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.

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.

Evaluation of bacterial contamination of platelet concentrates collected at Tehran regional blood center

In spite of major advances in the field of quality assurance in the process of collection, preparation and storage of platelets, bacterial infection following platelet transfusion remains a major problem in transfusion medicine. The present study was carried out in order to evaluate bacterial contamination of platelet concentrates collected at Tehran Regional Educational Blood Transfusion Center. Bacterial growth of samples of platelet concentrates was studied in blood agar, EMB and thioglycollate broth after 48 hours at 37°C. The use of differentiation tests was made when any bacterial growth was observed. Simultaneously, the samples were also cultured in thioglycollate broth and studied for any turbidity or color change within 7 days. Any changes made the samples to be cultured in blood agar and EMB. Finally, the contamination rate and the ratio of contaminating bacteria were determined. Out of 7700 samples, three fourth [5775 samples] were taken from the cord and one fourth [1925] from both the bag and the cord. Out of 7700 samples of platelet concentrates studied, 14 [0. 18%] were found positive for bacterial contamination. The contamination rate was estimated to be one in every 550 tested platelets [0. 18%]. Since in cases of blood bag contamination, the cord had been contaminated as well, there was then no difference on whether the sample was taken from the bag or cord. The bacteria identified were as follows: Staph. epidermidis [n=4], Staph. saprophyticus [n=2], Acinetobacter [n=5], Bacillus sp. [n=3]. The results show that screening platelet concentrates for bacterial contamination is necessary for blood transfusion centers and hospital blood banks

Effect of pre-storage gamma irradiation on red blood cells.

BACKGROUND & OBJECTIVE: The irradiation of blood components has received increased attention due to increasing categories of patients eligible to receive such blood to prevent transfusion-associated graft versus host disease. Irradiation leads to enhancement of storage lesions, which could have deleterious effects when such blood is transfused. The aim of the present study was to assess the biochemical changes during conventional preservation of irradiated and non-irradiated whole blood. METHODS: Ten units of whole blood were taken from healthy donors and divided into two parts. One aliquot was subjected to gamma irradiation and then stored under conventional blood banking conditions. Sampling was done from these irradiated and non-irradiated blood bags and tests for free plasma haemoglobin, plasma potassium and lactate dehydrogenase (LDH) were performed. RESULTS: A progressive increase in the mean values of plasma Hb, K+ and LDH was seen in both the groups. The increase was statistically significant. INTERPRETATION & CONCLUSION: Our findings indicated that the gamma irradiation of blood resulted in increased plasma haemoglobin, potassium and LDH. These biochemical changes might not have clinical significance when irradiated blood is transfused to a select group of patients. There is a need for further in vivo studies to follow up the consequences of transfusion of irradiated blood in patients.

Evaluation of platelet storage lesions in platelet concentrates stored for seven days.

BACKGROUND & OBJECTIVES: Platelet storage lesions (PSL) are detrimental to the post transfusion functional capacity of platelets. As EDTA enhances the storage-induced changes, changes in platelet indices with and without EDTA incubation are promising new tests to monitor the PSL. The present study was undertaken to monitor the PSL in 40 units of pooled platelet concentrates harvested by platelet rich plasma (PRP) method and stored for seven days in second generation platelet storage containers using conventional in vitro tests and platelet indices. METHODS: Morphological changes in platelets were monitored by automated haematological cell counter for platelet count and mean platelet volume (MPV). Samples were incubated with K2EDTA for 1 h and platelet indices were repeated on the EDTA incubated samples. Difference between pre-and post-EDTA incubation of platelet count (dPLT) and MPV (dMPV) were calculated. Metabolic parameters such as pH, pO2, pCO2 and ATP were measured. RESULTS: There was no significant change in the indices without EDTA during storage, however, after EDTA incubation, significant changes were noted in dPLT and dMPV. The mean dPLT on day 0 was 75.15 x 10(3)/microl decreasing to 44.4 x 10(3)/microl on day 7, while dMPV from 0.76 fl on day 0 increased to 1.34 fl on day 7 (P < 0.05). Metabolic parameters showed a significant decrease in pH and pCO2 concurrent with increasing pO2 during storage (P < 0.05). Average ATP level on day 0 was 21.09 micromol/dl falling to 10.59 micromol/dl on day 7. INTERPRETATION & CONCLUSION: The results indicate that storage induced lesions take place even in second generation platelet storage containers under recommended conditions of storage. Platelet indices especially after EDTA incubation are useful in monitoring PSL. However, how much these changes contribute to poor post transfusion survival and haemostatic function of platelets need to be investigated.

Effect of temperature on functional response of platelet concentrates stored in PVC bags.

Haemostatic efficacy of platelet concentrates prepared and stored in locally available PVC triple bags was compared against a Japanese bag. In vitro functional parameters studied included shape change, aggregation and secretion in response to ADP. We have observed remarkable difference in the aggregatory response of platelets stored at slightly varying temperatures. The stimulatory responses of platelets stored with constant agitation at 70 strokes per min and 23 +/- 2 degrees C, deteriorated drastically by the time platelets were stored for 48 h. Both the rate and the extent of aggregation were affected showing no response to ADP at 72 h. However, when platelets were stored in a BOD incubator, thermostated at 22 +/- 0.5 degrees C, with continuous horizontal agitation at 70 strokes per min, 50 per cent functional response was retained till 72 h. We also demonstrated fragmentation of platelet membrane during storage. The membrane fragments collected by high speed centrifugation, expressed PF3 activity. Shedding of microvesicles indicates alterations at the membrane level that possibly cause functional lesion during storage. Our data suggest the significance of controlling the storage temperature steadily, to get maximum post transfusion efficacy.

Effect of storage of blood samples on DNA yield, quality and fingerprinting: a forensic approach.

Various storage treatments on human blood samples have been described with respect to DNA yield, quality and fingerprinting. Blood samples were stored at room temperature, 4 degrees C, -20 degrees C and -70 degrees C for different duration varying from 1, 2, 4 and 8 weeks with or without anticoagulant/preservative (EDTA or Heparin). DNA was isolated from these stored samples, quantitated by spectrophotometry and subjected to DNA fingerprinting using a human alphoid satellite DNA sequence (TRF 5.6) and a 33 mer oligonucleotide (O-chi-1) as probes. The polymorphic DNA bands were apparent between 2 to 9 kb size range and the fingerprints were individual-specific. Our results suggest that higher amount of genomic DNA can be recovered from blood samples stored at temperatures 4 degrees C or below in the presence of EDTA or heparin.

Effect of storage duration on cell cycle kinetics and yield of chromosomal aberrations in PHA-responsive human peripheral blood lymphocytes.

Phytohaemagglutinin (PHA)-responsive lymphocytes from human peripheral blood samples, either irradiated or un-irradiated, showed increased frequency of first division metaphase cells (detected by fluorescence plus Giemsa (FPG) staining) as a function of duration of storage. Irradiated and subsequently stored samples showed small but significant increase for the yield of dicentrics. The yield of aberrant metaphases and deletions (excess acentrics) remained unchanged. Increasing Bromodeoxyuridine (BrdU) concentrations slowed down the cell cycle progression but did not influence the yield of aberrations including that of dicentrics.

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...