RESUMO
BACKGROUND: Dengue viruses (DENV 1-4) are emerging across the world, and these viruses pose a risk to transfusion safety. Pathogen inactivation may be an alternative approach for managing the risk of DENV transfusion transmission. This study aimed to investigate the ability of riboflavin and UV light to inactivate DENV 1-4 in platelet concentrates. MATERIALS AND METHODS: DENV 1-4 were spiked into buffy coat-derived platelet concentrates in additive solution (SSP+) before being treated with riboflavin and UV light. Infectious virus was quantified pre- and posttreatment, and the reduction in viral infectivity was calculated. RESULTS: All four DENV serotypes were modestly reduced after treatment. The greatest amount of reduction in infectivity was observed for DENV-4 (1·81 log reduction) followed by DENV-3 (1·71 log reduction), DENV-2 (1·45 log reduction) and then DENV-1 (1·28 log reduction). CONCLUSION: Our study demonstrates that DENV 1-4 titres are modestly reduced following treatment with riboflavin and UV light. With the increasing number of transfusion-transmitted cases of DENV around the globe, and the increasing incidence and geographical distribution of DENV, additional approaches for maintaining blood safety may be required in the future.
Assuntos
Vírus da Dengue/fisiologia , Fármacos Fotossensibilizantes/farmacologia , Riboflavina/farmacologia , Raios Ultravioleta , Inativação de Vírus/efeitos dos fármacos , Plaquetas/citologia , Plaquetas/virologia , Segurança do Sangue , Vírus da Dengue/genética , Vírus da Dengue/metabolismo , Humanos , Transfusão de Plaquetas , RNA Viral/metabolismo , Reação em Cadeia da Polimerase em Tempo Real , Sorogrupo , Inativação de Vírus/efeitos da radiaçãoRESUMO
BACKGROUND AND OBJECTIVES: Transfusion is associated with a risk of infection and alloimmunization. Pathogen reduction using riboflavin and UV light (Mirasol treatment) inactivates pathogens and leucocytes. With increasing adoption of the technology in clinical use, regulatory agencies have recommended the introduction of quality control measures to monitor pathogen reduction efficacy. We sought to develop a real-time PCR-based assay to document the impact of pathogen reduction on the mitochondrial genome in blood components. MATERIALS AND METHODS: DNA was extracted from platelet and plasma components before and after treatment with riboflavin and UV light. Inhibition of PCR amplification of mitochondrial DNA (mtDNA) in short- and long-amplicon target regions, ranging from under 200 base pairs (bp) to over 1800 bp, was measured in treated relative to untreated components. RESULTS: Pathogen reduction of platelets using riboflavin and UV light resulted in inhibition of PCR amplification of long-amplicon mtDNA targets, demonstrating approximately 1 log reduction of amplification relative to untreated products. Amplification of short-amplicon mtDNA targets was not affected by treatment. Evaluation of 110 blinded platelet samples from the PREPAReS clinical trial resulted in prediction of treatment status with 100% accuracy. Pathogen reduction of plasma components resulted in similar levels of PCR inhibition, while testing of 30 blinded plasma samples resulted in prediction of treatment status with 93% accuracy. CONCLUSION: A differential sized amplicon real-time PCR assay of mitochondrial DNA effectively documents nucleic acid damage induced by Mirasol treatment of platelets. The use of the assay for plasma product pathogen reduction requires further investigation.
Assuntos
Patógenos Transmitidos pelo Sangue/efeitos dos fármacos , Patógenos Transmitidos pelo Sangue/efeitos da radiação , DNA Mitocondrial/análise , Mitocôndrias/genética , Reação em Cadeia da Polimerase em Tempo Real , Riboflavina/farmacologia , Raios Ultravioleta , Plaquetas/metabolismo , Plaquetas/microbiologia , DNA Mitocondrial/normas , Humanos , Plasma/microbiologia , Controle de Qualidade , Reação em Cadeia da Polimerase em Tempo Real/normasRESUMO
BACKGROUND AND OBJECTIVES: Pathogen reduction technologies (PRT) for platelets are now compatible with both plasma and platelet additive solutions (PAS). The aim of this study was to examine the effect of PRT on the platelet storage lesion, in the presence of PAS with low plasma carryover. MATERIALS AND METHODS: PRT-treated (Mirasol) and untreated buffy coat-derived platelet concentrates prepared in 28% plasma/PAS-IIIM were evaluated using in vitro cell quality parameters on days 1, 2, 5, and 7 post-collection. RESULTS: At day 5, there were no significant differences between control and PRT treated platelets for swirl, viability, pO(2) , pCO(2) , mean platelet volume and adenosine diphosphate-induced aggregation. PRT treatment did not affect the functional integrity of the mitochondria. However, PRT resulted in a decrease in pH and enhancement of platelet glycolysis and activation, evidenced by increased glucose consumption and lactate production rates, increased expression of CD62P, CD63, annexin V staining and increased secretion of cytokines (P < 0.05). Hypotonic shock response and aggregation in response to collagen were also significantly reduced in PRT treated platelets (P < 0.05). CONCLUSION: Despite the observed differences in platelet metabolism and activation observed following PRT treatment in PAS and low plasma carryover, the results suggest that treatment and storage of platelets in PAS is no more detrimental to platelets than treatment and storage in plasma.
Assuntos
Plaquetas/citologia , Plaquetas/metabolismo , Preservação de Sangue , Segurança do Sangue/métodos , Desinfecção/métodos , Plasma , Antígenos de Plaquetas Humanas/metabolismo , Desinfecção/instrumentação , Humanos , Concentração de Íons de Hidrogênio , Pressão Osmótica , Soluções Farmacêuticas/farmacologia , Agregação Plaquetária/efeitos dos fármacos , Fatores de TempoRESUMO
BACKGROUND: The Mirasol Pathogen Reduction Technology System (PRT) for Plasma (CaridianBCT) is based on a riboflavin and UV light treatment process resulting in pathogen inactivation due to irreversible, photochemically induced damage of nucleic acids. This study evaluated the in vitro protein quality of plasma products treated with riboflavin and UV light following treatment and subsequent storage for up to 104 weeks at -30 degrees C. MATERIALS AND METHODS: Apheresis and whole blood-derived plasma products were combined with riboflavin solution and exposed to ultraviolet light. Treated plasma was then flash frozen, within 8 h of collection, stored at -30 degrees C for up to 104 weeks and analysed at different stages of storage using standard coagulation assays. Results were compared with paired, untreated units stored for the same intervals. RESULTS: The average percent protein retention for all time-points in PRT-treated plasma samples after 36, 69, 87 and 104 weeks of storage at -30 degrees C in comparison with controls held under similar conditions were: Total Protein, 101%, Factor VIII, 79%, Fibrinogen, 78%, Factor II, 87%, Factor XII, 86%, Factor X, 84% and Factor IX, 81%. Anticoagulant and inhibitor proteins showed between 90% and 100% retention after 1 year (52 weeks) and 69 weeks of storage. No clinically relevant complement activation was observed in treated and stored samples. CONCLUSION: Riboflavin and UV light-treated plasma demonstrates reductions in several plasma coagulation factors following treatment. This reduction in activity levels is noted immediately after treatment and remains relatively constant during 2 years of storage at -30 degrees C.
Assuntos
Remoção de Componentes Sanguíneos/métodos , Proteínas Sanguíneas/análise , Patógenos Transmitidos pelo Sangue/isolamento & purificação , Plasma/fisiologia , Riboflavina/farmacologia , Preservação de Sangue/métodos , Criopreservação/métodos , Humanos , Plasma/efeitos dos fármacos , Plasma/efeitos da radiação , Raios UltravioletaRESUMO
Several derivatives of cholesterol containing oxyethylene headgroups with and without a terminal galactose have been synthesized in order to examine the effects of immobilizing a cryoprotectant at a membrane surface. In this work, we have studied the ability of the triethoxycholesterol (TEC) and triethoxycholesterol galactose (TEC-Gal) derivatives to modulate the phase behavior of phosphatidylcholine and phosphatidylethanolamine membranes. Methods of fluorescence polarization, 31P-NMR and freeze-fracture electron microscopy were employed to monitor these changes in lipid phase behavior. Fluorescence polarization data demonstrated the ability of the derivatives to fluidize gel state and rigidify liquid-crystalline state phosphatidylcholines in a manner similar to that observed for cholesterol. Unlike cholesterol, however, the Tm of dipalmitoylphosphatidylcholine (DPPC) was reduced in a concentration-dependent manner with each of the derivatives. Freeze-fracture electron microscopy and 31P-NMR of DOPE dispersions indicate an increase in the lamellar to hexagonal phase-transition temperature on the order of 10-20 C degrees above room temperature for mixtures with 20 mol% of the derivatives. These results are discussed in terms of the properties exhibited by compounds such as carbohydrates, which are known to serve as cryoprotectants for synthetic and biological membranes.
Assuntos
Crioprotetores , Lipossomos , 1,2-Dipalmitoilfosfatidilcolina , Colesterol/análogos & derivados , Técnica de Fratura por Congelamento , Galactosídeos , Espectroscopia de Ressonância Magnética/métodos , Microscopia Eletrônica , Fosfatidilcolinas , Fosfatidiletanolaminas , Espectrometria de Fluorescência/métodos , TermodinâmicaRESUMO
The photochemistry and photophysics of 3-amino-6-iodoacridine (Acr-I) was studied. Photolysis (350 nm) of Acr-I (free base) generates products consistent with a free radical intermediate in methanol, benzene and carbon tetrachloride. The Acr-I hydrochloride is shown to bind to calf thymus DNA and to the self-complementary dinucleotide cytidylyl-(3'-5')-guanosine (CpG) miniduplex in a manner similar to that of proflavine (Acr-NH2), a known DNA intercalator. The Acr-I is shown to more efficiently nick supercoiled plasmid DNA pBR322 upon 350 nm or 420 nm photolysis than Acr-NH2. The efficiency of Acr-I-sensitized DNA nicking is not oxygen dependent. Photolysis of the Acr-I/(CpG)2 complex leads to cleavage of the dinucleotide and to cytidine base release by selective damage to a specific ribose moiety. Dinucleotide cleavage occurs equally well in the presence or absence of oxygen, thereby eliminating a singlet oxygen- or peroxyl radical-mediated process. Photolysis of Acr-I in the presence of a mononucleotide (GMP) or a non-self-complementary dinucleotide (uridylyl-[3'-5']-cytidine-UpC) does not lead to fragmentation and base release. Similarly, photolysis of the Acr-NH2/(CpG)2 complex does not lead to fragmentation and base release. The data indicate that photolysis of an iodinated intercalator bound to CpG or plasmid DNA generates an intercalated aryl radical and that the reactive intermediate initiates a sequence of reactions that efficiently nick nucleic acids. The inactivation of lambda phage sensitized by Acr-I with UV (350 nm) light is oxygen independent but with visible (420 nm) light is strongly oxygen dependent. The Acr-I fluoresces more intensely when excited at 446 than at 376 nm. Thus, UV photolysis may lead to C-I bond homolysis and free radical formation, a process that is not energetically feasible with visible light. The results demonstrate the difficulty of extrapolating model studies involving simple molecules and DNA to understanding the mechanism of viral inactivation with a particular sensitizer.
Assuntos
Aminoacridinas/química , Bacteriófago lambda/efeitos dos fármacos , Bacteriófago lambda/crescimento & desenvolvimento , Ativação Viral/efeitos dos fármacos , Aminoacridinas/metabolismo , Animais , Bovinos , Fenômenos Químicos , Físico-Química , DNA/efeitos dos fármacos , DNA/metabolismo , Fotoquímica , Fotólise , Ativação Viral/efeitos da radiaçãoRESUMO
Amino or polyamino derivatives of naphthalene (N-H), anthracene (A-H) and 8-alkoxypsoralen (PSR-H) were prepared along with their monobrominated analogs (N-Br, A-Br and PSR-Br). The ammonium salts of these compounds are all water soluble and bind strongly to calf thymus DNA and to lambda phage, a double-helical DNA, protein-coated virus. Binding of the sensitizer to DNA occurs, presumably by a mixture of hydrophobic, intercalative and electrostatic interactions. Relative binding constants to calf thymus DNA and to lambda phage were measured by the ethidium bromide fluorescence quenching assay. In general the brominated analogs bind more tightly to calf thymus DNA and to the virus than to the nonhalogenated analogs. It is demonstrated that the brominated aromatics are much more effective at inactivating lambda phage upon photoactivation (lambda approximately 310 or 350 nm) than are their nonbrominated analogs. At identical sensitizer concentrations (by weight) and light flux N-Br, A-Br, and PSR-Br produce 5-6 more logs of viral inactivation than their nonbrominated counterparts (N-H, A-H and PSR-H, respectively). The bromine effect may originate from light-induced electron transfer and subsequent cleavage of the C-Br bond of the sensitizer radical anion bonds to form aryl radicals. Singlet oxygen cannot be responsible for the viral inactivation because the brominated sensitizers are equally effective in the presence and absence of oxygen. Dithiothreitol does not protect lambda phage from light-induced inactivation by the brominated sensitizer thereby demonstrating that the photogenerated reactive intermediates responsible for the effect are complexed to the virus and are not generated free in solution.
Assuntos
Bacteriófago lambda/efeitos dos fármacos , Fármacos Fotossensibilizantes/farmacologia , Animais , Antracenos/farmacologia , Bacteriófago lambda/efeitos da radiação , Bovinos , Dano ao DNA , Furocumarinas/farmacologia , Técnicas In Vitro , Naftalenos/farmacologia , FotoquímicaRESUMO
To assess the viability of human red blood cells that have been lyophilized and reconstituted to the hydrated state, we phlebotomized a unit of whole blood from six healthy male volunteers. Their packed red blood cells were lyophilized at -40 degrees C and stored at 4 degrees C. Upon rehydration, recovery of erythrocytes was 85.2 +/- 2.79%. Aliquots of 20 ml were labeled with 51Cr and re-infused into the original donors for red cell survival studies. The red cells retained ABO and Rh identity upon rehydration. There were no adverse clinical affects of re-infusion. The half time of 51Cr disappearance from the circulation was 31 +/- 8.19 days, and there was no evidence of significant splenic sequestration on the day of reinfusion. Red cell indices of the rehydrated erythrocytes were normal, oxyhemoglobin content was 98.58 +/- 1.46%, and P50 was 27.25 +/- 1.84 mmHg. Although deformability was slightly decreased, the osmotic fragility and filterability of the red cells were normal. These data demonstrate that human erythrocytes can be lyophilized and reconstituted to the hydrated state and survive normally in the circulation. Metabolic, osmotic, hematological and rheological function remains intact.
Assuntos
Transfusão de Eritrócitos , Eritrócitos/citologia , Adulto , Sobrevivência Celular/fisiologia , Estudos de Avaliação como Assunto , Liofilização , Humanos , Masculino , Valores de ReferênciaRESUMO
BACKGROUND AND OBJECTIVES: In recent years, the desire to develop methods to inactivate pathogens in blood components has continued to grow. Several of these proposed approaches have been introduced or are currently in clinical studies. The use of chemical inactivating agents must be considered in terms of the current safety of the blood supply and the potential risks that the introduction of new chemical entities into blood components may carry. The impact which these treatment procedures have on the in vitro and in vivo performance of these products must also be considered relative to the potential benefit of the pathogen inactivation potential they offer. This paper will discuss one possible approach for inactivating pathogens in blood using vitamin B2, Riboflavin, and light. MATERIALS AND METHODS: We have used Riboflavin for treating plasma and platelets and evaluated protein quality and platelet function in vitro. Initial toxicology tests to assess the impact of infusion of photoproducts generated in these processes have also been conducted in rodents. Cytotoxicity evaluations have been used to assess the possible impact of photoproduct toxicity in vivo. Virus and bacteria spiking studies using a variety of human and animal model pathogens have been conducted in order to asses the efficacy of this process. RESULTS: Initial toxicology assessment of the photoproducts of Riboflavin generated under the proposed treatment conditions have been favorable. Virus and bacteria clearance studies have demonstrated efficacy of the procedure against a wide range of human and animal pathogens, including intracellular HIV-1. Studies with platelet and plasma function indicated reductions in vitro comparable to other proposed treatment approaches. CONCLUSION: The use of Riboflavin in a photochemical decontamination process for blood components shows promise.
Assuntos
Patógenos Transmitidos pelo Sangue , Riboflavina/farmacologia , Plaquetas/efeitos dos fármacos , Plaquetas/microbiologia , Plaquetas/efeitos da radiação , Proteínas Sanguíneas/metabolismo , Humanos , Luz , Fotoquímica , Riboflavina/metabolismo , Riboflavina/efeitos da radiação , EsterilizaçãoRESUMO
Egg PC vesicles frozen and thawed in the presence of carbohydrate derivatives do not incur damage normally associated with freeze thawing. Treated vesicles maintain membrane integrity as evidenced by the lack of lipid intermixing and maintenance of vesicle size following freezing and thawing. This protection is conferred at a derivative:lipid ratio of 0.4 mol/mol, significantly lower than the amount of carbohydrate required when not attached directly to the vesicle. This result indicates that only the carbohydrate at the vesicle surface is responsible for imparting stability to the membrane. This effect can be modulated by variations in the nature of the surrounding medium or alterations in the structure of the carbohydrate, suggesting that direct interactions between the carbohydrate and membrane occur at the membrane interface which are sensitive to bulk phase properties.
Assuntos
Crioprotetores , Glicolipídeos , Transferência de Energia , Congelamento , Luz , Fosfatidilcolinas , Espalhamento de RadiaçãoRESUMO
This study was designed to establish an alternative technique to the morphology scoring method described by Kunicki et al. [Transfusion 1975;15:414-421] using light transmission. A reference scale for platelet quality (expressed as a percentage of fresh cells) was established with a mixture of fresh platelets and damaged platelets frozen without cryoprotectant. The transmittance, read against autologous plasma, was compared to morphology, hypotonic stress response (HSR) and ADP-induced aggregation. The results showed that transmittance, morphology and HSR were related linearly to the percentage of fresh platelets. There was a significant correlation between relative transmittance (RT) and morphology (r = 0.932), and RT and HSR (r = 0.91), but a very poor correlation was found between transmittance and aggregation. Measurements of transmittance provided an estimation of the global aspect of a population of cryopreserved platelets in terms of shape, size and composition, and the relative transmittance of a sample to that of the fresh population could predict morphology and HSR. It is concluded that these measurements offer a valid alternative method to the subjective morphology scoring obtained with cryopreserved platelets.
Assuntos
Plaquetas/citologia , Preservação de Sangue , Criopreservação , Nefelometria e Turbidimetria , Difosfato de Adenosina/farmacologia , Plaquetas/efeitos dos fármacos , Calibragem , Sobrevivência Celular , Crioprotetores/farmacologia , Humanos , Soluções Hipotônicas/farmacologia , Pressão Osmótica , Agregação Plaquetária/efeitos dos fármacosRESUMO
The desirability of extending the therapeutic benefits of erythrocyte transfusions beyond the currently available options is well established. Extended frozen storage of rare blood types, hemoglobin-based or artificial blood substitutes for treatment of short-term deficits in oxygen-carrying capacity, novel approaches to tissue oxygenation for cancer therapy, and efficient storage and availability of erythrocytes in military field installations are all potential applications of ongoing research and development efforts. This review explains the need for improvement in erythrocyte storage and development of blood substitutes, the current problems in the field, and the progress made toward solving these problems in the past year.
Assuntos
Preservação de Sangue/métodos , Substitutos Sanguíneos , Eritrócitos , Animais , HumanosRESUMO
Normal human erythrocytes (RBC) were freeze-dried under conditions that caused minimal modification in normal RBC metabolic activities. Because of the known effects of long-term storage on metabolic activities, we studied the effects of our lyophilization process on RBC metabolism. Of all the metabolic enzymes studied, only triosephosphate isomerase (D-glyceraldehyde-3-phosphate ketol-isomerase, EC 5.3.1.1), enolase (2-phospho-D-glyceratehydro-lyase, EC 4.2.1.11), and pyruvate kinase (ATP:pyruvate O2-phosphotransferase, EC 2.7.1.40) were decreased when compared with fresh control nonlyophilized RBC. The activities of these enzymes did not differ significantly from those of blood bank RBC. Concentrations of high-energy intermediates, ATP, and 2,3-diphosphoglycerate, along with lactate and ATP production were decreased in lyophilized RBC. No enzymes of the pentose phosphate shunt were altered during lyophilization. In addition, our data show that lyophilized RBC possess an intact capacity to (i) synthesize adenine nucleotides and (ii) reduce MetHb to Hb and, thus, maintain the Hb in a functional physiologic state similar to fresh nonlyophilized RBC. The present study demonstrates the possibility of lyophilizing RBC in a manner that maintains normal metabolic and enzymatic function upon rehydration.
Assuntos
Preservação de Sangue/métodos , Eritrócitos/metabolismo , Nucleotídeos de Adenina/sangue , Liofilização , Glutationa/sangue , Glicólise , Humanos , Técnicas In Vitro , Metemoglobina/metabolismoRESUMO
We have examined the behavior of the dry phospholipid dipalmitoylphosphatidylcholine (DPPC) in the presence of several carbohydrate derivatives. These carbohydrate derivatives possess a hydrophobic portion which is incorporated directly into the DPPC membrane and a hydrophilic portion which places the carbohydrate structure at the membrane interface with the surrounding matrix. In the presence of these derivatives, the physical properties of the membrane are altered. These alterations are evident in changes observed in the phosphate and carbonyl vibrational modes of the phospholipid portion of the membrane. In addition, the phase transition behavior of the lipid is significantly altered as evidenced by a reduction in the gel to liquid-crystalline phase transition temperature. These results are consistent with those previously reported for free carbohydrates interacting with membranes in which a water replacement hypothesis has been used to explain the behavior. The attachment of carbohydrates to the membrane enhances these effects by localizing the agent responsible for these alterations at the membrane interface.
Assuntos
1,2-Dipalmitoilfosfatidilcolina/química , Carboidratos/química , Lipídeos de Membrana/química , Varredura Diferencial de Calorimetria , Colesterol/análogos & derivados , Colesterol/química , Técnicas In Vitro , Maltose/análogos & derivados , Maltose/química , Espectrofotometria Infravermelho , Análise Espectral Raman , TemperaturaRESUMO
Human red cells (RBCs) were collected in CPDA-1 and then freeze-dried in lyoprotective solution. The lyophilized RBCs were then stored at -20 degrees C for 7 days. At the end of the storage period, the lyophilized RBCs were rehydrated and washed in dextrose saline. The washed, reconstituted, lyophilized RBCs were resuspended in final wash solutions of ADSOL, CPDA-1, or a special additive solution containing glucose, citrate, phosphate, adenine, and mannitol, and then they were stored at 4 degrees C for an additional 7 days. The main purpose of this study was to determine whether human RBCs can be lyophilized in such a manner that normal metabolic, rheologic, and cellular properties are maintained during rehydration and subsequent storage in standard blood bank preservative solutions. Our results show that reconstituted, lyophilized RBCs maintained levels of ATP, 2,3 DPG, lactate, and cellular properties that are equal to or better than those in control nonlyophilized RBCs stored for a comparable period in CPDA-1. Reconstituted, lyophilized RBCs stored at 4 degrees C after rehydration also show better maintenance of ATP, 2,3 DPG, and lactate than do control RBCs stored in the same preservative solutions for comparable periods.
Assuntos
Preservação de Sangue/métodos , Eritrócitos/citologia , 2,3-Difosfoglicerato , Adenina , Difosfato de Adenosina/sangue , Monofosfato de Adenosina/sangue , Trifosfato de Adenosina/sangue , Citratos , Temperatura Baixa , Crioprotetores , Ácidos Difosfoglicéricos/sangue , Deformação Eritrocítica , Índices de Eritrócitos , Eritrócitos/química , Liofilização , Glucose , Humanos , Lactatos/sangue , Manitol , Fragilidade Osmótica , Fosfatos , Soluções para Reidratação , Cloreto de Sódio , SoluçõesRESUMO
BACKGROUND AND OBJECTIVES: A pathogen-reduction technology (PRT) system using riboflavin and light has been developed for the treatment of platelet concentrates (PC) obtained by either buffy coat preparation (BCPC) or apheresis procedures (APPC). The aim of this study was to evaluate the effects of the treatment process on in vitro cell quality and on riboflavin conversion in PC. MATERIALS AND METHODS: BCPC were prepared with the Compomat G4 from whole blood which had been stored overnight after collection. APPC were obtained using the TRIMA apheresis procedure. Both PC products had been stored for 18-24 h prior to PRT treatment. BCPC and APPC were treated with PRT on day 2 and day 1 of shelf-life, respectively. The treated PCs were then maintained for an additional 5 days after the PRT treatment. A panel of cell quality assays and high-performance liquid chromatography (HPLC) analysis were performed. RESULTS: Cell counts and plasma lactate dehydrogenase (LDH) levels during storage indicated that PRT did not induce significant cell lysis. Acceleration of a decrease in glucose and an increase in lactate was observed for treated PCs, but no significant differences were observed between treated BCPC and APPC. The pH of treated samples remained above 7.0, although was lower than that of the control. Platelet morphology of BCPC and APPC was well preserved. P-selectin expression indicated significant platelet activation when compared with control PC (BCPC on day 6: 39% vs. 12%; APPC on day 5: 35% vs. 18%). Both P-selectin expression and microparticle formation were not significantly different between treated BCPC and APPC during storage. The JC-1 assay also displayed no loss of mitochondria integrity during the storage of treated products. Approximately 20% of riboflavin converted into photoproducts, including lumichrome. CONCLUSIONS: PRT treatment had an effect on the development of the normal platelet storage lesion at a level which seems tolerable for clinical usage.
Assuntos
Remoção de Componentes Sanguíneos , Plaquetas/microbiologia , Luz , Riboflavina/farmacologia , Remoção de Componentes Sanguíneos/métodos , Plaquetas/química , Plaquetas/efeitos dos fármacos , Plaquetas/fisiologia , Preservação de Sangue , Cromatografia Líquida , Glucose/análise , Humanos , Concentração de Íons de Hidrogênio , L-Lactato Desidrogenase/sangue , Ácido Láctico/análise , Selectina-P/sangue , Ativação Plaquetária/fisiologia , Contagem de Plaquetas , Preservação de TecidoRESUMO
Inactivation of viruses in blood products requires that the method employed display selectivity in its action for viral elements while not affecting the biological entity of interest. Several methods have been developed for the treatment of human plasma or products derived from human plasma. An effective technique for the treatment of the cellular components of blood has been lacking, in part due to the inability to develop agents capable of selectively targeting viral agents in the milieu of cellular material. In this paper, we examine the behavior of a group of viral sensitizers designed to be added to cellular samples and be activated upon exposure to UVA light. Upon activation, these agents are capable of disrupting nucleic acids of the virus in a manner that renders them inactive for proliferation. The selectivity observed in this inactivation is determined by the chemical structure of the sensitizer, which can be varied to increase viral killing capacity while diminishing collateral damage to cellular and protein constituents.