bluuebunny/crossref_metadata_2025 huggingface dataset:
Higher optical density of an antigen assay predicts thrombosis in patients with heparin‐induced thrombocytopenia
Authors: Altuntas Fevzi , Matevosyan Karen , Burner James , Shen Yu‐Min , Sarode Ravindra
Published: March 2008

Objectives:  To correlate optical density and percent inhibition of a two‐step heparin‐induced thrombocytopenia (HIT) antigen assay with thrombosis; the assay utilizes reaction inhibition characteristics of a high heparin concentration.

Patients and Methods:  Patients with more than 50% decrease in platelet count or thrombocytopenia (<150 × 109/L) after exposure to heparin, who had a positive two‐step antigen assay [optical density (OD) >0.4 and >50 inhibition with high concentration of heparin] were included in the study.

Results:  Forty of 94 HIT patients had thrombosis at diagnosis; 54/94 had isolated‐HIT without thrombosis. Eight of the isolated‐HIT patients developed thrombosis within the next 30 d; thus, a total of 48 patients had thrombosis at day 30. At diagnosis there was no significant difference in OD between HIT patients with thrombosis and those with isolated‐HIT. However, OD was significantly higher in all patients with thrombosis (n = 48, 1.34 ± 0.89), including isolated‐HIT patients who later developed thrombosis within 30 d (n = 8, 1.84 ± 0.64) as compared to isolated‐HIT patients who did not develop thrombosis (0.96 ± 0.75; P = 0.011 and P = 0.008). The Receiver Operative Characteristic Curve showed that OD >1.27 in the isolated‐HIT group had a significantly higher chance of developing thrombosis by day 30. None of these groups showed significant difference in percent inhibition. Multivariate analysis showed a 2.8‐fold increased risk of thrombosis in females. Similarly, thrombotic risk increased with age and OD values.

Conclusion:  Higher OD is associated with significant risk of subsequent thrombosis in patients with isolated‐HIT; percent inhibition, however, was not predictive.
Development of a non‐invasive measurement system to the thickness of the subcutaneous adipose tissue layer
Authors: Choi Yeon Shik , Hong Hyuck Ki , Kim Beom Joon , Kim Myeung Nam , Park Hyo Derk
Published: March 2008

Background/purpose:  It is important to measure the thickness of hypodermis, including the subcutaneous fat layer in several fields such as global assessment of nutritional status and monitoring of dietary manipulation. It also provides useful information concerning the amount of peripheral adipose tissue and can be used as an index of obesity.

Methods:  To measure the thickness of the subcutaneous adipose tissue layer, the optical properties of tissue components, several lipids and animal tissue samples in vitro were measured over the wavelength range from 1000 to 1700 nm, using the transmittance measurement system and the diffuse reflectance measurement system. A wavelength range for the thickness measurements was selected from the experimental result. The corresponding signal according to the changes of adipose tissue thickness was measured.

Results:  Experimental data measured by the non‐invasive measurement system were compared with the actual thickness of animal samples, and the results could be explained by the three‐layered tissue model. The result showed high correlation to the thickness changes (R2 = 0.9954).

Conclusions:  The experimental result implies positive possibility for the development of a system to measure the thickness of subcutaneous adipose tissue using near infrared in vivo.
Are growth and recapture of hatchery‐reared and resident brown trout (Salmo truttaL.) density dependent after stocking?
Authors: Baer J. , Brinker A.
Published: March 2008

Abstract – The population density of brown trout (Salmo trutta) in a small natural system was manipulated in six equal‐length stream sections by stocking hatchery‐reared 1+ brown trout (unstocked, tripled and quintupled) over two consecutive years. The results showed that hatchery‐reared trout grew more slowly and were more mobile than resident trout, and that their growth was inversely density dependent. In contrast, growth of the resident trout was density independent. The recapture of 1+ resident and hatchery‐reared trout was inversely density dependent. This is most likely a consequence of increased competition. However, after a single winter the population density returned to its base level prior stocking and older resident trout showed no density‐dependent recapture. Thus, the advantage of stocking, here higher biomass, may have the detrimental effect of decreasing resident stocks of the same size class during summer.
FK506 and Erectile Function Preservation in the Cavernous Nerve Injury Model: Optimal Dosing and Timing
Authors: Mulhall John P. , Müller Alexander , Donohue John F. , Golijanin Dragan , Tal Raanan , Akin-Olugbade Yemi , Kobylarz Keith , Cohen-Gould Leona , Bennett Nelson E. , Scardino Peter
Published: March 2008

Introduction

The immunophilin-ligand FK506 has been shown to ameliorate erectile function and preserve cavernous nerve (CN) architecture in short-term-studies using rat models of CN injury.

Aim

The aim of this series was to ascertain the optimal dose and timing of FK506 administration in this animal model.

Methods

Rats underwent bilateral CN crush and were treated with FK506 at different time points. There were control (C) and sham groups for each time point. Based on preliminary experiments, the CN-crush rats had no treatment (C) or either FK506 1 mg/kg (BL) or 3.2 mg/kg (BH) for 3 days prior to and the day of CN crush (PRE), on the day of and for 3 days following CN crush (POST) and for 3 days pre-, on the day of, and 3 days post-CN crush (PP).

Main Outcome Measurements

All animals had measurement of intracavernosal pressure/mean arterial blood pressure (ICP/MAP) ratios at 28 days post-CN crush. Structural analysis was conducted in the POST groups. Penile tissue was assessed for apoptosis with terminal deoxynucleotidyl transferase biotin-dUTP nick end labeling assay and immunohistochemically for neural factors (growth associated protein 43 [GAP43], nerve growth factor [NGF], and neural nitric oxide synthase [nNOS]). The CN architecture was examined by transmission electron microscopy (TEM).

Results

Sham animals had an ICP/MAP ratio of 70%. Only the BH-POST group revealed an improved ICP/MAP ratio compared with C (50 ± 9% vs. 32 ± 8%, P <0.01). nNOS staining was significantly restored reaching sham levels in BL-POST and BH-POST groups vs. C (P <0.05). NGF and GAP43 staining displayed no significant differences between C and treatment groups (P <0.05). Apoptosis was significantly reduced in BL-POST and BH-POST groups compared with C (16 ± 4%, 21 ± 9%, and 63 ± 7%, P <0.001). TEM exhibited preservation of CN architecture for BH-POST compared with C.

Conclusion

These results suggest that short-term treatment with doses of FK506 higher than previously utilized preserves erectile function in the rat CN-injury model. Pretreatment appears to offer no advantage. However, FK506 administration just prior to CN injury and for a short-time post-injury achieves the best functional and structural preservation outcomes.
Iodine biofortification and antioxidant capacity of lettuce: potential benefits for cultivation and human health
Authors: Blasco B. , Rios J.J. , Cervilla L.M. , Sánchez‐Rodrigez E. , Ruiz J.M. , Romero L.
Published: March 2008

Iodine is considered an essential trace element for mammals, and its deficiency is related to numerous pathologies as severe as goitre, reproductive failure, mental retardation and brain damage, among others. Currently, about 30% of the world’s population are affected by this deficiency, and thus, in an attempt to ameliorate these nutritional disorders, we propose a biofortification programme with iodine by an application of different dosages and forms of this element (iodide versus iodate) in lettuce plants. In this work, a study has been made of the iodine concentration in roots and edible leaves and their influence on nutritional quality through an analysis of its antioxidant capacity. The results showed that the most appropriate application rates in hydroponic cultivation were 40 μM or lower in the form I− because these concentrations did not reduce biomass in the treated plants with respect to control plants and caused a foliar accumulation of this element that guarantees the viability of this type of programmes. Furthermore, these data are novel, given that the treated plants show a significant increase in antioxidant compounds after the application of iodine.
Mechanism of strong resistance to Potato leafroll virus infection in a clone of potato (Solanum tuberosum)
Authors: Solomon‐Blackburn R.M. , Nikan J. , Barker H
Published: March 2008

The Solanum tuberosum clone G8107(1) is highly resistant to Potato leafroll virus (PLRV) accumulation following graft inoculation and to PLRV infection by aphids. In glasshouse/insectary experiments, plants were inoculated with viruliferous aphids (Myzus persicae); infection was detectable in some inoculated leaves (by enzyme‐linked immunosorbent assay or reverse transcription polymerase chain reaction) but not in petioles, stems or non‐inoculated leaves of inoculated plants. Secondary infection in tuber‐progeny plants of inoculated plants was rare in G8107(1) (detected in only one of 86 tuber‐progeny plants of 24 inoculated plants), while universal in susceptible controls. This resistance did not break down when tuber sprouts were inoculated or when a more efficient M. persicae vector clone was used. Aphids were able to acquire and transmit PLRV infection from infected plants of G8107(1) to test plants but at a lower frequency than from the susceptible cv. Maris Piper. We suggest that an important element of the very strong resistance of this clone to PLRV infection is resistance to virus movement within leaves or from leaves to petioles, thus preventing spread to the rest of the plant.
Sustained response to interferon‐α plus ribavirin therapy for chronic hepatitis C is closely associated with increased dynamism of intrahepatic natural killer and natural killer T cells
Authors: Yamagiwa Satoshi , Matsuda Yasunobu , Ichida Takafumi , Honda Yutaka , Takamura Masaaki , Sugahara Satoshi , Ishikawa Toru , Ohkoshi Shogo , Sato Yoshinobu , Aoyagi Yutaka
Published: March 2008

Aim:  Previous studies have revealed that functional impairment of innate immune cells, including natural killer (NK) and natural killer T (NKT) cells, might be associated with the persistence of hepatitis C virus (HCV) infection. However, the involvement of innate immune cells, which predominate in the liver, in therapeutic HCV clearance is still unclear.

Methods:  To clarify the role of intrahepatic innate immune cells in the clinical outcome of patients with chronic hepatitis C (CHC) treated with interferon‐α plus ribavirin (IFN/RBV), we prospectively investigated the status of NK and NKT cells in paired liver biopsy and peripheral blood (PB) samples obtained from 21 CHC patients before and immediately after IFN/RBV treatment by flow cytometry. Normal liver and PB samples were obtained from 10 healthy donors for living donor liver transplantation.

Results:  Before treatment, intrahepatic NK and NKT cells constituted a significantly lower proportion in CHC patients than in healthy individuals (P < 0.05). After IFN/RBV treatment, the proportions and absolute numbers of CD3‐CD161+ NK and CD3+CD56+ NKT cells in the liver, but not in PB, were significantly increased in sustained responders (SR) as compared with poor responders (P < 0.05). The proportion of CD3+CD161+ NKT cells was also increased in the liver of SR after the treatment. Moreover, there was a striking increase of activated CD152+ cells among CD3+CD56+ NKT cells in the liver of SR (P = 0.041).

Conclusion:  These findings demonstrate that sustained response to IFN/RBV treatment for patients with CHC is closely associated with increased dynamism of NK and NKT cells in the liver.
Prevalence of esophagogastric varices in patients with non‐alcoholic steatohepatitis
Authors: Nakamura Shinichi , Konishi Hiroyuki , Kishino Maiko , Yatsuji Satoru , Tokushige Katsutoshi , Hashimoto Etsuko , Shiratori Keiko
Published: March 2008

Aim: In non‐alcoholic steatohepatitis (NASH), fibrosis begins around the central veins, as also happens with alcoholic liver disease, so the symptoms of portal hypertension may be due to central vein occlusion. The aim of this study was to define the prevalence of esophagogastric varices and the clinical outcome after endoscopic treatment in NASH patients with severe fibrosis.

Methods: The subjects were 72 patients with clinicopathologically confirmed NASH who had bridging fibrosis (F3) or cirrhosis (F4) determined by the examination of liver biopsy specimens, and who underwent upper gastrointestinal endoscopy. The prevalence and pattern of endoscopically detected varices at the time of liver biopsy were evaluated. The results of NASH patients (n = 11) with endoscopically treated esophageal varices were compared to those with alcoholic (n = 67) and hepatitis C virus‐associated cirrhosis (n = 152).

Results: Esophagogastric varices were detected in 34 out of the 72 (47.2%) patients; esophageal varices in 25 (34.7%) and gastric varices in nine (12.5%), while six of these patients had variceal bleeding. In NASH patients, the cumulative recurrence‐free probability at 24 months after endoscopic treatment was 63.6%, the bleeding‐free probability was 90.9%, and the 5‐year survival was 100%. Only one out 11 patients died of liver failure at 70 months after treatment.

Conclusion: About half of NASH patients with severe fibrosis had esophagogastric varices. The clinical status and course of the varices do not necessarily improve after endoscopic treatment. NASH patients with esophagogastric varices need to be followed up carefully, like patients with other chronic liver diseases.
Fulminant hepatitis and late onset hepatic failure in Japan
Authors: Fujiwara Kenji , Mochida Satoshi , Matsui Atsushi , Nakayama Nobuaki , Nagoshi Sumiko , Toda Gotaro ,   Intractable Liver Diseases Study Group of Japan
Published: March 2008

Aim:  A nationwide survey was performed to clarify the present state of fulminant hepatitis and late onset hepatic failure (LOHF) between 1998 and 2003 in Japan.

Methods:  Three hundred and sixteen, 318 and 64 patients, respectively, with acute and subacute types of fulminant hepatitis and LOHF, in which grade II or more severe hepatic encephalopathy occurred within 10 days, between 11 days and 8 weeks and between 8 and 24 weeks, respectively, after the onset of disease symptoms, were analyzed.

Results:  Complications such as metabolic syndrome were underlying in 41.5% of patients with subacute fulminant hepatitis and 51.6% of patients with LOHF, and most of such patients had received daily medications. The etiology of fulminant hepatitis was viral infection in 71.2% of the acute type and 31.8% in the subacute type. Hepatitis B virus (HBV) infection was found in most of these patients; transient infection prevailed in the acute type; and HBV carrier prevailed in the subacute type. The etiology was unknown in 42.8% and 53.1% of the subacute type and LOHF, respectively. Autoimmune hepatitis and drug allergy‐induced liver injury were found in 10.7% and 11.3%, respectively, of the subacute type. Artificial liver support with plasma exchange and/or hemodiafiltration took place in more than 90% of all patients. The survival rates of the patients without liver transplantation were 53.7% in the acute and 24.4% in the subacute type, and 11.5% in LOHF. The prognosis was especially poor in HBV carriers and patients with autoimmune hepatitis. The survival rates of those who underwent liver transplantation were 56.3%, 39.3% and 23.4% in the acute type, subacute type and LOHF, respectively.

Conclusion:  The etiology and prognosis differed in patients with fulminant hepatitis and LOHF depending on the disease types in Japan, and liver transplantation improved the prognosis of the patients irrespective of the disease type and etiology.
Preventive effects of ME3738 on hepatic fibrosis induced by bile duct ligation in rats
Authors: Maeda Kazunori , Koda Masahiko , Matono Tomomitsu , Sugihara Takaaki , Yamamoto Satoru , Ueki Masaru , Murawaki Yoshikazu , Yamashita Nobuyuki , Nishiyama Shoji
Published: March 2008

Aim:  The aim of this study was to examine the preventive effects of ME3738 on hepatic fibrosis induced by bile duct ligation (BDL) in rats.

Methods:  ME3738 (20 mg/day) was administered orally for 21 days immediately after BDL. Fibrosis was assessed by measuring hepatic hydroxyproline (Hyp) content. Activated hepatic stellate cells (HSCs) were assessed by α‐smooth muscle actin (α‐SMA) immunostaining. Hepatic thiobarbituric acid‐reactive substance (TBARS), 4‐hydroxy‐2‐nonenal (4‐HNE) and 8‐hydroxy‐2‐deoxyguanosine (8‐OHdG) immunostaining were used to analyze oxidative stress. The gene expressions of collagen‐I, transforming growth factor‐β1 (TGF‐β1), tissue inhibitor of metalloproteinases‐1 (TIMP‐1), interleukin‐6 (IL‐6) and heme oxygenase‐1 (HO‐1) in the liver were examined by real‐time reverse transcriptase polymerase chain reaction (RT–PCR).

Results:  Hepatic Hyp content and the area of hepatic fibrosis in BDL rats treated with ME3738 were reduced by 24% and 39% compared with non‐treated BDL rats (hepatic Hyp, 9.40 ± 2.85 vs. 12.39 ± 3.91 mg/liver; P = 0.036; area of hepatic fibrosis, 13.1 ± 3.8 vs. 21.5 ± 10.9; P = 0.045). Furthermore, α‐SMA‐positive cells were significantly reduced by 40% (22.3 ± 14.8 vs. 37.6 ± 14.2; P = 0.011), collagen‐I mRNA by 83% (6.5 ± 2.2 vs. 38.3 ± 9.1; P = 0.002), HO‐1 mRNA by 58% (4.13 ± 1.22 vs. 9.73 ± 1.80; P = 0.018) and hepatic HO‐1 content by 26% (2.13 ± 0.80 vs. 2.87 ± 0.19; P = 0.01) following ME3738 treatment. The hepatic expression of TBARS, 4‐HNE, 8‐OHdG and mRNA levels of TGF‐β1, TIMP‐1 and IL‐6 in the liver were unchanged by ME3738 treatment.

Conclusion:  Oral ME3738 administration may prevent the progression of hepatic fibrosis in BDL rats through suppression of the activation and collagen synthesis of HSC and, in part, oxidative stress. ME3738 has potential as a therapeutic drug for cholestatic liver fibrosis.
Clinical Assessment of Acid‐Base Status: Comparison of the Henderson‐Hasselbalch and Strong Ion Approaches
Authors: Constable Peter D.
Published: March 2008

Abstract:The traditional approach for clinically assessing acid‐base status uses the Henderson‐Hasselbalch equation to categorize 4 primary acid‐base disturbances: respiratory acidosis (increased PCO2), respiratory alkalosis (decreased PCO2), metabolic acidosis (decreased extracellular base excess or actual HCO3−concentration), and metabolic alkalosis (increased extracellular base excess or actual HCO3−concentration). The anion gap is calculated to detect unidentified anions in plasma. This approach works well clinically and is recommended for use whenever serum total protein, albumin, and phosphate concentrations are approximately normal. However, because the Henderson‐Hasselbalch approach is more descriptive than mechanistic, when these concentrations are markedly abnormal the Henderson‐Hasselbalch equation frequently provides erroneous information as to the cause of an acid‐base disturbance. The new quantitive physicochemical approach to evaluating acid‐base balance uses the simplified strong ion model to categorize 6 primary acid‐base disturbances: respiratory acidosis (increased PCO2), respiratory alkalosis (decreased PCO2), strong ion acidosis (decreased strong ion difference), strong ion alkalosis (increased strong ion difference), nonvolatile buffer ion acidosis (increased plasma concentrations of albumin, globulins, or phosphate), and nonvolatile buffer ion alkalosis (decreased plasma concentrations of albumin, globulins, or phosphate). The strong ion gap is calculated to detect unidentified anions in plasma. The simplified strong ion approach works well clinically and is recommended for use whenever serum total protein, albumin, or phosphate concentrations are markedly abnormal. The simplified strong ion approach is mechanistic and is therefore well suited for describing the cause of any acid‐base disturbance.
Validation of the Coulter AcT Diff Hematology Analyzer for Analysis of Blood of Common Domestic Animals
Authors: Dawson Hilary , Hoff Brent , Grift Evert , Tvedten Harold , Shoukri Mohammed
Published: March 2008

Abstract: The objective of this study was to compare and assess the agreement between the Coulter AcT Diff hematology analyzer (CAD) and the Bayer Technicon H1 (H1) using blood samples from 391 animals of 4 species. The H1 has been used in veterinary laboratories for many years. Recently, Coulter modified the CAD and added veterinary software for hematologic analysis of feline, canine, and equine samples. A comparison of hemograms from dogs, cats, horses, and cattle was made using EDTA‐anticoagulated blood samples. Both instruments were calibrated using human blood products. Performance characteristics were excellent for most values. The exceptions were MCV in canine samples (concordance correlation of .710), platelet counts for feline and equine samples (.258 and .740, respectively), feline and bovine WBC counts (.863 and .857, respectively), and bovine hemoglobin (.876).
Mixed Venous and Arterial Blood in Bovine Coccygeal Vessel Samples for Blood Gas Analysis
Authors: Tvedten Harold , Kopcia Michelle , Haines Carolyn
Published: March 2008

Abstract: Bovine coccygeal (median, caudal) vessel samples are not always venous in origin but may be arterial or a mixture of venous and arterial blood. Results of blood gas analysis of blood samples collected from 39 cows were consistent with typical venous, mixed venous‐arterial, or typical arterial blood. This observation was made while establishing reference intervals for a new blood gas instrument. The pO2 and the oxygen saturation of hemoglobin (SO2) were most reflective of the amount of arterial blood contamination in the sample, but pCO2 and pH also were altered. The pO2 values ranged from 21 to 127 mm Hg, and SO2 ranged from 35% to 100%. The mean pCO2 in venous type samples was 46.3 mm Hg compared to 36.7 mm Hg in arterial type samples. The mean pH in venous type samples was 7.42 compared to 7.47 in arterial type samples.
Effect of Temperature, Storage Time, and Sample Type on Sorbitol Dehydrogenase Activity in Llama Serum and Plasma
Authors: Tornquist Susan J. , Dodson Linda , Lanning Dirk V.
Published: March 2008

Abstract:Serum and heparinized plasma samples were collected from 11 adult, clinically healthy llamas. Aliquots were assayed for sorbitol dehydrogenase (SDH) activity after storage at room temperature (20°C), 4°C, or −20°C for defined time intervals up to 1 week postcollection. Sorbitol dehydrogenase activity in all samples was within reference intervals for our laboratory. No difference was found between serum and plasma SDH activity when measured immediately (within 1 hour) after collection. Sorbitol dehydrogenase activity decreased to 79% of initial activity by 24 hours in serum stored at room temperature; plasma had 94% of initial SDH activity under the same conditions. Sorbitol dehydrogenase activity was stable in both plasma and serum stored for up to 1 week at 4°C or −20°C. With the exception of serum stored at 20°C for > 8 hours, in vitro stability of llama SDH was adequate for its use in diagnostic testing.
Effect of Specimen Collection and Storage on Blood Glucose and Lactate Concentrations in Healthy, Hyperthyroid and Diabetic Cats
Authors: Mary M. Christopher DVM, PhD  , O'Neill Sharron
Published: March 2008

Abstract:The objective of this study was to compare and investigate differences in glucose and lactate concentrations in sodium fluoride/potassium oxalate (NaF/Ox) plasma and serum in healthy cats and cats with metabolic disease. Glucose and lactate concentrations were determined in routinely processed serum and NaF/Ox plasma obtained from healthy (n = 30), hyperthyroid (n = 27) and diabetic (n = 30) cats, and in samples from 6 healthy cats stored at 25°C or 4°C for 0,1, 2, 4, or 8 hours. The packed cell volume (PCV) of blood collected in NaF/Ox was compared with that of blood collected in EDTA. Mean glucose concentration was significantly (P < .05) lower in NaF/Ox plasma than in serum in all groups of cats, by 0.7–2.5 mmol/L (11–45 mg/dL); the difference was greater in cats with hyperglycemia. Mean lactate concentration was significantly higher in serum than in NaF/Ox plasma in all groups of cats, by 0.4–1.2 mmol/L (3.6–10.8 mg/dL); the difference was greater in hyperthyroid and diabetic cats. In vitro, only serum stored on the clot for ≥ 1hour at 25°C had significantly lower glucose and higher lactate concentrations. The PCV of NaF/Ox‐anticoagulated blood was lower than that of EDTA‐anticoagulated blood, by 7.0%± 1.4% (P<.01). In conclusion, collection of feline blood in NaF/Ox was necessary to prevent in vitro increases in lactate concentration; however, NaF/Ox artifactually decreased plasma glucose concentration because of RBC shrinkage. The PCV should not be determined on blood collected in NaF/Ox.
Immunohistochemical Characterization of Estrogen and Progesterone Receptors in Lymphoma of Horses
Authors: Henson Kristin L. , Alleman A. Rick , Kelley Lynda C. , Mahaffey Edward A.
Published: March 2008

Abstract:Immunohistochemical techniques were used to examine 29 cases of equine lymphoma for estrogen receptor (ER) and progesterone receptor (PR) expression. The lymphomas examined included T‐cell‐rich large B‐cell lymphomas, B‐cell neoplasms, and T‐cell lymphomas. The individual cases were also classified according to the anatomic location of the tumors. One normal equine lymph node was also examined for ER and PR expression. All of the cases of equine lymphoma and the normal lymph node were negative for ER. A total of 16/29 (55%) PR‐positive lymphomas were identified. Seven of the 12 (58%) T‐cell‐rich large B‐cell lymphomas were positive, 7/11 (64%) B‐cell tumors were positive, and 2/6 (33%) T‐cell neoplasms were positive. Anatomically, 6/9 (66%) subcutaneous lymphomas were PR positive, 3/5 (60%) intrathoracic lymphomas were positive, 1/4 (25%) intra‐abdominal lymphomas were positive, 2/5 (40%) intra‐abdominal/intrathoracic lymphomas were positive, 1/2 (50%) upper airway lymphomas were positive, and 3/3 (100%) splenic lymphomas were positive. One case involving abdominal and thoracic tumors and leukemia was negative for PR expression. The normal lymph node contained a low percentage (1.9%) of PR‐positive lymphocytes. The presence of PR in neoplastic equine lymphoid tissue indicates that these tumors may be responsive to serum progesterone. Also, identification of PR‐positive cells in the normal lymph node suggests that PR may be constitu‐tively expressed in normal equine lymphocytes. Further studies are needed to quantify PR levels in normal and malignant equine lymphoid tissue and to determine the usefulness of either progestin or antiprogestin drugs in the management of equine lymphoma.
Validation of an Immunoturbidimetric D‐Dimer Assay in Canine Citrated Plasma
Authors: Caldin Marco , Furlanello Tommaso , Lubas George
Published: March 2008

Abstract: D‐dimer is a neoantigen formed when thrombin initiates the transformation of fibrinogen to fibrin; it is derived from plasmin digestion of cross‐linked fibrin. In human medicine, the usefulness of this analyte in diagnosing disseminated intravascular coagulation (DIC) has been assessed in patients fulfilling the clinical and laboratory requirements for this disorder. In canine medicine, the use of D‐dimer is relatively new. Detailed studies are needed to understand the relationship between D‐dimer concentration in plasma and DIC status in dogs. We validated a D‐dimer immunoturbidimetric assay (Tina‐quant [a] D‐Dimer, Boehringer Mannheim) in canine citrated plasma samples. Intra‐assay and interassay variability (coefficient of variation) was 5.63% and 8.82%, respectively. The assay was linear, using 2 samples with low and high D‐dimer concentrations (r = .996 and .998). Accuracy was 102.2% and 95.7% based on a recovery study in which 2 samples were assessed. Reference values for D‐dimer were established using 70 healthy dogs that were assessed clinically and evaluated on the basis of a complete laboratory workup. The reference range was set between 0.02 and 0.28 μg/mL (chi‐square test for normal distribution, P > .05).
Mechanism‐Based Diagnostic Reasoning: Thoughts on Teaching Introductory Clinical Pathology
Authors: Bender Holly S. , Lockee Barbara B. , Danielson Jared A. , Mills Eric M. , Boon G. Daniel , Burton John K. , Vermeer Pamela J. , Zimmerman Kurt L. , Hilmer Kelly M.
Published: March 2008

Abstract: Teaching introductory clinical pathology to veterinary students is a challenging endeavor that requires a shift in learning strategies from rote memorization to diagnostic reasoning. Educational research has identified discrete cognitive stages required to achieve the automated, unconscious thinking process used by experts. Building on this knowledge, we developed a case‐based approach to clinical pathology instruction that actively engages students in the learning process and links performance with positive reward. Simulated cases provide context and create a structure, or “schema”, which enhances the learning process by enabling students to synthesize facts and link them with their causal mechanism to reach a defensible diagnostic conclusion. Web‐based tools, including the “Problem List Generator” and tutorials, have been developed to facilitate this process. Through the collaborative Biomedical Informatics Research Group, we are working to further develop and evaluate Web‐based instructional tools and new educational methods, to clarify the diagnostic reasoning processes used by experienced clinical pathologists, and, ultimately, to better educate our future students to be effective diagnosticians.
Flow Cytometric Evaluation of Canine Bone Marrow Differential Cell Counts
Authors: Weiss Douglas J. , Blauvelt Melissa , Sykes Jane , McClenahan David
Published: March 2008

Abstract: Three flow cytometric techniques were evaluated for determination of differential cell counts on canine clinical bone marrow specimens. Techniques included staining bone marrow specimens with 2′7′‐dichlo‐rofluorescein (DCF) or 3,3′‐dihexyloxacarbocyanine iodide (DiOC6) and evaluation of forward‐angle light scatter vs. side‐angle light scatter plots. Flow cytometric evaluation of bone marrow cells stained with DCF failed to separate bone marrow cells into distinct cell populations. Staining with DiOC6 resulted in separation of bone marrow cells into populations of mature and immature erythroid cells, mature and immature myeloid cells, and lymphocytes. The scatter plot method resulted in identification of mature and immature erythroid cells, immature myeloid cells, metamyelocytes, and bands and segmenters. Lymphocytes could not be differentiated from mature erythroid cells by the scatter plot method. When the results of the DiOC6 method and the scatter plot method were compared with manual bone marrow differential cell counts, the scatter plot method had more similar mean values and higher correlation coefficients. The scatter plot method has the potential of providing rapid semiquantitative assessment of bone marrow differential cell counts in dogs for specimens that contain low numbers of lymphocytes.
Age‐Related Changes in Hematology and Plasma Chemistry Values of Hybrid Striped Bass (Morone chrysops × Morone saxatilis)
Authors: Hrubec Terry C. , Smith Stephen A. , Robertson John L.
Published: March 2008

Abstract:Hybrid striped bass (Morone chrysops × Morone saxatilis) are an important aquaculture species yet there are few diagnostic tools available to assess their health. Hematology and clinical chemistry analyses are not used extensively in fish medicine due to the lack of reference intervals for various fish species, and because factors such as age can affect blood values. There is little published information regarding age‐related changes in blood values of juvenile fish. It is important to evaluate juvenile fish, as this is the time they are raised in aquaculture settings. Determining age‐related changes in the blood values of fishes would further develop clinical pathology as a diagnostic tool, enhancing both fish medicine and the aquaculture industry. The results of standard hematology and clinical chemistry analysis were evaluated in juvenile hybrid striped bass at 4, 6, 9, 15, and 19 months of age. Values for PCV and RBC indices were significantly lower, and plasma protein concentration was significantly higher in younger fish. Total WBC and lymphocyte counts were significantly higher in fish at 6 and 9 months of age, while neutrophil and monocyte counts were higher at 6, 9, and 15 months. Eosinophil counts were significantly higher in 9‐month‐old fish. The majority of hematologic values fell within previously established reference intervals, indicating that only slight modification to the intervals is necessary for evaluating hematologic results of hybrid striped bass at different ages. The following analytes deviated sufficiently from adult reference intervals to warrant separate reference values: plasma protein concentration at 4 months, WBC and lymphocyte counts at 15 and 19 months, and thrombocyte‐like‐cells at 9 months of age. Values for most biochemical analytes were significantly different among age groups except for creatinine and potassium concentrations. Comparisons with reference intervals were not made for biochemical analytes, because established reference intervals were not available. Age‐related changes in hematologic and biochemical values of striped bass were similar to those reported for rainbow trout and mammals.
Comparison of Refractometer and Biuret Methods for Total Protein Measurement in Body Cavity Fluids
Authors: George Jeanne W. , O'Neill Sharron L.
Published: March 2008

Abstract: Most hand‐held medical refractometers have internal scales that limit protein measurement to results ≥ 2.5 g/dL. Tables for conversion of refraction (r) to protein concentration for values as low as 0.1 g/dL were published in the 1960s, but their accuracy for use on body fluids has not been established. The purpose of this study was to assess the reliability of body cavity fluid protein determination by refractometry. We compared the protein concentration of 25 body cavity fluids as determined by 2 Goldberg type hand‐held refractometers with results obtained by the biuret method. Published charts converting refraction (r) to protein concentration were used to determine protein concentration in samples with protein <2.5 g/dL. Higher protein values were read directly from the instruments. The range of comparison was limited to ≥ 0.6 g/dL, the lowest concentration of the biuret method's standard curve. Twenty‐one peritoneal fluid, 2 pleural fluid and 2 pericardial fluid samples from 16 horses, 5 cattle, 3 dogs, 2 llamas and 1 cat were tested. The results obtained by the two refractometers were closely and linearly related to biuret results (P<.001), with slopes by linear regression analysis close to 1, and correlation coefficients >0.977. Based on this study, the range for quantification of body cavity fluid protein concentration by refractometry can be extended below 2.5 g/dL, allowing for quantitative assessment of most clinical samples.
Comparison of Liver Cytology and Biopsy Diagnoses in Dogs and Cats: 56 Cases
Authors: Roth Lois
Published: March 2008

Abstract: The results of liver cytology and corresponding biopsy specimens submitted to the Pathology Department at Angell Memorial Animal Hospital during 1998 were retrospectively reviewed to objectively assess the diagnostic value of liver cytology. A “corresponding” biopsy was defined as a biopsy specimen obtained within 1 day of the cytology specimen. Fifty‐six cases were reviewed, including 25 from dogs and 31 from cats. Results were in complete agreement in 34 cases, in partial agreement in 11 cases, and in disagreement in 11 cases. Agreement occurred most often in cases of fatty change (10 cases) and neoplasia (7 cases of lymphoma and 3 cases of epithelial tumor). Disagreement was most common in cases of hepatitis (6 cases) in which inflammation was not seen in cytologic specimens. Other causes for disagreement included fibrosis (2 cases), and 1 case each of amyloidosis, hemangiosarcoma, and lymphoma. Three cytologic specimens were considered suggestive of hepatitis because of leukocytosis in the background, although clinicians were advised to compare cytologic observation to CBC results. Cytologic evaluation of the liver was useful for determining disease processes, especially when disease distribution was diffuse.
Stability of Hemostatic Proteins in Canine Fresh Frozen Plasma Units
Authors: Wardrop K. Jane , Brooks Marjory B.
Published: March 2008

Abstract: The stability of hemostatic proteins, including coagulation factors II, VII, VIII, IX, and X and von Willebrand factor (vWf), in canine fresh frozen plasma (FFP) units stored for up to 1 year was studied. Plasma units from 7 donor dogs were subjected to 4 treatments following collection, including storage at −30°C for 3 months, 6 months, and 1 year and storage at −20°C for 6 months. Coagulant factor activity and vWf concentrations were measured at these times. Significant differences between prestorage and poststorage values were noted for factors VIII, IX, and X, and vWf. Differences seemed to be least pronounced for plasma stored for 3 months; however, a significant interaction between prestorage and poststorage differences and the 4 treatment groups could not be demonstrated. On the basis of factor content in the present study, 15–20 mL/kg of FFP stored for up to 1 year was capable of providing approximately 10–15 U/kg of vWf and factors VIII, IX, X, and II, whereas 10 mL/kg FFP provided at least 10 U/kg of factor VII.
Alterations in Normal Canine Platelets During Storage in EDTA Anticoagulated Blood
Authors: Wilkerson Melinda J. , Shuman Wilma
Published: March 2008

Abstract:Flow cytometric detection of platelet surface‐associated IgG (PSAIgG) can be used to determine whether immunologic factors are contributing to thrombocytopenia in dogs. In vitro alterations in platelet activation and morphology, however, could impact the results of this test. The purpose of this study was to determine whether the PSAIgG test for immune‐mediated thrombocytopenia was valid on whole blood in EDTA anticoagulant after 24–72 hours of storage, and to characterize other alterations in canine platelets that could impact immunologic testing. Platelets were harvested and analyzed immediately after blood collection and after 24, 48, and 72 hours of storage at 4°C. Spontaneous and thrombin‐induced changes in the following platelet parameters were evaluated using flow cytometric techniques: PSAIgG, platelet microparticle formation, membrane expression of P‐selectin and glycoprotein CD61, exogenous IgG binding, surface‐exposed phosphatidylserine, and fibrinogen binding. The amount of PSAIgG increased 6‐to 9‐fold in stored samples compared with fresh samples. Platelet microparticle formation was spontaneous in stored samples and increased significantly over time. Membrane phosphatidylserine, P‐selectin, and fibrinogen binding were not altered by storage, indicating that platelet activation was minimal in stored samples. Although storage decreased the percentage of platelets positive for CD61 by 8‐to 10‐fold compared with fresh samples, activation by high‐dose thrombin partially restored the percentage of CD61‐positive platelets in 24‐hour‐old samples. In conclusion, even though platelets stored in EDTA for up to 72 hours remain in a resting state, aged platelets have an increased tendency to form microparticles and have increased surface IgG and decreased surface CD61, which may contribute to false‐positive results for tests of immune‐mediated thrombocytopenia.
Platelet Size, Platelet Surface‐Associated IgG, and Reticulated Platelets in Dogs with Immune‐Mediated Thrombocytopenia
Authors: Wilkerson Melinda J. , Shuman Wilma , Swist Shannon , Harkin Kenneth , Meinkoth James , Kocan A. Alan
Published: March 2008

Abstract:Immune‐mediated thrombocytopenia (IMT) is a disorder in which bound IgG on the surface of platelets results in platelet removal and alterations in mean platelet volume. Using flow cytometry, alterations in platelet size, platelet surface‐associated IgG (PSAIgG), and numbers of reticulated platelets were determined in 13 dogs with primary IMT and 4 dogs with secondary IMT induced by experimental infection withBabesia gibsoni. Effects of sample age on platelet parameters also were determined, using samples from 20 dogs with normal platelet counts analyzed within 4 hours and after 24, 48, and 72 hours of storage in EDTA. No significant changes in platelet count, platelet size, or reticulated platelet percentage were observed in samples assayed within 4 and 24 hours of blood collection; whereas PSAIgG values increased 3 to 7 fold in samples stored for 24–72 hours. Using reference values for freshly collected or 24‐hour‐old samples, 10 of 13 (77%) dogs with primary IMT and allB gibsoni‐infected dogs had increased PSAIgG levels. In 12 (75%) of the 16 dogs with thrombocytopenia the percentage of reticulated platelets was increased; however, absolute numbers of reticulated platelets were within reference values. Moreover, PSAIgG level and the percentage of reticulated platelets were not always increased concurrently in dogs with primary and secondary IMT. Platelet microparticles were detected in allB gibsoni‐infecteddogs, 8 of 13 (62%) dogs with primary IMT, and transiently in a dog that responded to immunosuppressive treatment. The results of this study indicate that sample age and time of sampling during disease affect interpretation of platelet parameters in dogs with IMT.
Overestimation of Canine Albumin Concentration with the Bromcresol Green Method in Heparinized Plasma Samples
Authors: Stokol Tracy , Tarrant Jacqueline M. , Scarlett Janet M.
Published: March 2008

Abstract: Albumin concentrations are routinely measured in dogs with bromcresol green (BCG)‐binding assays on automated chemistry analyzers. Several variables affect this assay, including the length of reaction time, sample type, and lack of specificity of BCG for albumin. We observed that albumin concentrations measured with BCG appeared higher in heparinized plasma samples in sick dogs. The objective of this study was to determine the effect of anticoagulant and assay procedure on BCG albumin concentrations in clinically ill dogs. We hypothesized that albumin concentrations would be overestimated in heparinized plasma compared with serum because of the combination of heparin and fibrinogen. Furthermore, we hypothesized that the overestimation would be influenced by assay parameters. Blood was collected from 32 clinically ill dogs into tubes containing heparin, citrate, or no anticoagulant. Citrate was chosen to assess the effect of fibrinogen in the absence of heparin. Albumin concentration was measured in all 3 sample types from each dog using 2 different BCG procedures on an automated chemistry analyzer. The BCG procedures (standard and modified) differed in the wavelengths used for absorbance readings (standard, 600/700; modified, 570/505) and the time point at which absorbance was measured (standard, 100 seconds; modified, 40 seconds). In addition, the modified method incorporated a sample blank. Globulin fractions, fibrinogen concentration, and indices of lipemia, hemolysis, and icterus were evaluated for their contribution to the overestimation of albumin concentration in heparinized plasma compared with serum samples. Albumin concentrations were significantly higher (P≤ .05) in heparinized plasma (mean±SE, 3.8 ± 0.1 g/dL) than in serum (3.6 ± 0.2 g/dL) or citrated plasma (3.2 ± 0.1 g/dL). Overestimation was evident only with the standard BCG procedure. Multiple linear regression analysis indicated that fibrinogen was largely responsible for the higher albumin concentration in heparinized plasma with the standard method. Based on these results, heparin is not recommended as an anticoagulant for albumin measurement in dogs when using a BCG method unless the assay uses a sample blank and a reaction time of <1 minute.
Babesia gibsoniInfection in a Dog from Indiana
Authors: Irizarry‐Rovira Armando R. , Stephens Julie , Christian John , Kjemtrup Anne , DeNicola Dennis B. , Widmer William R. , Conrad Patricia A.
Published: March 2008

Abstract:A 10‐year‐old spayed female mixed‐breed dog was presented to the Purdue University Veterinary Teaching Hospital (PUVTH) with complaints of persistent anemia with occasional exacerbations, anorexia, and lethargy. The dog had been presented to the referring veterinarian 2 months prior with multiple bite wounds received during a fight with 3 Pit Bull Terriers. The dog was discharged after the wounds were cleaned and surgically closed. Upon admission to the PUVTH, blood was collected for a complete blood count and biochemical analysis. Microscopic examination of peripheral blood smears revealed intraerythrocytic protozoal parasites consistent withBabesia gibsoni. Molecular analysis confirmed that the organism wasB gibsoniand that its 18S ribosomal RNA sequence was identical to that of otherB gibsoniisolates from Oklahoma, North Carolina, and Okinawa, Japan. Hematologic changes included moderately severe, regenerative, macrocytic, normochromic anemia, with poikilocytosis, polychromasia, anisocytosis, and a marked increase in nucleated RBCs. Biochemical changes included increased serum alanine aminotransferase, alkaline phosphatase, and gamma glutamyl‐transferase activities. The dog was treated with imidocarb, but despite initial clinical improvement, the dog died 2 weeks after the first dose. A necropsy was not performed. The infection in this dog is the first reported case ofB gibsoniinfection in Indiana. Because of the widespread geographical distribution of the organism, veterinarians and veterinary clinical pathologists throughout the United States should carefully examine Romanowsky‐stained blood smears from patients with acute hemolytic anemia for small intraerythrocytic babesial parasites.