bluuebunny/crossref_metadata_2025_split huggingface dataset:
Effects of Blood Processing Techniques on Sodium and Potassium Values: A Comparison Between Aldabra Tortoises (Geochelone gigantea) and Burmese Mountain Tortoises (Manouria emys)
Authors: Abou‐Madi Noha , Jacobson Elliott R. 
Published: March 2008

Background:Hematologic and plasma biochemical evaluations are routinely used in evaluating the chelonian patient, but appropriate processing techniques have been minimally defined.Objectives:This study was designed to compare the effects of temperature, time, anticoagulant, and species on sodium and potassium values in the Aldabra tortoise (Geochelone gigantea) and the Burmese mountain tortoise (Manouria emys).Methods:Blood samples from 7 Aldabra tortoises and 8 Burmese mountain tortoises were collected into tubes without anticoagulant and tubes containing lithium heparin. Sodium and potassium concentrations were measured by flame photometry in serum and plasma harvested immediately after collection and from aliquots of whole blood stored at 4°C and 25°C for 5 to 120 minutes.Results:In Aldabra tortoises, storage time and temperature had no significant effect on potassium concentrations in heparinized blood and in blood without anticoagulant. However, sodium concentrations in serum and plasma decreased significantly in samples without anticoagulant stored at 4°C and 25°C and in heparinized samples stored at 4°C. In Burmese mountain tortoises, potassium concentrations in serum and plasma increased significantly with time in samples without anticoagulant and in heparinized samples stored at 4°C and 25°C, but the increases were less at 4°C. Sodium concentrations in serum and plasma decreased significantly in blood without anticoagulant and heparinized blood stored at 4°C and 25°C.Conclusions:Storage of blood samples with and without anticoagulant at 4°C significantly improved the stability of potassium and sodium concentrations in both species of tortoises. Early separation of red cells from serum or plasma after blood collection is especially important to ensure the reliability of potassium measurements.
A Protocol to Guide Development of a Sensitive ELISA for Canine Erythropoietin
Authors: Giampaoli Saverio , Facciabene Andrea , Mennuni Carmela 
Published: March 2008

Background:The determination of canine erythropoietin (EPO) concentration is crucial for monitoring the effect of human recombinant (hr) EPO therapy in dogs with chronic renal failure. Current assays are not specific for canine EPO and not sensitive enough to detect physiologic EPO levels in dogs.Objective:The objective of this study was to develop a simple and sensitive ELISA for canine EPO that could serve as a starting point for developing a commercially available assay.Methods:The ELISA was based on a mouse monoclonal antibody (mAb) and a rabbit polyclonal antibody (pAb) using 2 different immunization techniques: gene electrotransfer (GET) to generate the pAb and multiple antigen peptides (MAPs) to generate the mAb. The ELISA was performed using both EPO obtained from HeLa cells transfected with an expression plasmid encoding canine EPO and canine plasma with known concentrations of EPO.Results:The ELISA standard curve was linear for canine EPO concentrations of 7–66 mU/ml. Coefficients of variation were about 10%. No cross‐reactivity between canine EPO and hrEPO was detected.Conclusions:Using novel GET and MAP technology, we developed a sensitive and specific ELISA for canine EPO that can be used to guide future clinical applications for EPO detection and monitoring in dogs.
Evaluation of variation in serum globulin concentrations in dairy cattle
Authors: Chorfi Younès , Lanevschi‐Pietersma Anne , Girard Vincent , Tremblay Armand 
Published: March 2008

Background:Several factors may influence the concentration of serum globulins in healthy cows and thereby affect clinical interpretation; however, few studies have addressed sources of variation in globulin values.Objectives:The purpose of this study was to compare colorimetry‐based total serum globulin values with electrophoretically‐determined serum globulin fractions and with IgG concentration, and to evaluate diurnal and long‐term physiological variation and the effects of lactation and venipuncture site on serum globulin concentrations in Holstein dairy cattle.Methods:Serum total globulin and albumin concentrations were analyzed by colorimetry and electrophoresis in 86 lactating cows; IgG concentrations were determined by radial immunodiffusion in 41 dry and 34 lactating cows. Serum globulins were analyzed hourly for 24 hours in 8 lactating cows and weekly for 15 weeks in 6 additional cows. Globulin concentrations were compared in samples obtained from jugular and coccygeal venipuncture sites in 4 cows. Results were analyzed using parametric statistical tests.Results:Colorimetry‐based total serum globulin concentrations correlated well with y‐globulin fractions (r2= 0.87) and IgG concentrations (r2= 0.91). Diurnal variation of total serum globulins concentration was significant (P= .01); however, globulins did not vary significantly over a 15‐week period. Mean serum globulins concentration in samples obtained from the jugular vein was 2.35 g/L higher than that in samples obtained by coccygeal venipuncture (P< .0001).Conclusions:The colorimetric method used widely in routine laboratory analyses remains a useful test for globulins determination in dairy cattle. However, time of sampling and venipuncture site should be considered in the interpretation of serum globulins on serial or interindividual specimens.
Inaccuracy of routine creatinine measurement in canine urine
Authors: Trumel Catherine , Diquélou Armelle , Lefebvre Hervé , Braun Jean‐Pierre 
Published: March 2008

Background:Urine creatinine concentration often is used in ratios such as urine protein:creatinine to compensate for dilution or concentration of spot urine samples. Objective: The purpose of this study was to compare the accuracy of different techniques of urine creatinine measurement currently available for veterinary practitioners.Methods:In 104 samples of canine urine diluted 1:20 with distilled water, creatinine concentration was measured using a kinetic Jaffé reaction assay, and an enzymatic technique on an automatic analyzer (Elimat) and 3 benchtop analyzers (Reflovet, Scil; Vitros DT2, Ortho‐Clinical Diagnostics; Vettest 8008, IDEXX) used in veterinary practice.Results:The Jaffé and enzymatic techniques on the Elimat were not significantly different, and their inaccuracy tested with human control urines was <5%. The benchtop analyzers underestimated creatinine concentration, especially at concentrations >2000 mg/L. Inaccuracy was higher with multilayer slide technology systems (Vitros and Vettest) than with the Reflovet system. Results were approximately 25% and 2% lower, respectively, than with the Elimat at urine creatinine concentrations about 2000 mg/L. Conclusion: Inaccuracy in urine creatinine measurements using benchtop analyzers should be taken into account when defining decision thresholds, which should be corrected according to the method used to avoid misinterpretations.
Comparison of amylase and lipase activities in serum and plasma of dogs
Authors: Médaille C. , Briend‐Marchal A. 
Published: March 2008

Background: Amylase and lipase activities are most often determined in serum, although heparinized plasma is more convenient to obtain and is used for many routine biochemical analyses. Objective: The purpose of this study was to compare amylase and lipase activities in serum and plasma of dogs and to determine whether either specimen type is acceptable for analysis.Methods: Serum and heparinized plasma were obtained from 101 randomly selected dogs and analyzed in parallel for α‐amylase and lipase. Results were compared using Passing‐Bablock regression, Bland‐Altman difference plots, and correlation analysis.Results: There was a high correlation between the results obtained from serum and those from plasma. Regressions (with 95% confidence intervals in parentheses) were as follows: lipaseplasma= 0.984 (0.976/0.995) × lipaseSerum ‐ 0.9 (2.9/0.7) (r= .999); α‐amylaseplasma= 1.003 (0.977/1.032) ×α‐amylaseserum ‐ 1.9 (‐20.7/23.3) (r = .991). Mean differences (serum ‐ plasma) were 8 U/L and 4 U/L for lipase and α‐amylase, respectively. Classification of results as normal or abnormal did not differ according to specimen type.Conclusion: In dogs, lipase and α‐amylase activites can be determined with the same level of accuracy in serum and in heparinized plasma.
Evaluation of a chromogenic assay to measure the factor Xa inhibitory activity of unfractionated heparin in canine plasma
Authors: Brooks Marjory B. 
Published: March 2008

Background: Unfractionated heparin (UFH) has a complex pharmacologic profile that necessitates patient monitoring to prevent inadequate anticoagulation or overdosage and hemorrhage. Factor Xa inhibitory assays (to measure anti‐Xa activity) are used to adjust UFH dosage and define safe and effective regimens for specific thrombotic disorders in humans. Objective: In this study, the accuracy, linearity, and clinical utility of a chromogenic assay were assessed for monitoring UFH anti‐Xa activity in canine plasma samples.Methods: A commercial assay (Rotachrom Heparin, Diagnostica Stago, Parsippany, NJ, USA) was used to measure anti‐Xa activity in canine plasma samples spiked with different concentrations of UFH. Background absorbance and assay linearity were compared for canine and human plasmas. Percentage recovery of UFH anti‐Xa activity and intra‐ and interassay imprecisions were investigated by multiple measurements of canine plasma to which known amounts of UFH were added. The spiked plasma samples also were used to determine the heparin sensitivity of an activated partial thromboplastin time (aPTT) test.Results: Canine plasma samples were assayed at a higher dilution than were human plasma samples (3:8 versus 4:8) to eliminate higher background anti‐Xa activity in canine plasma. Using this modification, the recovery of anti‐Xa activity in canine plasma was linear (R2 > .9) at concentrations of 0–0.75 U/mL UFH. Intra‐ and interassay imprecisions for plasma samples containing 0.5 U/mL UFH were <10%, whereas samples containing 0.25 U/mL UFH had imprecisions of 13% and 24%, respectively. The anti‐Xa activity range of 0.5–0.75 U/mL caused prolongation of aPTTs to 1.5–2.5 times the assay mean.Conclusions: Plasma anti‐Xa activity of dogs treated with UFH can be accurately monitored using this commercially‐available chromogenic assay.
Molecular cloning and expression of canine hepcidin
Authors: Fry Michael M. , Liggett Jason L. , Baek Seung Joon 
Published: March 2008

Background: Hepcidin is a recently identified acute phase protein with antimicrobial and iron regulatory functions. It has been suggested that hepcidin may be the key mediator of anemia of chronic disease. Our research group is interested in developing a diagnostic assay to measure hepcidin in dogs.Objectives: The objectives of this study were to clone and sequence the canine hepcidin gene and to gather preliminary data about tissue expression of hepcidin in dogs.Methods: RNA was extracted from fresh canine liver tissue and cDNA was generated and amplified. Standard reverse transcription polymerase chain reaction techniques were used with degenerate primers based on sequence homo logy between several other species. The amino acid (AA) sequence was compared with known sequences in other species. Tissue expression of canine hepcidin was determined by Western blot.Results: The canine hepcidin cDNA sequence encoded a highly conserved protein of 85 AAs with 8 cysteine residues at the C‐terminal end. This protein was likely the precursor form (pro‐hepcidin) of a smaller secreted peptide. Phylogenetic analysis showed that human hepcidin was more homologous with canine than with rodent hepcidin. In dogs, as in people, hepcidin was expressed most strongly in the liver. Western blotting showed a clear band of approximately 9 kDa, consistent with pro‐hepcidin. Weak expression was also detected in canine kidney and lung tissues. Conclusion: The results of this study establish the basis for future investigation involving canine hepcidin and suggest that the dog may be a suitable model for studying the role of hepcidin in human health and disease.
Determination of free and total cyst(e)ine in plasma of dogs and cats
Authors: Tôrres Cristina L. , Miller Joshua W. , Rogers Quinton R. 
Published: March 2008

Background: In human blood, the amino acid cysteine forms disulfide bonds with itself and with other sulfhydryl compounds in their free form and with sulfhydryls in protein. Protein‐bound cysteine is lost when plasma proteins are removed before amino acid analysis. Objective: The purpose of this study was to assess the time course and extent of cyst(e)ine (cysteine + half‐cystine) loss in dog and cat plasma.Methods: An equal volume of 6% sulfosalicylic acid was added to plasma aliquots at 0,2,4,10,16,24, 36, 48, 60, and 72 hours after separation of blood cells. Tris‐2‐carboxyethyl‐phosphine hydrochloride (TCEP‐HC1), a reducing agent, was used to regenerate total plasma cyst(e)ine after 3 months of sample storage (−20°C).Results: Initial free cyst(e)ine concentrations (mean ± SEM) were higher in canine plasma (77 ± 4 μmol/L) than in feline plasma (37 ± 3 μmol/L). Free plasma cyst(e)ine concentrations in dogs and cats decreased after first‐order kinetics, with a half‐life of 23 and 69 hours, respectively. Total plasma cysteine after TCEP‐HC1 treatment was similar for dogs (290 μmol/L) and cats (296 μmol/L), but the percentage of free cysteine was higher (P = .02) in dogs (27%) than in cats (13%). Over half of the cyst(e)ine, homocysteine, cysteinylglycine, and glutathione were bound in vivo to plasma proteins. Conclusion: These results emphasize the importance of removing plasma proteins within 1 hour after blood collection for reliable assay of free plasma cyst(e)ine.
Artifactual hypoglycemia associated with hemotrophic mycoplasma infection in a lamb
Authors: Burkhard Mary Jo , Garry Franklyn 
Published: March 2008

Background:A 35‐day‐old male lamb withMycoplasma ovisinfection (previouslyEperyihrozoon avis) was evaluated because of severe hypoglycemia (serum glucose 4 mg/dL, Hitachi 704 automated chemistry analyzer) inconsistent with the animal's condition. Whole blood glucose concentration measured with a glucometer was 74 mg/dL. Objective: The purpose of this study was to investigate this discrepancy through in vitro evaluation of the patient's blood.Methods:Blood was incubated alone, with increasing concentrations of plasma, or with equine serum of known glucose concentration for 0,15, 30, and 60 minutes at room temperature; end‐point glucose concentrations were compared with blood from a control sheep handled similarly.Results:A rapid decline in glucose concentration was observed in heparinized or EDTA anticoagulated whole blood from the infected lamb incubated alone or with the equine serum. Glucose concentrations in incubated samples from a control sheep remained stable. Incubation of increasing concentrations of heparinized blood with autologous plasma resulted in decreased glucose concentrations in patient, but not control, blood. As parasitemia decreased after treatment, serum glucose concentration increased, serum lactate concentration decreased, and in vitro glucose concentration stabilized.Conclusions:These findings are consistent with parasite‐associated in vitro glucose consumption. An increase in the lamb's plasma glucose concentration associated with reduction of parasite load suggested excess glucose consumption also may have occurred in vivo.
The Membrane-Proximal External Region of the Human Immunodeficiency Virus Type 1 Envelope: Dominant Site of Antibody Neutralization and Target for Vaccine Design
Authors: Montero Marinieve , van Houten Nienke E. , Wang Xin , Scott Jamie K. 
Published: March 2008

SUMMARYEnormous efforts have been made to produce a protective vaccine against human immunodeficiency virus type 1; there has been little success. However, the identification of broadly neutralizing antibodies against epitopes on the highly conserved membrane-proximal external region (MPER) of the gp41 envelope protein has delineated this region as an attractive vaccine target. Furthermore, emerging structural information on the MPER has provided vaccine designers with new insights for building relevant immunogens. This review describes the current state of the field regarding (i) the structure and function of the gp41 MPER; (ii) the structure and binding mechanisms of the broadly neutralizing antibodies 2F5, 4E10, and Z13; and (iii) the development of an MPER-targeting vaccine. In addition, emerging approaches to vaccine design are presented.
Cohesion Group Approach for Evolutionary Analysis of TyrA, a Protein Family with Wide-Ranging Substrate Specificities
Authors: Bonner Carol A. , Disz Terrence , Hwang Kaitlyn , Song Jian , Vonstein Veronika , Overbeek Ross , Jensen Roy A. 
Published: March 2008

SUMMARY
Many enzymes and other proteins are difficult subjects for bioinformatic analysis because they exhibit variant catalytic, structural, regulatory, and fusion mode features within a protein family whose sequences are not highly conserved. However, such features reflect dynamic and interesting scenarios of evolutionary importance. The value of experimental data obtained from individual organisms is instantly magnified to the extent that given features of the experimental organism can be projected upon related organisms. But how can one decide how far along the similarity scale it is reasonable to go before such inferences become doubtful? How can a credible picture of evolutionary events be deduced within the vertical trace of inheritance in combination with intervening events of lateral gene transfer (LGT)? We present a comprehensive analysis of a dehydrogenase protein family (TyrA) as a prototype example of how these goals can be accomplished through the use of cohesion group analysis. With this approach, the full collection of homologs is sorted into groups by a method that eliminates bias caused by an uneven representation of sequences from organisms whose phylogenetic spacing is not optimal. Each sufficiently populated cohesion group is phylogenetically coherent and defined by an overall congruence with a distinct section of the 16S rRNA gene tree. Exceptions that occasionally are found implicate a clearly defined LGT scenario whereby the recipient lineage is apparent and the donor lineage of the gene transferred is localized to those organisms that define the cohesion group. Systematic procedures to manage and organize otherwise overwhelming amounts of data are demonstrated.
Fe-S Cluster Assembly Pathways in Bacteria
Authors: Ayala-Castro Carla , Saini Avneesh , Outten F. Wayne 
Published: March 2008

SUMMARYIron-sulfur (Fe-S) clusters are required for critical biochemical pathways, including respiration, photosynthesis, and nitrogen fixation. Assembly of these iron cofactors is a carefully controlled process in cells to avoid toxicity from free iron and sulfide. Multiple Fe-S cluster assembly pathways are present in bacteria to carry out basal cluster assembly, stress-responsive cluster assembly, and enzyme-specific cluster assembly. Although biochemical and genetic characterization is providing a partial picture of in vivo Fe-S cluster assembly, a number of mechanistic questions remain unanswered. Furthermore, new factors involved in Fe-S cluster assembly and repair have recently been identified and are expanding the complexity of current models. Here we attempt to summarize recent advances and to highlight new avenues of research in the field of Fe-S cluster assembly.
The cultural context of visual hallucinations
Authors: Knight W D , Fox N C , Rossor M N , Warren J D 
Published: March 2008

Abstract
Visual hallucinations (VH) are a cardinal neuropsychiatric symptom and often have important diagnostic implications. The interpretation of VH is influenced by the patient's social and cultural milieu, but the impact of socio-cultural factors on the interpretation, presentation and detection of VH has been little studied. When patients exhibit VH and other neuropsychiatric phenomena, appropriate sensitivity to the role of cultural factors is an important determinant of the success of the medical consultation. We discuss this issue using three illustrative cases.
Antibiotic-resistant Campylobacter: an increasing problem
Authors: Kent A J , Farouk L , Main J , Hoare J M 
Published: March 2008

Abstract
The case is described of a 27-year-old woman who presented with an acute diarrhoeal illness. She was initially poorly responsive to antibiotics and developed lymphocytic ascites. Diagnosis was difficult to establish, and peritoneal tuberculosis was considered to be the most likely cause of her symptoms. Serological tests eventually confirmed Campylobacter jejuni infection. Campylobacter is one of the most common bacterial diarrhoeal infections, and complications, except for colitis, are rare except in specific disease states—for example, patients with cirrhosis or undergoing peritoneal dialysis. Antibiotic resistance is an increasing problem, and this may potentially lead to a greater incidence of complications in the future.
Ellipticine derivative NSC 338258 represents a potential new antineoplastic agent for the treatment of multiple myeloma
Authors: Tian Erming , Landowski Terry H. , Stephens Owen W. , Yaccoby Shmuel , Barlogie Bart , Shaughnessy John D. 
Published: March 2008

Abstract
High-risk multiple myeloma can be correlated with amplification and overexpression of the cell cycle regulator CKS1B. Herein, we used the COMPARE algorithm to correlate high expression of CKS1B mRNA in the NCI-60 cell line panel with the concentration causing 50% growth inhibition (GI50) of &gt;40,000 synthetic compounds. This led to the identification of NSC 338258 (EPED3), a highly stable, hydrophilic derivative of the plant alkaloid ellipticine. In vitro, this synthetic anticancer compound exhibits dramatic cytotoxic activity against myeloma cells grown in suspension or in coculture with stromal cells. EPED3-induced cell cycle arrest and an apoptotic progression that appear to be a consequence of the instantaneous effect of the drug on cytoplasmic organelles, particularly mitochondria. Disruption of mitochondria and cytoplasmic distribution of cytochrome c initiated the intracellular proteolytic cascade through the intrinsic apoptotic pathway. EPED3 is able to induce apoptosis in myeloma cells with de novo or acquired resistance to commonly administered antimyeloma agents. Collectively, our data suggest that EPED3 targets mitochondrial function to rapidly deplete chemical energy and initiate apoptosis in myeloma cells at nanomolar concentrations while leaving stromal cells unharmed. [Mol Cancer Ther 2008;7(3):500–9]