Cold Ringer Research Articles

Mesenchymal stromal cell treatment of donor kidneys during ex vivo normothermic machine perfusion: A porcine renal autotransplantation study.

Normothermic machine perfusion (NMP) of injured kidneys offers the opportunity for interventions to metabolically active organs prior to transplantation. Mesenchymal stromal cells (MSCs) can exert regenerative and anti-inflammatory effects in ischemia-reperfusion injury. The aims of this study were to evaluate the safety and feasibility of MSC treatment of kidneys during NMP using a porcine autotransplantation model, and examine potential MSC treatment-associated kidney improvements up to 14days posttransplant. After 75min of kidney warm ischemia, four experimental groups of n=7 underwent 14h of oxygenated hypothermic machine perfusion. In three groups this was followed by 240min of NMP with infusion of vehicle, 10million porcine, or 10million human adipose-derived MSCs. All kidneys were autotransplanted after contralateral nephrectomy. MSC treatment did not affect perfusion hemodynamics during NMP or cause adverse effects at reperfusion, with 100% animal survival. MSCs did not affect plasma creatinine, glomerular filtration rate, neutrophil gelatinase-associated lipocalin concentrations or kidney damage assessed by histology during the 14days, and MSCs retention was demonstrated in renal cortex. Infusing MSCs during ex vivo NMP of porcine kidneys was safe and feasible. Within the short posttransplant follow-up period, no beneficial effects of ex vivo MSC therapy could be demonstrated.

CONFERENCIA PLENARIA: Tecnología aplicada a la insulinoterapia: 100 años después

1921 fue el hito histórico en la diabetes. Y aunque habían existido intentos previos, el cirujano ortopedista Frederick Banting y el estudiante Charles Best, con tenacidad e ingenio en el área de Fisiología de la Universidad de Toronto bajo la dirección de JJR MacLeod (quien aparentemente no creía demasiado en el proyecto), lograron extraer una sustancia de páncreas de perros a los que ligaron los ductus hasta que el páncreas se redujo a 1/3; quedó fibrótico, se trozó, molió con arena, colocó tolueno y extrajo con Ringer helado con lo que se obtuvo una solución. El Doctor Joseph Gilchrist (con diabetes desde 1917), amigo de Banting, bebió el preparado sin ningún resultado, pero luego al inyectar a la perrita Marjory con diabetes por pacreatectomía, su glucosa primero subió, pero luego comenzó a descender y permaneció viva y vivaz, hasta que un tiempo después se la sacrificó para estudiar los efectos del preparado, que se llamó isletin.

Regulation of Prdx6 by Nrf2 Mediated Through aiPLA2 in White Matter Reperfusion Injury.

Hypoxia and reperfusion produces overproduction of ROS (reactive oxygen species), which may lead to mitochondrial dysfunction leading to cell death and apoptosis. Here, we explore the hypothesis that Prdx6 protects the spinal cord white matter from hypoxia-reperfusion injury and elucidate the possible mechanism by which Prdx6 elicits its protective effects. Briefly, rats were deeply anesthetized with isoflurane. A 30-mm section of the spinal cord was rapidly removed and placed in cold Ringer's solution (2-4°C). The dissected dorsal column was exposed to hypoxia with 95% N2 and 5% CO2 and reperfusion with 95% O2 and 5% CO2. The expression of Prdx6 significantly upregulated in white matter after hypoxia compared to the sham group, whereas reperfusion caused a gradual decrease in Prdx6 expression after reperfusion injury. For the first time, our study revealed the novel expression and localized expression of Prdx6 in astrocytes after hypoxia, and possible communication of astrocytes and axons through Prdx6. The gradual increase in Nrf2 expression suggests a negative regulation of Prdx6 through Nrf2 signaling. Furthermore, inhibition of aiPLA2 activity of Prdx6 by MJ33 shows that the regulation of Prdx6 by Nrf2 is mediated through aiPLA2 activity. The present study uncovers a differential distribution of Prdx6 in axons and astrocytes and regulation of Prdx6 in hypoxia-reperfusion injury. The low levels of Prdx6 in reperfusion injury lead to increased inflammation and apoptosis in the white matter; therefore, the results of this study suggest that Prdx6 has a protective role in spinal hypoxia-reperfusion injury.

Early Postoperative Renal Dysfunction Predicts Long-Term Renal Function Degradation after Type IV Thoracoabdominal Aortic Aneurysm Surgical Repair

Type IV thoracoabdominal aortic aneurysm surgical repair is often complicated by postoperative acute kidney injury. The aim was to evaluate early renal injury influence on long-term renal function. All type IV thoracoabdominal surgical repair performed between January 2000 and January 2014 in our tertiary hospital were included in this retrospective observational study. All procedures were performed through a retroperitoneal approach with at least suprarenal aortic cross-clamping. Cold Ringer Lactate was used to perfuse the kidneys. Serum creatinine (Scr.) and glomerular filtration rate (GFR) were recorded preoperatively, daily until discharge and at least annually during follow-up. Postoperative renal dysfunction was classified using the RIFLE score. Predictors of long-term renal decline were identified by logistic regression and a Cox model. Of total, 80 patients were included. Aortic clamping level was suprarenal (10%), supramesenteric (37%) or supracoeliac (53%). Ischemic durations were 29±9min for the gastrointestinal tract and the right kidney, 54±28min for the left kidney. Three patients died postoperatively. At discharge, 31 (38.8%) patients did not have a postoperative renal impairment (RIFLE-), compared with 49 (61.2%) who had a renal dysfunction (RIFLE+). GFR was 89±29ml/min vs 68±37ml/min, respectively (P<0.01). In the RIFLE+group, Scr. was increased by x1.5 (Risk) for 22 patients, x2 (Injury) for 19 patients, and ×3 (Failure) for 8 patients. Mean follow-up was 59months. Eighteen patients died, and 2 patients started permanent dialysis at 46 and 118months during follow-up. The only predictive factor of long-term GFR degradation was a postoperative GFR below 45ml/min (OR: 16.5; 95%; P<0.001). Postoperative renal dysfunction was a frequent complication, associated with long-term renal function degradation.

Protective Effects of Cold Ringer's Solution Perfusion in Abdominal Aortic Surgery Requiring Renal Artery Clamp

Treating abdominal aortic aneurysms (AAA) of the juxtarenal artery with renal artery clamps burdens the kidneys. We investigated the outcomes of intra-operative renal artery perfusion using the cold Ringer's solution method for renal protection. We enrolled 290 AAA patients who underwent open aortic repair. Surgical outcomes were investigated based on renal protection. We evaluated 231 patients requiring infrarenal artery clamp (Group I), and 59 patients requiring perfusion in addition to the clamp (Group J). Patient demographics, acute kidney injury (AKI) incidence (Group I: 11.7% and Group J: 20.3%), hospital mortality (Group I: 1.3% and Group J: 1.7%), and 30-day mortality (Group I: 0.4% and Group J: 0%) were not different between the groups. The AKI incidence was low (13%) in cases requiring a renal artery clamp for ≥45 min (n=40). Perfusion with cold Ringer's solution offers renal protection and may improve surgical outcomes.

Significance of Cold Renal Perfusion on Renal Function and Clinical Outcomes When Renal Ischemia Time Exceeded 30 Min during Pararenal and Juxtarenal Abdominal Aortic Aneurysm Surgery

Objectives: To investigate the influence of cold renal perfusion on renal function and clinical outcomes in cases where the renal ischemia time exceeded 30 min during pararenal and juxtarenal abdominal aortic aneurysm (P/JAAA) surgery. Methods and Results: Fifty-four patients who underwent open repair for P/JAAAs were retrospectively analyzed. Thirty-nine patients received renal perfusion with cold Ringer’s solution (perfusion group) and 15 patients did not receive renal perfusion (non-perfusion group). There were no significant differences in preoperative serum creatinine level (Cr) (1.08 ± 0.42 vs. 1.35 ± 0.71 mg/dL, p = 0.09), percentage of patients with Cr > 2 mg/dL [2/38 (5%) vs. 2/15 (13%), p = 0.8], and renal ischemia time during proximal aortic clamping (49 ± 21 vs. 47 ± 11 min; p = 0.8) between the groups. Postoperative Cr was significantly lower in the perfusion group than in the non-perfusion group (1.48 ± 0.76 vs. 2.23 ± 1.21 mg/dL, p 2 mg/dL was also significantly lower in the perfusion group than in the non-perfusion group [5 (13%) vs. 7 (47%), p Conclusion: Renal artery perfusion with cold Ringer’s solution clearly reduced the deterioration of postoperative renal function compared to non-renal perfusion.

Model of Short Bowel Syndrome and Segmental Intestinal Autotransplantation in Swine

Introduction: Intestinal transplantation using living-related donors could minimize the shortage of donor grafts. Nevertheless, the number of living donor small bowel transplantation in the clinical practice is small, probably due to lack of experimental models of this procedure. Objective: To describe a swine model of segmental intestinal autotransplantation producing short bowel syndrome and simulating living donor transplantation, based in our previous technique of liver autotransplantation1. Methods: Six males Landrace swine weighting 25 to 40 Kg were used. A midline incision with the exposition of the entire small bowel was performed (figure 1). The middle part of the mesenteric artery and vein were dissected to identify the pedicle (figures 2 and 3) from a segment in the middle of the small bowel with approximately 300 centimeters length that represents about 1/7 of the swine’s entire small bowel. These mesenteric artery and vein branches were sectioned near the main mesenteric artery and vein (figures 3 and 4). The graft was flashed with cold lactate ringer by the arterial branch (5). The remaining small bowel was removed from the proximal jejune to the distal ileum. A segment of the infrarenal aorta and inferior vena cava was dissected, cross clamped and anastomosed to the mesenteric artery and vein branches (figure 6). Following reperfusion, ileum and jejune anastomoses were performed. The grafts were monitored macroscopically. Animal were sacrificed after two hours of the reperfusion and samples from the graft were removed for histological analysis. Results: All animals survived until the end of the experiment. After reperfusion all grafts rapidly turned in a normal color (figure 7) and no major anastomosis complications were observed. The median period of total operation time, graft cold ischemia and vascular anastomosis were 104, 52 and 33 minutes respectively. The histological analysis observed mild reperfusion injury in two cases and normal in the other grafts. Conclusion: This model of segmental intestinal transplantation is feasible and simulates short bowel syndrome and living donation transplantation. This model is cheaper than allotransplantation and optimal for intestinal transplantation training and to study segmental intestinal graft physiology.Figure

Native Spleen Removal and Modified Multivisceral Transplantation in Rats

[1]Aim: Various techniques and several modifications have been described for experimental intestinal transplantation. Modified multivisceral (MMV) graft offers the advantage of including all abdominal viscera with the corresponding immunologic load without the potential liver inclusion benefits. Our aim was to describe some technical modifications of MMV transplantation in rats with the addition of native splenectomy. Methods: Heterotopic syngeneic (Lewis-Lewis) MMV transplantations were performed in 5 pair of rats. Average weight of donors and recipients was 198± 10g and 250±14g respectively. Animals were operated under isoflurane inhalation anesthesia. Each graft consisted of the distal half of the stomach, duodenum, pancreas, spleen and the small bowel of the donor. The graft spleen was removed in the back table in two of them. Native intestine was preserved, avoiding the need of biliary reconstruction. The native spleen was removed. The vascular pedicle consisted of the celiac trunk and superior mesenteric artery with a cuff of aorta and the portal vein. Cold (4°C) Ringer’s solution with heparin (10 units/ml) was used to perfuse the vessels and irrigate the intestinal lumen. Venous outflow of the graft was reestablished first with end-to-side anastomoses between graft portal vein and recipient vena cava. Arterial revascularization was achieved by end-to-side arterial anastomoses between the cuff of the donor aorta and the recipient infrarenal aorta. Results: 4/5 recipients survived more than 1h after the procedure. One died due to arterial bleeding. All grafts showed adequate initial reperfusion, one showed mild congestion due to portal estenosis. Donor and recipient time were 37±7.7 min and 66.6±3.8 min respectively; venous anastomosis time was 14±3 min and arterial anastomosis time was 12±1 min. Conclusions: MMV offers an excellent experimental model. The lymphoid load can be modified with the inclusion of the graft spleen, and the removal of the native spleen is an original modification that improves the interest of this model for GVHD and other immunological studies. Reference: 1. Galvao F, et al. Transplant. 2013; 96 (2):e4-5.

Low Rate of Intraoperative Seizures During Awake Craniotomy in a Prospective Cohort with 374 Supratentorial Brain Lesions: Electrocorticography Is Not Mandatory

Awake craniotomy (AC) in brain lesions has allowed an improvement of both oncologic and functional results. However, intraoperative seizures (IOSs) were reported as a cause of failure of AC. Here, we analyze the incidence, risk factors, and consequences of IOSs in a prospective cohort of 374 ACs without electrocorticography (ECoG). We performed a prospective study including all patients who underwent AC for an intra-axial supratentorial cerebral lesion from 2009-2014 in our department. Occurrence of IOS was analyzed with respect to medical and epilepsy history, tumor characteristics, operative technique, and postoperative outcomes. The study comprised 374 patients with a major incidence of low-grade glioma (86%). Most of the patients (83%) had epilepsy history before surgery (20% had intractable seizures). Preoperative mean Karnofsky performance scale (KPS) score was 91. IOSs occurred in 13 patients (3.4%). All IOSs were partial seizures, which quickly resolved by irrigation with cold Ringer lactate. No procedure failed because of IOS, and the rate of aborted AC whatever the cause was nil. Mean stimulation current intensity for cortical and subcortical mapping was 2.25 ± 0.6 mA. Presurgical refractory epilepsy was not associated with a higher incidence of IOS. Three months after surgery, no patients had severe or disabling permanent worsening, even within the IOS group (mean KPS score of 93.7). AC for intra-axial brain lesion can be safely and reproducibly achieved without ECoG, with a low rate of IOS and excellent functional results, even in patients with preoperative intractable epilepsy.

EVAR: is Suprarenal Fixation Really without Consequence on the Renal Function?

Objectives: To determine if renal perfusion with a cold crystalloid solution enriched with histidine-tryptophanketoglurarate (Custodiol, Dr. Franz-Kohler Chemie GmbH, Bensheim, Germany) offers a better protection against renal ischemic lesions compared to cold Ringer lactate in patients treated by open surgery for a pararenal aortic aneurysms (PAA). Materials and Methods: We studied 256 consecutive patients treated by open surgery for PAA between 1993 and 2013. In181cases thedirect perfusionof at least one renal artery was carried out. Among these patients, 87 had renal perfusion with Ringer lactate solution and 94 with a Custodiol solution. A score of propensity, based on clinical variables which would probably have influenced the renal function, was carried out to correct skews which could be associated with the use of Custodiol. Acute postoperative renal failure (ARF)was stratified in five classes according to the postoperative serum creatinine values and the need for dialysis, and wascompared in the twogroups, and the independent predictive factors ofARFwere identifiedwith amultivariate analysis. Results: After the score of propensity we could compare 74 patients perfused with Custodiol with those receiving a Ringer lactate perfusion. Overall mortality at 30 days was 3.4%,with a need for temporary hemodialysis or continuous venovenous hemofiltration in 4.7% of cases, and dialysis at discharge in 2.7% of the cases. Absence of ARF > 2 (> 100% rise in the baseline level of creatinine) and the need for dialysis were significantly better in the Custodiol group (P1⁄4.007; and P1⁄4.04, respectively). With the multivariate analysis the Custodiol perfusion and the duration of clamping were the predictive factors independent of non-IRA> 2. Conclusion: In our series of PAA repair, renal perfusion with 4 C Custodiol offers a better renal protection compared to Ringer lactate at 4 C. More series and/or randomized trials are necessary to confirm this conclusion.

Techniques and Strategies for Renal Protection in the Open Surgery of Pararenal Aneurysms

Objectives: To determine if renal perfusion with a cold crystalloid solution enriched with histidine-tryptophanketoglurarate (Custodiol, Dr. Franz-Kohler Chemie GmbH, Bensheim, Germany) offers a better protection against renal ischemic lesions compared to cold Ringer lactate in patients treated by open surgery for a pararenal aortic aneurysms (PAA). Materials and Methods: We studied 256 consecutive patients treated by open surgery for PAA between 1993 and 2013. In181cases thedirect perfusionof at least one renal artery was carried out. Among these patients, 87 had renal perfusion with Ringer lactate solution and 94 with a Custodiol solution. A score of propensity, based on clinical variables which would probably have influenced the renal function, was carried out to correct skews which could be associated with the use of Custodiol. Acute postoperative renal failure (ARF)was stratified in five classes according to the postoperative serum creatinine values and the need for dialysis, and wascompared in the twogroups, and the independent predictive factors ofARFwere identifiedwith amultivariate analysis. Results: After the score of propensity we could compare 74 patients perfused with Custodiol with those receiving a Ringer lactate perfusion. Overall mortality at 30 days was 3.4%,with a need for temporary hemodialysis or continuous venovenous hemofiltration in 4.7% of cases, and dialysis at discharge in 2.7% of the cases. Absence of ARF > 2 (> 100% rise in the baseline level of creatinine) and the need for dialysis were significantly better in the Custodiol group (P1⁄4.007; and P1⁄4.04, respectively). With the multivariate analysis the Custodiol perfusion and the duration of clamping were the predictive factors independent of non-IRA> 2. Conclusion: In our series of PAA repair, renal perfusion with 4 C Custodiol offers a better renal protection compared to Ringer lactate at 4 C. More series and/or randomized trials are necessary to confirm this conclusion.

Dantrolene in the treatment of malignant hyperthermia: a case report

Case report The patient, a 19 year old healthy male, who suffered childhood asthma, was anaesthetised for a planned mandible sagittal split and bimaxillar osteotomia due to prognathia. Anaesthesia was induced with propofol 140 mg and suxamethonium 100 mg was administered for muscle relaxation. Relaxation was not optimal but intubation was performed without problems. Anaesthesia was maintained with remifentanil (0.6-2.3μg/kg/min) and desflurane (4.97vol%). After 15 minutes the airway pressure was 30mbar, which was initially interpreted as broncho-obstructivity due to his childhood asthma, however, it was probably a sign of muscle rigidity. The patient was given minute ventilation at 8.2 litres, with pCO2 subsequently remaining stable for 2 hours. However, this increased to 10 kPa and the patient began to sweat. The patient’s temperature rose from 35.5°C 15 minutes after the start of the procedure to 37.9°C after 2.5 hours, but 15 minutes later the patient’s temperature was 38.9°C. MH was diagnosed and dantrolene 380 mg administered. Minute ventilation was increased to 12 litres FiO2 100%. Cooling was performed with cold intravenous Ringer Acetate and surface cooling with ice. After 25 minutes the temperature was 37.6°C. The patient remained on the ventilator for 11 hours after anaesthesia. There was mild recrudescence at approximately 6 hours, with blood pH gradually falling from 7.43 to 7.31, and pCO2 increasing from 4.9 to 6.8. A 200 mg dantrolene infusion was therefore administered over 2 hours, prior to extubation of the patient, such that the total dose of dantrolene administered was 580 mg (29 vials). When the patient came round he reported severe muscle pain. With regard to surgery, the mandible sagittal split was performed, but the maxillary surgery was cancelled, leaving the patient unable to close his mouth or bite. Over the following 2 weeks, prior to the second surgery, the patient was only able to drink and lost 6 kg. Second surgery was performed in trigger free anaesthesia without problems. In vitro contracture test revealed MH susceptible results (halothane contracture 2.8g and caffeine 1.3 g). The patient has p. 552 Arg>Trp mutation in RYR1.

Aortoiliac aneurysm with congenital right pelvic kidney

The association of congenital pelvic kidney with abdominal aortoiliac aneurysm is an extremely rare clinical finding. Previous reports have described various methods of aneurysm repair with successful preservation of the function of pelvic kidney. However, to our knowledge, reconstruction of more than two renal arteries has not been established. We report a case of abdominal aortic aneurysm complicated by congenital right pelvic kidney in a 72-year-old man. Computed tomography (CT) revealed an abdominal aortic aneurysm with a maximum diameter of 54mm and a right common iliac aneurysm of 45mm. In addition, he had a congenital right pelvic kidney and CT angiography identified three right pelvic renal arteries. The upper artery originated from the bifurcation of the terminal aorta and the lower two originated from the right common iliac artery. Three-dimensional CT was helpful for the accurate planning of the operation. Open surgical repair of the aortoiliac aneurysm with a Dacron bifurcated graft replacement was decided and reimplantation of all three right pelvic kidney arteries to the right limb of the graft was also performed. For renal preservation, the right pelvic kidney arteries were perfused with cold Ringer's lactate using a rapid infusion pump and coronary perfusion cannula. The patient's postoperative course was uneventful, and worsening of renal function was not observed. The perfusion of renal arteries with cold Ringer's solution was thought to be a simple and appropriate procedure for renal protection.

Improved arterial blood oxygenation following intravenous infusion of cold supersaturated dissolved oxygen solution.

BACKGROUNDOne of the primary goals of critical care medicine is to support adequate gas exchange without iatrogenic sequelae. An emerging method of delivering supplemental oxygen is intravenously rather than via the traditional inhalation route. The objective of this study was to evaluate the gas-exchange effects of infusing cold intravenous (IV) fluids containing very high partial pressures of dissolved oxygen (>760 mm Hg) in a porcine model.METHODSJuvenile swines were anesthetized and mechanically ventilated. Each animal received an infusion of cold (13 °C) Ringer’s lactate solution (30 mL/kg/hour), which had been supersaturated with dissolved oxygen gas (39.7 mg/L dissolved oxygen, 992 mm Hg, 30.5 mL/L). Arterial blood gases and physiologic measurements were repeated at 15-minute intervals during a 60-minute IV infusion of the supersaturated dissolved oxygen solution. Each animal served as its own control.RESULTSFive swines (12.9 ± 0.9 kg) were studied. Following the 60-minute infusion, there were significant increases in PaO2 and SaO2 (P < 0.05) and a significant decrease in PaCO2 (P < 0.05), with a corresponding normalization in arterial blood pH. Additionally, there was a significant decrease in core body temperature (P < 0.05) when compared to the baseline preinfusion state.CONCLUSIONSA cold, supersaturated dissolved oxygen solution may be intravenously administered to improve arterial blood oxygenation and ventilation parameters and induce a mild therapeutic hypothermia in a porcine model.

Hyperacute rejection in the xenogenic transplanted rat liver is triggered by the complement system only in the presence of leukocytes and free radical species

Reactive oxygen species (ROS) and nitric oxide species (NOS) are pivotal after ischemia-reperfusion. However, the role of different cells on the formation of free radical species after xenotransplantation remains elusive. We hypothesized that ROS and NOS formed during hyperacute rejection are dependent on leukocytes, erythrocytes, activated thrombocytes, and Kupffer cells (KCs). To address this issue, we developed a model of xenoperfused rat liver and assessed the relationship between free radical production and graft dysfunction. Livers from Sprague-Dawley rats were isolated, flushed with cold Ringer solution, and perfused at physically flow rates for 120 min after 1 h of ischemia. The control group was perfused with rat whole blood (n = 9). In the study groups, the livers were perfused with human whole blood, human plasma with erythrocytes, and plasma with erythrocytes and isolated thrombocytes (n = 9/group). In an additional group, gadolinium chloride (GdCl3), a selective Kupffer cell (KC) toxic agent, was applied. Liver damage, hyperacute rejection, and the depletion of KCs were monitored histologically. Liver damage and function were determined by means of liver enzymes, portal pressure, and bile production. Malondialdehyde (MDA), nitric oxide formation, and peroxynitrite concentration, as well as total glutathione (tGSH) level, were measured as indicators for free radical formation and anti-oxidative status. Significant differences in the MDA, NO, peroxynitrite levels, and GSH levels after reperfusion with various cell populations were observed. Markedly high ROS/RNS production was evident in the KCs and the xenogeneic whole-blood group. The oxidative stress was mainly caused by leukocytes and to lower extent by KCs, but only in combination with leukocytes. Neither erythrocytes, thrombocytes, nor hepatocytes had an effect on the release of ROS and RNS, as we could not observe significant differences in the MDA, peroxynitrite, and NO levels in these groups compared with control. Tissue injury and hyperacute rejection were more evident in the KC and whole-blood livers. No sign of damage was observed for the control, erythrocyte, and thrombocyte group. Removal of leukocytes from the perfusate by filtration had a major protective effect on the liver function and the grade of hyperacute rejection, whereas KC depletion reduced the ROS production, but did not have an impact on the hyperacute rejection and liver damage. In all xenogeneic perfused groups, the activation of the complement was histologically observed by positive C3c and C9b. Neither KC depletion nor the removal of leukocytes or thrombocytes from the perfusate had an effect on the activation of the complement system. Damage of the rat liver by the complement system was only observed in association with leukocytes. Our data revealed that various cell populations contribute to the formation of free radicals in our model. The production of free radicals was mainly linked to leukocytes and to a minor extent to KCs, but only in combination with leukocytes. Free radicals critically contribute to injury, rejection, and dysfunction of the xenotransplanted liver. Furthermore, hyperacute rejection in the xenogeneic perfused liver is triggered by the complement system only in the presence of leukocytes and free radical formation.

Pulmonary embolism as a cause of cardiac arrest: Hypothermia in post-resuscitation period (cooling therapy)

Pulmonary embolism as a possible cause of acute heart failure is a potentially fatal condition that can cause death in all age groups. Patients successfully resuscitated after cardiac arrest have a high risk of increased mortality and their poor long-term outcome is often associated with severe neurological complications. This is a case report of a 67-year-old man after a successful cardiopulmonary resuscitation (CPR) which was followed by therapeutic hypothermia (TH). The patient visited the dermatological outpatients' department with clinical presentation of pain and swelling of the right leg, shortness of breath and chest pain. During examination the patient lost consciousness, stopped breathing and had cardiac arrest. ECG was done which registered asystole. We began CPR. After 59 minutes of resuscitation return of heartbeat was achieved. The patient was transported to the Emergency Department. On admission, after computerized tomography (CT) of the chest confirmed massive pulmonary embolism (PE), the patient was administered thrombolytic therapy with Metalyse (tenecteplase) and anticoagulation therapy (heparin). After stabilization, therapeutic hypothermia was applied. Combination of EMCOOLSpad on the chest and abdomen and cold Ringer lactate 500 ml at 4 degrees C was flushed. Temperature was decreased to 33 degrees C and kept stabile for 24 hours. After eight days the patient was conscious with a minimal neurological deficit. As shown in this case report, and according to the rich experience elsewhere, cooling therapy after out-of-hospital cardiac arrest and successful CRP may be useful in preventing neurological complications.

Correcting the axial shrinkage of skeletal muscle thick sections visualized by confocal microscopy

Confocal microscopy is a suitable method for measurements and visualization of skeletal muscle fibres and the neighbouring capillaries. When using 3D images of thick sections the tissue deformation effects should be avoided. We studied the deformation in thick sections of the rat skeletal muscle from complete stacks of images captured with confocal microscope. We measured the apparent thickness of the stacks and compared it to the slice thickness deduced from calibrated microtome settings. The ratio of both values yielded the axial scaling factor for every image stack. Careful sample preparation and treatment of the tissue cryosections with cold Ringer solution minimize the tissue deformation. We conclude that rescaling by the inverse of the axial scaling factor of the stack of optical slices in the direction of the microscope optical axis satisfactorily corrects the axial deformation of skeletal muscle samples.

Oral intake of hydrogen-rich water inhibits intimal hyperplasia in arterialized vein grafts in rats

Arterialized vein grafts often fail due to intimal hyperplasia. Hydrogen potently protects organs and cells from many insults via its anti-inflammatory and antioxidant properties. We investigated the efficacy of oral administration of hydrogen-rich water (HW) for prevention of intimal hyperplasia. The inferior vena cava was excised, stored in cold Ringer solution for 2 h, and placed as an interposition graft in the abdominal aorta of syngeneic Lewis rats. HW was generated by immersing a magnesium stick in tap water (Mg + 2H(2)O → Mg (OH)(2) + H(2)). Beginning on the day of graft implantation, recipients were given tap water [regular water (RW)], HW or HW that had been subsequently degassed water (DW). Six weeks after grafting, the grafts in the rats given RW or DW had developed intimal hyperplasia, accompanied by increased oxidative injury. HW significantly suppressed intimal hyperplasia. One week after grafting, the grafts in HW-treated rats exhibited improved endothelial integrity with less platelet and white blood cell aggregation. Up-regulation of the mRNAs for intracellular adhesion molecules was attenuated in the vein grafts of the rats receiving HW. Activation of p38 mitogen-activated protein kinase, matrix metalloproteinase (MMP)-2, and MMP-9 was also significantly inhibited in grafts receiving HW. In rat smooth muscle cell (A7r5) cultures, hydrogen treatment for 24 h reduced smooth muscle cell migration. Drinking HW significantly reduced neointima formation after vein grafting in rats. Drinking HW may have therapeutic value as a novel therapy for intimal hyperplasia and could easily be incorporated into daily life.

Oral Intake of Hydrogen-rich Water Inhibits Intimal Hyperplasia in Arterialized Vein Grafts in Rats

Introduction: Arterialized vein grafts often fail due to intimal hyperplasia (IH), which remains a serious concern for patients after arterial bypass surgery due to a lack of pharmacological agents that prevent IH. Hydrogen gas potently protects organs and cells from many insults via its anti-inflammatory, antiapoptotic, and antioxidant properties. We investigated the efficacy of oral administration of hydrogen-rich water (HW), a feasible approach for hydrogen intake, for prevention of IH. Methods: The inferior vena cava was excised from donor rats, stored in cold Ringer solution for 2 hours, and placed as an interposition graft in the abdominal aorta of syngeneic Lewis rats. HW for drinking was generated by immersing a magnesium stick in tap water (Mg + 2H2O → Mg (OH)2+ H2). Beginning on the day of graft implantation, recipients were given, as drinking water, either regular tap water (RW), HW, or HW that had been subsequently degassed (DW) (Table1). Smooth muscle cell migration in culture was assessed using A7r5 cells. Results: Six weeks after grafting, the grafts in the rats given RW or DW had developed IH, accompanied by increased expression of 4HNE and 8-OHdG, oxidative injury markers. HW significantly suppressed intimal hyperplasia (Table 1) and expression of 4HNE and 8-OHdG. One week after grafting, the mRNAs for ICAM and endothelin receptor –A were upregulated in the vein grafts of rats that received RW or DW. This upregulation was attenuated in grafts of rats that received HW. Activation of p38 MAPK, matrix metalloproteinase (MMP)-2 andMMP-9 was also significantly inhibited in grafts that received HW as compared with those in rats that received RW or DW. Scanning electron microscopy revealed endothelial denudation with deposition of platelets and microthrombi, 1 week after engrafting, in the vein grafts of rats that received RW or DW. HW attenuated these changes; the grafts exhibited improved endothelial integrity with less platelet and white blood cell aggregation.No adverse effects, such as diarrhea or body weight loss, were seen during the follow-up period. in rat smooth muscle cell (A7r5) cultures, hydrogen treatment for 24 hours reduced smooth muscle cell migration and did not affect cell proliferation. Conclusions: Drinking HW significantly reduced neointima formation after vein grafting in rats. Drinking HW may have therapeutic value as a novel and alternative therapy for IH, which could also easily be incorporated into daily life. the therapeutic effects of HW may be related to hydrogen's anti-inflammatory actions or ability to decrease cell adhesion. Hydrogen merits further investigation for therapeutic use in the patients who undergo vein grafting. Table 1Drinking Water for Each Experimental Group Drinking water Mg stick Mg conc (mg/L ) H2 Conc (mM) pH IH at 6 weeks (n=6) (Intima/intima+media) Regular water (RW) No <1 .0 0 6.8-7.1 49.0±10.4 Degassed water (DW) Yes 6.2-7.9 0 9.2-9.9 52.2 ± 11.1 ∗ p<0.05 vs RW. Hydrogen water (HW) Yes 6.1-7.9 0.55-0.65 9.3-9.8 21.0 ± 13.9 ∗ p<0.05 vs RW. ∗ p<0.05 vs RW. Open table in a new tab

Inherent rhythmcity and interstitial cells of Cajal in a frog vein

Interstitial cells of Cajal are responsible for rhythmic contractions of the musculature of the gastrointestinal tract and blood vessels.The existence of these cells and spontaneous rhythmicity were noticed in amphibian vein and the findings are reported in this paper.The postcaval vein was identified in the frog, Rana tigrina and was perfused with amphibian Ringer solution after isolation.Contractile activity was recorded through a tension transducer connected to a polygraph.The isolated postcaval vein showed spontaneous rhythmic activity. Addition of cold Ringer solution decreased, while warm Ringer increased, the rate of contraction. Adrenaline caused inhibition of rhythmic activity at a dosage that increased the rate of isolated sinus venosus.Sections of the postcaval vein,when stained supravitally with methylene blue, showed the presence of interstitial cells of Cajal. Photic stimulation of the vein in the presence of methylene blue led to a significant decrease in the rate of spontaneous beating of the vein.These findings indicate that the postcaval vein of frog is capable of inherent rhythmcity, which is dependent on the interstitial cells of Cajal but is independent of the sinus venosus.