المؤلفون: Yang, Jiangcun, Yin, Wen, Zhang, Yali, Sun, Yang, Ma, Ting, Gu, Shunli, Gao, Ying, Zhang, Xiaole, Yuan, Jun, Wang, Wenhua

المصدر: Transfusion; Mar2018, Vol. 58 Issue 3, p736-747, 12p, 1 Color Photograph, 1 Black and White Photograph, 2 Charts, 4 Graphs

مصطلحات موضوعية: BLOOD platelets, BACTERIAL contamination, LACTATE dehydrogenase, BLOOD transfusion, CELL morphology, HEMORRHAGE, BLOOD collection, BLOOD gases analysis, BLOOD platelet activation, BLOOD platelet aggregation, HEMOSTASIS, LACTIC acid, TEMPERATURE, PLATELET-rich plasma, PREVENTION

مستخلص: Background: Platelet (PLT) storage at cold temperatures (4°C) can reduce bacterial contamination and lower the risk of transfusion-related complications. We compared the effects of 22 and 4°C storage conditions for PLTs to further explore the efficiency of hemostasis in acute bleeding and extended PLT shelf life.Study Design and Methods: Manually prepared PLTs (PLT concentrates in plasma, not PLT additive solution) were stored at 4 and 22°C. The PLT counts, scanning electronic microscope observations, blood gas indices, biochemical indices, PLT aggregative function, and surface CD62P expression were monitored and compared between the groups.Results: There was no obvious change in PLT counts between Day 21 at 4°C and Day 5 at 22°C. PLTs stored at 4°C for 10 to 14 days were dramatically activated, had rough surfaces, and showed a significant degree of long pseudopodia formation. The pH of the PLTs on Day 5 was lower at 22°C than at 4°C, while the lactate dehydrogenase and lactic acid levels in the former group were significantly higher (p < 0.005). The maximum aggregation rates induced by collagen and arachidonic acid in the PLTs stored at 4°C for 5 days remained higher than 80%, while the rates induced by four inducers in the PLTs stored at 22°C were less than 5%. PLTs stored at 4°C for 10 to 14 days showed higher surface expression of PAC-1 and CD62P.Conclusion: PLT counts, cellular morphologies, PLT membranes, cytoplasmic structures, aggregation rates, and hemostatic PLT function stored at 4°C for 10 to 14 days were better than those stored at 22°C for 5 days. [ABSTRACT FROM AUTHOR]

: Copyright of Transfusion is the property of Wiley-Blackwell and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

المؤلفون: Jeannie Callum, Darinka Sakac, Nagina Parmar, Yulia Lin, Katerina Pavenski, Qi-Long Yi, Tik Nga Tong, Lani Lieberman, Jacob Pendergrast, Christine Cserti-Gazdewich, Donald R. Branch, Megan Blacquiere, Wendy Lau, Nadine Shehata, Emeralda Burke-Murphy

المصدر: Transfusion. 60:3010-3018

مصطلحات موضوعية: Adult, Male, medicine.medical_specialty, Immunology, 030204 cardiovascular system & hematology, Hemolysis, Gastroenterology, Monocytes, 03 medical and health sciences, chemistry.chemical_compound, 0302 clinical medicine, In vivo, hemic and lymphatic diseases, Lactate dehydrogenase, Internal medicine, medicine, Humans, Immunology and Allergy, Hematologic Tests, biology, business.industry, Monocyte, Autologous Monocytes, Haptoglobin, Immunoglobulins, Intravenous, food and beverages, Hematology, medicine.disease, medicine.anatomical_structure, chemistry, biology.protein, Hemoglobin, Antibody, business, 030215 immunology

الوصف: Background Hemolysis following the administration of intravenous immunoglobulin (IVIG) is an important adverse event (AE). While the monocyte monolayer assay (MMA) has been used to predict in vivo hemolysis when serologically incompatible blood may be transfused, it has also been shown to correlate with IVIG-associated hemolysis. In this study, the MMA was examined for its utility in assessing the risk of hemolysis after IVIG. Study design and methods Forty-two non-blood group O patients receiving high-dose IVIG (≥2 g/kg) were examined using an autologous and allogeneic MMA. Hemolysis was defined by a drop in hemoglobin of ≥1 g/L, a positive direct antiglobulin test (DAT) and eluate, and a decrease in haptoglobin or increase in lactate dehydrogenase and/or reticulocytes. Results Forty-two patients provided 50 assessable postinfusion samples, with hemolysis observed in 20 (40%) of cases. Autologous MMA using post-IVIG red blood cells significantly correlated with clinical outcomes when compared to allogeneic MMA (P = .0320 vs .5806, t test). No significant difference in receiver operating characteristics was observed when comparing autologous MMA testing against DAT for the diagnosis of IVIG-associated hemolysis. However, when using samples collected 5 to 10 days after receipt of high-dose IVIG, the autologous MMA had higher sensitivity than the DAT. Conclusion MMA testing with autologous monocytes collected 5 to 10 days after receipt of high-dose IVIG can be used for the diagnosis of IVIG-associated hemolysis and may be of particular value in cases in which the Day 5 to 10 DAT is negative.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::f6a767c7da2c52b549f00e393acc6f18Test
https://doi.org/10.1111/trf.16131Test

المؤلفون: Peters, Anna L., Kunanayagam, Renoja K., van Bruggen, Robin, de Korte, Dirk, Juffermans, Nicole P., Vlaar, Alexander P.J.

المصدر: Transfusion; Jan2017, Vol. 57 Issue 1, p53-59, 7p, 2 Graphs

مصطلحات موضوعية: RED blood cell transfusion, BLOOD transfusion, ERYTHROCYTES, BLOOD plasma, ENDOTOXEMIA, IRON in the blood, TREATMENT of endotoxemia, BILIRUBIN, BLOOD collection, AUTOTRANSFUSION of blood, BLOOD proteins, CLINICAL trials, COMPARATIVE studies, FERRITIN, GLOBULINS, HEMOGLOBINS, IRON, LACTATE dehydrogenase, RESEARCH methodology, MEDICAL cooperation, RESEARCH, TIME, EVALUATION research, LIPOPOLYSACCHARIDES

مستخلص: Background: Transfusion of a single unit of stored red blood cells (RBCs) has been hypothesized to induce supra-physiological levels of non-transferrin bound iron (NTBI), which may enhance inflammation and act as a nutrient for bacteria. We investigated the relation between RBC storage time and iron levels in a clinically relevant "two-hit" human transfusion model.Study Design and Methods: Eighteen healthy male volunteers (ages 18-35 years) were infused with 2 ng lipopolysaccharide (LPS)/kg to induce systemic inflammatory response syndrome. Two hours later, each participant received either 1 unit of 2-day stored (2D) autologous RBCs, 35-day stored (35D) autologous RBCs, or an equal volume of saline. Every 2 hours up to 8 hours after LPS infusion, hemoglobin, hemolysis parameters, and iron parameters, including NTBI, were measured.Results: Transfusion of both 2D and 35D RBCs caused increases in hemoglobin, plasma iron, and transferrin saturation; whereas levels remained stable in the saline group. Transfusion of 35D RBCs did not result in hemolysis nor did it lead to increased levels of NTBI compared with 2D RBCs or saline. LPS induced increases in ferritin, haptoglobin, bilirubin, and lactate dehydrogenase that were similar in all three groups.Conclusion: We conclude that 35D autologous RBCs do not cause hemolysis or increased levels of NTBI during human endotoxemia. [ABSTRACT FROM AUTHOR]

: Copyright of Transfusion is the property of Wiley-Blackwell and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

المؤلفون: Christine M. Leeper, Mark H. Yazer, Barbara A. Gaines, Katrina M Morgan, Stephen Strotmeyer, Darrell J. Triulzi

المصدر: Transfusion. 61

مصطلحات موضوعية: Male, medicine.medical_specialty, Adolescent, Bilirubin, Immunology, Hemolysis, ABO Blood-Group System, chemistry.chemical_compound, Lactate dehydrogenase, Internal medicine, Humans, Immunology and Allergy, Medicine, Blood Transfusion, Child, Adverse effect, Whole blood, Creatinine, biology, business.industry, Haptoglobin, Transfusion Reaction, Hematology, medicine.disease, chemistry, Child, Preschool, biology.protein, Female, business, Pediatric trauma

الوصف: BACKGROUND: Low-titer Group O Whole Blood (LTOWB) is used with increasing frequency in adult and pediatric trauma and massive bleeding transfusion protocols. There is a risk of acute hemolytic reactions in non-group O recipients due to the passive transfusion of anti-A and anti-B in the LTOWB. This study investigated the hemolysis risk among pediatric recipients of LTOWB. STUDY DESIGN AND METHODS: Blood bank records were queried for pediatric recipients of LTOWB between June 2016 and August 2020 and merged with clinical data. The primary outcome was laboratory evidence of hemolysis as manifested by changes in lactate dehydrogenase (LDH), haptoglobin, total bilirubin, reticulocyte count, potassium, and creatinine. Per protocol, these values were collected on hospital days 0-2 for recipients of LTOWB. Transfusion reactions were reported to the hospital's blood bank. RESULTS: Forty-seven children received LTOWB transfusion between 2016 and 2020; 21 were group O and 26 were non-group O. The groups were comparable in terms of the total volume of transfused blood products, demographics, and clinical outcomes. The most common indication for LTOWB transfusion was hemorrhagic shock due to trauma. There were no clinically or statistically significant differences in baseline, post-transfusion day 1, or post-transfusion day 2 hemolysis markers between the group O and non-group O LTOWB recipients. There were no adverse events or transfusion reactions reported. DISCUSSION: Use of up to 40 ml/kg of LTOWB appears to be serologically safe for children in hemorrhagic shock.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::c2aca65b83e2bd282c62f57ac721f90bTest
https://doi.org/10.1111/trf.16456Test

المؤلفون: Malla, Midhun, Seetharam, Mahesh

المصدر: Transfusion; Jan2016, Vol. 56 Issue 1, p160-163, 4p, 1 Chart

مصطلحات موضوعية: BLOOD coagulation disorders, THROMBOCYTOPENIA treatment, JEHOVAH'S Witnesses, ERYTHROCYTES, LACTATE dehydrogenase, IMMUNOGLOBULINS, BLOOD disease treatment, PERNICIOUS anemia diagnosis, THROMBOTIC thrombocytopenic purpura, THROMBOTIC thrombocytopenic purpura treatment, DIFFERENTIAL diagnosis, PLASMAPHERESIS, DIAGNOSIS

مستخلص: Background: Thrombotic thrombocytopenic purpura (TTP) is a rare microvascular occlusive disorder characterized by systemic intravascular aggregation of platelets, thrombocytopenia, and mechanical injury to red blood cells. We report a rare case of pernicious anemia presenting as TTP in a Jehovah's Witness.Case Report: A 46-year-old Jehovah's Witness female presented with epigastric pain, vomiting, and diarrhea for 2 days and fatigue and paresthesias for 4 weeks. Initial laboratory evaluation showed severe anemia and thrombocytopenia with elevated total bilirubin and lactate dehydrogenase. Peripheral blood smear showed schistocytes, macroovalocytes, and hypersegmented neutrophils. TTP was suspected and plasmapheresis was offered. The patient refused it due to her religious beliefs. Due to the presence of macroovalocytes and hypersegmented neutrophils, vitamin B12 level was checked and found to be extremely low. Anti-intrinsic factor antibodies and anti-parietal cell antibodies were also positive; hence a diagnosis of pernicious anemia was established. Treatment with intramuscular vitamin B12 was initiated, which resulted in dramatic neurologic and hematologic improvement.Discussion: Vitamin B12 deficiency can lead to elevated levels of homocysteine in the blood. Homocysteine can cause endothelial dysfunction, which can lead to formation of microvascular thrombi. Due to this phenomenon, vitamin B12 deficiency can rarely present with schistocytes and thrombocytopenia, which combined with other stigmata of vitamin B12 deficiency, can be misdiagnosed as TTP. [ABSTRACT FROM AUTHOR]

: Copyright of Transfusion is the property of Wiley-Blackwell and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

المؤلفون: F. Bernadette West, Jeanne E. Hendrickson, Raisa Balbuena-Merle, Christopher A. Tormey

المصدر: Transfusion. 59:1911-1915

مصطلحات موضوعية: Acute Hemolytic Transfusion Reaction, biology, business.industry, Bilirubin, Immunology, Hematology, 030204 cardiovascular system & hematology, medicine.disease, Hemolysis, 03 medical and health sciences, chemistry.chemical_compound, 0302 clinical medicine, Apheresis, chemistry, Lactate dehydrogenase, Anesthesia, medicine, biology.protein, Immunology and Allergy, Platelet, Hemoglobin, Antibody, business, 030215 immunology

الوصف: BACKGROUND Hemolytic transfusion reactions from out-of-group plasma in platelet (PLT) transfusions are uncommon, with most involving passive transfer of anti-A. Only rare reactions have ever been reported due to anti-B. STUDY DESIGN AND METHODS An apheresis PLT product was donated by a blood group O male, processed using PLT additive solution, and pathogen reduced. Postreaction recipient testing included an antibody screen using gel technology, a direct antiglobulin test (DAT) using immunoglobulin G and C3, and an eluate against group O and B cells. Postreaction donor testing included measuring anti-B titers in saline, with and without anti-human globulin. RESULTS A 60-year-old blood group B patient with relapsed acute myeloid leukemia developed confusion, fever, and hypotension within hours after a blood group O PLT transfusion. The posttransfusion reaction evaluation was remarkable for a positive DAT 3+ for C3; the eluate showed anti-B. Rapid extravascular hemolysis occurred, with a 50% decline in hemoglobin, a high lactate dehydrogenase, and a high bilirubin. She was resuscitated with fluids, blood products, pressors, and oxygen and died of asystole 60 hours later. The donor's anti-B titers were 128 by tube testing at immediate spin and 512 at the anti-human globulin phase. Notably, a group B patient at a different hospital received a split of the same apheresis unit, with no reaction. CONCLUSION To our knowledge, this is the first fatality reported from passively transfused anti-B. The fact that one transfusion recipient died whereas another did not have any reported reaction highlights the potential importance of recipient variables in isohemagglutinin-mediated hemolysis.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_________::67b4a4a5b86bdefac796ff92bb057e4aTest
https://doi.org/10.1111/trf.15240Test

المؤلفون: Angelo D'Alessandro, Andrew J. Dunham, Rachel Culp-Hill, Kirk C. Hansen, Travis Nemkov, Tamir Kanias, Monika Dzieciatkowska, Davide Stefanoni, Michael P. Busch, Elan Z. Eisenmesser, Tatsuro Yoshida, Julie A. Reisz, Larry J. Dumont, James C. Zimring, Ryan C. Hill

المصدر: Transfusion. 58:2978-2991

مصطلحات موضوعية: 0301 basic medicine, biology, Chemistry, Immunology, Aldolase A, Hematology, Methylation, 030204 cardiovascular system & hematology, medicine.disease, Hemolysis, 03 medical and health sciences, chemistry.chemical_compound, 030104 developmental biology, 0302 clinical medicine, Biochemistry, Lactate dehydrogenase, medicine, biology.protein, Protein methylation, Immunology and Allergy, Spectrin, Hemoglobin, Deamidation

الوصف: Background Being devoid of de novo protein synthesis capacity, red blood cells (RBCs) have evolved to recycle oxidatively damaged proteins via mechanisms that involve methylation of dehydrated and deamidated aspartate and asparagine residues. Here we hypothesize that such mechanisms are relevant to routine storage in the blood bank. Study design and methods Within the framework of the REDS-III RBC-Omics (Recipient Epidemiology Donor Evaluation Study III Red Blood Cell-Omics) study, packed RBC units (n = 599) were stored under blood bank conditions for 10, 23, and 42 days and profiled for oxidative hemolysis and time-dependent metabolic dysregulation of the trans-sulfuration pathway. Results In these units, methionine consumption positively correlated with storage age and oxidative hemolysis. Mechanistic studies show that this phenomenon is favored by oxidative stress or hyperoxic storage (sulfur dioxide >95%), and prevented by hypoxia or methyltransferase inhibition. Through a combination of proteomics approaches and 13 C-methionine tracing, we observed oxidation-induced increases in both Asn deamidation to Asp and formation of methyl-Asp on key structural proteins and enzymes, including Band 3, hemoglobin, ankyrin, 4.1, spectrin beta, aldolase, glyceraldehyde 3-phosphate dehydrogenase, biphosphoglycerate mutase, lactate dehydrogenase and catalase. Methylated regions tended to map proximal to the active site (e.g., N316 of glyceraldehyde 3-phosphate dehydrogenase) and/or residues interacting with the N-terminal cytosolic domain of Band 3. Conclusion While methylation of basic amino acid residues serves as an epigenetic modification in nucleated cells, protein methylation at carboxylate side chains and deamidated asparagines is a nonepigenetic posttranslational sensor of oxidative stress and refrigerated storage in anucleated human RBCs.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_________::67cd8d816aeeaa0c664013aafd68cce7Test
https://doi.org/10.1111/trf.14936Test

المؤلفون: Telen, Marilyn J., Afenyi‐Annan, Araba, Garrett, Melanie E., Combs, Martha R., Orringer, Eugene P., Ashley‐Koch, Allison E.

المصدر: Transfusion; Jun2015, Vol. 55 Issue 6pt2, p1378-1387, 10p, 6 Charts, 4 Graphs

مصطلحات موضوعية: SICKLE cell anemia treatment, BLOOD transfusion reaction, ANTIBODY specificity, LACTATE dehydrogenase, CHRONIC pain, BLOOD sampling, GENETIC polymorphisms, PATHOLOGICAL physiology

مستخلص: Background Alloimmunization remains a significant complication of transfusion and has been associated with multiple factors, including inflammation, an important pathophysiologic mechanism in sickle cell disease ( SCD). We explored whether alloimmunization is associated with disease severity in SCD. Study Design and Methods Adult SCD patients were enrolled in a study of outcome-modifying genes in SCD. Historical records of patients with SCD at two participating institutions were reviewed for data on antigen phenotype and alloimmunization. Differences in demographic, clinical, and laboratory findings; end-organ damage; and overall disease severity were then compared between alloimmunized and nonalloimmunized patients. Results Of 319 patients, 87 (27%) were alloimmunized. Alloantibody specificities differed from those previously described, especially due to the significantly higher frequency of anti- S. Although alloimmunization was not associated with frequency of vasoocclusive episodes, a higher percentage of alloimmunized patients had chronic pain, as defined by daily use of short-acting narcotics (p = 0.006), long-acting narcotics (p = 0.013), or both (p = 0.03). Additionally, alloimmunized patients had poorer survival (hazard ratio, 1.92; p = 0.01) and were more likely to have avascular necrosis (p = 0.024), end-organ damage (p = 0.049), and red blood cell autoantibodies (p < 0.001), even after controlling for the effects of age, sex, and hemoglobin diagnosis. Alloimmunization was not associated with other SCD-related complications, such as acute chest syndrome or stroke. Conclusion Alloimmunization in SCD may be associated with chronic pain, risk of end-organ damage, and shorter survival. These novel findings suggest new directions for the investigation of immune response-mediated pathways common to alloimmunization and chronic pain. [ABSTRACT FROM AUTHOR]

: Copyright of Transfusion is the property of Wiley-Blackwell and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

المؤلفون: Nicole P. Juffermans, Robin van Bruggen, Alexander P.J. Vlaar, Dirk de Korte, Anna L. Peters, Renoja K. Kunanayagam

المساهمون: Graduate School, Intensive Care Medicine, Landsteiner Laboratory, Amsterdam Cardiovascular Sciences, ACS - Pulmonary hypertension & thrombosis, ACS - Microcirculation

المصدر: Transfusion, 57(1), 53-59. Wiley-Blackwell

مصطلحات موضوعية: Adult, Lipopolysaccharides, Male, medicine.medical_specialty, Time Factors, Blood transfusion, Adolescent, Iron, medicine.medical_treatment, Immunology, 030204 cardiovascular system & hematology, Blood Transfusion, Autologous, Hemoglobins, 03 medical and health sciences, chemistry.chemical_compound, 0302 clinical medicine, Internal medicine, Lactate dehydrogenase, Humans, Immunology and Allergy, Medicine, chemistry.chemical_classification, Haptoglobins, L-Lactate Dehydrogenase, biology, business.industry, Transferrin saturation, Haptoglobin, Bilirubin, Hematology, medicine.disease, Endotoxemia, Hemolysis, Ferritin, Endocrinology, chemistry, Blood Preservation, Transferrin, Ferritins, biology.protein, Hemoglobin, Erythrocyte Transfusion, business, 030215 immunology

الوصف: BACKGROUND Transfusion of a single unit of stored red blood cells (RBCs) has been hypothesized to induce supra-physiological levels of non-transferrin bound iron (NTBI), which may enhance inflammation and act as a nutrient for bacteria. We investigated the relation between RBC storage time and iron levels in a clinically relevant “two-hit” human transfusion model. STUDY DESIGN AND METHODS Eighteen healthy male volunteers (ages 18-35 years) were infused with 2 ng lipopolysaccharide (LPS)/kg to induce systemic inflammatory response syndrome. Two hours later, each participant received either 1 unit of 2-day stored (2D) autologous RBCs, 35-day stored (35D) autologous RBCs, or an equal volume of saline. Every 2 hours up to 8 hours after LPS infusion, hemoglobin, hemolysis parameters, and iron parameters, including NTBI, were measured. RESULTS Transfusion of both 2D and 35D RBCs caused increases in hemoglobin, plasma iron, and transferrin saturation; whereas levels remained stable in the saline group. Transfusion of 35D RBCs did not result in hemolysis nor did it lead to increased levels of NTBI compared with 2D RBCs or saline. LPS induced increases in ferritin, haptoglobin, bilirubin, and lactate dehydrogenase that were similar in all three groups. CONCLUSION We conclude that 35D autologous RBCs do not cause hemolysis or increased levels of NTBI during human endotoxemia.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::53dd3b0edd58a9d83b4925c13633bb20Test
https://doi.org/10.1111/trf.13849Test

المؤلفون: Jason L. Sperry, Alain Corcos, Mark H. Yazer, Vincent Anto, Marshall P. Bahr, Darrell J. Triulzi, Jansen N. Seheult, Louis H. Alarcon

المصدر: Transfusion. 58:2280-2288

مصطلحات موضوعية: Creatinine, Resuscitation, biology, business.industry, Immunology, Haptoglobin, 030208 emergency & critical care medicine, Hematology, 030204 cardiovascular system & hematology, medicine.disease, Hemolysis, 03 medical and health sciences, chemistry.chemical_compound, 0302 clinical medicine, chemistry, Blood product, Anesthesia, Lactate dehydrogenase, medicine, biology.protein, Immunology and Allergy, Adverse effect, business, Whole blood

الوصف: BACKGROUND The use of cold-stored low-titer group O whole blood (LTOWB) for civilian trauma patients is gaining popularity. However, hemolysis might occur among non-group O recipients. This study evaluated the serologic safety of transfusing up to 4 units of LTOWB. STUDY DESIGN AND METHODS Hypotensive male and at least 50-year-old female trauma patients who received leukoreduced, uncrossmatched, group O+, low-titer (

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_________::26c2c7556b76df5643b7076d2b891b18Test
https://doi.org/10.1111/trf.14771Test