/- 1 to 45 +/- 2% which could be significantly reduced by (-)- or (+)-propranolol but not by acebutolol or timolol. 6. The protective effects of (-)- and (+)-propranolol, (-)- and (+)-penbutolol against the Ca(2+)-overload induced by LPC were not associated with the beta-adrenoceptor antagonistic action, but probably with an unknown action which is related to the preservation of membrane integrity. Further studies are necessary to clarify the exact mechanisms of the protective action of these beta-adrenoceptor antagonists against the Ca(2+)-overload induced by LPC.

Adrenergic beta-Antagonists↗

PubMed1996

Dot-immunogold filtration assay as a screening test for syphilis.

A dot-immunogold filtration assay (DIGFA) for the rapid detection of reaginic antibody in the serum of syphilitic patients was developed. The assay was simple, rapid, and reproducible. The test completion time was 2 min, and the assay required no equipment. The positive dot was very obvious, and the results could easily be determined with the naked eye. A total of 350 serum samples were examined by DIGFA, the rapid plasma reagin test, and the fluorescent treponemal antibody-absorption test. The levels of agreement between DIGFA and the rapid reagin test and between DIGFA and the fluorescent treponemal antibody-absorption test were 100 and 98%, respectively. The results of clinical application indicated that DIGFA could be used as a routine screening test for syphilis.

Antibodies, Bacterial↗

PubMed1996

Expression of an equine herpesvirus 1 ICP22/ICP27 hybrid protein encoded by defective interfering particles associated with persistent infection.

Defective interfering (DI) particles of equine herpesvirus type 1 (EHV-1) are capable of mediating persistent infection (S. A. Dauenhauer, R. A. Robinson, and D. J. O'Callaghan, J. Gen. Virol. 60:1-14, 1982; R. A. Robinson, R. B. Vance, and D. J. O'Callaghan, J. Virol. 36:204-219, 1980). Sequence analysis of cloned DI particle DNA revealed that portions of two regulatory genes, ICP22 (IR4) and ICP27 (UL3), are linked in frame to form a unique hybrid open reading frame (ORF). This hybrid ORF, designated as the IR4/UL3 gene, encodes the amino-terminal 196 amino acids of the IR4 protein (ICP22 homolog) and the carboxy-terminal 68 amino acids of the UL3 protein (ICP27 homolog). Portions of DNA sequences encoding these two regulatory proteins, separated by more than 115 kbp in the standard virus genome, were linked presumably by a homologous recombination event between two identical 8-bp sequences. Reverse transcriptase-PCR and S1 nuclease analyses revealed that this unique ORF is transcribed by utilizing the transcription initiation site of ICP22 and the polyadenylation signal of ICP27 in DI particle-enriched infection. Immunoprecipitation and Western blot (immunoblot) analyses with antisera to the ICP22 and ICP27 proteins demonstrated that a 31-kDa hybrid protein was synthesized in the DI particle-enriched infection but not in standard virus infection. This 31-kDa hybrid protein was expressed at the same time as the ICP22 protein in DI particle-enriched infection and migrated at the same location on polyacrylamide gel electrophoresis as the protein expressed from a cloned IR4/UL3 expression vector. These observations suggested that the unique IR4/UL3 hybrid gene is expressed from the DI particle genome and may play a role in DI particle-mediated persistent infection.

Amino Acid Sequence↗

PubMed1996

Effects of a naturally occurring mutation in the hepatitis B virus basal core promoter on precore gene expression and viral replication.

The basal core promoter (BCP) of hepatitis B virus (HBV) controls the transcription of both the precore RNA and the core RNA. The precore RNA codes for the secreted e antigen, while the core RNA codes for the major core protein and the DNA polymerase and also is the pregenomic RNA. The double mutation of nucleotides 1762 and 1764 in the BCP from A and G to T and A, respectively, is frequently observed in HBV sequences isolated from chronic patients. Several papers have reported conflicting results regarding whether this double mutation is important for e antigen expression. In order to address this issue, we have introduced this double mutation into the HBV genome and studied its effects on HBV gene expression and replication. Our results indicate that the mutated BCP can no longer bind a liver-enriched transcription factor(s) and that the transcription of only precore RNA and, consequently, the expression of e antigen were reduced. The reduction of precore gene expression was accompanied by an increase in progeny virus production. This increase was found to occur at or immediately prior to the encapsidation of the pregenomic RNA. Thus, the results of our in vitro study resolve the discrepancy of previous clinical observations and indicate that this double mutation suppresses but does not abolish the e antigen phenotype. The implications of these findings in the pathogenesis of HBV are discussed.

Animals↗

PubMed1996

Prostate carcinoma: MR imaging findings after cryosurgery.

PURPOSE: To determine the morphologic changes in the prostate gland after cryosurgery and to assess the value of magnetic resonance (MR) imaging in detecting residual tumor. MATERIALS AND METHODS: Twenty-seven patients with prostate carcinoma underwent endorectal MR imaging after cryosurgery. Eighteen patients also underwent MR imaging before cryosurgery. MR findings were correlated with posttreatment clinical data, prostate-specific antigen level, and biopsy results. RESULTS: Mean prostate volume had decreased by 52% in patients examined 8 weeks or more after cryosurgery. Zonal differentiation was lost in 22 of 27 patients (81%). Areas of intraprostatic necrosis were identified in 14 of 27 patients (52%). A thick capsule enveloped the gland and neurovascular bundles in 24 of 27 patients (89%). Osteonecrosis of the symphysis pubis was seen in six of 27 patients (22%). Positive and negative predictive values for MR assessment of recurrent tumor were 44% and 73%, respectively. CONCLUSION: Cryosurgery-induced changes in the prostate gland preclude reliable detection of residual prostate carcinoma at MR imaging.

Aged↗

PubMed1996

SERCA2a and mitochondrial cytochrome oxidase expression are increased in hearts of exercise-trained old rats.

Aging of rats results in slower activities of calcium transport by cardiac calcium adenosinetriphosphatase (ATPase) of the sarcoplasmic reticulum (SR) and mitochondrial cytochrome oxidase (COX). These enzyme activities are faster after exercise training of previously sedentary old rats. Our purpose was to determine whether the expression of the genes encoding SR calcium ATPase (SERCA2a) or COX is altered by exercise training. Old (24-mo-old) male Fischer 344 rats were assigned to SO (sedentary old) or EO (exercised old) groups and compared with younger (12-mo-old) sedentary rats (SM). EO rats were trained on a treadmill for 8-10 wk. SERCA2a and COX mRNAs were lower (P < 0.05) in SO compared with SM and EO, whereas glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and cardiac alpha-actin mRNAs were similar across groups. The immunoreactive protein contents of cardiac calcium ATPase, cytochrome c, sarcomeric actin, and GAPDH followed the changes, when observed, in mRNA contents. Thus pretranslational mechanisms may be modified in some genes during aging and exercise training of previously sedentary old rats.

Aging↗

PubMed1996

Synthesis and secretion of endothelin in a cortical collecting duct cell line.

Previous experiments have shown that epithelial cells in the renal medulla produce endothelin-1 (ET-1) and possess ETB receptors. It has been suggested that medullary ET-1 may affect water and sodium absorption along the collecting ducts in an autocrine fashion. To study possible mechanisms responsible for the regulation of medullary ET-1 production, experiments were performed in M-1 cells and mIMCD-K2 cells, cell lines derived from cortical and inner medullary collecting ducts of SV40 transgenic mice, grown to confluence on collagen-coated filter inserts. Both cell lines were found to express ET-1 mRNA and to secrete ET almost exclusively into the basolateral medium as long as the transepithelial resistance was high. Inhibition of transcription with actinomycin D was followed by a decline in both ET mRNA [halftime (t1/2) = 30 min] and ET secretion (t1/2 = approximately 90 min). The addition of arginine vasopressin (AVP, 10(-8) M; 2- or 4-h exposure) or incubation of M-1 cells in hypertonic media (+50 mM NaCl, 4- or 6-h exposure) did not significantly alter ET secretion or ET-1 mRNA expression. In contrast, simultaneously increasing AVP(10(-8) M in the basolateral medium) and tonicity (+50 mM NaCl) for 4 h increased ET secretion (from 28.9 +/- 3.9 to 41.8 +/- 3.8 pg.h-1.mg protein-1; P = 0.029, n = 10) and ET-1 mRNA (control = 2,138 cpm/microliter, log of 3.33 +/- 0.048, n = 4; AVP + NaCl = 3,548.1 cpm/microliter, log of 3.55 +/- 0.09; P = 0.045, n = 5). Exposure of M-1 cells to hypertonic media (+50 mM NaCl or 100 mM mannitol) for 24 h was associated with a marked reduction of ET secretion (-83.9% with NaCl and -78.4% with mannitol; P < 0.0001). This reduction was attenuated, but not prevented, by the presence of AVP in the basolateral medium (-40%). ET-1 mRNA, in contrast, did not change with 24-h exposure to hypertonic media and increased when AVP was present. Results are compatible with the concept that generation of ET by collecting duct cells may contribute in a complex and time-dependent fashion to the paracrine control of collecting duct cell function.

Animals↗

PubMed1996

Inducible nitric oxide synthase expression in cerebrovascular smooth muscle and neutrophils after traumatic brain injury in immature rats.

The inflammatory response after traumatic brain injury (TBI) includes cytokine production, leukocyte infiltration, and microglial activation. Production of nitric oxide by inducible nitric oxide synthase (iNOS) occurs during acute inflammation outside of the CNS and in models of cerebral ischemia, and therefore may contribute to the inflammatory response after TBI. The purpose of this study was to localize and define the time course of iNOS expression after TBI in the immature rat. Immature Wistar rats (age 3.5-4.5 wk) were anesthetized and subjected to percussive trauma to the right parietal cortex. Nontraumatized rats were used as controls (n = 7). At 2, 24, 48, or 168 h (n = 3/group) posttrauma rats were killed by perfusion fixation. Brains were removed, frozen, sectioned, immunostained with antibodies against iNOS and glial fibrillary acidic protein (GFAP, a marker specific for astrocytes), and imaged using fluorescent detection systems. There was no detectable expression of iNOS in control brains. At 2h, minimal cerebrovascular iNOS expression was seen in the peritrauma area. At 24 and 48 h, there was marked peritrauma cerebrovascular iNOS expression that appeared to be restricted to vascular smooth muscle cells and infiltrated leukocytes. Further dual-immunolabeling showed that the leukocytes expressing iNOS were predominantly neutrophils. At 168 h, iNOS expression was no longer detectable. iNOS was not detectable in GFAP-positive cells. The prominent expression of iNOS protein after TBI in cerebrovascular smooth muscle cells and infiltrated neutrophils suggests that iNOS may play a role in cerebrovascular disturbances and secondary brain injury after trauma.

Animals↗

PubMed1996

Identification of calcium-dependent phospholipase A2 isoforms in human and rat pancreatic islets and insulin secreting beta-cell lines.

Phospholipase A2 (PLA2) and its end product, arachidonic acid, are thought to be important signaling components in insulin secretion from pancreatic beta-cells. Because there are multiple Ca2+ -dependent and independent PLA2 biochemical activities in beta-cells, we have used a combination of molecular and immunological techniques to identify the isoforms of Ca2+ -dependent PLA2 present in pancreatic beta-cells. Total RNA extracted from the purified rat and human pancreatic islets and from insulin-secreting beta-TC3 and beta-HC6 cells was used as a template for complementary DNA (cDNA) synthesis. The RT-PCR was performed based on the oligonucleotide primers designed for the 14- kDa type II PLA2 and the 85-kDa cytosolic PLA2. The PCR products for both enzymes yielded single bands (375 bp and 910 bp for type II PLA2 and cytosolic PLA2, respectively). The PCR-generated cDNA fragments were confirmed to be identical to the type II and cytosolic PLA2 isoforms expressed in rat tissues, U937 cells, and A9 cells by DNA sequencing of the PCR products. The presence of these two isoforms of PLA2 was further confirmed by immunoblotting of extracts of pancreatic islets and beta-cells using specific antibodies directed toward each type of PLA2. Demonstration of the presence of type II and cytosolic PLA2 isoforms in islets provides the framework for further investigation of the regulation of PLA2 isoforms and their role in insulin secretion.

Animals↗

PubMed1996

Na concentration of CSF and cardiovascular adjustment in thermally dehydrated rats.

To assess the effect of the Na concentration of cerebrospinal fluid (CSF) on arterial pressure regulation under dehydration, the Na concentration in CSF ([Na]csf) was measured continuously with an Na-sensitive electrode in the lateral ventricle (LV) together with mean arterial pressure (MAP), central venous pressure (CVP) and cardiac output (CO) during and after hypotonic infusion (INF, approximately 200 mOsm/kg H2O, 5 microliters/min for 10 min) into the LV of dehydrated and pentobarbital anesthetized rats. Total vascular conductance (TVC) was calculated from CO/(MAP-CVP). Sino-aortic denervation was performed to eliminate any influence of arterial baroreflexes on MAP changed by [Na]csf. After dehydration, MAP increased by 20 mmHg (p < 0.001) despite a 9% decrease in blood volume (p < 0.001) while [Na]csf increased by 13 mEq/kg H2O (p < 0.001). This increase in MAP was attributed to the decrease in TVC. After hypotonic INF began, MAP and [Na]csf decreased and the maximal decreases of -13 mmHg (p < 0.001) and -28 mEq/kg H2O (p < 0.001) were found at the end of INF, respectively. The changes in MAP (r = 0.97, p < 0.001), CO (r = 0.97, p < 0.001), and TVC (r = -0.90, p < 0.001) were highly correlated with the change in [Na]csf during INF and the following 10-min recovery. These results suggest that the increase in [Na]csf is involved in the maintenance of arterial pressure by decreasing TVC and by attenuating the decrease in CO caused by dehydration-induced hypovolemia.

Adrenalectomy↗

PubMed1996

Hormonal ablation of prostatic cancer: effects on prostate morphology, tumor detection, and staging by endorectal coil MR imaging.

OBJECTIVE: The purpose of our study was to evaluate the effect of androgen deprivation therapy on the MR imaging findings of prostate gland anatomy and cancer pathology in men with prostatic cancer treated with hormonal ablation before radical prostatectomy. MATERIALS AND METHODS: Twenty-two patients (mean age, 66 years old) were divided into two groups: in group I (n=10), MR imaging studies were done before and after hormonal treatment; in group II (n=12), MR imaging studies were done only after hormonal treatment. MR imaging was performed on a 1.5 T-scanner (Signa; General Electric Medical Systems, Milwaukee, WI) and included transverse plane phased-array coil T1-weighted images (TR/TE, 600/12), combined endorectal phased-array coil transverse plane T1-weighted images, fast spin-echo T2-weighted (4000/102), and coronal plane fast spin-echo T2-weighted images. Image evaluation was by consensus and included assessment of the gland size, signal intensity, tumor depiction, extracapsular extension, seminal vesicle invasion, and overall staging accuracy (Jewett and Whitmore classification). MR imaging findings were correlated with pathologic findings of step section radical prostatectomy. RESULTS: After hormonal therapy, the volume of the prostate gland showed a mean decrease of 33.5% +/- 19.6% SD (range, 0-64%). Volume reduction in the transition zone (mean 29.2% +/- 22% SD) was less than in the peripheral zone (mean, 55.8% +/- 25.8% SD) (p < .05). On T2-weighted images, the peripheral zone showed homogeneous decreases in signal intensity in 13 of 22 (58%) patients. Compared with pathologic findings, the accuracy of tumor detection by MR imaging was 74% (98 of 132 sites). Tumor presence was overestimated in 32 of 132 (24%) sites. Overall staging accuracy after hormonal ablation was 68% (15 of 22). The positive predictive value and negative predictive value for extracapsular extension were 57% (13 of 23 sites) and 90% (19 of 21 sites), respectively, and for seminal vesicle invasion were 80% (8 of 10 sites) and 97% (33 of 34 sites), respectively. CONCLUSION: As detected by MR imaging, hormonal ablation caused a decrease in size and signal intensity of the prostate gland and seminal vesicles and overestimation of tumor presence and extracapsular extension.

Aged↗

PubMed1996

[Relationship between particle fluence and dose in cabin of recoverable satellite].

Radiation doses in the cabin of the recoverable satellite measured by GJ-III personal dosimeters and LiF thermoluminescent dosimeters were presented. The relationship between the particle fluence calculated from instrument display and the measured doses was studied. It shows that the particle fluence can be directly calculated from the measured dose with a transformation coefficient which is not obviously affected by shielding around the measuring site. The estimated average energy of the particles in the cabin is on the level of Galactic cosmic radiation.

Cosmic Radiation↗

PubMed1996

[A quantitative study on mandibular incisor point border movement between the immediate complete denture wearers and the second complete denture wearers]

The mandibular incisor point border movement orbits of 20 patients wearing immediate complete dentures for 3 months and the same patients wearing the second complete dentures for 3 months were raced by SGG then measured and analysed.The results showed that the ranges of mandibular incisor point border movement of patients wearing immediate complete dentures for 3 months were significantly narrower than those of the same patients wearing the second complete dentures for 3 months (P<0.01).The ratio of the former to the latter was only 59.00%-69.96%.It could be concluded that the ranges of mandibular incisor point border movement of immediate complete denture wearers were limited.Thus the immediate complete dentures were transitional dentures and only used for treating purpose.The patients must have another pair of complete dentures after 3 months interim.

Journal Article↗

PubMed1996

Influence of four diets on uric acid metabolism and endogenous acid production in healthy beagles.

OBJECTIVE: To evaluate the influence of 3 diets used to dissolve or prevent ammonium urate uroliths in dogs, and a diet formulated for growth, on 24-hour excretions of uric acid, ammonia, net acid, titratable acid, bicarbonate, and creatinine; 24-hour urine volumes; pH values of 24-hour urine samples; plasma uric acid concentration; serum creatinine concentration; and endogenous creatinine clearance values. DESIGN: Randomized block. ANIMALS: Six reproductively intact female Beagles, 3.9 to 4.2 years old, weighing 8.5 to 11.1 kg. PROCEDURES: Four diets were evaluated for their ability to dissolve magnesium ammonium phosphate hexahydrate (struvite) uroliths (diet S); to minimize uric acid excretion (diet U); to minimize clinical signs associated with renal failure (diet K); and to promote growth in pups (diet P). Each diet was fed for 14 days; then 24-hour urine samples were collected. An adult maintenance diet was fed during a 7-day washout period. RESULTS: Consumption of diet U was associated with lowest plasma uric acid concentration, lowest 24-hour urinary uric acid, ammonia, titratable acid, and net acid excretions, lowest endogenous creatinine clearance values, highest 24-hour urinary bicarbonate excretion and urine pH values, and highest 24-hour urine volumes. Consumption of diet P was associated with opposite results; results of consumption of diets S and K were intermediate between those for diets U and P. CONCLUSION: Consumption of diet U by healthy Beagles is associated with reduced magnitude of urinary excretion of uric acid and ammonia, with alkaluria, and with polyuria, which may be beneficial in the management of ammonium urate uroliths in dogs. CLINICAL RELEVANCE: Results support use of diet U for management of ammonium urate urolithiasis in dogs.

Ammonia↗

PubMed1996

Effects of various methods of preservation on the stability of uric acid in frozen canine urine.

OBJECTIVE: To evaluate the effects of dilution and alkalinization, separately and together, on the stability of uric acid in canine urine stored at -20 C. DESIGN: Prospective-controlled study. ANIMALS: 5 dogs with confirmed ammonium urate uroliths, 6 Beagles, and 6 mixed-breed dogs. PROCEDURE: Dogs were fed a 31.4% protein (dry weight), meat-based diet for 21 days, and urine samples were collected on day 22. Urine samples were preserved, using combinations of dilution and alkalinization, and divided into 1-ml aliquots for storage at -20 C for 1 to 12 weeks. Urine uric acid concentrations were measured, using high-performance liquid chromatography, on day of collection (baseline), and after 1, 2, 4, 8, and 12 weeks. RESULTS: Alkalinization did not have a significant effect on reproducibility of measurements of uric acid concentrations in urine; however, dilution did have a significant effect. Compared with baseline, uric acid concentrations in urine samples collected from dogs with ammonium urate uroliths and Beagles and diluted 1:10 or 1:20 with deionized water were not different after storage for 1 to 12 weeks. Uric acid concentrations in urine samples collected from mixed-breed dogs did not differ from baseline values during the 12-week storage period whether samples were undiluted or were diluted 1:10 or 1:20 with deionized water. CONCLUSIONS: Measurements of uric acid concentration are most reproducible in canine urine samples stored at -20 C for 1 to 12 weeks when samples are diluted 1:20 with deionized water. CLINICAL RELEVANCE: To ensure reproducibility of measurements of uric acid concentration in urine samples collected from dogs affected with urate uroliths, urine should be diluted 1:20 with deionized water. Alkalinization is not necessary, and is not recommended because of the additional step in processing and its potential to interfere with measurement of other urinary analytes.

Animals↗