Effects of dietary raffinose on growth, non-specific immunity, intestinal morphology and microbiome of juvenile hybrid sturgeon (Acipenser baeri Brandt ♀ × A. schrenckii Brandt ♂).

This study was performed to determine the efficacy of raffinose on the growth, non-specific immunity, intestinal morphology and microbiota of juvenile hybrid sturgeon, (Acipenser baeri Brandt ♀ × A. schrenckii Brandt ♂). Hybrid sturgeons were divided into 2 groups and each group was fed with diets supplemented with or without raffinose for 56 days. Hybrid sturgeon fed diet supplemented with raffinose had significantly higher final body weight (FBW), specific growth rate (SGR), and weight gain ratio (WGR) than fish fed the control diet (P < 0.05). Raffinose in diet had no negative effect on feed intake (FI) and feed conversion ratio (FCR) (P > 0.05). Compared with the control diet, the myeloperoxidase (MPO) and respiratory burst (NBT) activitives were significantly higher in sturgeon fed the raffinose supplemented diet (P < 0.05). The increasing of intestinal villi area and mucosal folds were observed in intestinal tract of sturgeon when they fed the raffinose supplemented diet. Meanwhile, the residual bait of intestinal tract was relatively lower in sturgeon with raffinose treatment. High-throughput sequencing revealed that majority of reads derived from the sturgeon digesta were constituted by members of Proteobacteria, Firmicutes, Fusobacteria and Actinobacteria. Shannon's diversity index existed significant difference among dietary treatments indicating that the overall microbial community was modified to a large extent by dietary raffinose. In conclusion, supplementation of the diet with raffinose is capable of improving hybrid sturgeon growth performances and intestinal morphology, modifying the intestinal microbial composition.

Protective role of normothermic machine perfusion during reduced-size liver transplantation in pigs.

The purpose of this study is to explore whether normothermic machine perfusion (NMP) preservation is superior to cold preservation during reduced-size liver transplantation (RSLT) in pigs. Twenty-four healthy Ba-Ma mini pigs were used (aged >13 months; weight 25-35 kg; regardless of sex). The animals were randomized into 2 groups. In group A (NMP), donor livers were harvested without warm ischemia time and heartbeats and then were connected to the NMP system to reduce the livers' size under the normothermic condition. In group B (University of Wisconsin [UW] solution), donor livers were harvested without warm ischemia time and heartbeats after being perfused by UW solution and were then preserved in 0°C-4°C UW solution to reduce the livers' size under cold conditions. After that, liver transplantation without venovenous bypass was performed. General RSLT information of the pigs from the 2 groups was recorded; the serological indices were measured; and routine pathological examination of liver tissue was observed. A significant difference was observed in the intraoperative bleeding between the 2 groups (P < 0.05), whereas no significant difference was found in the other indices (all P > 0.05). Significant differences of alanine aminotransferase levels, aspartate aminotransferase levels, and lactate dehydrogenase levels between the 2 groups were observed between postoperative days 3 and 5 (P < 0.05). Significant differences of lactic acid levels between the 2 groups were observed between postoperative days 2 and 5 (P < 0.05). Compared with the cold preservation group, the liver tissues of the NMP preservation group only rarely experienced liver cell necrosis and maintained integrities in the hepatic sinusoid spaces and endothelial cells. In conclusion, NMP preservation is superior to cold preservation during RSLT in pigs. Liver Transplantation 22 968-978 2016 AASLD.

Chitosan-based scaffold modified with D-(+) raffinose for cartilage repair: an in vivo study.

BACKGROUND: Osteochondral defects significantly affect patients' quality of life and represent challenging tissue lesions, because of the poor regenerative capacity of cartilage. Tissue engineering has long sought to promote cartilage repair, by employing artificial scaffolds to enhance cell capacity to deposit new cartilage. An ideal biomaterial should closely mimic the natural environment of the tissue, to promote scaffold colonization, cell differentiation and the maintenance of a differentiated cellular phenotype. The present study evaluated chitosan scaffolds enriched with D-(+) raffinose in osteochondral defects in rabbits. Cartilage defects were created in distal femurs, both on the condyle and on the trochlea, and were left untreated or received a chitosan scaffold. The animals were sacrificed after 2 or 4 weeks, and samples were analysed microscopically. RESULTS: The retrieved implants were surrounded by a fibrous capsule and contained a noticeable inflammatory infiltrate. No hyaline cartilage was formed in the defects. Although defect closure reached approximately 100% in the control group after 4 weeks, defects did not completely heal when filled with chitosan. In these samples, the lesion contained granulation tissue at 2 weeks, which was then replaced by fibrous connective tissue by week 4. Noteworthy, chitosan never appeared to be integrated in the surrounding cartilage. CONCLUSIONS: In conclusion, the present study highlights the limits of D-(+) raffinose-enriched chitosan for cartilage regeneration and offers useful information for further development of this material for tissue repair.

In-ovo injection of Bacillus subtilis, raffinose, and their combinations enhances hatchability, gut health, nutrient transport- and intestinal function-related genes, and early development of broiler chicks.

An experiment was conducted to assess the response of chicks to in-ovo injection of Bacillus subtilis (probiotic), raffinose (prebiotic), and their combinations. The study used 1,500 embryonated eggs allotted to 10 groups/ 6 replicates (150 eggs/group). The experimental treatments were: 1) un-injected control (NC); 2) sham (sterile distilled water) (PC); 3) probiotic 4 × 105CFU/egg (LBS); 4) probiotic 4 × 106CFU/egg (HBS); 5) prebiotic 2 mg/egg (LR); (6 prebiotic 3 mg/egg (HR); 7) probiotic 4 × 105CFU + prebiotic 2 mg/egg (LBS+LR); 8) probiotic 4 × 105CFU + prebiotic 3 mg/egg (LBS+HR); 9) probiotic 4 × 106CFU + prebiotic 2 mg/egg (HBS+LR); and 10) probiotic 4 × 106CFU + prebiotic 3 mg/egg (HBS+HR). Results showed that in-ovo inclusion of Bacillus subtilis, prebiotic, and their combinations improved hatchability, yolk-free chick weight, and chick weight compared to the control group. Moreover, the in-ovo treatment reduced residual yolk weight on the day of hatch compared to the control group. Different levels of in-ovo B. subtilis alone or combined with raffinose significantly (P ≤ 0.001) reduced total bacterial count and total yeast and mold count compared to the negative control group. Total coliform and E. coli decreased significantly (P ≤ 0.001) in groups treated with probiotics, prebiotics, and synbiotics with different doses during incubation compared to those in the control. Clostridium spp. was not detected in the groups injected with B. subtilis alone or combined with raffinose. In-ovo probiotics and synbiotics (LBS+LR & LBS+HR) significantly (P ≤ 0.001) increased ileal villus length compared to other groups. In-ovo treatment increased mRNA expression of JAM-2 compared to the control group. The fold change significantly increased in group LBS+HR for genes MUC-2, OCLN, VEGF, SGLT-1, and EAAT-3 compared to the negative control. In conclusion, in-ovo injection of a low dose of B. subtilis plus a high or low dose of raffinose can positively affect hatching traits, cecal microbial populations, intestinal histomorphometry, nutrient transport- and intestinal function-related genes, and chick quality of newly hatched broiler chicks.

Sodium thiosulfate-supplemented UW solution protects renal grafts against prolonged cold ischemia-reperfusion injury in a murine model of syngeneic kidney transplantation.

INTRODUCTION: Cold ischemia-reperfusion injury (IRI) is an inevitable event that increases post-transplant complications. We have previously demonstrated that supplementation of University of Wisconsin (UW) solution with non-FDA-approved hydrogen sulfide (H2S) donor molecules minimizes cold IRI and improves renal graft function after transplantation. The present study investigates whether an FDA-approved H2S donor molecule, sodium thiosulfate (STS), will have the same or superior effect in a clinically relevant rat model of syngeneic orthotopic kidney transplantation. METHOD: Thirty Lewis rats underwent bilateral nephrectomy followed by syngeneic orthotopic transplantation of the left kidney after 24-hour preservation in either UW or UW+STS solution at 4 °C. Rats were monitored to post-transplant day 14 and sacrificed to assess renal function (urine output, serum creatinine and blood urea nitrogen). Kidney sections were stained with H&E, TUNEL, CD68, and myeloperoxidase (MPO) to detect acute tubular necrosis (ATN), apoptosis, macrophage infiltration, and neutrophil infiltration. RESULT: UW+STS grafts showed significantly improved graft function immediately after transplantation, with improved recipient survival compared to UW grafts (p < 0.05). Histopathological examination revealed significantly reduced ATN, apoptosis, macrophage and neutrophil infiltration and downregulation of pro-inflammatory and pro-apoptotic genes in UW+STS grafts compared to UW grafts (p < 0.05). CONCLUSION: We show for the first time that preservation of renal grafts in STS-supplemented UW solution protects against prolonged cold IRI by suppressing apoptotic and inflammatory pathways, and thereby improving graft function and prolonging recipient survival. This could represent a novel clinically applicable therapeutic strategy to minimize the detrimental clinical outcome of prolonged cold IRI in kidney transplantation.

Soybean oligosaccharide, stachyose, and raffinose in broilers diets: effects on odor compound concentration and microbiota in cecal digesta.

Soybean oligosaccharides have been previously shown to be associated with the production of major odor-causing compounds in broilers, although little is known about the role of stachyose and raffinose, which are key components of soybean oligosaccharide, in broiler cecal microbiota and odor compound production. To this end, soybean oligosaccharide, stachyose, and raffinose were added to the birds' diets to investigate their effects on odor compound production and the microbial community characteristics of the cecum in broilers. A total of 300 one-day-old Arbor Acre broilers with similar initial live weight were randomly allocated into 5 dietary groups with 6 replicates of 10 birds. The diets included soybean meal (positive control), soybean meal-free (negative control), 0.6% soybean oligosaccharide, 0.6% stachyose, or 0.6% raffinose. After a 49-D feeding period, both ceca were aseptically removed postmortem, and the contents were collected and analyzed for skatole, indole, volatile fatty acids, and lactic acid by using high performance liquid chromatography. Bacterial communities were detected by using a high-throughput sequencing platform based on IlluminaMiSeq 2500. Levels of skatole and indole tended to be lower in the dietary supplementation of oligosaccharides. The lowest levels of skatole and indole were observed in the stachyose group (P < 0.05), while the highest levels were found in the negative control group (P < 0.05). Concentrations of acetic acid and propionic acid in the stachyose group were increased (P < 0.05) while those of butyric acid and lactic acid were decreased (P < 0.05) compared with the soybean oligosaccharide and raffinose groups. Firmicutes and Bacteroidetes were prevalent in all groups, the proportion of Bacteroidetes was slightly decreased in the stachyose group, and Verrucomicrobia was abundant in the raffinose group (P > 0.05). Bacterial genera Alistipes and Parabacteroides were comparably abundant in the stachyose group, while Bacteroides, Lactobacillus, and Akkermansia were more abundant in the negative control, stachyose, and raffinose groups, respectively. Collectively, these findings demonstrated that dietary oligosaccharide supplementation significantly reduced odor compound production by modulating the cecal microbial community. Compared with soybean oligosaccharide and raffinose, the addition of stachyose into diets may help improve gut fermentation and minimize odor compound generation in broilers.

Effects of Oxygen During Long-term Hypothermic Machine Perfusion in a Porcine Model of Kidney Donation After Circulatory Death.

BACKGROUND: Hypothermic machine perfusion (HMP) has become standard care in many center's to preserve kidneys donated after circulatory death (DCD). Despite a significant reduction in metabolism at low temperatures, the remaining cellular activity requires oxygen. Because of the role and safety of oxygen during HMP has not been fully clarified, its supply during HMP is not standard yet. This study investigates the effect of administering oxygen during HMP on renal function in a porcine DCD model. METHODS: After 30 minutes of warm ischemia, porcine slaughterhouse kidneys were preserved for 24 hours by means of cold storage (CS), or HMP with Belzer Machine Perfusion Solution supplemented with no oxygen, 21% or 100% oxygen. Next, kidneys were reperfused for 4 hours in a normothermic machine perfusion setup. RESULTS: HMP resulted in significantly better kidney function during normothermic machine perfusion. Thiobarbituric acid-reactive substances, markers of oxidative stress, were significantly lower in HMP preserved kidneys. HMP preserved kidneys showed significantly lower aspartate aminotransferase and lactate dehydrogenase levels compared with kidneys preserved by CS. No differences were found between the HMP groups subjected to different oxygen concentrations. Adenosine triphosphate levels significantly improved during HMP when active oxygenation was applied. CONCLUSIONS: This study showed that preservation of DCD kidneys with HMP is superior to CS. Although the addition of oxygen to HMP did not result in significantly improved renal function, beneficial effects were found in terms of reduced oxidative stress and energy status. Oxygen addition proofed to be safe and did not show detrimental effects.

Ductal injection of preservation solution increases islet yields in islet isolation and improves islet graft function.

For islet transplantation, it is important to obtain an available islet mass adequate for diabetes reversal from a single donor pancreas. A recent report demonstrated that the use of M-Kyoto solution instead of UW solution improved islet yields in the two-layer method for pancreas preservation. The present study investigated whether the ductal injection of a large volume of preservation solution (UW and M-Kyoto solution) before pancreas storage improves islet yields. Islet yield both before and after purification was significantly higher in the ductal injection (+) group compared with the ductal injection (-) group. TUNEL-positive cells in the ductal injection (+) group were significantly decreased in comparison to the ductal injection (-) group. The ductal injection of preservation solution increased the ATP level in the pancreas tissue and reduced trypsin activity during the digestion step. Annexin V and PI assays showed that the ductal injection prevents islet apoptosis. In a transplant model, the ductal injection improved islet graft function. These findings suggest that the ductal injection of preservation solution, especially the M-Kyoto solution, leads to improved outcomes for pancreatic islet transplantation. Based on these data, this technique is now used for clinical islet transplantation from non-heart-beating donor pancreata or living donor pancreas.

Acute and chronic effects of honey and its carbohydrate constituents on calcium absorption in rats.

The effects of honey and its carbohydrate constituents (glucose, fructose, and raffinose) on calcium absorption in rats were investigated in acute and chronic feeding studies. In the acute study, rats ( n = 120) were gavaged with an oral solution consisting of (a) 10 microCi (45)Ca, (b) 25 mg of calcium as calcium acetate, and (c) one of the following: 0 mg of honey (control), or 200, 500, or 800 mg of honey, a glucose-fructose mixture, 10.75 mg of raffinose, or 200 mg of raffinose. Another group received (45)Ca intraperitoneally. Femurs were collected 2 days later and analyzed for (45)Ca content. Rats given 500 and 800 mg of honey showed 25.5 and 33.6% increases in calcium absorption ( P<0.05), respectively, over the control group. Groups given the glucose-fructose mixture or 200 mg of raffinose had a significantly higher increase in calcium absorption than the control group (17.1 and 25.6%, respectively). In the chronic study, rats (n=96) were fed for 8 weeks with either 0% honey (control), 5% honey, 10% honey, or a glucose-fructose-raffinose (GFR) mixture. Femurs of GFR-fed rats had significantly lower calcium content, (45)Ca absorption, width, and BMD (at distal region) than control rats. Groups fed honey did not show the negative effects of GFR on bone, but had no advantage over the control group. No significant differences were observed in femur length, density, strength, or BMC among any treatment group compared to the control group. These results indicate that although a positive dose-response effect of honey and its carbohydrate constituents on calcium absorption was observed in the acute study, this effect disappeared upon long-term feeding in rats, implying adaptation had occurred.

Low density, good flowability cyclodextrin-raffinose binary carrier for dry powder inhaler: anti-hygroscopicity and aerosolization performance enhancement.

BACKGROUND: The hygroscopicity of raffinose carrier for dry powder inhaler (DPI) was the main obstacle for its further application. Hygroscopicity-induced agglomeration would cause deterioration of aerosolization performance of raffinose, undermining the delivery efficiency. METHODS: Cyclodextrin-raffinose binary carriers (CRBCs) were produced by spray-drying so as to surmount the above issue. Physicochemical attributes and formation mechanism of CRBCs were explored in detail. The flow property of CRBCs was examined by FT4 Powder Rheometer. Hygroscopicity of CRBCs was elucidated by dynamic vapor sorption study. Aerosolization performance was evaluated by in vitro deposition profile and in vivo pharmacokinetic profile of CRBC based DPI formulations. RESULTS: The optimal formulation of CRBC (R4) was proven to possess anti-hygroscopicity and aerosolization performance enhancement properties. Concisely, the moisture uptake of R4 was c.a. 5% which was far lower than spray-dried raffinose (R0, c.a. 65%). R4 exhibited a high fine particle fraction value of 70.56 ± 0.61% and it was 3.75-fold against R0. The pulmonary and plasmatic bioavailability of R4 were significantly higher than R0 (p < 0.05). CONCLUSION: CRBC with anti-hygroscopicity and aerosolization performance enhancement properties was a promising approach for pulmonary drug delivery, which could provide new possibilities to the application of hygroscopic carriers for DPI.

Hypolipidemic Effect of Red Gram (Cajanus cajan L.) Prebiotic Oligosaccharides in Wistar NIN Rats.

The hypolipidemic effect of red gram prebiotics of raffinose family oligosaccharides was studied in Wistar National Institute of Nutrition male rat strain. The study consisted of 36 rats randomly divided into three groups of 12 rats each. For 16 weeks, Group I was fed with the control diet; Group II was fed with a diet containing 3% standard raffinose as the reference group; Group III received the diet containing 3% red gram prebiotics. The results showed that the gain in body weight was low in the red gram prebiotics-supplemented group followed by the control group; highest increase of body weight was seen in the raffinose standard-fed group. Serum glucose levels of the red gram prebiotic-fed group decreased 14.92% compared to the control group and increased 2.07% compared to the reference group. The decrease in serum triglycerides (TG) levels of the red gram prebiotic-fed groups was 32.76% compared to the control group and 33.64% compared to the reference group. Decrease in the serum TC of the red gram-fed animals was 18.51% and 4.63% compared to the control group and the reference group, respectively. Increase in the level of serum high-density lipoprotein cholesterol (HDL-C) in the red gram-fed animals was 18.51% compared to the control group and 4.63% compared to the reference group. The present study can be a proof for the use of prebiotics as a preventive measure for overweight and obesity in humans, and legume prebiotics can be explored as a novel prebiotic product in the consumer market.

Combined Flush With Histidine-Tryptophan-Ketoglutarate and University of Wisconsin Solutions in Liver Transplantation: Preliminary Results.

INTRODUCTION: Ischemia reperfusion injury (IRI) is the main cause of early allograft dysfunction (EAD) and subsequent primary allograft failure (PAF). OBJECTIVES: The purpose of this study is to compare IRI, EAD, and PAF in liver transplantation in a cohort of patients perfused with histidine-tryptophan-ketoglutarate (HTK) solution and University of Wisconsin (UW) solution versus HTK alone. METHODS: A randomized trial was performed to compare outcomes in liver recipients who underwent transplantation surgery in the University Regional Hospital of Malaga, Spain. Forty patients were randomized to two groups. Primary endpoints included IRI, EAD, PAF, re-intervention, acute cellular rejection, retransplantation, arterial complications, and biliary complications at postoperative day 90. RESULTS: Postoperative glutamic oxaloacetic transaminase (1869.15 ± 1559.75 UI/L vs. 953.15 ± 777.27 UI/L; P = .004) and glutamic pyruvic transaminase (1333.60 ± 1115.49 U/L vs. 721.70 ± 725.02 U/L; P = .023) were significantly higher in patients perfused with HTK alone. A clear tendency was observed in recipients perfused with HTK alone to present moderate to severe IRI (7 patients in the HTK + UW solution group vs. 15 patients in the HTK-alone solution group; P = .06), EAD (0 patients in the HTK + UW solution group vs. 0 patients in the HTK-alone solution group; P = .76), and PAF (3 patients in the HTK + UW solution group vs. 8 patients in the HTK-alone solution group; P = .15). CONCLUSIONS: Initial perfusion with HTK solution followed by UW solution in liver transplantation improves early liver function as compared to perfusion with HTK alone.

Retrograde Flushing of Living Donor Renal Allografts via the Renal Vein: A Simple, Effective Technique.

BACKGROUND: Prograde flushing (PF) of living donor renal allografts with preservation solution via the renal artery or arteries is standard practice. PF may be difficult and potentially injurious to the donor kidney, especially in grafts with small or multiple arteries. In this report, we present our experience with retrograde flushing (RF) of 7 living donor kidneys via the renal vein. METHODS: Retrospective review of 7 consecutive living donor renal transplants performed using the RF technique was performed. The 7 preceding living donor renal transplants performed using the standard arterial PF technique served as a control group. RESULTS: All 7 recipients of RF kidneys experienced immediate graft function. At postoperative days 3 and 30, there was no difference in estimated glomerular filtration rate between the RF study group and PF controls. CONCLUSIONS: The RF technique is simple and safe, with results equivalent to the PF technique. The RF technique may be especially useful after recovering kidneys with small and/or multiple arteries.

Effect of in ovo injection of raffinose on growth performance and gut health parameters of broiler chicken.

The effects of in ovo injection of raffinose (RFO) as a prebiotic on growth performance, relative weight of proventriculus, gizzard, drumstick and breast muscles, and ileum mucosa morphology were examined in Cobb 500 broilers. A total of 240 fertilized eggs were divided into 4 groups: a non-injected with intact shell and 3 levels of RFO solution (1.5, 3.0, and 4.5 mg in 0.2 mL of an aqueous diluents). The RFO solution was injected into the air sac on d 12 of incubation. In total 144 birds were fed a standard diet and management and sacrificed at d 21 post hatch for collection of samples. Total RNA was extracted from the small intestine, and RT-qPCR was performed to quantify mRNA levels of marker genes of immune cells. Injection of RFO had no significant effect (P > 0.05) on d one body weight of chicks. On d 21, the relative weight of the proventriculus, drumstick, breast, and gizzard was not affected (P > 0.05) by RFO. On hatch d, the villus height increased linearly (P < 0.01) with an increasing dose of RFO. Also, an increasing dose of RFO increased the villus height and villus height:crypt depth ratio (P < 0.05) but did not affect the crypt depth on d 21. The expression levels of CD3 and chB6, which are T cell and B cell marker genes, respectively, were significantly enhanced by high dose RFO (4.5 mg). In conclusion, although an increasing dose of RFO in ovo injection did not significantly influence growth performance or slaughter yield of broilers, RFO has the potential of enhancing ileum mucosa morphology and improving immunity in the small intestine, which are indicators of improved gut health.

Intra Amniotic Administration of Raffinose and Stachyose Affects the Intestinal Brush Border Functionality and Alters Gut Microflora Populations.

This study investigates the effectiveness of two types of prebiotics-stachyose and raffinose-which are present in staple food crops that are widely consumed in regions where dietary Fe deficiency is a health concern. The hypothesis is that these prebiotics will improve Fe status, intestinal functionality, and increase health-promoting bacterial populations in vivo (Gallus gallus). By using the intra-amniotic administration procedure, prebiotic treatment solutions were injected in ovo (day 17 of embryonic incubation) with varying concentrations of a 1.0 mL pure raffinose or stachyose in 18 MΩ H2O. Four treatment groups (50, 100 mg·mL-1 raffinose or stachyose) and two controls (18 MΩ H2O and non-injected) were utilized. At hatch the cecum, small intestine, liver, and blood were collected for assessment of the relative abundance of the gut microflora, relative expression of Fe-related genes and brush border membrane functional genes, hepatic ferritin levels, and hemoglobin levels, respectively. The prebiotic treatments increased the relative expression of brush border membrane functionality proteins (p < 0.05), decreased the relative expression of Fe-related proteins (p < 0.05), and increased villus surface area. Raffinose and stachyose increased the relative abundance of probiotics (p < 0.05), and decreased that of pathogenic bacteria. Raffinose and stachyose beneficially affected the gut microflora, Fe bioavailability, and brush border membrane functionality. Our investigations have led to a greater understanding of these prebiotics' effects on intestinal health and mineral metabolism.

Effects of N-Acetylcysteine Addition to University of Wisconsin Solution on the Rate of Ischemia-Reperfusion Injury in Adult Orthotopic Liver Transplant.

OBJECTIVES: One of the main concerns in liver transplant is the prolonged ischemia time, which may lead to primary graft nonfunction or delayed function. N-acetylcysteine is known as a hepato-protective agent in different studies, which may improve human hepatocyte viability in steatotic donor livers. This study investigated whether N-acetylcysteine can decrease the rate of ischemia-reperfusion syndrome and improve short-term outcome in liver transplant recipients. MATERIALS AND METHODS: This was a double-blind, randomized, control clinical trial of 115 patients. Between April 2012 and January 2013, patients with orthotopic liver transplant were randomly divided into 2 groups; in 49 cases N-acetylcysteine was added to University of Wisconsin solution as the preservative liquid (experimental group), and in 66 cases standard University of Wisconsin solution was used (control group). We compared postreperfusion hypotension, inotrope requirement before and after portal reperfusion, intermittent arterial blood gas analysis and potassium measurement, pathological review of transplanted liver, in-hospital complications, morbidity, and mortality. RESULTS: There was no significant difference between the groups regarding time to hepatic artery reperfusion, hospital stay, vascular complications, inotrope requirement before and after portal declamping, and blood gas analysis. Hypotension after portal reperfusion was significantly more common in experimental group compared with control group (P = .005). Retransplant and in-hospital mortality were comparable between the groups. CONCLUSIONS: Preservation of the liver inside Univer-sity of Wisconsin solution plus N-acetylcysteine did not change the rate of ischemia reperfusion injury and short-term outcome in liver transplant recipients.

Pancreatic L-Glutamine Administration Protects Pig Islets From Cold Ischemic Injury and Increases Resistance Toward Inflammatory Mediators.

The isolation and transplantation of porcine islets represent a future option for the treatment of type 1 diabetic patients. Stringent product release criteria and limited availability of transgenic and specific pathogen-free pigs will essentially require processing of explanted pig pancreata in specialized, possibly remote isolation facilities, whereby pancreata are exposed to cold ischemia due to prolonged tissue transit time. In the present study we investigated whether pancreas oxygenation can be efficiently combined with an antioxidant strategy utilizing intraductal L-glutamine administration. Pig pancreata were intraductally perfused after retrieval and after cold storage in oxygen-precharged perfluorohexyloctane utilizing University of Wisconsin solution supplemented with (n = 16) or without (n = 14) 5 mmol/L L-glutamine. After isolation purified islets were subjected to extensive quality assessment. Islet recovery postpurification was significantly higher in glutamine-treated pancreata (77.0 ± 3.3% vs. 60.3 ± 6.0%, p < 0.05). Glutamine administration increased intraislet content of reduced glutathione (117.8 ± 16.5 vs. 15.9 ± 2.8 ng/ng protein, p < 0.001) associated with increased islet recovery after culture (65.8 ± 12.1% vs. 40.3 ± 11.7%, p < 0.05), enhanced glucose stimulation index (1.82 ± 0.16 vs. 1.38 ± 0.10, p < 0.05), and improved posttransplant function in diabetic nude mice (p < 0.05). Furthermore, intraductally administered glutamine increased pig islet resistance toward reactive oxygen species, nitric oxide, and high-dose proinflammatory cytokines. The present study demonstrates that quality and function of pig islets exposed to warm and cold ischemia can significantly be improved using intraductal l-glutamine administration. As the efficiency of the intraductal route may be inferior compared to intravascular administration further studies should aim on assessment of l-glutamine as supplement for pancreas perfusion during organ procurement.

A phase II dose-response study of hemoglobin raffimer (Hemolink) in elective coronary artery bypass surgery.

BACKGROUND: We performed this study to determine the dose-response of hemoglobin raffimer administered in conjunction with intraoperative autologous donation in patients undergoing coronary artery bypass grafting surgery. A secondary objective was to evaluate hemoglobin raffimer for reducing the incidence of allogeneic red blood cell transfusions. METHODS: This was a phase II, single-blind, multicenter, placebo-controlled, open-label study. Patients undergoing coronary artery bypass grafting with cardiopulmonary bypass and intraoperative autologous donation were randomized to receive a single dose of hemoglobin raffimer or control (10% pentastarch). Patients were sequentially enrolled in a dose block of 250, 500, 750, and 1000 mL. RESULTS: Sixty patients received hemoglobin raffimer (n = 30) or control (n = 30). Hemoglobin raffimer was well tolerated. Most (98%) adverse events were mild or moderate in severity. There was an expected dose-dependent increase in the incidence of blood pressure increases and jaundice in hemoglobin raffimer-treated patients. In a dose-pooled analysis of hemoglobin raffimer versus control, increased blood pressure (43% vs 17%), nausea (37% vs 33%), and atrial fibrillation (37% vs 17%) were the most frequently reported adverse events. All serious adverse events were considered unrelated or unlikely to be related to study drug. No hemoglobin raffimer-treated patient required an intraoperative allogeneic red blood cell transfusion, compared with 5 (17%) pentastarch-treated patients (P =.052). This advantage of hemoglobin raffimer was maintained at 24 hours after surgery (7% vs 37%; P =.010) and up to 5 days after surgery (10% vs 47%; P =.0034). CONCLUSIONS: Hemoglobin raffimer was not associated with any serious adverse events in patients undergoing primary coronary artery bypass grafting with cardiopulmonary bypass and intraoperative autologous donation in a dose-response study up to 1000 mL. Hemoglobin raffimer was effective in facilitating decreased exposure or avoidance of allogeneic red blood cell transfusions when used in conjunction with intraoperative autologous donation.

Celsior solution provides superior post-ischemic right ventricular function as compared with UW solution in a porcine heart transplantation model.

BACKGROUND: Use of the new cardioprotective Celsior solution has been suggested for organ preservation in cardiac transplantation, but selective data for right ventricular function, of special interest in the clinical setting, have not been evaluated. METHODS: Celsior solution was compared with the clinical standard University of Wisconsin solution (UW) in a porcine allogenic heart transplantation model with accurate isovolumic measurement of right ventricular (RV) function. RESULTS: Maximum RV developed pressures were significantly different between Celsior and UW groups (51.1 +/- 9.6 mm Hg vs 42.2 +/- 15.4 mm Hg after 1 hour, respectively, and 55.6 +/- 7.8 mm Hg vs 45.1 +/- 16.2 mm Hg after 2 hours, respectively; p = 0.02, 2-way analysis of variance). CONCLUSIONS: Celsior significantly improves post-ischemic right ventricular function when compared with UW solution in an experimental heart transplantation model.

Pulmonary graft preservation: a worldwide survey of current clinical practice.

BACKGROUND: Flush perfusion of pulmonary grafts with cold modified EuroCollins solution supplemented by prostaglandin treatment was introduced clinically 10 years ago. Primary graft failure remains a major cause of morbidity and death after lung transplantation. During the last decade, much experimental work has led to reports of alternative storage solutions, differing storage conditions, and pharmacologic interventions that improve pulmonary graft performance. It is unclear how these findings have influenced current clinical practice. METHODS: A worldwide survey of the 125 centers performing lung transplantation was conducted by questionnaire. RESULTS: One hundred twelve replies were received (90%). Most centers (n = 86) continue to use EuroCollins solution (77%), of whom 69% include prostaglandin therapy and 32% donor steroid treatment. University of Wisconsin solution (UW) is used by 15 centers (13.5%), of which 10 (67%) use prostaglandin and seven (47%) use donor steroids. Nine centers use Papworth solution and one uses donor core cooling. The volume of flush used varied widely, from 20 to 120 ml/kg, with median volumes of 60, 60, and 30 ml/kg in centers using EuroCollins, UW, and Papworth solutions, respectively. Two thirds of centers using EuroCollins solution store grafts at 0 degrees to 5 degrees C, and one third at 5 degrees to 10 degrees C. One center that uses EuroCollins solution stores grafts at 10 degrees to 15 degrees C. Centers using UW solution are evenly split at 0 degrees to 5 degrees C and 5 degrees to 10 degrees C. Most centers that use Papworth solution store grafts at 5 degrees to 10 degrees C. Only six centers use superoxide radical scavengers. The maximum ischemic period accepted by centers varies from 4 to 12 hours, with median periods of 8, 7, 6, and 6 hours for the UW, EuroCollins, Papworth, and donor core cooling centers, respectively. All but one of the UW centers (93%) expressed satisfaction with the quality of graft preservation achieved by UW solution. Only 58 of the 86 centers using EuroCollins solution (67%) were satisfied. Six of nine centers using Papworth solution were satisfied. CONCLUSIONS: There has been a trend toward the use of UW solution and a slightly warmer storage temperature. However, for most centers, graft storage techniques have changed little over the last decade.