Effects of aged stored autologous red blood cells on human plasma metabolome.

Cold storage of blood for 5 to 6 weeks has been shown to impair endothelial function after transfusion and has been associated with measures of end-organ dysfunction. Although the products of hemolysis, such as cell-free plasma hemoglobin, arginase, heme, and iron, in part mediate these effects, a complete analysis of transfused metabolites that may affect organ function has not been evaluated to date. Blood stored for either 5 or 42 days was collected from 18 healthy autologous volunteers, prior to and after autologous transfusion into the forearm circulation, followed by metabolomics analyses. Significant metabolic changes were observed in the plasma levels of hemolytic markers, oxidized purines, plasticizers, and oxidized lipids in recipients of blood stored for 42 days, compared with 5 days. Notably, transfusion of day 42 red blood cells (RBCs) increased circulating levels of plasticizers (diethylhexyl phthalate and derivatives) by up to 18-fold. Similarly, transfusion of day 42 blood significantly increased circulating levels of proinflammatory oxylipins, including prostaglandins, hydroxyeicosatrienoic acids (HETEs), and dihydroxyoctadecenoic acids. Oxylipins were the most significantly increasing metabolites (for 9-HETE: up to ∼41-fold, P = 3.7e-06) in day 42 supernatants. Measurements of arginine metabolism confirmed an increase in arginase activity at the expense of nitric oxide synthesis capacity in the bloodstream of recipients of day 42 blood, which correlated with measurements of hemodynamics. Metabolic changes in stored RBC supernatants impact the plasma metabolome of healthy transfusion recipients, with observed increases in plasticizers, as well as vasoactive, pro-oxidative, proinflammatory, and immunomodulatory metabolites after 42 days of storage.

Short-term effects of stored homologous red blood cell transfusion on cardiorespiratory function and inflammation: an experimental study in a hypovolemia model

The pathophysiological mechanisms associated with the effects of red blood cell (RBC) transfusion on cardiopulmonary function and inflammation are unclear. We developed an experimental model of homologous 14-days stored RBC transfusion in hypovolemic swine to evaluate the short-term effects of transfusion on cardiopulmonary system and inflammation. Sixteen healthy male anesthetized swine (68±3.3 kg) were submitted to controlled hemorrhage (25% of blood volume). Two units of non-filtered RBC from each animal were stored under blood bank conditions for 14 days. After 30 min of hypovolemia, the control group (n=8) received an infusion of lactated Ringer's solution (three times the removed volume). The transfusion group (n=8) received two units of homologous 14-days stored RBC and lactated Ringer's solution in a volume that was three times the difference between blood removed and blood transfusion infused. Both groups were followed up for 6 h after resuscitation with collection of hemodynamic and respiratory data. Cytokines and RNA expression were measured in plasma and lung tissue. Stored RBC transfusion significantly increased mixed oxygen venous saturation and arterial oxygen content. Transfusion was not associated with alterations on pulmonary function. Pulmonary concentrations of cytokines were not different between groups. Gene expression for lung cytokines demonstrated a 2-fold increase in mRNA level for inducible nitric oxide synthase and a 0.5-fold decrease in mRNA content for IL-21 in the transfused group. Thus, stored homologous RBC transfusion in a hypovolemia model improved cardiovascular parameters but did not induce significant effects on microcirculation, pulmonary inflammation and respiratory function up to 6 h after transfusion.

Short-term effects of stored homologous red blood cell transfusion on cardiorespiratory function and inflammation: an experimental study in a hypovolemia model.

The pathophysiological mechanisms associated with the effects of red blood cell (RBC) transfusion on cardiopulmonary function and inflammation are unclear. We developed an experimental model of homologous 14-days stored RBC transfusion in hypovolemic swine to evaluate the short-term effects of transfusion on cardiopulmonary system and inflammation. Sixteen healthy male anesthetized swine (68±3.3 kg) were submitted to controlled hemorrhage (25% of blood volume). Two units of non-filtered RBC from each animal were stored under blood bank conditions for 14 days. After 30 min of hypovolemia, the control group (n=8) received an infusion of lactated Ringer's solution (three times the removed volume). The transfusion group (n=8) received two units of homologous 14-days stored RBC and lactated Ringer's solution in a volume that was three times the difference between blood removed and blood transfusion infused. Both groups were followed up for 6 h after resuscitation with collection of hemodynamic and respiratory data. Cytokines and RNA expression were measured in plasma and lung tissue. Stored RBC transfusion significantly increased mixed oxygen venous saturation and arterial oxygen content. Transfusion was not associated with alterations on pulmonary function. Pulmonary concentrations of cytokines were not different between groups. Gene expression for lung cytokines demonstrated a 2-fold increase in mRNA level for inducible nitric oxide synthase and a 0.5-fold decrease in mRNA content for IL-21 in the transfused group. Thus, stored homologous RBC transfusion in a hypovolemia model improved cardiovascular parameters but did not induce significant effects on microcirculation, pulmonary inflammation and respiratory function up to 6 h after transfusion.

Influência da temperatura e do tempo de armazenamento sobre amostras de soro e plasma caninos na análise da enzima alanina aminotransferase (ALT) / Effect of temperature and storage time on canine serum and plasma samples in the analysis of enzyme alanine aminotransferase (ALT)

Este trabalho teve como objetivo avaliar a influência do tempo e da temperatura de armazenamento sobre os valores das dosagens bioquímicas de ALT em 60 amostras de soro e plasma de cães, divididas em dois grupos: GR (grupo refrigerado, a 5 °C) e GC (grupo congelado, a -20 °C), por até 96 horas analisadas em seis tempos e até 60 dias, analisadas em cinco tempos, respectivamente. O estudo mostrou que o congelamento e a refrigeração não provocaram variações entre as amostras (p< 0,005). Os dados obtidos nas dosagens bioquímicas revelaram que o soro e o plasma com EDTA mantiveram-se próximos dos valores basais, ou seja, não se alteraram significantemente ao longo do tempo de conservação e nas diferentes temperaturas estudadas.(AU)

This study aimed to evaluate the effect of time and storage temperature on the values of ALT biochemical measurements in 60 samples of serum and plasma of dogs divided into two groups: RG (refrigerated group, at 5 °C) and FG (frozen group, at -20 °C) for 96 hours analyzed at six moments and up to 60 days examined at five moments, respectively. The study showed that the effect of freezing and cooling caused no variation between the samples (p <0.005). The data showed that the biochemical serum and EDTA plasma remained near baseline, i.e ., they did not change significantly over time and at different storage temperatures.(AU)

Influência da temperatura e do tempo de armazenamento sobre amostras de soro e plasma caninos na análise da enzima alanina aminotransferase (ALT) / Effect of temperature and storage time on canine serum and plasma samples in the analysis of enzyme alanine aminotransferase (ALT)

Este trabalho teve como objetivo avaliar a influência do tempo e da temperatura de armazenamento sobre os valores das dosagens bioquímicas de ALT em 60 amostras de soro e plasma de cães, divididas em dois grupos: GR (grupo refrigerado, a 5 °C) e GC (grupo congelado, a -20 °C), por até 96 horas analisadas em seis tempos e até 60 dias, analisadas em cinco tempos, respectivamente. O estudo mostrou que o congelamento e a refrigeração não provocaram variações entre as amostras (p< 0,005). Os dados obtidos nas dosagens bioquímicas revelaram que o soro e o plasma com EDTA mantiveram-se próximos dos valores basais, ou seja, não se alteraram significantemente ao longo do tempo de conservação e nas diferentes temperaturas estudadas.

This study aimed to evaluate the effect of time and storage temperature on the values of ALT biochemical measurements in 60 samples of serum and plasma of dogs divided into two groups: RG (refrigerated group, at 5 °C) and FG (frozen group, at -20 °C) for 96 hours analyzed at six moments and up to 60 days examined at five moments, respectively. The study showed that the effect of freezing and cooling caused no variation between the samples (p <0.005). The data showed that the biochemical serum and EDTA plasma remained near baseline, i.e ., they did not change significantly over time and at different storage temperatures.

Efeito da refrigeração sobre o exame hemogasométrico em sangue venoso de ovinos / Effect of refrigeration on the hemogasometric examination of venous blood in sheep

Com o propósito de avaliar o efeito da refrigeração sobre o exame hemogasométrico, foram utilizados 12 ovinos machos, hígidos, da raça Santa Inês, com cerca de quatro meses de idade e peso variando entre 30 e 45 kg. As amostras de sangue destinadas ao exame hemogasométrico foram coletadas em duplicata utilizando-se agulhas descartáveis acopladas à seringas plásticas contendo cerca de 1000UI de heparina sódica. Durante e após a coleta tomou-se o cuidado de evitar a entrada de bolhas de ar no interior da seringa. As amostras não conservadas foram mantidas a temperatura ambiente, entre 23 e 25°C, e aquelas destinadas à refrigeração foram acondicionadas em isopor contendo 3kg de água gelada e 3kg de gelo, mantendo-se assim uma temperatura entre 0 e 4° C. As análises hemogasométricas foram determinadas imediatamente após coleta e com 1, 2, 3, 4, 5, 6, 8, 10, 12 e 24 horas. As análises dos resultados indicaram alterações significativas nas amostras mantidas a temperatura ambiente, caracterizado-se por diminuições, a partir da 4, 8 e 10 horas após coleta, para os valores do pH, BE e StB, respectivamente, e por aumento, a partir da 6 hora, para os valores da PCO(2 subscrito).Com relação as amostras conservadas, não foram evidenciadas variações significativas dos parâmetros ao longo dos tempos de análise. Conclui-se, portanto, que amostras de sangue venoso de ovinos são viáveis, para a realização do exame hemogasométrico, até 24 horas após coleta, desde que mantidas sob adequada refrigeração.

With the objective of evaluating the effect of refrigeration on the hemogasometric exam, venous blood samples were collected from 12 healthy male sheep, Santa Ines breed, with a mean age of 4 months old, and body weight raging from 30 to 45 kg. The blood samples for the hemogasometric examination were collected in two aliquots from each animal, using dispensable needles connected to plastic syringes containing about 1000 IU sodium heparin. During and after the sampling, the care of avoiding the presence of air bubbles in the syringe was attempted. The samples without conservation were kept at room temperature, between 23 an 25°C, and the samples under refrigeration were kept in an box containing 3 L of cold water and 3Kg of ice, to maintain a temperature between 0 and 4°C. The hemogasometric analyses were made immediate1y after collection an after 1, 2, 3, 4, 5, 6, 8, 10, 12 and 24 hours. The results indicated significant alteration in samples kept at room temperature, characterized by decline, starting at 4, 8 and 10 hour post-collection, of the values f pH, BE and StB, respectivelly, and by a raise, since the 6th hour, of the values of PCO(2subscript). No significant variations of the parameters were seen in the refrigerated samples during the study. So, the conclusion is that ovine venous blood samples are viable, for the determination of the hemogasometric evaluation, until 24 hours after the collection, when kept (maintained) under adequate refrigeration.

Efeito da refrigeração sobre o exame hemogasométrico em sangue venoso de ovinos / Effect of refrigeration on the hemogasometric examination of venous blood in sheep

Com o propósito de avaliar o efeito da refrigeração sobre o exame hemogasométrico, foram utilizados 12 ovinos machos, hígidos, da raça Santa Inês, com cerca de quatro meses de idade e peso variando entre 30 e 45 kg. As amostras de sangue destinadas ao exame hemogasométrico foram coletadas em duplicata utilizando-se agulhas descartáveis acopladas à seringas plásticas contendo cerca de 1000UI de heparina sódica. Durante e após a coleta tomou-se o cuidado de evitar a entrada de bolhas de ar no interior da seringa. As amostras não conservadas foram mantidas a temperatura ambiente, entre 23 e 25°C, e aquelas destinadas à refrigeração foram acondicionadas em isopor contendo 3kg de água gelada e 3kg de gelo, mantendo-se assim uma temperatura entre 0 e 4° C. As análises hemogasométricas foram determinadas imediatamente após coleta e com 1, 2, 3, 4, 5, 6, 8, 10, 12 e 24 horas. As análises dos resultados indicaram alterações significativas nas amostras mantidas a temperatura ambiente, caracterizado-se por diminuições, a partir da 4, 8 e 10 horas após coleta, para os valores do pH, BE e StB, respectivamente, e por aumento, a partir da 6 hora, para os valores da PCO(2 subscrito).Com relação as amostras conservadas, não foram evidenciadas variações significativas dos parâmetros ao longo dos tempos de análise. Conclui-se, portanto, que amostras de sangue venoso de ovinos são viáveis, para a realização do exame hemogasométrico, até 24 horas após coleta, desde que mantidas sob adequada refrigeração.(AU)

With the objective of evaluating the effect of refrigeration on the hemogasometric exam, venous blood samples were collected from 12 healthy male sheep, Santa Ines breed, with a mean age of 4 months old, and body weight raging from 30 to 45 kg. The blood samples for the hemogasometric examination were collected in two aliquots from each animal, using dispensable needles connected to plastic syringes containing about 1000 IU sodium heparin. During and after the sampling, the care of avoiding the presence of air bubbles in the syringe was attempted. The samples without conservation were kept at room temperature, between 23 an 25°C, and the samples under refrigeration were kept in an box containing 3 L of cold water and 3Kg of ice, to maintain a temperature between 0 and 4°C. The hemogasometric analyses were made immediate1y after collection an after 1, 2, 3, 4, 5, 6, 8, 10, 12 and 24 hours. The results indicated significant alteration in samples kept at room temperature, characterized by decline, starting at 4, 8 and 10 hour post-collection, of the values f pH, BE and StB, respectivelly, and by a raise, since the 6th hour, of the values of PCO(2subscript). No significant variations of the parameters were seen in the refrigerated samples during the study. So, the conclusion is that ovine venous blood samples are viable, for the determination of the hemogasometric evaluation, until 24 hours after the collection, when kept (maintained) under adequate refrigeration.(AU)

Contribuiçäo ao conhecimento das lesöes de estocagem: estudo do sistema de óxido-reduçäo glutationa dependente em eritrócitos coletados em CPDA-1 / Contribution on knowledge of storage lesions: glutathione anti-oxidant system study

A presente investigaçäo teve como objetivo o estudo do sistema de óxido-reduçäo glutationa-dependente em 34 CGV coletados em CPDA-1, nos dias 0 e 35 de estocagem. Amostras de 12 CGV foram examinadas logo após a coleta em heparina e CPDA-1, constituindo o grupo dia 0. Os valores do dia 0 foram considerados controles para 12 CGV examinados no 35§ dia de estocagem. Foram efetuados testes de reduçäo da MetaHb pelo Am (atividade G6PD) e pela CIS (atividade GSSG-Rd), dosagens de GSHt, GSSG, MDA total e eritrocitário, MetHb e Hb plasmática. Os resultados do dia 0 demonstraram preservaçäo da capacidade antioxidante da via das pentoses. Foi também observada uma lesäo incipiente da membrana, principalmente após centrifugaçäo refrigerada em GV heparinizados. No dia 35, constatou-se atividade deficiente da G6PD, deficiência de GSSG-Rd (funcional, dependente da atividade G6PD), diminuiçäo da GSHt, da GSSG, manutençäo da relaçäo [GHS] / [GSSG], refletindo uma lentidäo na capacidade antioxidante da vida das pentoses. Pelos altos níveis de Hb plasmática, ficou evidenciada uma importante lesäo da membrana, associada entretanto, a níveis pouco elevados de MDA. Estes CGC que sofreram manipulaçäo eventual e freqüênte, durante a estocagem, foram comparados a 4 unidades de CGV estocados imóveis pelo mesmo período, para GSHt e atividades G6PD e GSSG-Rd. As unidades agitadadas apresentaram o dobro das taxas de GSHt, sugerindo que a agitaçäo favorece maior síntetse e também maior reciclagem da GSH. A atividade G6PD foi menor, embora näo significante, nas unidades agitadas, provavelmente pela oxidaçäo dos seus grupamentos SH, pois um maior fluxo de oxigênio ocorre no interior da bolsa. A GSSG-Rd acompanhou funcionalmente a deficiência em G6PD. Foram estudados 10 CGV estocados imóveis por 35 dias, 6 de 150 ml e 4 de 300 ml, comparando-se amostras de GV do topo, centro, fundo da bolsa e após mistura completa do conteúdo, para alguns índices hematimétricos, GSHt e atividades G6PD e GSSG-Rd. A camada do fundo dos CGV demonstrou ser a mais compacta e menos preservada, com GV de menor VCM, maior CHCM e menores níveis de GSHt. A camada do centro foi a melhor preservada, tanto quanto a índices hematimétricos, quanto a taxas de regeneraçäo de GSHt e níveis mais homogêneos de G6PD. Nestas unidades imóveis, parece ocorrer um "choque metabólico", quando da mistura dos GV com plasma, proteases, oxigênio e espécies oxigênio reativas, pois a regeneraçäo da GSHt é semelhante...