raffinose and Hypothermia

Perfluorocarbons (PFCs) are inert solutions that have a high capacity for dissolving oxygen. There has been a continuing level of research into the delivery of oxygen during solid organ preservation with the use of PFCs. The one- and two-layer methods have been used as static storage techniques, proving particularly successful for pancreas preservation. They can also be formulated as an emulsion for continual perfusion or as a simple flush solution. The success of PFCs in organ preservation seems to be somewhat organ and species dependant, and further experimental evidence is needed to establish their application.

Topics: Adenosine; Allopurinol; Animals; Cells; Dogs; Emulsions; Fluorocarbons; Glutathione; Heart; Humans; Hypothermia; Insulin; Liver; Lung; Organ Preservation; Organ Preservation Solutions; Perfusion; Rabbits; Raffinose; Rats; Temperature

Functional recovery following heart transplantation mainly depends on the ability of preservative solution in providing the physical and biochemical environment so as to maintain the viability of the tissue during preservation and in reperfusion. Here we demonstrate the protective effects of a novel bisindolylmaleimide derivative, MS1, on enhancing the functional recovery of the heart following long-term hypothermic preservation when added to the preservative solution.. After anesthesia and artificial ventilation, the hearts were rapidly isolated and perfused with Kreb's Henseleit buffer at 37 degrees C in working mode. After 30 minutes of perfusion, the hearts were arrested with cardioplegic solution and preserved in University of Wisconsin solution with (UW-MS1 group) or without MS1 (UW-Vehicle group) for 12 h at 4 degrees C. After 12 hours, the hearts were reperfused for 60 minutes.. MS1 treated hearts showed: a) significant recovery of cardiac functions (P<0.001), b) well-preserved myocardial ATP levels (P<0.001), c) less myocardial water content (P<0.01), d) reduced oxidative stress (P<0.001), e) less intracellular swelling and well-preserved mitochondria, and g) activation of cell survival cascades compared to the control hearts preserved in UW solution without MS1. In contrast, these protective effects of MS1 were abolished on opening the permeability transition pore before MS1 treatment.. These results altogether indicate the efficacy of this compound in protecting the myocardium against reperfusion injury and thus making this drug a clinically useful tool in patients undergoing reperfusion after cardiac surgeries.

Topics: Adenosine; Allopurinol; Animals; Glutathione; Heart; Hypothermia; Indoles; Insulin; Lipid Peroxidation; Male; Maleimides; Mitochondria, Heart; Myocardium; Organ Preservation; Organ Preservation Solutions; Protein Kinase C; Raffinose; Rats; Rats, Wistar

Redox-active iron, catalyzing the generation of reactive oxygen species, has been implicated in experimental renal ischemia-reperfusion injury. However, in clinical transplantation, it is unknown whether redox-active iron is involved in the pathophysiology of ischemic injury of non-heart-beating (NHB) donor kidneys. We measured redox-active iron concentrations in perfusate samples of 231 deceased donor kidneys that were preserved by machine pulsatile perfusion at our institution between May 1998 and November 2002 using the bleomycin detectable iron assay. During machine pulsatile perfusion, redox-active iron was released into the preservation solution. Ischemically injured NHB donor kidneys had significantly higher perfusate redox-active iron concentrations than heart-beating (HB) donor kidneys that were not subjected to warm ischemia (3.9 +/- 1.1 vs. 2.8 +/- 1.0 micromol/L, p = 0.001). Moreover, redox-active iron concentration was an independent predictor of post-transplant graft viability (odds ratio 1.68, p = 0.01) and added predictive value to currently available donor and graft characteristics. This was particularly evident in uncontrolled NHB donor kidneys for which there is the greatest uncertainty about transplant outcomes. Therefore, perfusate redox-active iron concentration shows promise as a novel viability marker of NHB donor kidneys.

Topics: Adenosine; Allopurinol; Cadaver; Cell Survival; Glutathione; Graft Survival; Heart Arrest; Humans; Hypothermia; Insulin; Iron; Ischemia; Kidney; Kidney Transplantation; Organ Preservation Solutions; Oxidation-Reduction; Perfusion; Predictive Value of Tests; Raffinose; Reactive Oxygen Species; Tissue Donors; Treatment Outcome

The chronic organ shortage has led to the reconsideration of marginal donor pools such as non-heart-beating donors (NHBD). The use of these livers is limited due to their minimal tolerance for cold preservation. The aim of this study was to examine the combination of two different therapeutic strategies for the preservation of livers from NHBD. The livers of male Wistar rats were harvested after the induction of cardiac arrest via phrenotomy (30, 90 minutes). Livers were perfused with 10 mL of UW solution (UW), followed by hypothermic preservation with or without insufflation of gaseous oxygen (O2). In one group a fibrinolytic preflush (10 mL of Ringer's containing 7500 IU of streptokinase) was performed with subsequent preservation with O2 (O2+SK). After storage (24 h/4 degrees C/UW) livers were reperfused in vitro. Livers retrieved from heart beating donors served as controls. The results showed that even after only 30 minutes of warm ischemia livers displayed a serious disturbance in vascular perfusion (portal venous pressure, PVP = 7.4 +/- 0.2* versus control: 4.1 +/- 0.5 mmHg), associated with a more than 10-fold increase in enzyme release (ALT: 26819 +/- 513* versus control 683 +/- 152 mU/g/L), which was consistent with a significant depression in bile synthesis (1.21 +/- 0.35* versus 19.36 +/- 2.16 microL/g/45 min). However, these impairments could be prevented with O2. Even after 90 minutes of WI, the function was significant better using aerobic preservation (ALT: 3204 +/- 549 mU/g/L). With a supplementary fibrinolytic preflush, we effectively preserved livers up to 90 minutes of WI with results comparable to livers from heart beating donors with no WI (ALT: 1623 +/- 432 mU/g/L). The combination of these two techniques represents a new therapeutic approach for livers with extended or unclear WI periods in non-heart-beating donors (*P <.05 versus control).

Topics: Adenosine; Aerobiosis; Allopurinol; Animals; Bile; Cell Death; Disease Models, Animal; Fibrinolysis; Glutathione; Heart Arrest; Hypothermia; Insulin; Liver; Male; Organ Preservation; Organ Preservation Solutions; Raffinose; Rats; Rats, Wistar; Vascular Resistance

Topics: Adenosine; Allopurinol; Animals; Carbon Dioxide; Cold Temperature; Dogs; Glutathione; Hypothermia; Insulin; Lung; Lung Transplantation; Organ Preservation; Organ Preservation Solutions; Oxygen; Postoperative Complications; Potassium; Pulmonary Embolism; Raffinose; Reperfusion; Sodium Oxybate

Altered cellular calcium (Ca) homeostasis may be important in mediating hypothermic injury in preserved kidneys. In this study the effect of hypothermic (5 degrees C) storage on ionized intracellular Ca concentration ([Ca]i) in rabbit tubules was examined using Indo-1. Tubules were stored up to 250 min in UW-gluconate solution containing either 0.0, 0.5, 1.5, or 5.0 mM Ca (yielding about 3.6, 62, 371, and 1,010 microM ionized solution Ca (Ca2+) at 5 degrees C, respectively). [Ca]i increased to about 1,600 nM within 1 min after suspension in UW solution followed by a decrease in [Ca]i during the subsequent 60 min in all groups, suggesting mitochondrial Ca sequestration. Thereafter, [Ca]i either 1) increased in tubules incubated with 1.5 and 5.0 mM Ca to levels greater than 2,500 nM; 2) decreased to about 800 nM in tubules incubated with 0.5 mM Ca and then remained stable; or 3) continued to decrease in tubules incubated with 0.0 mM added Ca to reach an apparent steady-state concentration of about 175 nM after 180 min of incubation. The early spike in [Ca]i was unaffected by adding EGTA (solution Ca2+ = 50 nM). Ryanodine eliminated the [Ca]i spike, indicating that cooling in UW-gluconate solution caused release of endoplasmic reticulum Ca. This study shows that [Ca]i initially increases after exposure to UW-gluconate solution and appears to be transiently buffered through intracellular, probably mitochondrial, sequestration. Saturation of cellular buffer mechanisms resulted in a sustained dependence of [Ca]i on extracellular Ca2+. These results support the hypothesis that the effect of Ca on kidney viability is related to solution-induced alterations in [Ca]i.

Topics: Adenosine; Allopurinol; Animals; Calcium; Cattle; Cold Temperature; Gluconates; Glutathione; Homeostasis; Hypothermia; In Vitro Techniques; Insulin; Intracellular Fluid; Kidney Tubules; Kinetics; Organ Preservation Solutions; Raffinose; Reperfusion Injury; Tissue Preservation

Topics: Adenosine; Allopurinol; Animals; Cold Temperature; Glutathione; Graft Survival; Hypothermia; Insulin; Mice; Mice, Inbred Strains; Organ Culture Techniques; Organ Preservation; Organ Preservation Solutions; Oxygen; Raffinose; Solutions; Thyroid Gland; Time Factors; Transplantation, Homologous; Transplantation, Isogeneic