Age of blood and recipient factors determine the severity of transfusion-related acute lung injury (TRALI)

Introduction Critical care patients frequently receive blood transfusions. Some reports show an association between aged or stored blood and increased morbidity and mortality, including the development of transfusion-related acute lung injury (TRALI). However, the existence of conflicting data endorses the need for research to either reject this association, or to confirm it and elucidate the underlying mechanisms. Methods Twenty-eight sheep were randomised into two groups, receiving saline or lipopolysaccharide (LPS). Sheep were further randomised to also receive transfusion of pooled and heat-inactivated supernatant from fresh (Day 1) or stored (Day 42) non-leucoreduced human packed red blood cells (PRBC) or an infusion of saline. TRALI was defined by hypoxaemia during or within two hours of transfusion and histological evidence of pulmonary oedema. Regression modelling compared physiology between groups, and to a previous study, using stored platelet concentrates (PLT). Samples of the transfused blood products also underwent cytokine array and biochemical analyses, and their neutrophil priming ability was measured in vitro. Results TRALI did not develop in sheep that first received saline-infusion. In contrast, 80% of sheep that first received LPS-infusion developed TRALI following transfusion with "stored PRBC." The decreased mean arterial pressure and cardiac output as well as increased central venous pressure and body temperature were more severe for TRALI induced by "stored PRBC" than by "stored PLT." Storage-related accumulation of several factors was demonstrated in both "stored PRBC" and "stored PLT", and was associated with increased in vitro neutrophil priming. Concentrations of several factors were higher in the "stored PRBC" than in the "stored PLT," however, there was no difference to neutrophil priming in vitro. Conclusions In this in vivo ovine model, both recipient and blood product factors contributed to the development of TRALI. Sick (LPS infused) sheep rather than healthy (saline infused) sheep predominantly developed TRALI when transfused with supernatant from stored but not fresh PRBC. "Stored PRBC" induced a more severe injury than "stored PLT" and had a different storage lesion profile, suggesting that these outcomes may be associated with storage lesion factors unique to each blood product type. Therefore, the transfusion of fresh rather than stored PRBC may minimise the risk of TRALI.

Packed red blood cells suppress myeloid dendritic cell and monocyte inflammatory responses in a whole blood model of transfusion

Purpose/Objective: The basis for poor outcomes in some patients post transfusion remains largely unknown. Despite leukodepletion, there is still evidence of immunomodulatory effects of transfusion that require further study. In addition, there is evidence that the age of blood components transfused significantly affects patient outcomes. Myeloid dendritic cell (DC) and monocyte immune function were studied utilising an in vitro whole blood model of transfusion. Materials and methods: Freshly collected (‘recipient’) whole blood was cultured with ABO compatible leukodepleted PRBC at 25% blood replacement-volume (6hrs). PRBC were assayed at [Day (D) 2, 14, 28and 42 (date-of expiry)]. In parallel, LPS or Zymosan (Zy) were added to mimic infection. Recipients were maintained for the duration of the time course (2 recipients, 4 PRBC units, n = 8).Recipient DC and monocyte intracellular cytokines and chemokines (IL-6, IL-10, IL-12,TNF-a, IL-1a, IL-8, IP-10, MIP-1a, MIP-1b, MCP-1) were measured using flow cytometry. Changes in immune response were calculated by comparison to a parallel no transfusion control (Wilcoxin matched pairs). Influence of storage age was calculated using ANOVA. Results: Significant suppression of DC and monocyte inflammatory responses were evident. DC and monocyte production of IL-1a was reduced following exposure to PRBC regardless of storage age (P < 0.05 at all time points). Storage independent PRBC mediated suppression of DC and monocyte IL-1a was also evident in cultures costimulated with Zy. In cultures co-stimulated with either LPS or Zy, significant suppression of DC and monocyte TNF-a and IL-6 was also evident. PRBC storage attenuated monocyte TNF-a production when co-cultured with LPS (P < 0.01 ANOVA). DC and monocyte production of MIP-1a was significantly reduced following exposure to PRBC (DC: P < 0.05 at D2, 28, 42; Monocyte P < 0.05 all time points). In cultures co-stimulated with LPS and zymosan, a similar suppression of MIP-1a production was also evident, and production of both DC and monocyte MIP-1b and IP-10 were also significantly reduced. Conclusions: The complexity of the transfusion context was reflected in the whole blood approach utilised. Significant suppression of these key DC and monocyte immune responses may contribute to patient outcomes, such as increased risk of infection and longer hospital stay, following blood transfusion.

Neutrophil oxidative burst is primed by supernatant from stored red cells : implications for Transfusion-Related Acute Lung Injury (TRALI)

Aim/Background: Transfusion-related acute lung injury (TRALI) is a potentially fatal adverse transfusion reaction. It is hypothesised to occur via a two-insult mechanism: the recipient’s underlying co-morbidity in addition to the transfusion of blood products activate neutrophils in the lung resulting in damaged endothelium and capillary leakage. Neutrophil activation may occur by antibody or non-antibody related mechanisms, with the length of storage of cellular blood products implicated in the latter. This study investigated non-antibody mediated priming and/or activation of neutrophil oxidative burst. Methods: A cytochrome C reduction assay was used to assess priming and activation of neutrophil oxidative burst by pooled supernatant (SN) from day 1 (D1; n=75) and day 42 (D42; n=113) packed red blood cells (PRBC). Pooled PRBC-SN were assessed in parallel with PAF (priming), fMLP (activating), PAF + fMLP (priming + activating) and buffer only (negative) controls. Cytochrome C reduction was measured over 30min at 37oC (inclusive of 10min priming). Neutrophil activation by PRBC-SN was assessed cf. buffer only and neutrophil priming by PRBC-SN was assessed by co-incubation with fMLP cf. fMLP alone. One-way ANOVA; Newman-Keuls post-test; p<0.05; n=10 independent assays. Results: Neither D1- nor D42- PRBC-SN alone activated neutrophil oxidative burst. In addition, D1-PRBC-SN did not prime fMLP-activated neutrophil oxidative burst. D42-PRBC-SN did, however, prime neutrophils for subsequent activation of oxidative burst by fMLP, the magnitude of response being similar to PAF (a known neutrophil priming agonist). Conclusion: These findings are consistent with the two-insult mechanism of TRALI. Factors released into the SN during PRBC storage contributed to neutrophil priming synergistically with other neutrophil stimulating agonists. This implicates PRBC storage duration as a key factor contributing to non-immune neutrophil activation in the development of TRALI in patients with pre-disposing inflammatory conditions.

Stored blood products augment inflammation in a two-event model of Transfusion Related Acute Lung Injury (TRALI)

Aim/Background TRALI is hypothesised to develop via a two-event mechanism involving both the patieint's underlying morbidity and blood product factors. The storage of cellular products has been implicated in cases of non-antibody mediated TRALI, however the pathophysiological mechanisms are undefined. We investigated blood product storage-related modulation of inflmmatory cells and medicators involved in TRALI. Methods In an in vitro mode, fresh human whole blood was mixed with culture media (control) or LPS as a 1st event and "transfused" with 10% (v/v) pooled supernatant (SN) from Day 1 (d1, n=75) or Day 42 (D42, n=113) packed red blood cells (PRBCs) as a 2nd event. Following 6hrs, culture SN was used to assess the overall inflammatory response (cytometric bead array) and a duplicate assay containing protein transport inhibitor was used to assess neutrophil- and monocyte-specific inflmamatory responses using multi-colour flow cytometry. Panels: IL-6, IL-8, IL-10, IL-12, IL-1, TNF, MCP-1, IP-10, MIP-1. One-way ANOVA 95% CI. Results In the absence of LPS, exposure to D1 or D42 PRBC-SN reduced monocyte expression of IL-6, IL-8 and Il-10. D42 PRBC-SN also reduced monocyte IP-10, and the overall IL-8 production was increased. In the presence of LPS, D1-PRBC SN only modified overall IP-10 levels which were reduced. However, cf LPS alone, the combination of LPS and D42 PRBC-SN resulted in increased neutrophil and monocyte productionof IL-1 and IL-8 as well as reduced monocyte TNF production. Additionally, LPS and D42 PRBC-SN resulted in overall inflmmatory changes: elevated IL-8, -1, IL-10 and MIP-1, and reduced TNF and IP-10. Conclusion In the absence of LPS, exposure to both fresh and stored PRBC-SN predominantly resulted in an anti-imflammatory response. However, in the presence of LPS, stored but not fresh PRBC-SN augmented a pro-inflammatory profile. These data support the two-event mechanism of TRALI pathogenesis, and highlights blood product storage duration as a factor contributing to the development of inflmamatory responses in non-antibody mediated TRALI.

Potential role of bioactive lipids in the development of non-antibody mediated transfusion-related acute lung injury (TRALI)

Transfusion-related acute lung injury (TRALI) has been the leading cause of transfusion-related morbidity and mortality in the UK and the USA in recent years. A threshold mechanism of TRALI has been proposed in which both patient factors (type and/or severity of clinical insult) and blood product factors (strength and/or concentration of antibodies or biological response modifiers) interact to surpass a threshold for TRALI development (Bux et al. Br J Haematol; 2007; 136: 788-99). The risk of developing antibody-mediated TRALI has been minimised by the introduction of risk-reduction strategies such as limiting the use of plasma from female donors. In contrast, there are no strategies currently in place to mitigate the development of non-antibody mediated TRALI as the mechanisms remain largely undefined. Previous studies have implicated non-polar lipids such as arachidonic acid and various species of hydroxyeicosatetranoic acid (HETE) in the development of non-antibody mediated TRALI (Silliman et al. Transfusion; 2011; 51: 2549-54), however the contribution of these lipids to the development of an inflammatory response in TRALI is poorly understood.

Soluble mediators in platelet concentrates modulate dendritic cell inflammatory responses in an experimental model of transfusion

The transfusion of platelet concentrates (PCs) is widely used to treat thrombocytopenia and severe trauma. Ex vivo storage of PCs is associated with a storage lesion characterized by partial platelet activation and the release of soluble mediators, such as soluble CD40 ligand (sCD40L), RANTES, and interleukin (IL)-8. An in vitro whole blood culture transfusion model was employed to assess whether mediators present in PC supernatants (PC-SNs) modulated dendritic cell (DC)-specific inflammatory responses (intracellular staining) and the overall inflammatory response (cytometric bead array). Lipopolysaccharide (LPS) was included in parallel cultures to model the impact of PC-SNs on cell responses following toll-like receptor-mediated pathogen recognition. The impact of both the PC dose (10%, 25%) and ex vivo storage period was investigated [day 2 (D2), day 5 (D5), day 7 (D7)]. PC-SNs alone had minimal impact on DC-specific inflammatory responses and the overall inflammatory response. However, in the presence of LPS, exposure to PC-SNs resulted in a significant dose associated suppression of the production of DC IL-12, IL-6, IL-1a, tumor necrosis factor-a (TNF-a), and macrophage inflammatory protein (MIP)-1b and storage-associated suppression of the production of DC IL-10, TNF-a, and IL-8. For the overall inflammatory response, IL-6, TNF-a, MIP-1a, MIP-1b, and inflammatory protein (IP)-10 were significantly suppressed and IL-8, IL-10, and IL-1b significantly increased following exposure to PC-SNs in the presence of LPS. These data suggest that soluble mediators present in PCs significantly suppress DC function and modulate the overall inflammatory response, particularly in the presence of an infectious stimulus. Given the central role of DCs in the initiation and regulation of the immune response, these results suggest that modulation of the DC inflammatory profile is a probable mechanism contributing to transfusion-related complications.

A comprehensive ovine model of blood transfusion

Background The growing awareness of transfusion-associated morbidity and mortality necessitates investigations into the underlying mechanisms. Small animals have been the dominant transfusion model but have associated limitations. This study aimed to develop a comprehensive large animal (ovine) model of transfusion encompassing: blood collection, processing and storage, compatibility testing right through to post-transfusion outcomes. Materials and methods Two units of blood were collected from each of 12 adult male Merino sheep and processed into 24 ovine-packed red blood cell (PRBC) units. Baseline haematological parameters of ovine blood and PRBC cells were analysed. Biochemical changes in ovine PRBCs were characterized during the 42-day storage period. Immunological compatibility of the blood was confirmed with sera from potential recipient sheep, using a saline and albumin agglutination cross-match. Following confirmation of compatibility, each recipient sheep (n = 12) was transfused with two units of ovine PRBC. Results Procedures for collecting, processing, cross-matching and transfusing ovine blood were established. Although ovine red blood cells are smaller and higher in number, their mean cell haemoglobin concentration is similar to human red blood cells. Ovine PRBC showed improved storage properties in saline–adenine–glucose–mannitol (SAG-M) compared with previous human PRBC studies. Seventy-six compatibility tests were performed and 17·1% were incompatible. Only cross-match compatible ovine PRBC were transfused and no adverse reactions were observed. Conclusion These findings demonstrate the utility of the ovine model for future blood transfusion studies and highlight the importance of compatibility testing in animal models involving homologous transfusions.

Utilizacion del zumo de jicaro (Crescentia cujete) en el tratamiento de la dermatomicosis en terneros de la raza Reina en la Finca Santa Rosa de la UNA

El presente estudio se realizó con el objetivo de Determinar la efectividad fungicida del zumo de jícaro (Crescentia cujete) en el tratamiento de la Dermatomicosis en terneros de la raza Reina de la Finca Santa Rosa de la UNA. En el trabajo experimental se utilizó un diseño completamente al azar (D.C.A) el que estuvo compuesto por un lote de 15 terneros divididos en 3 grupos, cada grupo formado por 5 animales eleccionados al azar y sometidos a tratamientos distintos, aplicando una sola dosis cada 24 horas durante tres días. Tratamiento I: Tintura de yodo al 5%. Tratamiento II: Zumo de jícaro al 50%. Tratamiento III: Zumo jícaro al 100%. Obteniéndose los siguientes resultados, el microorganismos causantes de la Dermatomicosis en terneros de la raza Reina en la Finca Santa Rosa de la UNA es el Trichophyton verrucosum. Los tratamientos II y III tuvieron las mejores respuestas en el control, de la Dermatomicosis , con un porcentaje de efectividad del 82%, y 78% respectivamente y con un 40.% para el tratamiento I, se encontró diferencia significativa p< 0.05 entre los tratamientos, siendo el Zumo de jícaro al 50% tienden a curarse mejor los animales que con los otros tratamiento, seguido del Zumo jícaro al 100%).Según el análisis del costo de la dosis se puede decir que el zumo de jícaro es un funguicida económico para los productores.

Estudio preliminar de la utilizacion del anamu (Petiveria alliacea) en la reduccion del puerperio bovino en la Finca El Rosario, Municipio de la Trinidad, departamento de Esteli

El presente estudio se realizó con el objetivo de determinar el grado de efectividad del anamú (Petiveria alliacea) en la reducción del puerperio bovino, en la finca El Rosario, Municipio de La Trinidad, Departamento de Esteli, ubicado entre las coordenadas 12 ° 58’ de latitud norte y 86 ° 14' de longitud oeste con una altura sobre el nivel del mar 601.22 m.s.n.m., con una temperatura promedio anual 23° C. y con precipitación anual de entre 800 y 2000 m.m. El clima es tropical seco con poca precipitación pluvial.Se tomaron 30 vacas paridas al azar y se dividieron en dos grupos de 15 vacas, el tratamiento I aplicación de anamú(Petiveria alliacea)al 20% y el tratamiento II el control sin tratamiento. Los resultados obtenidos, indican que la principal causa del puerperio prolongado es la hipofunción ovárica presente en 20 vacas que representa un 83%, seguido de los quistes folicular 4 animales con un 17%. Los animales tratados con Anamú (Petiveria alliacea)presentan celos a los 79 días, mientras que los animales no tratados presentan celos a los 100 días, obteniéndose que con este tratamiento se reduce el puerperio en 21 días por animal. Al analizar el IPC entre tratamiento, los animales tratados con Anamú presentaron duración de 55-90 días, mientras los no tratado presentaron duración de 81-120 días.

Estudio preliminar de la utilizacion del aceite y resina de Neem (Azadirachta indica A. Juss) en el tratamiento de la sarna Psoroptica equina en la comarca Pilar del municipio de Santo Domingo, departamento de Chontales

El presente estudio se realizó en la finca "El Porvenir" ubicada en los 12°15' de latitud norte y 85"04' de longitud oeste, en el municipio de Santo Domingo departamento de Chontales, el clima es de sabana tropical con temperaturas entre 25 a 27"C. La duración del experimento fue de 18 días a partir de la selección e identificación de los animales bajo los tratamientos evaluados. Con el objetivo de comparar el efecto del aceite y resina de Neem al 40 % versus lvermectina pasta al 1% en el control de la sama Psoróptica equina. Se utilizó un Diseño Completamente al Azar (D.C.A.). Se seleccionaron 21 equinos con edades promedios de 1O a l 5 años, estos se dividieron en 3 grupos, cada grupo formado por 7 equinos por tratamiento, Tratamiento P: Ivermectina pasta 1% a razón de lg por cada 100 kg de peso vivo, vía oral. Tratamiento A:aceite de Neem acaricida tópico botánico a base de aceite de Neem, se aplicó vía tópica un promedio de 20 mi por cada animal. Tratamiento R: resina de Neem, acaricida tópico botánico a base de resina de Neem, se aplicó vía tópica, un promedio de 20 mi por cada equino. A los equinos se les realizó un examen clinico al inicio del ensayo y a los 14 días post aplicación, se utilizó el diagnóstico clínico y sintomatológico para evaluar el comportamiento de los tratamientos. Para cuantificar las reducciones de las lesiones se utilizaron tres variables: detención, reducción y control de las lesiones. Los tratamientos con Neem aceite y resina al 400/o arrojaron los siguientes resultados: Aceite 40% logró alcanzar un 100% del control de las lesiones y la resina alcanzó 86%. La Ivermectina pasta 1% sólo alcanzó la reducción de las lesiones en un 100%. Entre los tres tratamientos, el que dio mejores resultados fue el aceite de Neem al 40% con un porcentaje de eficiencia en el control de la sama equina en un 100%.

Estudio preliminar de la utilizacion del agua de mar como suplemento nutritivo de sales minerales, como alternativa para la ganancia de peso en terneros al destete en la Finca "Santa Rita", comarca El Castillo, del municipio de Mulukuku, RAAN

El presente trabajo se realizó con el objetivo de evaluar la utilización del Agua de Mar como suplemento nutritivo de sales minerales, como alternativa en la ganancia de peso en terneros al destete en la Finca Sta. Rita, comarca el Castillo, municipio de Mulukukú, Región Autónoma del Atlántico Norte (RAAN), ubicada en las coordenadas: 13° 10 ́08 ́ ́ N y 085° 05 ́ 18 ́ ́ W, elevación sobre el nivel del mar de 137m. Se empleó un diseño completamente al azar (D.C.A), donde fueron utilizados 30 animales distribuidos aleatoriamente en tres grupos de 10 animales. Tratamiento I: 1 000 ml de agua de mar 1 vez al día por 30 días ; Tratamiento II 1 000 ml de agua de mar 2 veces al día por 30 días; Tratamiento III: Tratamiento testigo (no se aplicó ningún suplemento mineral). Los resultados obtenidos en ganancia de peso vivo, demostraron que no hubo diferencia significativa entre tratamientos, a pesar de esto se observó una pequeña diferencia teniendo los mejores resultados el Tratamiento I seguido del Tratamiento II y por último el Tratamiento III. De igual forma no se obtuvieron diferencias significativas en la ganancia media diaria entre tratamientos, obteniendo el Tratamiento I 0.4089 kg/Animal/dia, para el Tratamiento II fue de 0.3717 kg y para el Tratamiento III fue de 0.3585. Al realizar el análisis financiero observamos que el tratamiento que mayor rentabilidad nos proporcionaría es el Tratamiento I, obteniendo una utilidad neta de $894.96 dólares, el que se emplea en la finca es el Tratamiento III y este nos deja una utilidad neta de $751.02 dólares, al comparar estas utilidades encontramos una diferencia de $143.94 dólares.