bluuebunny/crossref_metadata_2025 huggingface dataset:
Suppressive effect of oral administration of branched‐chain amino acid granules on oxidative stress and inflammation in HCV‐positive patients with liver cirrhosis
Authors: Ohno Tomoyoshi , Tanaka Yasuhito , Sugauchi Fuminaka , Orito Etsuro , Hasegawa Izumi , Nukaya Haruhiko , Kato Atsunaga , Matunaga Seijiro , Endo Masayuki , Tanaka Yoshito , Sakakibara Kenji , Mizokami Masashi
Published: March 2008

Aim:  In chronic hepatitis C virus (HCV) infection, it is thought that both chronic persistent inflammation and oxidative stress contribute to the development of hepatocellular carcinoma (HCC), and it has been reported that long‐term oral supplementation with branched‐chain amino acid (BCAA) granules could inhibit liver carcinogenesis. However, the extent of the involvement of these factors remains obscure.

Methods:  To clarify the involvement of inflammation and oxidative stress in the inhibition of liver carcinogenesis, we evaluated the effect of oral administration of BCAA granules on oxidative stress and inflammation in HCV‐positive patients with liver cirrhosis.

Results:  Twenty‐seven patients were enrolled in the study: 18 of the patients were treated with BCAA granules (administered group) and nine were observed without BCAA granules (non‐administered group). In the non‐administered group, the production of oxidative stress, as indicated by urine 8‐hydroxydeoxyguanosine (8‐OHdG) and 15‐F2t‐Isoprostane (8‐IsoPs), significantly increased with time, while in the administered group the levels of ferritin and 8‐OHdG decreased significantly. Comparison of the two groups demonstrated that highly sensitive CRP, ferritin, 8‐OHdG and 8‐IsoPs were significantly reduced by taking BCAA granules. The time‐course analysis showed that ferritin and highly sensitive CRP seemed to decrease first, followed by a decrease of 8‐OHdG and 8‐IsoPs.

Conclusion:  These findings indicated that the administration of BCAA granules influenced microinflammation and the metabolism of iron in HCV‐positive patients with liver cirrhosis, and subsequently seemed to reduce the production of oxidative stress, possibly leading to a decrease in the occurrence of HCC.
HCV selection and HVR1 evolution in a chimpanzee chronically infected with HCV‐1 over 12 years
Authors: Lu Ling , Tatsunori Nakano , Li Chunhua , Waheed Sana , Gao Fengxiang , Robertson Betty H.
Published: March 2008

Aim:  To study hepatitis C virus (HCV) selection and hypervariable region‐1 (HVR1) evolution in a chimpanzee chronically infected with HCV‐1 over 12 years after inoculation with a human factor VIII concentrate contaminated with HCV.

Methods:  From the inoculum, the earliest chimpanzee plasma and 12 annual plasma samples, HCV fragments including HVR1 were amplified followed by cloning and sequencing.

Results:  Five HCV subtypes – 1a, 1b, 2a, 2b, 3a – and multiple 1a strains were identified in the inoculum. Two 1a strains were found in the earliest chimpanzee sample, while a single HCV‐1 strain was detected in the 12 annual samples. None of the chimpanzee sequences were identical to those found in the inoculum. Over 12 years, HVR1 patterns changed irregularly, but a few patterns showed identical nucleotide or amino acid sequences. In the last three years, the variety of HVR1 patterns decreased, while the proportion of major patterns increased. These corresponded to a higher virus load and a lower number of amino acid substitutions. Simultaneously, the HVR1 sequences became more similar to the consensus sequence of the 1a subtype.

Conclusion:  HCV selection was observed from the inoculum to the inoculated chimpanzee and from the early acute hepatitis to the persistent chronic infection. The selection occurred at three levels: among subtypes after transmission, among isolates during acute hepatitis and among quasispecies in chronic infection.
Changes in intrahepatic gene expression profiles from chronic hepatitis to hepatocellular carcinoma in patients with hepatitis C virus infection
Authors: Furuta Keiko , Sato Sayaka , Yamauchi Taeko , Kakumu Shinichi
Published: March 2008

Aim:  DNA microarray technology has enabled genome‐wide analysis of gene transcript levels, which has yielded insight into the molecular nature of hepatitis C virus infection. However, little insight into the molecular nature of the early to advanced stages of chronic liver disease has as yet been obtained.

Methods:  We compared the gene expression profiles of liver biopsy specimens from 14 patients at different stages of chronic hepatitis C. We also evaluated the liver tissue of hepatocellular carcinoma and its surrounding region obtained surgically in seven patients with hepatitis C virus infection.

Results:  Of 21 073 genes, 582 genes showed significant changes in expression levels across the disease group. Twenty‐eight samples from six disease groups clustered according to the histological classification except for 4 samples. A heat map produced by hierarchical clustering revealed nine clusters where gene expression profiles were changed in abundance. Among 44 genes which changed twofold or more in transcript abundance, transcripts from chronic hepatitis tended to be upregulated, and gradually downregulated according to disease progression toward hepatocellular carcinoma in five of nine clusters. In chronic hepatitis, transcripts relating to metabolism and immune response were upregulated, while in hepatocellular carcinoma, transcripts associated with cell cycle, growth, proliferation, apoptosis and signaling pathway were upregulated.

Conclusion:  Disease progression in hepatitis C virus‐infected patients appeared to be associated with changes in gene expression profiles in the liver consistent with plausible functional categories, although we should confirm these findings using larger samples.
A model for estimating cirrhosis in patients with type 1 autoimmune hepatitis
Authors: Miyake Yasuhiro , Iwasaki Yoshiaki , Terada Ryo , Nagano Takuya , Kobashi Haruhiko , Sakaguchi Kohsaku , Shiratori Yasushi
Published: March 2008

Aim:  Longstanding cirrhosis has been recognized as a risk factor for the development of hepatocellular carcinoma in patients with autoimmune hepatitis (AIH). Thus, the accurate determination of cirrhosis is important for prognostication, decisions regarding treatment and monitoring of disease progression. The aim of this study was to identify independent predictors of cirrhosis and to develop a model for estimating cirrhosis in patients with type 1 AIH.

Methods:  Using the training sample, consisting of 121 patients with type 1 AIH, we retrospectively examined independent predictors of cirrhosis and constructed a model for estimating cirrhosis. Validation was prospectively performed in the validation sample, consisting of 35 patients.

Results:  Using a stepwise multiple linear regression analysis, three predictors of serum immunoglobulin A level, ratio of aspartate aminotransferase to alanine aminotransferase, and platelet count were elicited, and a model for estimating cirrhosis was determined as follows: risk score = −0.113 + 0.0006056 × immunoglobulin A (mg/dL) + 0.155 × ratio of aspartate aminotransferase to alanine aminotransferase − 0.007079 × platelet (×104/mm3). In the training sample, the sensitivity and specificity were 90% and 83%, respectively, when patients presenting a risk score ≥0.20 were estimated to be cirrhotic. When this model was applied to the validation sample, the sensitivity, specificity, positive predictive value, negative predictive value and accuracy were 83%, 97%, 83%, 97% and 94%, respectively.

Conclusion:  It is suggested that this model could be useful for the estimation of cirrhosis in patients with type 1 autoimmune hepatitis.
Hematologic Evaluation of Normal and Anemic Lambs with the Technicon H*1 Using EDTA or Heparin as Anticoagulants
Authors: Vatn Synnøve , Framstad Tore , Torsteinbø Wenche O.
Published: March 2008

Abstract: A study was performed to evaluate blood from young lambs using the Technicon H*1 hematology analyzer, with emphasis on RBC parameters, comparison of tripotassium EDTA and heparin, and the effects of storage on heparinized blood. Blood samples from lambs 2 days to 18 weeks of age were analyzed within 6 hours, revealing a high precision, except for WBC counts in heparinized blood. The HCT values estimated by the H*1 correlated well (r2= .90) with those obtained by the microhematocrit method. Mean hematologic values obtained for heparinized blood differed by up to 4% from values obtained for blood collected into EDTA. WBC counts decreased 8.4% in heparinized blood stored at 4°C for 1 day, but differences observed in RBC counts were ≤ 2%. Problems occurred when analyzing blood from young lambs with low hemoglobin values because the H*1 incorrectly counted microcytes with volumes of < 10 fL as platelets. When the necessary corrections were performed, the H*1 was useful for analyzing RBC parameters in lamb blood collected both into EDTA and into heparin.
Discriminant and Multiple Regression Analysis of Anemia and Opportunistic Infection in Fell Pony Foals
Authors: Dixon John B. , Savage Malcolm , Wattret Andrew , Taylor Peter , Ross Gordon , Carter Stuart D. , Kelly Donald F. , Haywood Susan , Phythian Clare , Macintyre Andrew R. , Bell Susan C. , Knottenbelt Derek C. , Green Jack R.
Published: March 2008

Abstract: A condition resembling acquired immunodeficiency with anemia has been described in British Fell pony foals. The pathogenesis is unknown. The present study involved 322 foals of which 164 were of Fell breed and 12 were affected. Discriminant and multiple regression analyses were used to test the results of all clinical pathology parameters from these foals for statistical association with (1) breed origin and (2) presence of disease. Only hematocrit associated significantly with presence of disease. However, significant associations were found with Fell breed origin, including serum albumin concentration and lymphocyte count. The unexpected discovery of sporadic lymphocytosis in the breed indicates the value of this kind of analysis and may be evidence that Fell foal immunodeficiency is associated with retroviral infection.
Diagnosis of Chronic Myelogenous Leukemia in a Dog Using Morphologic, Cytochemical, and Flow Cytometric Techniques
Authors: Tarrant Jacqueline M. , Stokol Tracy , Blue Julia T. , McDonough Sean P. , Farrell Peter
Published: March 2008

Abstract:Chronic myelogenous leukemia was diagnosed in a 3.5‐year‐old neutered male Golden Retriever. The diagnosis was based on persistent leukocytosis (>73.0×103/μL), composed of a proportionate left shift to pro‐granulocytes with no evidence of underlying inflammation, infection, or neoplasia. Marked dysplasia was evident in neutrophils and platelets in peripheral blood. Bone marrow and splenic aspirates were dominated by mature and immature neutrophils with < 2% myeloblasts. Cytochemical and flow cytometric assays confirmed that cells in the peripheral blood and spleen were of committed neutrophil lineage. The dog responded initially to treatment with hydroxyurea, but developed acute undifferentiated leukemia approximately 83 days after initial presentation.
Blood Chemistry Values of Juvenile Red Pacu (Piaractus brachypomus)
Authors: Sakamoto Kaori , Lewbart Gregory A. , Smith Thomas M.
Published: March 2008

Abstract:Blood samples were collected from 29 juvenile red pacu (Piaractus brachypomus), ornamental freshwater fish, to establish baseline blood chemistry values. Mean (minimum‐maximum) values, obtained by automated bichromatic analysis and ion selective electrode analysis, were as follows: sodium, 150.4 (146–159) mmol/L; potassium, 3.93 (2.7–5.0) mmol/L; chloride, 138.7 (128–150) mmol/L; total CO2, 7.5 (6–10) mmol/L; albumin, 0.86 (0.5–1.0) g/dL; lactate dehydrogenase, 237.8 (65–692) IU/L; aspartate aminotransferase, 49.1 (0–125) IU/L; creatinine, 0.31 (0.2–0.4) mg/dL; calcium, 10.80 (9.5–12.5) mg/dL; anion gap, 6.89 (1.2–12.5) mmol/L; and phosphorus, 7.29 (4.1–8.9) mg/dL.
Immunohistochemical Study of a Granular Cell Tumor on the Tongue of a Dog
Authors: Rallis T.S. , Tontis D.K. , Soubasis N.H. , Patsiaura K.K. , Papazoglou L.G. , Adamama‐Moraitou K.K.
Published: March 2008

Abstract: A granular cell tumor (GCT; myoblastoma) was diagnosed on the tongue of a 12‐year‐old English Pointer with clinical signs of mild oral dysphagia. The diagnosis was confirmed by histopathologic examination and immunohistochemistry. The tumor was positive for S‐100 protein, but also was positive for desmin, and was only weakly positive for PAS, which is unusual for GCTs. An epithelioid type of leiomyoma (leiomyo‐blastoma) was considered less likely on the basis of negative staining for smooth muscle actin. Treatment consisted of surgical resection of the tumor. The animal was in excellent clinical condition 1 year after surgery. Although GCT of the tongue has been reported previously in the dog, determining the cell of origin is still problematic. Immunohistochemistry is helpful for histogenetic classification and necessary for differentiation from leiomyoblastoma.
Cytologic Evaluation of Primary and Secondary Myelodysplastic Syndromes in the Dog
Authors: Weiss Douglas J. , Aird Betsy
Published: March 2008

Abstract:Myelodysplastic syndromes are a heterogeneous group of acquired primary and secondary alterations of hematopoietic stem cells that result in cytopenias in blood and cytologic features of dysplasia in blood and/or bone marrow. To better understand the cytologic features that would permit differentiation of primary and secondary forms of myelodysplasia, we reviewed 267 consecutive bone marrow reports from dogs. These reports indicated that 34 dogs (12.7%) had dysgranulopoiesis, dyserythropoiesis, and/or dysthrombopoiesis in >10% of granulopoietic cells, erythroid cells, and/or megakaryocytes, respectively. Thirteen dogs had primary myelodysplastic syndromes, and 21 had secondary myelodysplastic syndromes. Of the 13 dogs with primary myelodysplasia, 4 were subclassified as myelodysplastic syndrome with refractory anemia (MDS‐RA), and 9 were subclassified as myelodysplastic syndrome with excess blasts (MDS‐EB). Secondary conditions associated with dysplasia in the bone marrow included malignant lymphoma (n = 5), myelofibrosis (n = 3), immune‐mediated thrombocytopenia (n = 4), immune‐mediated hemolytic anemia (n = 5), multiple myeloma with mel‐phalan administration (n = 1), pyometra with estrogen administration (n = 1), polycythemia vera (n = 1), and thrombopathia (n = 1). MDS‐RA was characterized by <5% myeloblasts in bone marrow, normal granulocyte maturation ratio, increased erythroid maturation ratio, and dysplastic changes in >15% of erythroid cells. MSD‐EB was characterized by > 5% myeloblasts in bone marrow, high granulocyte maturation and erythroid maturation ratios, >32% dysplastic granulocytes, and the presence of small atypical immature myeloid cells. Secondary myelodysplastic syndromes were characterized by <5% myeloblasts in bone marrow, variable granulocyte maturation and erythroid maturation ratios, and variable dysplastic features. These results indicate that morphology alone cannot be used to distinguish primary and secondary myelodysplastic syndromes in dogs.
Vortex Mixing of Feline Blood to Disaggregate Platelet Clumps
Authors: Tvedten Harold , Korcal David
Published: March 2008

Abstract: Platelet clumping is a common cause of erroneous platelet counts in cats. Mixing of blood with a vortex mixer was evaluated as a method to disaggregate platelet clumps in feline blood and thus obtain accurate platelet counts. Whole blood samples from 42 cats with platelet clumping and 10 control cats without platelet clumping were mixed for 1 minute at the maximal setting using a standard vortex mixer. Blood smears (for subjective assessment of the type and amount of platelet clumping), platelet counts, and total leukocyte counts were evaluated before and after mixing. Vortex treatment of blood samples with platelet clumps caused an increased platelet count in all but 1 sample. Although most samples had strong increases in platelet counts after mixing, only a minority of samples (5 of 42) appeared to have all platelet clumps dispersed. Of 39 feline blood samples with platelet counts initially <200×109 cells/L, 23 counts increased to >200×109 cells/L and 34 counts increased to >100×109 cells/L. Overall, mixing gave inconsistent and partial improvement in platelet counts. Total leukocyte counts were not significantly affected by vortex mixing. Vortex mixing of 10 feline blood samples without platelet clumping had no consistent effect on platelet or WBC counts. In conclusion, vortex mixing of feline blood does not appear to be a consistent means of correcting the problem of feline platelet clumping.
Hyperlipasemia in 6 Dogs with Pancreatic or Hepatic Neoplasia: Evidence for Tumor Lipase Production
Authors: Quigley Kelley A. , Jackson Marion L. , Haines Deborah M.
Published: March 2008

Abstract:A marked increase in serum pancreatic lipase (PL) activity with minimal concurrent increase in serum α‐amylase activity was observed in 6 dogs with pancreatic or hepatic neoplasia. Serum PL activity ranged from 5410 U/L to 42,900 U/L, 11 to 93 times the upper reference limit for our laboratory. Neoplasms included pancreatic adenocarcinoma (n=3), endocrine carcinoma (n=2), and hepatic carcinoma of unknown origin (n=1). Tumor tissue and normal canine pancreatic tissue were stained histochemically with diastase periodic acid‐Schiff to demonstrate zymogen granule content, and immunohistochemically with human monoclonal antibody for PL. Normal canine pancreatic tissue stained positive for both zymogen granule content and PL. Five of the 6 tumors stained positive for zymogen granule content, and 2 of the 6 tumors stained positive for PL. Together, the his‐tochemical and immunohistochemical staining patterns suggested tumor PL production in 5 of the 6 dogs. Marked, unexplained hyperlipasemia may be a noninvasive indicator and biochemical marker for neoplasia of the pancreas and liver in dogs.
Evaluation of a Citrate‐Based Anticoagulant with Platelet Inhibitory Activity for Feline Blood Cell Counts
Authors: Norman Elizabeth J. , Barron Ronnie C. J. , Nash Andrew S. , Clampitt Roger B.
Published: March 2008

Abstract: Aggregation of feline platelets in vitro results in difficulty assessing platelet number. A citrate‐based anticoagulant containing the platelet inhibitors theophylline, adenosine, and dipyridamole (CTAD; Diatube‐H, Becton Dickinson, Oxford, UK) has been developed for use in human platelet studies and heparin assays. To evaluate the efficacy of CTAD in reducing platelet aggregation in feline blood samples, aliquots of blood from 51 cats were anticoagulated with EDTA, CTAD, and for 12 samples, citrate solution. Samples preserved in CTAD had significantly higher (P ≤ .001) platelet counts, as determined by an impedance counter, hemacy‐tometer, and smear estimation, than samples preserved in EDTA. In addition, subjective assessment of blood smears showed significantly fewer platelet aggregates (P<.001) in CTAD‐treated samples compared with EDTA samples. Although values were similar, automated platelet counts and smear estimates of platelet number were significantly higher (P < .05) and platelet aggregation was significantly less (P < .05) in CTAD samples than in citrate samples. These results suggest that the platelet inhibitory activity of CTAD reduced feline platelet aggregation. Automated total WBC counts in CTAD samples were significantly lower (P<.001) than automated counts in EDTA samples but were similar to manual WBC counts in EDTA samples. Differences in both platelet and WBC counts between CTAD and EDTA or citrate samples were clinically relevant. Mean platelet volume and MCV were significantly lower (P< .05) in CTAD samples than in EDTA samples. No effect was seen on cell morphology or staining characteristics. The anticoagulant CTAD offers an advantage over both EDTA and citrate for feline hematologic analysis, by decreasing pseudothrombocytopenia and pseudoleukocytosis.
The Usefulness and Limitations of Hand‐held Refractometers in Veterinary Laboratory Medicine: An Historical and Technical Review
Authors: George Jeanne W.
Published: March 2008

Abstract:Medical hand‐held refractometers have been used in veterinary practice since their development in the 1960s. They have become ubiquitous for the measurement of protein and urine solute concentrations because of their rapidity of analysis, ease of use, and relatively low cost. Refraction of light offers advantages for the determination of solute concentrations because the measurement requires no chemical alteration of the specimen. Numerous authors have reported that the results of protein estimation by refractometry for domestic mammals correlate well with those obtained by the biuret method, although others have reported both higher and lower refractometric results compared with biuret results. Major discrepancies between biuret and refractometric results have been reported for avian samples. Some of the variation in reported results may be due to differences in design by refractometer manufacturers. Another possible source may be variation in the biuret reagent mixture and assay conditions. Refractometers also can be used to calculate serum water concentration. A table that converts index of refraction to serum water concentration can be used to convert electrolyte concentration from mmol/L of serum to mmol/L of serum water, a more accurate indicator of effective electrolyte concentration. Refractometers are especially useful for determining urine specific gravity on veterinary samples because they require relatively small sample volumes. Specific gravity continues to be the most common unit for reporting total solids concentration. Some solutes, such as acetone, may cause false increases in specific gravity by refractometry, as they increase refraction but are less dense than water.
Biochemical and Antioxidant Changes in Plasma and Erythrocytes of Pentathlon Horses Before and After Exercise
Authors: Balogh N. , Gaál T. , Ribiczeyné P. Sz. , Petri Á.
Published: March 2008

Abstract:Physical exercise in the horse induces a series of normal physiological and biochemical adaptations. Increasing metabolism and oxygen uptake may induce oxidative stress in various organs. The aim of this study was to examine exercise‐induced changes in some plasma and RBC biochemical and antioxidant variables in pentathlon horses. Blood samples were taken from 14 horses before, immediately after, and 24 hours after competing in two 1‐minute runs of intense exercise over jumps. The peak intensity periods were preceded by a 20‐minute warm‐up and separated by a 20‐minute break. The following plasma biochemical analytes were determined: total protein, uric acid, and lactate concentrations, and lactate dehydrogenase (LDH) and creatine kinase (CK) activities. Total antioxidant status (TAS) and the ferric reducing ability of plasma (FRAP) also were measured. Thiobarbituric acid‐reactive substances (TBARS), reduced glutathione (GSH), and total protein concentrations, and glutathione peroxidase (GSHPx) and superoxide dismutase (SOD) activities were determined in RBC hemolysates. Significantly increased concentrations of total protein, lactate, and FRAP, and increased activities of CK and LDH were observed immediately postexercise compared with pre‐exercise samples (P < .05). All results returned to approximately initial values after 24 hours of rest. RBC GSH and TBARS concentrations did not change immediately after exercise, but decreased after 24 hours of rest (P < .05). Plasma uric acid and FRAP values were positively correlated in a linear model (r= .78). In summary, the type of exercise applied in this study, which can be considered quite usual for pentathlon horses, caused detectable biochemical and lipid peroxidative changes in plasma and RBCs. FRAP and TAS values changed in opposite directions, indicating that when antioxidant capacity is assessed using different methods, highly different results may be obtained.
Fine Needle Aspiration Cytology of Pilomatricoma in Three Dogs
Authors: Masserdotti Carlo , Ubbiali FedericaAnnamaria
Published: March 2008

Background — Fine needle aspiration cytology is being used for the diagnosis of various neoplasms, but we are unaware of reports dealing with the cytologic features of canine pilomatricoma.

Objective — The purpose of this report was to describe the cytologic features of pilomatricoma in 3 dogs.

Methods — Fine‐needle aspirates were obtained from cutaneous masses using a 25‐ga needle. Smears were prepared and stained with May‐Grünwald‐Giemsa and hematoxylin and eosin. The cutaneous masses were excised and routinely processed for histologic examination.

Results — Cytologic features of all 3 tumors included high cellularity and numerous clusters of tightly arranged of basaloid cells with evenly distributed chromatin and small distinct nucleoli surrounding sheets of “ghost cells”. Ghost cells were characterized by a central unstained zone corresponding to the site previously occupied by the nucleus. Amorphous keratinized material was observed in 1 tumor. Histopathologic findings confirmed the diagnosis of pilomatricoma in all 3 cases.

Conclusions — Basaloid cells in association with ghost cells are important cytopathologic criteria that may provide a definitive cytologic diagnosis of pilomatricoma in dogs, and may help avoid a false diagnosis of malignancy.
Biclonal Gammopathy Associated with Immunoglobulin A in a Dog with Multiple Myeloma
Authors: Ramaiah Shashi K. , Seguin M. Alexis , Carwile Henry F. , Raskin Rose E.
Published: March 2008

A 10‐year‐old neutered male Airedale Terrier was evaluated for inappetance, weight loss, and lameness. Multiple myeloma was diagnosed based on bone marrow plasmacytosis, multiple lytic bone lesions, and hyperglobulinemia with a clonal gammopathy on serum protein electrophoresis. Splenic plasmacytosis, and retinal lesions consistent with hyperviscosity syndrome also were found. Temporary responses to 2 different chemotherapy protocols (melphalan and prednisone, and cyclophosphamide and prednisone) were seen, with remission of clinical signs and a decrease in the biclonal gammopathy but no resolution of the splenic mass. Eventual return of clinical signs led to euthanasia at 175 days postdiagnosis. Necropsy examination confirmed multiple myeloma involving bone marrow and spleen, and glomerulonephritis. An immunoglobulin‐A (IgA) gammopathy was demonstrated by immunoelectrophoresis; biclonality was ascertained by immunofixation electrophoresis. The clonal components consisted of intact Ig with a heavy chain of the a class and a light chain of undetermined class. To our knowledge, this is the first report of undimerized biclonal gammopathy in a dog caused by a single heavy chain class involving IgA.
Use of the Vettest 8008 and Refractometry for Determination of Total Protein, Albumin, and Globulin Concentrations in Feline Effusions
Authors: Papasouliotis Kostas , Murphy Kate , Dodkin Steve , Torrance Andy G.
Published: March 2008

Background— Pleural and peritoneal effusion is a common clinical finding in feline practice. Determination of fluid albumin (ALB) and globulin (GLOB) concentrations in addition to total protein (TP) concentration can be helpful in diagnosing or ruling out certain diseases in cats, especially feline infectious peritonitis (FIP).

Objective— The objective of this study was to compare effusion TP, ALB, and GLOB results obtained by a refractometer and a bench‐top dry chemistry analyzer with those results obtained by a reference method.

Methods— Twenty‐six pleural and 14 peritoneal effusion samples were analyzed from 40 cats with various diseases. TP and ALB concentrations were determined by a reference automated wet chemistry analyzer (Kone Specific, Kone Instruments, Espoo, Finland), a bench‐top dry chemistry analyzer (Vettest 8008, IDEXX Laboratories Ltd, Chalfont St Peter, UK), and a refractometer (Atago SPR‐T2, Atago Co, Tokyo, Japan). GLOB, albumin to globulin (A/G) ratio, and globulins as a percentage of total proteins (GLOB%) were calculated. Results were analyzed by paired t tests, difference plots, and Deming's regression analysis.

Results— Correlation coefficients (r) for TP with Vettest versus Kone and refractometer versus Kone methods were .97 and .94, respectively. GLOB and GLOB% values were significantly higher and A/G ratios were significantly lower with Vettest versus Kone methods. Correlation coefficients for ALB, GLOB, GLOB% and A/G ratio with Vettest versus Kone methods were .86, .93, .82, and .73, respectively. Although correlation with other methods was good, the refractometer underestimated TP concentrations in 3 samples.

Conclusions— The refractometer is an acceptable method for determination of TP concentration in feline effusions. The Vettest 8008 also is an acceptable method for the determination of TP and ALB concentrations, however, calculated A/G ratios obtained with the Vettest are unacceptable.
Use of the Cell‐Dyn 3500 to Predict Leukemic Cell Lineage in Peripheral Blood of Dogs and Cats
Authors: Fernandes Peter J. , Modiano Jaime F. , Wojcieszyn John , Thomas Jennifer S. , Benson Patricia A. , III Smith Roger , Avery Anne C. , Burnett Robert C. , Boone Laura I. , Johnson Mark C. , Pierce Kenneth R.
Published: March 2008

Background—Morphology and cytochemistry are the foundation for classification of leukemias in dogs and cats. Advances in automated hematology instrumentation, immunophenotyping, cytogenetics, and molecular biology are significantly improving our ability to recognize and classify spontaneous myeloproliferative and lymphoproliferative disorders.

Objective—The purpose of this study was to assess the utility of flow cytometry‐based light scatter patterns provided by the Cell‐Dyn 3500 (CD3500) automated hematology analyzer to predict the lineage of leukemic cells in peripheral blood of dogs and cats.

Methods—Leukemic cells from 15 dogs and 6 cats were provisionally classified using an algorithm based on the CD3500 CBC output data and were subsequently phenotyped by enzyme cytochemistry, immunocytochemistry, indirect flow cytometry, and analysis of antigen receptor gene rearrangement.

Results—The algorithm led to correct predictions regarding the ontogeny of the leukemic cells (erythroid/megakaryocytic potential, myeloid leukemia, monocytic leukemia, chronic granulocytic leukemia, lymphoid leukemia) in 19/21 animals. Mismatches in the WBC impedance count and the WBC optical count in conjunction with microscopic assessment of blasts in the blood were useful for predicting myeloproliferative disorders with erythroid or megakaryocytic potential. The leukocyte light scatter patterns enabled distinction among myeloid leukemias (represented by acute myelomonocytic leukemia, acute monocytic leukemia, chronic granulocytic leukemia) and lymphocytic leukemias (including acute and chronic lymphocytic leukemias). One case of acute lymphocytic leukemia was misidentified as chronic lymphocytic leukemia.

Conclusions—Algorithmic analyses can be applied to data generated by the CD3500 to predict the ontogeny of leukemic cells in the peripheral blood of dogs and cats. This rapid and quantitative technique may be used to improve diagnostic decisions, expand therapeutic choices, and increase prognostic accuracy.
Ehlers‐Danlos—Like Syndrome in 2 Dogs: Clinical, Histologic, and Ultrastructural Findings
Authors: Paciello O. , Lamagna F. , Lamagna B. , Papparella S.
Published: March 2008

Background: Ehlers‐Danlos syndrome comprises a group of rare inherited connective tissue diseases characterized by skin hyperextensibility, joint laxity, skin and vessel fragility, and poor wound healing.

Objective: The purpose of this report was to describe the clinical, histologic, and ultrastructural findings in 2 dogs with collagenopathies consistent with Ehlers‐Danlos syndrome.

Methods: Two dogs were examined clinically; skin extensibility index was calculated. Skin biopsies obtained from the dorsum were examined by light and electron microscopy.

Results: Both dogs had clinical signs of skin hyperextensibility and fragility, lower skin elasticity, vessel fragility, and poor wound healing. One dog had a hip dislocation, and the other had bilateral medial patellar luxation (grade II), subcutaneous hematomas produced by minimal trauma, and generalized periodontitis. Histologic and ultrastructural examination confirmed abnormalities in the structure and arrangement of collagen fibrils. Fibroblasts were characterized by variable dilatation of the rough endoplasmic reticulum, and anomalous elastic fibers (elaunin fibers) were present in the dermis.

Conclusion: Although the primary defects underlying collagenopathies in animals are still unknown, analysis of the ultrastructural changes in collagen fibrils and clinical findings could facilitate better characterization of these disorders in dogs.
Metastatic Balloon Cell Melanoma in a Dog
Authors: Wilkerson Melinda J. , Dolce Karen , DeBey Brad M. , Heeb Heather , Davidson Harriet
Published: March 2008

Background: Balloon cell melanoma is a rare variant of amelanotic melanoma that is difficult to differentiate from sebaceous cell carcinoma, liposarcoma, and other clear cell neoplasms without immunohistochemistry or ultrastructural evidence of melanin or melanosomes.

Objective: The purpose of this report was to describe the clinical, cytologic, histologic, immunohistochemical, and ultrastructural findings in a dog with metastatic balloon cell melanoma.

Methods: A 6‐year‐old female Golden Retriever was evaluated for a white, flocculent infiltrate in the anterior chamber of the left eye and an enlarged left prescapular lymph node. Cytologic evaluation of the eye and lymph node were performed following aqueocentesis and fine‐needle aspiration, respectively. The affected lymph node was examined histologically and stained for cytokeratin, vimentin, S‐100, and Melan A. Following euthanasia a necropsy was performed and samples of the affected lymph node were examined by electron microscopy.

Results: Cytologic examination of the lymph node and aqueocentesis sample revealed round neoplastic cells that had abundant clear vacuolated cytoplasm. A tentative diagnosis of metastatic sebaceous cell carcinoma or clear cell neoplasm was made. Histologically, the affected lymph node had similar polygonal clear cells arranged in sheets and packets divided by delicate fibrovascular stroma. Immunohistochemical staining of the cells was negative for cytokeratin but positive for vimentin, weakly positive for S‐100, and strongly positive for Melan A. At necropsy, metastatic lesions were identified in the diaphragm, heart, lung, kidneys, left eye, prescapular and sublumbar lymph nodes, and multiple skin sites. Ultrastructural examination of neoplastic lymph nodes revealed many membrane‐bound vacuoles, myelinlike figures, and rare melanosomes.

Conclusion: Immunohistochemical staining and ultrastructural features of the neoplastic cells supported a diagnosis of metastatic balloon cell melanoma.
Cytologic Diagnosis of Peritoneal Cestodiasis in Dogs Caused byMesocestoidessp
Authors: Caruso Kimberly J. , James Michael P. , Fisher David , Paulson Roger L. , Christopher Mary M.
Published: March 2008

Background:Canine peritoneal larval cestodiasis caused by tapeworm larvae of the genusMesocestoidesis an uncommon and possibly fatal disease that can result in ascites and peritonitis. Although abdominal fluid analysis is recommended for dogs with ascites, the cytologic features ofMesocestoidesinfection have not been described fully.

Objective:Our goal was to describe the cytologic features ofMesocestoideslarvae and of associated ascitic fluid that was collected from the peritoneal cavity of 4 infected dogs.

Methods:Abdominal fluid was obtained from 4 dogs withMesocestoidessp infection. Gross, chemical, and microscopic evaluations of the fluid were performed using standard techniques.

Results:Cytologic findings in 1 dog included intact acephalic metacestodes (larvae without 4 suckers, not tetrathyridia) in various stages of asexual development, whereas fluid from the other 3 dogs contained primarily calcareous corpuscles, remnant tissue specific to cestodes. Abdominal fluid typically was an exudate, with suppuration, hemorrhage, and evidence of necrosis. Total protein concentrations ranged from 2.4 to 5.3 g/dL.

Conclusions:Abdominal fluid cytology was useful in the diagnosis ofMesocestoideslarval infections in the peritoneal cavity of dogs with ascites. Observation of characteristic calcareous corpuscles or intact metacestodes can provide a defininitive diagnosis of canine peritoneal larval cestodiasis in dogs.
Pneumonia in a Paso‐Fino Mare
Authors: MacNeill Amy L. , Alleman A. Rick , Franklin Robert P. , Long Maureen , Giguère Steeve , Uhl Elizabeth , López‐Martinez Alric , Wilkerson Melinda
Published: March 2008

A 5‐year‐old Paso‐Fino mare was presented for severe respiratory distress. The mare had foaled 2 months prior to presentation. The horse was in poor body condition with a dull hair coat. A mild fever was noted during physical examination and increased bronchovesicular sounds were auscultated. Thoracic radiographs showed an interstitial pattern and an alveolar infiltrate with distinct air bronchograms. Moderate purulent inflammation with increased mucus was observed in tracheal wash fluid, but no infectious agents were identified. A bronchoalveolar lavage (BAL) contained a large amount of mucus and reactive mononuclear phagocytes with variable numbers of intracellular fungal organisms morphologically consistent with Pneumocystis carinii. The mare had undetectable levels of immunoglobulin M (IgM) and decreased IgG levels in the serum. Immunophenotyping revealed decreased expression of major histocompatability complex (MHC) class II molecules. Moderate to marked hyperplasia of type II epithelial cells was present throughout histologic sections of lung, but the fungal organisms were not observed. A culture system has not been developed for diagnosis of P carinii infection. Instead, diagnosis of P carinii pneumonia is achieved by microscopic identification of characteristic morphologic features of the pathogen. Cytologic examination of BAL fluid is the preferred method used to diagnose human infection with P carinii. In humans, the diagnostic sensitivity of cytology is significantly higher than the sensitivity of histopathologic examination of lung biopsies. The difference in sensitivity between BAL cytology and lung histopathology may also apply to the diagnosis of P carinii pneumonia in horses. (MacNeill AL, Alleman AR, Franklin RP, Long M, Giguère S, Uhl E, López‐Martinez A, Wilkerson M. Pneumonia in a Paso‐Fino mare [Pneumocystis carinii pneumonia].
Abdominal Fluid from a Dog
Authors: Fry Michael M. , DeCock Hilde E. V. , Greeley Melanie A. , Vernau William
Published: March 2008

A 2‐year‐old intact female mixed‐breed dog with a 1‐month history of lethargy and anorexia was evaluated for abdominal distension and an abdominal mass. The dog's last heat cycle, her third, was 1 month prior to presentation, and no reproductive cycle abnormalities were noted at any time. Hematologic and serum biochemical abnormalities were consistent with hemorrhage and inflammation. Ultrasonographic examination confirmed a large midabdominal mass and a moderate amount of abdominal fluid. Cytologically, the fluid showed evidence of pyogranulomatous inflammation, hemorrhage, and mesothelial reactivity, as well as ciliated columnar cells and free cilia that were interpreted as likely of oviductal origin. The mass was removed surgically, and the histopathologic interpretation was oviductal hamartoma with marked stroma formation and acute hemorrhage. To the authors' knowledge, this is the first reported case of oviductal hamartoma in any species and the first reported case detailing the finding of ciliated columnar epithelial cells in the abdominal fluid of a dog. Ciliated columnar epithelial cells in abdominal fluid should be considered indicative of a likely underlying oviductal lesion.
Extensive Cutaneous Metastases in a Dog with Duodenal Adenocarcinoma
Authors: Juopperi Tarja A. , Cesta Mark , Tomlinson Lindsay , Grindem Carol B.
Published: March 2008

Abstract: A 6‐year‐old Rottweiler was presented to the North Carolina State University College of Veterinary Medicine for evaluation of multiple cutaneous nodules. The dog had a history of anorexia, vomiting, and hind‐limb paraplegia. Results of cytologic examination of the cutaneous nodules were consistent with a round cell tumor. At necropsy, primary tumors were found coalescing in the duodenum and the pancreas and extending into the associated mesentery. Numerous masses also were found throughout the skin, abdominal and thoracic viscera, and lumbar spinal cord. Histologically, the duodenal tumor had variable morphology, with some areas resembling adenocarcinoma and others resembling anaplastic round cell neoplasia; the skin and other metastatic lesions resembled round cell neoplasia. Immunohistochemistry of the cutaneous, duodenal, and pancreatic masses showed the neoplastic cells were positive for pancytokeratin, supporting an epithelial origin. In addition, low numbers of neoplastic cells were positive for periodic acid‐Schiff and Alcian blue, consistent with acid mucin production by duodenal epithelium. These findings confirmed that the cutaneous nodules were metastatic lesions originating from the duodenal adenocarcinoma. Cutaneous metastasis of intestinal carcinoma is rare in domestic animals. This case demonstrates the potential difficulty in diagnosing metastatic lesions based on cytologic and histologic morphology alone, because the cutaneous metastases may not resemble the primary neoplasm morphologically.
Three Cases of Canine Bile Peritonitis with Mucinous Material in Abdominal Fluid as the Prominent Cytologic Finding
Authors: Owens Sean D. , Gossett Ruanna , McElhaney M. Rebecca , Christopher Mary M. , Shelly Sonjia M.
Published: March 2008

Background: Bile peritonitis is a severe, nonseptic inflammatory response to bile in the peritoneal cavity. It may result from generalized or localized leakage of bile due to spontaneous rupture of the biliary system or as a complication of biliary tract inflammation, obstruction, manipulation, or trauma. Cytologically, bile in abdominal fluid appears as golden‐green granular pigment.

Objective: The purpose of this report is to describe the atypical cytologic features of abdominal fluid in 3 dogs with bile peritonitis.

Methods: As part of a diagnostic workup, abdominal fluid was obtained from 3 dogs with bile peritonitis and analyzed. In 2 dogs, fluid bilirubin concentration was determined and Hall's bile stain, Alcian blue‐periodic acid‐Schiff stain, and Mayer's mucicarmine stain were applied to direct smears of the fluid.

Results: Acellular mucinous fibrillar material in clumps and lakes was the prominent cytologic finding in the abdominal fluid from all 3 dogs. Bile pigment was not observed. Fluid from the 3 dogs contained increased numbers of inflammatory cells, predominantly neutrophils. Total protein concentration ranged from 2.9 to 5.6 g/dL. Fluid total bilirubin concentration was greater than twice that of the concurrent serum bilirubin concentration. Based on results of the special stains, the amorphous material was positive for mucosubstances, but was negative for bilirubin. In all dogs, bile peritonitis originated from a rent in the common bile duct.

Conclusions: Bile peritonitis with fibrillar mucinous material in abdominal fluid has not been described previously in dogs. The material was similar to “white bile” observed in humans and experimentally in dogs as a sequela to extrahepatic biliary tract obstruction. When mucinous material is observed in abdominal fluid from dogs and the fluid bilirubin concentration is greater than twice the serum bilirubin concentration, rupture of the extrahepatic biliary tract should be suspected.
Creatinine in the Dog: A Review
Authors: Braun J. P. , Lefebvre H. P. , Watson A. D. J.
Published: March 2008

Creatinine is the analyte most frequently measured in human and veterinary clinical chemistry laboratories as an indirect measure of glomerular filtration rate (GFR). Although creatinine metabolism and the difficulties of creatinine measurement have been reviewed in human medicine, similar reviews are lacking in veterinary medicine. The aim of this review is to summarize information and data about creatinine metabolism, measurement, and diagnostic significance in the dog. Plasma creatinine originates from the degradation of creatine and creatine phosphate, which are present mainly in muscle and in food. Creatinine is cleared by glomerular filtration with negligible renal secretion and extrarenal metabolism, and its clearance is a good estimate of GFR. Plasma and urine creatinine measurements are based on the nonspecific Jaffé reaction or specific enzymatic reactions; lack of assay accuracy precludes proper interlaboratory comparison of results. Preanalytical factors such as age and breed can have an impact on plasma creatinine (P‐creatinine) concentration, while many intraindividual factors of variation have little effect. Dehydration and drugs mainly affect P‐creatinine concentration in dogs by decreasing GFR. P‐creatinine is increased in renal failure, whatever its cause, and correlates with a decrease in GFR according to a curvilinear relationship, such that P‐creatinine is insensitive for detecting moderate decreases of GFR or for monitoring progression of GFR in dogs with severely reduced kidney function. Low sensitivity can be obviated by determining endogenous or exogenous clearance rates of creatinine. A technique for determining plasma clearance following IV bolus injection of exogenous creatinine and subsequent serial measurement of P‐creatinine does not require urine collection and with additional studies may become an established technique for creatinine clearance in dogs.
Guidelines for Resident Training in Veterinary Clinical Pathology. I. Clinical Chemistry
Authors: Christopher Mary M. , Stokol Tracy , Sharkey Leslie
Published: March 2008

Background: The Education Committee of the American Society for Veterinary Clinical Pathology identified a need for improved structure and guidance of clinical pathology resident training in clinical chemistry.

Objectives: The committee's goal was to develop learning objectives and competencies in knowledge, abilities, and skills in clinical chemistry; provide options and ideas for training activities; and identify clinical chemistry resources useful for clinical pathology faculty, training program coordinators, and residents.

Methods: Guidelines were developed and written with the input of Education Committee members and peer experts.

Results: The primary objectives of clinical chemistry training are: 1) to accrue a thorough, extensive, and relevant knowledge base of the types, principles, and properties of clinical chemistry tests and concepts of pathophysiology in animals; 2) to develop abilities to reason, think critically, and exercise judgment in clinical chemistry data interpretation, investigative problem‐solving, and hypothesis‐driven research; and 3) to acquire technical and statistical skills important in clinical chemistry and laboratory operations.

Conclusions: These guidelines define expected competencies that will help ensure proficiency, leadership, and the advancement of knowledge in veterinary clinical chemistry and provide a useful framework for didactic and clinical activities in resident training programs. The learning objectives can readily be adapted to institutional and individual needs, interests, goals, and resources.