bluuebunny/crossref_metadata_2025 huggingface dataset:
Effect of sublethal 6‐hydroxydopamine on the response to subsequent oxidative stress in dopaminergic cells: evidence for preconditioning
Authors: Leak Rehana K. , Liou Anthony K. F. , Zigmond Michael J.
Published: September 2006

Exposure to sublethal stress can trigger endogenous protection against subsequent, higher levels of stress. We tested for this preconditioning phenomenon in a model of Parkinson's disease by applying 6‐hydroxydopamine to the dopaminergic MN9D cell line. Exposure to sublethal concentrations of 6‐hydroxydopamine (5–10 µm) protected against the toxic effects of a subsequent exposure to a higher concentration (50 µm), as measured by the Hoechst assay for nuclear viability. This was accompanied by little or no protection against 6‐hydroxydopamine‐induced lactate dehydrogenase release, decline in ATP, or reduction in 3H‐dopamine uptake. The antioxidant, N‐acetyl cysteine (20 mm), when applied during preconditioning, abolished protection, as did the protein synthesis inhibitor, cycloheximide (0.2 µm). Preconditioning did not affect superoxide dismutase or glutathione peroxidase enzymes, or levels of heat shock protein‐72. However, Bcl‐2 protein levels rose with preconditioning. Preconditioning rapidly increased phosphorylation of kinases ERK1/2, Akt and JNK, and was abolished by pharmacological inhibitors of their activity. Finally, sublethal 6‐hydroxydopamine preconditioned against the toxicity of proteasome inhibitor, MG‐132 (1 µm). Thus, exposure of a dopaminergic cell line to sublethal oxidative stress can protect against additional oxidative stress due to translational and post‐translational modifications, as well as confer ‘cross‐tolerance’ against a different insult, proteasome inhibition.
Identifying autumn migration routes for the globally threatened Aquatic Warbler Acrocephalus paludicola
Authors: JULLIARD R. , BARGAIN B. , DUBOS A. , JIGUET F.
Published: September 2006

The migration strategy of the globally threatened Aquatic Warbler Acrocephalus paludicola needs to be known to ensure that effective action is taken to conserve its principal stopover sites. Using data from ringing stations during the autumn migration of reedbed warblers, we developed an index that allowed countries and sites to be compared in terms of their numerical importance to migrating Aquatic Warblers. According to this index, there was no significant decline in the number of Aquatic Warblers migrating through Europe during the 1990s. France appears to receive the largest numbers of this species in autumn compared with other European countries. Within France, only coastal regions are visited by the species (but not in the Mediterranean), with the largest numbers occurring in Normandy and Loire, while Brittany receives large numbers in some years but not in others. We also identified some French sites that are of major importance to migrating Aquatic Warblers depositing reserves on migration. France therefore plays a very important part in providing stopover sites for Aquatic Warblers, which means that France should play a major role in undertaking conservation measures for this threatened species.
Expression of receptor tyrosine kinases VEGFR‐1 (FLT‐1), VEGFR‐2 (KDR), EGFR‐1, PDGFRa and c‐Met in canine primary brain tumours
Authors: Dickinson P. J. , Roberts B. N. , Higgins R. J. , Leutenegger C. M. , Bollen A. W. , Kass P. H. , LeCouteur R. A.
Published: September 2006

Inhibition of tumour growth and angiogenesis by targeting key growth factor receptors is a promising therapeutic strategy for central nervous system tumours. Characterization of these growth factor receptors in canine primary brain tumours has not been done. Using quantitative real‐time TaqMan polymerase chain reaction (PCR), we evaluated the expression of messenger RNA (mRNA) for five tyrosine kinase growth factor receptors (vascular endothelial growth factor receptor [VEGFR]‐1, VEGFR‐2, endothelial growth factor receptor [EGFR]‐1, platelet‐derived growth factor receptor a [PDGFRa], and c‐Met) relative to normal cerebral cortex in 66 spontaneous canine primary brain tumours. Increased expression of VEGFR‐1 and VEGFR‐2 mRNA was greatest in grade IV astrocytomas (glioblastoma multiforme) and grade III (anaplastic) oligodendrogliomas. EGFR‐1 mRNA expression was more consistently increased than the other receptors in all tumour types, while increased PDGFRa mRNA expression was mostly restricted to oligodendrogliomas. The similarities in increased expression of these tyrosine kinase growth factor receptors in these canine tumours, as compared to data from their human counterparts, suggest that common molecular mechanisms may be present.
Mohs micrographic surgery: a technique for total margin assessment in veterinary cutaneous oncologic surgery
Authors: Bernstein J. A. , Hodgin E. C. , Holloway H. W. , Hedlund C. S. , Storey E. S. , Hubert J. D.
Published: September 2006

Mohs micrographic surgery (MMS) is the gold standard for the excision of locally invasive cutaneous malignancies in human dermatological surgery. Using a unique horizontal sectioning technique, MMS enables 100% surgical margin assessment and provides the lowest recurrence rates for locally invasive tumours. The purposes of this preliminary study were to explore the feasibility of application of MMS in the veterinary setting and to establish practical advantages and limitations of its use in a pilot programme. It was hypothesized that MMS technique could provide 100% tumour margin assessment using frozen and/or formalin‐fixed horizontal histopathologic sections. Tumour excision and colour‐coded mapping were performed, and specimen tissue was fixed using either frozen sections or formalin‐fixed sections. Horizontal sections were assessed for quality and presence and location of neoplastic cells based on the mapped orientation. The MMS technique was used in the excision of six squamous cell carcinomas and five mast cell tumours. In all cases, the MMS permitted 100% tumour margins examination.
Expression and sequence of canine SIRT2 and p53 genes in canine mammary tumour cells – effects on downstream targets Wip1 and p21/Cip1
Authors: DeInnocentes P. , Li L. X. , Sanchez R. L. , Bird R. C.
Published: September 2006

We have characterized gene dysfunction in a cellular model of spontaneous canine mammary cancer by investigating specific gene defects in SIRT2 and p53 genes for comparative studies among canine tumour‐derived cell lines. These genes and their downstream targets are involved in regulating gene silencing, cell cycle progression and prevention of senescence and apoptosis. Canine SIR2 reverse transcriptase–polymerase chain reaction amplicons were most homologous to human SIRT2 and revealed detectable transcripts in all cell lines. Canine SIRT2 contained non‐conserved amino acid substitutions, representing mutations or allelic differences and interspecies differences. Sequence differences between individuals in p53 and SIRT2 were found in two cell lines including a stop codon in p53 and substitutions of conserved cysteine residues in the Zn2+‐binding motif in SIRT2. Mutations in SIRT2 were coincident with expression of the p53 modulator, Wip1; a failure to activate p21/Cip1 and extended G2/M phase. A third cell line appeared to function normally in these two pathways and likely possesses other defects in proliferation‐control genes. This data identify potentially important defects in pathways regulated by p53 and SIRT2 that modulate cell proliferation and integrate development, apoptosis and proliferative lifespan. These genes offer promising therapeutic targets, contributing to the transformed/immortalized phenotype in spontaneous canine mammary cancer.
Telomerase activity in canine osteosarcoma
Authors: Kow K. , Bailey S. M. , Williams E. S. , Withrow S. , Lana S. E.
Published: September 2006

Appendicular osteosarcoma (OSA) is the most common primary bone tumour in dogs, and the prognosis with standard of care therapy of amputation and adjunctive chemotherapy is generally poor, with median survival times of 1 year. The ability of neoplastic cells to maintain their telomere length, by either telomerase activity or alternate methods, is an important step in tumour development and malignancy. The purpose of this study was to determine the presence of telomerase activity in canine OSA. To evaluate the frequency of alternative lengthening of telomeres in canine OSA, we have used the telomeric repeat amplification protocol in five canine cell lines and in six samples taken from clinical patients at the time of amputation. Our results reveal the presence of telomerase activity in 100% of canine OSA cell lines and 83% of clinical samples evaluated. This is in contrast to human OSA where 25–40% expression levels of telomerase are reported. Importantly, our results not only suggest that canine OSA may serve as a good model for aggressive telomerase‐positive forms of human OSA but also that antitelomerase therapy strategies for treatment of canine OSA may be more successful than in the treatment of majority of human patients with OSA.
Cytological comparison of fine‐needle aspirates of liver and spleen of normal dogs and of dogs with cutaneous mast cell tumours and an ultrasonographically normal appearing liver and spleen
Authors: Finora K. , Leibman N. F. , Fettman M. J. , Powers B. E. , Hackett T. A. , Withrow S. J.
Published: September 2006

Staging of dogs with cutaneous mast cell tumours (MCTs) is an important diagnostic step. Aspiration of the liver and the spleen is often part of routine staging. This study cytologically compared mast cell numbers in fine‐needle aspirates of liver and spleen of clinically normal unaffected dogs with those of dogs with cutaneous MCT and an ultrasonographically normal appearing liver and spleen. The unaffected dogs (n = 32) were selected from humane society dogs, and the affected dogs (n = 51) were selected from hospital cases. There were no statistically significant differences in each of the parameters evaluated for the liver aspirates. For splenic aspirates, affected dogs showed significantly more mast cells per cluster (P = 0.04) and more isolated mast cells per slide (P = 0.03) compared with unaffected dogs. However, no clinically important difference existed between the unaffected and affected dogs; thus, routine aspiration of an ultrasonographically normal appearing liver and spleen of dogs with cutaneous MCT does not appear to be a clinically useful staging tool.
A Case Study in the Use of Community‐Based Participatory Research in Public Health Nursing
Authors: Savage Christine L. , Xu Yin , Lee Rebecca , Rose Barbara L. , Kappesser Mary , Anthony Jean Spann
Published: September 2006

ABSTRACT
There is growing demand for research using a community‐based participatory (CBPR) approach. CBPR requires that the academic research team actively partner with community members and stakeholders in the entire research process. The community members are full partners with the researchers in relation to the development and implementation of the study, analysis of the data, and dissemination of the findings. The purpose of this article is to review four basic principles of CBPR and provide an example of how these CBPR principles were used in an ethnographic study related to the culture of African American infant health. In the pilot study, CBPR provided the framework for recruitment and retention of participants, ongoing data analysis, and dissemination of findings. Using CBPR provided the researchers an introduction into the selected community. Community members served as key informants about the culture of the community and provided access to potential participants. The community partners contributed to analysis of emerging themes and in the dissemination of findings to the community, stakeholders, and the scientific community. CBPR provides opportunities for community health nurse researchers to conduct research with vulnerable populations and sets the stage for implementing evidenced‐based nursing interventions in the community.
An Innovative Approach to Chairside Provisional Replacement of an Extracted Anterior Tooth: Use of Fiber‐Reinforced Ribbon‐Composites and a Natural Tooth
Authors: Eminkahyagil Neslihan , Erkut Selim
Published: September 2006

Immediate chairside replacement of an extracted anterior tooth may contribute to a patient's comfort, treatment acceptance, and expectations of treatment; however, fabrication of a custom restoration in the anterior region of the mouth may result in an esthetic compromise for patients during the fabrication period. Chairside tooth replacement is an excellent application of fiber‐reinforced composite resin technology. This article presents an innovative, affordable chairside procedure in which Ribbond Multi‐Purpose Bondable Reinforcement Ribbon is used to replace a single extracted anterior tooth using the patient's own tooth.
The Effect of Complete Dentures with a Metal Palate on Candida Species Growth in HIV‐Infected Patients
Authors: Perezous Leticia F. , Stevenson Gene C. , Flaitz Catherine M. , Goldschmidt Millicent E. , Engelmeier Robert L. , Nichols C. Mark
Published: September 2006

Purpose: This in vivo investigation compared the oral candidal population between heat‐cured acrylic resin and nickel‐chromium‐beryllium alloy in maxillary complete dentures in HIV‐infected patients.

Materials and Methods: Split‐palate maxillary complete dentures were fabricated for 19 HIV‐infected patients: one‐half of the palate was made in acrylic resin and the other half in nickel‐chromium‐beryllium. Patients were divided into low or high CD4+ lymphocyte count groups. Dentures were worn for 5 months. Palatal mucosa was clinically evaluated at baseline, 1, 3, and 5 months after denture insertion. Specimens were collected at 1, 3, and 5 months using a modified imprint culture method. Speciation of Candida was performed using a chromogenic culture medium. Two‐sample t‐test was employed to determine effects and significant interactions between the control and test groups and the low and high CD4+ lymphocyte groups. A χ2 test analyzed and compared the results of the clinical evaluation (p < 0.05).

Results: Significant differences were observed in the colony counts between both materials during the third (p= 0.046) and fifth months (p= 0.039). The low CD4+ group demonstrated significant differences during the third (p= 0.03) and fifth months (p= 0.05). There were no significant differences between the species of Candida that colonized either material with the exception of Candida dubliniensis (p < 0.001) and “Others” (p < 0.001) during the first and fifth months. There were no significant differences on the clinical appearance of the palatal mucosa between both materials (p= 1.00).

Conclusions: The metal base proved to be effective in decreasing the fungal growth typically present in complete dentures. Although overt clinical manifestations were not present, colony counts of Candida species were high in the acrylic resin denture bases of these patients. This investigation demonstrated that metal base complete dentures provide an important alternative dental service for edentulous HIV‐positive and other patients who are particularly prone to higher incidences of fungal infections.
Calcium Phosphate Cement: Review of Mechanical and Biological Properties
Authors: Ambard Alberto J. , Mueninghoff Leonard
Published: September 2006

Purpose: Calcium phosphate cement is a bioactive and biodegradable grafting material in the form of powder and liquid, which when mixed, sets as primarily hydroxyapatite, sometimes mixed with unreacted particles and other phases. This material has been extensively investigated due to its excellent biological properties, potential resorbability, molding capabilities, and easy manipulation. Because the material can potentially be replaced with bone after a period of time, it could retain the short‐term biological advantages of hydroxyapatite without the long‐term disadvantages. Although little is known about this material in the dental community, in vivo and in vitro studies show calcium phosphate cement as a promising material for grafting applications. In the following article, the authors review the biological and mechanical properties of calcium phosphate cement, as well as its potential use in clinical applications.

Materials and Methods: A Medline search was performed (timeline: 1980 to 2003) using the following keywords: calcium phosphate cement, hydroxyapatite cement, HA cement, and hydroxyapatite. The search was limited to the English language. The patent literature as well as a limited number of master's theses and books were reviewed after using the electronic database search service from a dental school library.

Results: Calcium phosphate cement appears to have excellent biological properties. At only 2 weeks, spicules of living bone with normal bone marrow and osteocytes can be seen. Excellent moldability is a desired clinical characteristic; however, further research is necessary in order to improve the mechanical properties of the cement. The resorption/replacement by bone capability of the cement remains controversial. Further research is needed to clarify this issue. Due to poor mechanical properties, clinical applications are currently limited to craniofacial applications. Further research is necessary to take advantage of the excellent biological properties of this cement under clinical applications.

Conclusion: Further research is necessary to understand and improve the behavior of this type of cement under clinical situations.
Effect of the Association of Nystatin with a Tissue Conditioner on its Ultimate Tensile Strength
Authors: Urban Vanessa Migliorini , De Souza Raphael Freitas , Galvao Arrais Cesar Augusto , Borsato Karina Tostes , Vaz Luís Geraldo
Published: September 2006

Purpose: This study evaluated the ultimate tensile strength of a tissue conditioner without nystatin incorporation (GI—control group) and the same tissue conditioner modified by the addition of nystatin in two concentrations: GII—500,000 International Units (U) and GIII—1,000,000 U, in which each milligram of the medicament corresponded to 6079 U.

Materials and Methods: Dumbbell‐shaped specimens (N= 7) with a central cross‐sectional area of 33 × 6 × 3 mm were produced for the three experimental groups. After polymerization following manufacturer's instructions, specimens were immersed in distilled water at 37°C for either 24 hours or 7 days and then tested in tension in the MTS 810 at 40 mm/minute. Data were analyzed by two‐way ANOVA followed by Tukey's test, at 95% level of confidence.

Results: The means (force‐grams (gf) ± standard deviation) of the ultimate tensile strength were: GI—634.29 ± 122.80; GII—561.92 ± 133.56; and GIII—547.30 ± 73.47 for 24‐hour storage, and GI—536.68 ± 54.71; GII—467.50 ± 143.51; and GIII—500.62 ± 159.76 for 7‐day storage. There were no statistically significant differences among the three experimental groups (p > 0.05). The ultimate tensile strength means of all experimental groups after 7 days were significantly lower than those observed after 24 hours (p= 0.04).

Conclusions: The results of this study suggest that the addition of nystatin into the tissue conditioner investigated in concentrations below 1,000,000 U did not affect its ultimate tensile strength.
Effect of Chemical Disinfectants and Repair Materials on the Transverse Strength of Repaired Heat‐Polymerized Acrylic Resin
Authors: Ellakwa Ayman E. , El‐Sheikh Ali M.
Published: September 2006

Purpose: The purpose of this study was to evaluate both the effects of immersion in different chemical disinfectant solutions and the type of repair material on the transverse strength of repaired heat‐polymerized acrylic resin.

Materials and Methods
: A total of 110 rectangular specimens (65 × 10 × 3 mm) of heat‐polymerized acrylic resin (Triplex) were fabricated. After polymerization, the specimens were polished, then stored in distilled water at 37°C for 1 week. The specimens were divided into 11 groups (n = 10) coded A to K. Specimens of Group A remained intact (control). The specimens of Groups C to F and Groups H to K were immersed in the following chemical disinfectant solutions (1%, 2.5%, and 5.25% sodium hypochlorite and 2% glutaraldehyde, respectively) for 10 minutes. The specimens of all groups except those of Group A were sectioned in the middle to create 10 mm gaps and repaired with the same resin (Groups B to F) and autopolymerizing acrylic resin (Groups G to K). The specimens of Groups C to F and Groups H to K were again immersed in the disinfectant solutions in the same sequence. The transverse strength (N/mm2) was tested for failure in a universal testing machine, at a crosshead speed of 5 mm/min. Two‐way analysis of variance (ANOVA) was performed to evaluate the effects of both the disinfectant solutions and repair materials on the transverse strength of repaired specimens. All data were statistically analyzed using one‐way analysis of variance followed by Tukey's test at 95% confidence level.

Results
: The repaired specimens treated with/without disinfectant solutions showed similar (p > 0.05) transverse strength values. No differences (p > 0.05) were detected among the repaired specimens either with heat‐polymerized or autopolymerizing acrylic resins. The intact specimens showed transverse strength values (86.9 ± 11.8) significantly higher (p < 0.05) than the values of the repaired specimens.

Conclusions
: Among the repaired specimens, transverse strength was not affected after immersion in the disinfectants for the immersion period tested (10 min). The repair material, either heat‐polymerized or autopolymerizing acrylic resin, had no effect on the transverse strength of the repaired acrylic resin specimens.
Prospective comparison of high‐dose plateletpheresis with the latest apheresis systems on the same donors
Authors: Picker Susanne Maria , Radojska Stela Marinova , Gathof Birgit Sybille
Published: September 2006

BACKGROUND: To improve productivity of automated platelet (PLT) collection, the industry has introduced new instruments or modifications to existing equipment.

STUDY DESIGN AND METHODS: With the same 8 donors for double (DDC) and triple‐dose PLT collection (TDC), the Baxter Amicus (AM), the Haemonetics MCS Plus (MCS+), and the Gambro Trima Accel (TA) were evaluated focusing on yield, duration, and citrate donor load. Target endpoints were set at 5.5 × 1011 to 6.0 × 1011 PLTs (DDC) and 7.5 × 1011 to 8.0 × 1011 PLTs (TDC) in up to 100 and 120 minutes’ donation time, respectively.

RESULTS: TA was the most efficient system (74.5 ± 3.9%) with significant differences from AM (71.1 ± 3.9%; p = 0.028) and MCS+ (64.0 ± 7.7%; p = 0.002). TA had advantages over AM for collection rate (10.9 × 109 ± 2.2 × 109 vs. 10.1 × 109 ± 1.5 × 109 PLTs/min; p = 0.382), whole blood processed (3928 ± 611 mL vs. 4219 ± 727 mL; p = 0.382), and time to obtain an established standard dose (TSD 2.5EU, 30.2 ± 5.6 vs. 37.7 ± 5.5 min; TSD 3.5US, 42.2 ± 7.8 min vs. 52.7 ± 7.7 min; p = 0.015), whereas AM was slightly superior in PLT yield (2.81 × 1011 ± 0.21 × 1011 vs. 2.76 × 1011 ± 0.31 × 1011/unit; p = 0.645). Owing to the lowest draw (42.3 ± 3.2 mL/min; p < 0.001) and collection rates (6.0 × 1011 ± 1.5 × 1011/min; p = 0.021), MCS+ was the slowest significantly (p < 0.001) but compensated with fewer citrate reactions owing to lower citrate infusion rates (0.78 ± 0.11 mL/min/L; p = 0.028).

CONCLUSION: High‐dose plateletpheresis was performed efficiently and safely with all three instruments. AM had advantages in PLT yield, and MCS+, in donor comfort. TA was the fastest in obtaining an established standard dose and, because of this advantage, the machine with the highest practical impact in routine use.
Transfusion‐related acute lung injury and pulmonary edema in critically ill patients: a retrospective study
Authors: Rana Rimki , Fernández‐Pérez Evans R. , Khan S. Anjum , Rana Sameer , Winters Jeffrey L. , Lesnick Timothy G. , Moore S. Breanndan , Gajic Ognjen
Published: September 2006

BACKGROUND: Using the recent Consensus Panel recommendations, we sought to describe the incidence of transfusion‐related acute lung injury (TRALI) and transfusion‐associated circulatory overload (TACO) in critically ill patients.

STUDY DESIGN AND METHODS: Consecutive patients at four intensive care units (ICUs) who did not require respiratory support at the time of transfusion were identified with custom electronic surveillance system that prospectively tracks the time of transfusion and onset of respiratory support. Respiratory failure was defined as the onset of noninvasive or invasive ventilator support within 6 hours of transfusion. Experts blinded to specific transfusion factors categorized the cases of pulmonary edema as permeability edema (suspected or possible TRALI) or hydrostatic edema (TACO) according to predefined algorithm. In a nested case‐control design, transfusion variables and lung injury risk factors were compared between the TRALI cases and controls matched by age, sex, and admission diagnosis.

RESULTS: There were 8902 units transfused in 1351 patients of whom 94 required new respiratory support within 6 hours of transfusion. Among 49 patients with confirmed acute pulmonary edema, experts identified 7 cases with suspected TRALI, 17 patients with possible TRALI, and 25 cases with TACO. The incidence of suspected TRALI was 1 in 1271 units transfused; possible TRALI, 1 in 534 per unit transfused; and TACO, 1 in 356 per unit transfused. When adjusted for sepsis and fluid balance in a stepwise conditional logistic regression analysis, patients who developed acute lung injury (suspected or possible TRALI) received larger amount of plasma (odds ratio 3.4, 95% confidence interval 1.2‐10.2, for each liter infused; p = 0.023).

CONCLUSION: In the ICU, pulmonary edema frequently occurs after blood transfusion. The association between infusion of plasma and the development of suspected or possible TRALI may have important implications with regards to etiology and prevention of this syndrome.
Arsine toxicity treated with red blood cell and plasma exchanges
Authors: Danielson Constance , Houseworth Julie , Skipworth Elaine , Smith Daniel , McCarthy Leo , Nanagas Kristine
Published: September 2006

BACKGROUND:  Acute toxicity due to inhalation of arsine gas (AsH3) has no known antidote. Exchange transfusion may be beneficial, and dialysis is often required because arsine may cause acute intravascular hemolysis and renal failure. A patient with arsine toxicity has recently been treated by both red blood cell exchange (RBC‐E) and plasma exchange (PE) therapy and our experience is reported.

CASE REPORT:  A 46‐year‐old man was accidentally and unknowingly exposed to arsine gas while observing an industrial procedure. Within 6 hours he developed fatigue, nausea, vomiting, and tingling in his extremities and voided dark urine. He quickly developed renal failure secondary to acute arsine toxicity (arsenic level, 1250 µg/L). Laboratory findings were a hematocrit level of 24 percent; blood urea nitrogen and creatinine, 84 and 5.5 mg per dL, respectively; bilirubin, 9.1 mg per dL; indirect bilirubin, 6.8 mg per dL; haptoglobin, less than 6 (normal, 30‐200); and lactic dehydrogenase, 10,413 units per L (normal, 265‐580). An emergent 1‐vol RBC‐E transfusion by continuous‐flow method revealed dramatic black, grossly hemolyzed plasma. After two additional RBC‐E and two PE and daily hemodialysis, he completely recovered over the course of 1 month.

CONCLUSION:  Patients with arsine toxicity resulting in intravascular hemolysis should receive RBC‐E as soon as possible. In addition, PE may be beneficial in removing the components of RBC lysis and further reducing arsenic levels.
Blood volume regulation in donors undergoing intermittent‐flow plasmapheresis involving a high extracorporeal blood volume
Authors: Karger Ralf , Halbe Monika , Dinges Gerhard , Wulf Hinnerk , Kretschmer Volker
Published: September 2006

BACKGROUND: Intermittent‐flow plasmapheresis (IFP) often involves a large extracorporeal blood volume (ECV) of donors during donation. Depending on equipment and donor characteristics, ECV can exceed 20 percent of a donor’s blood volume (BV). It was the aim of this study to delineate mechanisms of BV regulation associated with these volume shifts.

STUDY DESIGN AND METHODS: Parameters of BV regulation were recorded in 60 donors (30 men, 30 women) undergoing IFP, who were randomly selected after stratification for sex and BV. Shock index (SI), stroke volume (SV), cardiac index (CI), and thoracic fluid content (TFC) were determined at the beginning of the procedure and when maximum ECV (ECVmax) was reached with noninvasive techniques. In a control investigation, donors were kept in reclined position for the duration of an IFP session without actually donating.

RESULTS: SI increased significantly during IFP (+0.18; p < 0.0001). SV decreased significantly (−14.3 mL/stroke; p < 0.0001). CI did not decrease significantly (−0.07 L/min/m2 body surface area; p = 0.33). Preservation of CI was due to a significant rise in heart rate (+13.4 beats/min; p < 0.0001). TFC decreased significantly during IFP (−0.77 kΩ−1; p < 0.0001), indicating the development of an intravascular volume deficit. The changes of SI, heart rate, and TFC weakly correlated with ECVmax.

CONCLUSION: The hemodynamic response during IFP is consistent with a hypovolemic challenge of the donors and is sufficient to maintain cardiac function. ECVmax during donation does not reliably predict the degree of hypovolemic stress, as long as it remains below 20 percent. This might warrant reevaluation of collection limits based on ECVmax.
Life, death, and the risk of transfusion: a university hospital experience
Authors: Hay Shauna N. , Scanga Lori , Brecher Mark E.
Published: September 2006

BACKGROUND: Frequently one encounters donor recruitment statements like “. . . about 90 percent of American adults will need at least one blood transfusion in the course of a lifetime.” To assess the validity of such statements, the lifetime transfusion rate of patients dying in our tertiary‐care hospital was determined. It would be expected that patients dying in a tertiary‐care, Level 1 trauma center university hospital would have one of the highest transfusion rates.

STUDY DESIGN AND METHODS: A retrospective review of blood bank and electronic clinical records was conducted for all patients who expired at our institution from January 1, 2004, through December 31, 2004. Stillborn infants were excluded from analysis. The rate of transfusion was stratified by age and sex.

RESULTS: A total of 906 patients expired in 2004 at our institution. Of these, 35 were excluded (stillborn infants), leaving 871 patients for analysis. After review of blood bank records, 591 patients received blood products (67.9%). An additional 27 patients (3.1%) had a possible transfusion at another institution as the clinical record indicated complex surgical history. Only 253 patients (29.0%) did not have a documented transfusion event. Individuals who died before the age of 31 had the highest rates of transfusion (p  < 0.001, Fisher’s exact test, two‐sided).

CONCLUSIONS: The majority of patients who expire at our institution have a history of transfusion (67.9%‐71.0%). Patients who die before the age of 31 have the highest rate of transfusion.
Absence of detectable viremia in plasma and peripheral blood mononuclear cells from smallpox vaccinees: implications for blood safety
Authors: Srinivasan Kumar , Akolkar Pradip N. , Taffs Rolf E. , Hewlett Indira K.
Published: September 2006

BACKGROUND:  Mass smallpox vaccination with live vaccinia virus has been considered as a preventive measure to counter bioterrorism involving smallpox. This has raised concerns about the possibility of vaccinia virus being transmitted from vaccinated blood donors to recipients. The results of this study could be used to define an appropriate deferral period for blood donors (vaccinated against smallpox) to ensure safety of the blood supply.

STUDY DESIGN AND METHODS:  A procedure was developed to culture vaccinia virus from plasma and peripheral blood mononuclear cells (PBMNCs) of vaccinees enrolled in three smallpox vaccine clinical trials. A total of 665 plasma and PBMNC samples were obtained from 95 vaccinated subjects.

RESULTS:  Vaccinia viremia was not detected by virus culture from plasma and PBMNC samples of healthy vaccinees 3 to 56 days after vaccination under our assay conditions. Plasma viremia assay had a sensitivity of approximately 66 plaque‐forming units per mL with a Vero cell culture assay.

CONCLUSION:  The results of this study present evidence that in the case of mass vaccination, the risk of transmission of vaccinia virus by blood transfusion would likely be low.
KEL6 and KEL7 genotyping with sequence‐specific primers
Authors: Renoud Keli J. , Barracchini Kathleen , Byrne Karen M. , Adams Sharon , Pickett Angela , Caruccio Lorraine , Stroncek David F.
Published: September 2006

BACKGROUND:  Although antibodies to Jsa and Jsb are clinically significant, reagent‐quality anti‐Jsa and anti‐Jsb are not readily available. A sequence‐specific primer–polymerase chain reaction (SSP‐PCR) genotyping assay was tested that makes use of two single‐nucleotide polymorphisms (SNPs) at positions 1910 and 2019 of KEL. These SNPs distinguish the gene encoding Jsa, KEL6; and Jsb, KEL7.

STUDY DESIGN AND METHODS:  Four primer sets that selectively amplified KEL6 and KEL7 from genomic DNA were developed. Two sets detected the SNP at bp 1910 and two sets detected the bp 2019 SNP. KEL6 and KEL7 genotyping and Jsa and Jsb phenotyping results were compared among 64 subjects.

RESULTS:  The SSP‐PCRs were specific for KEL6 and KEL7 when testing DNA for three donors of known Js phenotype: Js(a+b–), Js(a–b+), and Js(a+b+). Genotyping results for the 1910 SNP were identical to the phenotyping results in all 64 subjects, but for the 2019 SNP, the genotyping and phenotyping results were identical for only 49 subjects. In 12 subjects with the Js(a–b+) phenotype, the 2019 SNP was heterozygous KEL6, KEL7; in 2 with Js(a–b+) and in 1 with Js(a+b+), the 2019 SNP was homozygous KEL6.

CONCLUSION:  KEL 2019‐bp SNP does not always correlate with the Js phenotype owing to the presence of an atypical KEL gene with a KEL7 polymorphism at 1910 and a KEL6 polymorphism at 2019. The KEL polymorphism at 2019 is silent and this allele yields a Js(a–b+) phenotype. Only analysis of the 1910‐bp SNP can be used to genotype KEL6 and KEL7.
Mirasol PRT treatment of donor white blood cells prevents the development of xenogeneic graft‐versus‐host disease in Rag2–/–γc–/– double knockout mice
Authors: Fast Loren D. , DiLeone Gilbert , Cardarelli Gene , Li Junzhi , Goodrich Raymond
Published: September 2006

BACKGROUND: Mirasol PRT (Navigant Biotechnologies) treatment utilizes exposure to light in the presence of riboflavin to introduce irreparable lesions to nucleic acids thereby inhibiting pathogen and WBC replication. The ability of Mirasol PRT–treated mononuclear cells (MNCs) to generate xenogeneic graft‐versus‐host disease (GVHD) responses was used to model transfusion‐associated GVHD (TAGVHD).

STUDY DESIGN AND METHODS: Pairs of human MNCs from five different individual donors that had or had not received Mirasol PRT treatment and contained 30 × 106 CD3+ cells were injected intraperitoneally into sublethally irradiated (350 cGy) Rag2–/–γc–/– double‐knockout mice. Recipient mice were weighed and observed regularly and euthanized when they exhibited symptoms of GVHD or at termination of the experiment. Recipient lymphoid compartments were collected and phenotyped for the presence of human lymphoid cells. The presence of human cytokines and/or immunoglobulins in the recipient plasma was also used to detect the presence of human cells.

RESULTS: Twelve of 14 mice injected with untreated cells developed xenogeneic GVHD, whereas 0 of 14 mice injected with Mirasol PRT–treated cells developed xenogeneic GVHD. End‐stage xenogeneic GVHD in the recipients of untreated cells was characterized by the presence of splenic human cytolytic CD4+ and CD8+ cells, with high levels of interferon‐γ, interleukin‐10, and xenoreactive antibodies in the plasma.

CONCLUSION: Mirasol PRT treatment of the donor MNCs abolished xenogeneic GVHD responses, indicating that the use of Mirasol PRT treatment of blood products should prevent the development of TAGVHD.
Leishmania: risk to the blood supply
Authors: Cardo Lisa J.
Published: September 2006

Leishmania infection is most often transmitted to humans via the bite of the phlebotomine sandfly, but transmission of Leishmania by transfusion has also been reported. There has been a huge increase in the incidence of cutaneous and visceral leishmaniasis in Iraq and Afghanistan. The deployment of US troops to these countries and published case reports of transmission to soldiers in endemic areas, by transfusion to infants with immature immune systems, and to individuals immunocompromised by disease or immunosuppressive therapy beckon a reexamination of blood donor deferral procedures. The length of the ongoing military conflict and the nature of exposure indicate that prior decisions regarding blood donor deferral made during the first Gulf War may no longer apply. Operation Iraqi Freedom and Operation Enduring Freedom present a much greater Leishmania threat than did Operation Desert Storm. Because most transmission by transfusion occurs in endemic areas, and visceral infection is asymptomatic in healthy individuals such as blood donors, it is difficult to determine the absolute risk of transmission by transfusion, but review of the literature provides many clues as to the appropriate measures to be taken for blood donor deferral.
Optimal prewarming conditions for Rh antibody testing
Authors: Endoh Teruo , Kobayashi Daisuke , Tsuji Naoki , Tanabe Hiromi , Koshida Saori , Yagihashi Atsuhito , Watanabe Naoki
Published: September 2006

BACKGROUND: Although prewarming (PW) can reduce the confounding agglutination of cold‐reactive antibodies when testing for warm‐reactive antibodies, PW also can adversely affect the detection of warm‐reactive antibodies. This study was conducted to determine PW conditions that optimized Rh antibody detection.

STUDY DESIGN AND METHODS: In 38 plasma samples with Rh system antibodies detected by any of four methods, and in 8 other samples with cold‐reactive antibodies, reactivity was assessed by titer and score.

RESULTS: PW to 37°C often reduced reaction scores by all methods especially the low‐ionic‐strength saline indirect antiglobulin test and bromelin methods. In analyses warming red blood cell (RBC) suspensions or plasma, the reaction scores were decreased only when the RBC suspension was warmed, suggesting that RBC PW decreased Rh antibody reactivity. The warming period ranging from 5 to 30 minutes all decreased Rh system reaction scores, an effect persisting up to 120 minutes. At lower temperatures (27, 30, and 33°C), numbers of samples showing decreased reaction score for Rh system antibodies decreased in a temperature‐dependent manner. Testing at 27°C also permitted some agglutination involving cold‐reactive antibodies, whereas higher temperatures successfully suppressed their agglutination.

CONCLUSION: PW at 30°C minimizes problems in accurately detecting Rh system antibodies.
Gene frequencies of the HNA‐4a and ‐5a neutrophil antigens in Brazilian persons and a new polymerase chain reaction‐restriction fragment length polymorphism method for HNA‐5a genotyping
Authors: Cardone José Daniel Braz , Bordin José Orlando , Chiba Akemi Kuroda , Norcia Angela Maria Matico Ikeda , Vieira‐Filho João Paulo Botelho
Published: September 2006

BACKGROUND: The HNA‐4a (Mart) and HNA‐5a (Ond) antigens are polymorphic variants of αM (CD11b) and αL (CD11a) subunits of the β2‐integrin, and are associated with single nucleotide polymorphisms (SNP) leading to amino acid dimorphisms. HNA‐4a has been linked to alloimmune neonatal neutropenia, but the HNA‐5a clinical significance is unclear.

STUDY DESIGN AND METHODS: Using a polymerase chain reaction (PCR) with sequence‐specific primers, the frequency of HNA‐4a among 121 Brazilian blood donors and 114 Amazon Indians was determined. A PCR‐restriction fragment length polymorphism (RFLP) method for HNA‐5a genotyping was developed, and the gene frequencies of this antigen were determined among 123 blood donors and 114 Indians. To validate the genotyping method, the amplified DNA from six previously obtained samples (two of each genotype) was sequenced.

RESULTS: The HNA‐4a (+/+), HNA‐4a (+/−), and HNA‐4a (−/−) genotype frequencies of blood donors (0.686, 0.273, 0.041) and Indians (1.000, 0.000, 0.000) were different (p < 0.01). The frequencies of HNA‐5a (+/+), HNA‐5a (+/−), and HNA‐5a (−/−) genotypes among blood donors (0.512, 0.399, 0.089) and Indians (0.746, 0.219, 0.035) also differed (p < 0.01). Sequencing demonstrated concordance with PCR‐RFLP genotyping in all six evaluated samples.

CONCLUSION: Comparing to another populations, Brazilians present a higher frequency of HNA‐4a‐negative allele, suggesting that Brazilians would be more susceptible to HNA‐4a alloimmunization. Moreover, the distribution of the HNA‐4 alleles observed in Amazon Indians is quite similar to that reported among Koreans. Besides that, a new effective and efficient HNA‐5a genotyping technique is now available for population studies.
Storage duration and white blood cell content of red blood cell (RBC) products increases adhesion of stored RBCs to endothelium under flow conditions
Authors: Anniss Angela M. , Sparrow Rosemary L.
Published: September 2006

BACKGROUND: Adherence of red blood cells (RBCs) to vascular endothelium impairs blood flow and decreases oxygen delivery. Although RBCs may be stored for up to 42 days before transfusion under current blood banking guidelines, little is known of how changes to RBCs during storage may affect their adherence properties. The influence of RBC product storage time and white blood cell (WBC) burden on the adherence of RBCs for transfusion to vascular endothelium under conditions of continuous flow was investigated in this study.

STUDY DESIGN AND METHODS: RBC samples were collected from nonleukoreduced (S‐RBC), buffy coat–poor (BCP‐RBC), and leukofiltered (LF‐RBC) products at fixed time points during storage. Samples were perfused, at controlled shear stress and temperature, across a confluent endothelial cell (EC) monolayer with a parallel‐flow chamber mounted to an inverted microscope. RBC‐EC interactions were recorded with a digital camera attached to the microscope.

RESULTS: The number of RBCs adhering to the EC layer increased significantly with storage time in all RBC products; however, WBC reduction delayed this increase. LF‐RBCs were also significantly less adherent than S‐RBC or BCP‐RBC products on Day 1 of storage (p  < 0.05). The strength of RBC attachment to vascular endothelium was significantly stronger in S‐RBC products compared to BCP‐RBC and LF‐RBC products.

CONCLUSION: Our findings indicate that product storage time and WBC burden increase the number and strength of adhesion of RBCs to vascular endothelium. These results may lead to greater understanding of the interaction of transfused RBCs with recipient endothelium and the biologic consequences of this adherence.
Recipient inflammation affects the frequency and magnitude of immunization to transfused red blood cells
Authors: Hendrickson Jeanne E. , Desmarets Maxime , Deshpande Seema S. , Chadwick Traci E. , Hillyer Christopher D. , Roback John D. , Zimring James C.
Published: September 2006

BACKGROUND: Most alloantigens on transfused red blood cells (RBCs) are weakly immunogenic, with only a 2 to 6 percent overall immunization rate even in patients receiving multiple transfusions. Although recipient genetics may contribute to responder and/or nonresponder status, in most cases HLA type does not predict humoral response to RBC antigens. In contrast, rates of alloimmunization do correspond to the underlying disease status of transfusion recipients, suggesting that acquired host factors may play an important role. In this context, it was hypothesized that the inflammatory status of a transfusion recipient would influence immunization to transfused RBCs.

STUDY DESIGN AND METHODS: A novel murine model for alloimmunization to RBC antigens was developed with the mHEL mouse, which expresses hen egg lysozyme (HEL) as a model blood group antigen. Leukoreduced mHEL RBCs were transfused into wild‐type recipient mice, and anti‐HEL responses were monitored. To test the stated hypothesis, some recipient animals were injected with polyinosinic polycytidylic acid (poly(I:C)), a synthetic double‐stranded RNA molecule that induces viral‐like inflammation.

RESULTS: Similar to the immunogenicity of most RBC antigens in humans, transfusion of mHEL RBCs into uninflamed mice was only a weak immunogen. In contrast, poly(I:C)‐treated mice had a significant increase in both the frequency and the magnitude of alloimmunization to the mHEL antigen.

CONCLUSIONS: These findings demonstrate that recipient inflammation with poly(I:C) significantly enhances humoral immunization to transfused alloantigens in a murine model. Moreover, these data suggest that the inflammatory status of human transfusion recipients may regulate the immunogenicity of transfused RBCs.
Intergovernmental Management: A View from the Bottom
Authors: McGuire Michael
Published: September 2006

Intergovernmental management is more than just intergovernmental relationships involving the federal government. Though Professor McGuire agrees that the federal government has become more intrusive and opportunistic since the mid‐20th century—resulting in less cooperative intergovernmental relations—he argues that collaborative management is much more prevalent than it is depicted in Dr. Conlan’s analysis. A bottom‐up view suggests that local and regional activity can be both opportunistic and collaborative, and such a perspective must be considered in any discussion of managing federalism.
From Virtue to Competence: Changing the Principles of Public Service
Authors: Macaulay Michael , Lawton Alan
Published: September 2006

Virtue has long been a central principle in the tradition of public service—to what extent is it still relevant today? Focusing on the role of the monitoring officer, a key official in the ethical framework of local government in the United Kingdom, this essay asks which virtues, if any, are still needed for public service and whether these virtues have been displaced by managerial notions of technical competence as the principles of public service delivery. The authors draw an initial distinction between virtue and competence that, upon further investigation, does not appear to be sustainable. Despite being drawn from two different academic perspectives—moral philosophy and management development—the concepts of virtue and competence are, in practice, very similar. This theoretical convergence is reflected in the practical concerns of monitoring officers and their perspective on public service ethics.