Comparison of Tube, Gel, and Immunochromatographic Strip Methods for Evaluation of Blood Transfusion Compatibility in Horses.

BACKGROUND: Assessment of blood compatibility, typically by tube agglutination (TUBE) and hemolysis crossmatch or, less commonly, by blood typing and alloantibody screening, often is performed before blood transfusion in horses. In contrast, gel column (GEL) and immunochromatographic strip (STRIP) techniques are preferred for compatibility testing in dogs and cats. OBJECTIVE: To determine the accuracy of novel and standard crossmatch and typing methods. ANIMALS: Thirty-eight healthy horses, previously blood typed and alloantibody screened. METHODS: TUBE and GEL crossmatches were performed on 146 different recipient-donor pairs with 56 incompatible TUBE crossmatches. Crossmatches were compared by nonparametric area under the curve of receiver operating characteristic (AUC-ROC) analyses. Horses also were blood typed by the novel immunochromatographic Ca typing STRIP. RESULTS: Compared to TUBE crossmatch, GEL had excellent accuracy for agglutination (AUC-ROC = 0.903), but marginal accuracy for hemolysis (AUC-ROC = 0.639). Compared to macroscopic TUBE, microscopic TUBE had excellent accuracy for agglutination (AUC-ROC = 0.912). The predicted crossmatch compatibility based on blood type and alloantibody assay showed excellent accuracy compared to TUBE and GEL (AUC-ROC = 0.843 and 0.897, respectively). However, there were more recipient-donor pairs identified as incompatible by both TUBE and GEL than predicted by blood type and antibody screen, suggesting the presence of unidentified alloantibodies. A Ca typing STRIP exhibited 100% sensitivity and specificity for the 35 Ca+ and 3 Ca- horses tested. CONCLUSIONS AND CLINICAL RELEVANCE: Gel column crossmatch and Ca typing immunochromatographic strip are simple and accurate methods to evaluate clinical blood compatibility.

Survival Time of Cross-Match Incompatible Red Blood Cells in Adult Horses.

BACKGROUND: There is a markedly reduced half-life of transfused RBCs when donor and recipient cats or humans are cross-match incompatible. Only 10-20% of horses have naturally occurring alloantibodies. Therefore, cross-match testing before blood transfusion is not always performed. HYPOTHESIS: Cross-match incompatibility predicts shortened RBC survival time as compared to that of compatible or autologous blood. ANIMALS: Twenty healthy adult horses. METHODS: Prospective trial. Blood type, anti-RBC antibody screen (before and 1 month after transfusion) and major and minor cross-match determined 10 donor-recipient pairs. Two pairs were cross-match compatible, the remainder incompatible. Donor blood (4 L) was collected into citrate phosphate dextrose adenine-1, labeled with NHS-biotin, and transfused into recipients. Samples were collected at 1 hour and 1, 2, 3, 5, 7, 14, 21, 28, and 35 days after transfusion, and biotinylated RBCs were detected by flow cytometry. Horses were monitored for transfusion reaction during transfusion and daily for 5 days. RESULTS: Cross-match incompatibility was significantly associated with decreased RBC survival time (P < .001). The half-life of transfused incompatible (cross-match >1+) allogenic equine RBCs was 4.7 (95% CI, 3.2-6.2) days versus 33.5 (24-43) days for compatible pairings. Cross-match incompatibility was associated with acute febrile transfusion reaction (P = .0083). At day 30, only 1 horse had developed novel anti-RBC antibodies. CONCLUSIONS AND CLINICAL IMPORTANCE: Cross-match incompatibility was predictive of febrile transfusion reaction and shortened transfused RBC survival, but did not result in production of anti-RBC antibodies at 30 days. Cross-match testing before transfusion is recommended.

Degenerative left shift as a prognostic tool in cats.

BACKGROUND: A degenerative left shift (DLS) is reported to be a poor prognostic indicator in dogs and cats. Limited data in dogs and no studies in cats have been published to investigate this claim. HYPOTHESIS/OBJECTIVES: To characterize the feline population affected by DLS and to determine if the presence and severity of DLS are associated with increased risk of euthanasia or death. ANIMALS: One hundred and eight cats with DLS (cases) and 322 cats without DLS (controls) presented to the University of California, Davis Veterinary Medical Teaching Hospital between April 1, 1995 and April 1, 2010. METHODS: Retrospective case-control study. All cases had a CBC performed within 24 hours of presentation in which immature granulocytic precursors exceeded mature neutrophils. Controls were matched by year of presentation and primary diagnosis. Survival analysis was used to determine risk of death or euthanasia from DLS and other potential predictors of outcome. RESULTS: Cases were more likely to die or be euthanized in hospital compared to controls (60/108 [56%] versus 107/322 [33%]). DLS was a significant predictor of death or euthanasia in hospitalized cats in both univariate and multivariate analysis (hazard ratio, 1.57; 95% confidence interval, 1.13-2.18). Trend analysis showed an increasing trend in the hazard of euthanasia or death with increasing severity of DLS. CONCLUSIONS AND CLINICAL IMPORTANCE: Cats with DLS are 1.57 times more likely to die or be euthanized in hospital than cats without DLS. In addition, increasing severity of DLS is associated with increased likelihood of death or euthanasia.

Diagnostic yield of cytologic analysis of pericardial effusion in dogs.

BACKGROUND: Pericardial effusion cytology is believed by many to be of limited value, yet few studies have evaluated its diagnostic utility. OBJECTIVES: To determine the diagnostic utility of cytologic analysis of pericardial effusion in dogs and to determine if consideration of additional data could improve the diagnostic yield. ANIMALS: Two hundred and fifty-nine dogs with cytologic analysis of pericardial effusion performed between April 1990 and June 2012. METHODS: Electronic medical records from a university teaching hospital were retrospectively reviewed; signalment, complete blood count, serum biochemistry, cytologic analysis of pericardial effusion, and echocardiographic data were recorded. Cytology was classified as diagnostic (infectious or neoplastic) or nondiagnostic (hemorrhagic or other) and groups were compared with multiple Student's t-tests. RESULTS: Cytology was grouped as nondiagnostic (92.3%) or diagnostic (7.7%) and characterized as hemorrhagic (90%), neoplastic (4.6%), infectious (3.1%), or other (2.3%). Overall cytologic analysis of pericardial effusion diagnostic utility was 7.7% and increased to 20.3% if the effusion hematocrit (HCT) <10%; echocardiographic evidence of a mass did not result in a significant increase in the diagnostic utility. CONCLUSIONS AND CLINICAL IMPORTANCE: The diagnostic utility of cytologic analysis of canine pericardial effusion is variable depending on the underlying etiology. In this group of dogs, the diagnostic yield of cytologic analysis was greater for pericardial effusion samples in which the HCT was less than 10%.

The prognostic utility of degenerative left shifts in dogs.

BACKGROUND: A degenerative left shift (DLS) in dogs is reported to be a poor prognostic indicator, but no studies have been reported to verify this claim. HYPOTHESIS/OBJECTIVES: To characterize the canine population affected by DLS and to determine if the presence and severity of the DLS are associated with increased risk of euthanasia or death. ANIMALS: Three-hundred and nineteen dogs with DLS (cases) and 918 dogs without DLS (controls) presented to the University of California, Davis Veterinary Medical Teaching Hospital between April 1, 1995 and April 1, 2010. METHODS: Retrospective case-control study. All cases had a CBC performed within 24 hours of presentation that showed an immature neutrophil count higher than the mature neutrophil count. Controls were matched by year of presentation and primary diagnosis. Survival analysis was used to determine the risk of death or euthanasia associated with DLS and other potential predictors. RESULTS: Half of cases versus 76% of controls were alive at discharge. Median in-hospital survival time was 7 days for cases and 13 days for controls. DLS was a significant predictor of death or euthanasia in both univariate and multivariate analysis (hazard ratio, HR, 1.9; 95% CI 1.54-2.34). CONCLUSIONS AND CLINICAL IMPORTANCE: DLS in dogs is associated with an increased risk of death or euthanasia. This finding, however, varies with disease diagnosis and should be interpreted in light of the individual patient.

The effects of therapeutic concentrations of gentamicin, amikacin and hyaluronic acid on cultured bone marrow-derived equine mesenchymal stem cells.

REASONS FOR PERFORMING STUDY: Joint inflammation and septic arthritis are both potential complications of intra-articular injections of bone marrow-derived mesenchymal stem cells (BM-MSCs). Clinicians may prophylactically co-inject BM-MSCs admixed with either antimicrobials or hyaluronic acid; however, the effect of these agents on cultured BM-MSCs is unknown. OBJECTIVE: To determine the effects of therapeutic levels of gentamicin, amikacin and hyaluronic acid on cultured equine BM-MSCs in vitro. STUDY DESIGN: In vitro experimental study. METHODS: Equine BM-MSCs from 4 healthy mature horses were isolated. Cultured BM-MSCs from each donor were incubated with gentamicin (150 mg), amikacin (250 mg), hyaluronic acid (22 mg) or 1% penicillin/streptomycin (control) under sterile conditions. Mesenchymal stem cells viability, proliferation, mediator secretion and culture media pH were measured. RESULTS: Incubation of BM-MSCs with gentamicin resulted in >95% MSC death after 45 min, and incubation of BM-MSCs with amikacin resulted in >95% MSC death after 2 h. Incubation of BM-MSCs with hyaluronic acid or penicillin/streptomycin (control) for up to 6 h resulted in sustained BM-MSC viability of 80% and >93%, respectively. All additives resulted in decreased media pH in the first minute; however, the pH then remained constant over the 6 h incubation period. No significant differences in BM-MSC proliferation or mediator secretion between the penicillin/streptomycin (control) and cells treated with hyaluronic acid were observed. CONCLUSION: Therapeutic concentrations of aminoglycoside antimicrobials are toxic to cultured equine BM-MSCs. The effects of hyaluronic acid on cultured MSC viability, proliferation and mediator secretion are minimal. POTENTIAL RELEVANCE: Based on these findings, the mixing of aminoglycoside antimicrobials and cultured equine BM-MSCs prior to therapeutic use is not recommended.

Collection of equine cord blood and placental tissues in 40 thoroughbred mares.

REASONS FOR PERFORMING STUDY: Stem cells derived from umbilical cord tissue (UCT) and umbilical cord blood (UCB) in human subjects and horses can be obtained in a minimally invasive fashion with successful propagation of mesenchymal stem cells (MSCs). Currently there are no detailed protocols documenting a procedure to harvest UCB and UCT safely for equine stem cell propagation. HYPOTHESIS: UCB and UCT could be collected without harm to mare or foal. OBJECTIVES: To develop a standard and safe method for UCB and UCT collection, and prospectively to compare foal and mare health between groups of animals where tissue was and was not collected. METHODS: This study was conducted at a Thoroughbred breeding facility in central California in 2008. UCB and UCT were collected from 40 mare and foal pairs. Clinical parameters including time for foal to stand and nurse, time for mare to pass the placenta, and foal haematology data at age 24 h were documented and compared to a control group, consisting of the succeeding 40 mare and foal pairs. RESULTS: UCB was obtained successfully from 36 of 40 (90%) mares and UCT from 38 of 40 (95%) mares. Bacterial contamination was documented in 6 out of 36 (16.6%) UCB samples. There were no significant differences in time to stand or nurse for foals or time to pass the placenta for mares, between the experimental and control groups. There were no clinically relevant differences identified in haematological data obtained from foals with and without UCB collection. CONCLUSIONS: UCB and UCT can be harvested safely without harm to mares or foals. POTENTIAL RELEVANCE: UCB and UCT samples collected in an inherently contaminated environment can be successfully disinfected and transported with minimal bacterial overgrowth for use in cell culture to isolate MSCs.

Transmission of visceral leishmaniasis through blood transfusions from infected English foxhounds to anemic dogs.

OBJECTIVE: To conduct serologic surveillance for Leishmania spp in English foxhounds from a kennel, as well as recipients of blood from these dogs, and determine whether L infantum organisms could be transmitted via blood transfusion. DESIGN: Serologic prevalence survey. ANIMALS: 120 English foxhounds and 51 dogs of various breeds receiving blood from these donors. PROCEDURE: Foxhound blood donors, foxhound nondonors, and nonfoxhound blood recipient dogs were evaluated serologically for Leishmania spp by indirect fluorescent antibody testing. Dogs that received packed RBC (PRBC) transfusions from foxhound donors from mid-1996 through mid-2000 were identified. Furthermore, dogs were serologically evaluated if they had received fresh frozen plasma (FFP) transfusions in 1999 and 2000 from seropositive foxhound blood donors. RESULTS: Thirty percent of the English Foxhounds were seropositive for Leishmania spp (titer > or = 1:16), although the degree of seropositivity varied considerably during the period. Furthermore, 57 foxhounds had been used as donors from 1996 to 2000, and 342 units of PRBC had been transfused to at least 227 patients. All 25 dogs screened that received PRBC from seronegative foxhound donors tested negative, whereas 3 of 7 dogs that received PRBC from seropositive donors tested positive. All 9 dogs that received FFP from seropositive foxhound donors remained seronegative. CONCLUSIONS AND CLINICAL RELEVANCE: To our knowledge, this report documents the first transmission of Leishmania spp by blood transfusion. The use of foxhounds as blood donors may not be advisable in North America.

A double-blind placebo-controlled case study of the use of donepezil to improve cognition in a schizoaffective disorder patient: functional MRI correlates.

Cognitive impairment in multiple domains is common in patients with schizophrenia and may be a powerful determinant of poor functional ability and quality of life. We report a double-blind, placebo-controlled, cross-over study of donepezil augmentation in a schizoaffective disorder patient stabilized on olanzapine pharmacotherapy. The patient showed significant improvements in several cognitive measures and increased activation of prefrontal cortex and basal ganglia on functional MRI during the donepezil augmentation. In addition, the donepezil augmentation resulted in a reduction of depressive symptoms and in significant improvements in functional abilities and quality of life. Further studies of donepezil augmentation of neuroleptics in schizophrenia are warranted.

Hyperthermic therapy for HIV infection.

The objective of this paper is to review what is known about the antiviral effects of fever and to highlight the scientific evidence supporting the hypothesis that hyperthermic therapy may prove to be a beneficial treatment modality for persons infected with HIV. Our hyperthermic hypothesis is based upon the mutant escape, quasispecies theory of HIV antigenic diversity. We propose that, if initiated during the asymptomatic stage of HIV infection, hyperthermia may prove to decrease the number of mutant HIV strains arising due to evolutionary pressures created by the patient's immune system, with a resultant prolongation of the asymptomatic period of infection. A review of the literature from three areas of investigation: the immune response to fever, heat as a tumor killing agent, and preliminary studies with fever and retroviral infections, strongly suggests that there is a good scientific basis for the use of hyperthermic therapy in a multimodal treatment approach to HIV infection.

Esmolol in the treatment of multifocal atrial tachycardia.

Multifocal atrial tachycardia (MAT) is a supraventricular tachydysrhythmia precipitated by a number of pharmacologic and physiologic disturbances. Corrections of these disturbances should take precedence in the treatment of MAT.

Endophthalmitis due to Sporothrix schenckii after penetrating ocular injury.

A 13-year-old boy developed endophthalmitis from traumatic inoculation of vegetable material penetrating his right eye. Sporothrix schenckii was morphologically identified from the intraocular fluids. However, repeated attempts to culture the organism proved negative. Seventeen sporotrical endophthalmitis cases have so far been reported, and before this report, only one of them was secondary to trauma. This is the sole eye to our knowledge not only to have escaped enucleation after sporotrical endophthalmitis but to have achieved useful (20/50) vision.

Efficacy and safety of guanadrel in elderly hypertensive patients.

Hypertension is common in the elderly and is associated with higher morbidity and mortality, which may be decreased by effective blood pressure control. Many antihypertensive drugs, however, are not well tolerated by the aged. We treated 21 patients (ten men and 11 women) between ages 65 and 84 years (mean, 73.6 years) with guanadrel sulfate. All patients had received prior antihypertensive therapy, which either was ineffective or caused undesirable side effects. Average follow-up time was 17 months. Mean systolic pressure on enrollment was 188 +/- 17 mm Hg and mean diastolic pressure was 100 +/- 10 mm Hg. After treatment, the mean systolic pressure was 139 +/- 15 mm Hg and mean diastolic pressure was 82 +/- 8 mm Hg. Dosage varied from 5 to 30 mg/d with a mean of 16 mg/d. The only significant side effects were fatigue, dizziness, and dyspnea reported in four patients. Eleven patients took the medication as monotherapy and ten received diuretics or diuretics and beta-blockers as additional therapy. Our conclusion is that guanadrel is an effective, well-tolerated medication for treatment of hypertension in the elderly.