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- The effects of conjugated linoleic acid isomers cis-9,trans-11 and trans-10,cis-12 on in vitro bovine embryo production and cryopreservation. [JOURNAL ARTICLE]
- J Dairy Sci 2014 Jul 30.
Conjugated linoleic acid (CLA) isomers can affect the lipid profile and signaling of cells and thereby alter their function. A total of 5,700 bovine oocytes were used in a structured series of experiments to test the effects of cis-9,trans-11 CLA and trans-10,cis-12 CLA in vitro. In experiment 1, high doses of each CLA isomer during in vitro maturation (IVM) were compared with high or low doses during the entire in vitro culture (IVC) of parthenogenetic embryos. High doses of the CLA isomers ranged from 50 to 200 μM and low doses were 15 and 25 μM. In experiment 2, the low doses of each CLA isomer were tested during IVM/IVC on embryos produced by in vitro fertilization (IVF). Experiment 3 compared the effects of 15 μM doses of each CLA isomer during IVM or IVC of IVF embryos. In experiment 4, post-rewarming survival rates and blastomere counts were assessed for embryos supplemented with each CLA isomer during IVM or for 36 h before vitrification. In experiment 1, when either CLA isomer was provided only during IVM, we observed no effects on overall rates of maturation, cleavage, or blastocysts (92.2 ± 1.6%, 78.3 ± 4.1%, and 28.9 ± 5.1%, respectively). However, high doses of each CLA isomer, but not low doses, during the entire embryo culture period decreased blastocyst rates (5-20%) in a dose-dependent manner. Cleavage rates improved with 15 or 50 μM CLA trans-10,cis-12. Progesterone concentrations in maturation media were significantly increased by high doses of each CLA isomer compared with control, but low doses of CLA isomers had no effect. In experiment 2 with IVF embryos, low doses of each CLA isomer did not alter cleavage rates (average 84.9 ± 1.9%) and only 25 μM CLA trans-10,cis-12 during IVC reduced blastocyst rates below those of controls (25.5 ± 2.1 vs. 38.2 ± 2.3%). The lipid content of embryos was increased and relative expression of the BIRC5 (baculoviral IAP repeat containing 5) gene was depressed by CLA trans-10,cis-12. In experiment 3, low doses (15 μM) of each CLA isomer during IVC significantly reduced blastocyst rates (20.6 ± 2.4% and 27.7 ± 1.2% vs. 34.18 ± 1.2% for CLA trans-10,cis-12 and CLA cis-9,trans-11 compared with control, respectively) with less effect of each CLA during IVM. In experiment 4, adding 100 μM CLA cis-9,trans-11 during the final 36 h of culture resulted in a high survival rate after rewarming and culture, and the higher embryo blastomere count was comparable to that of control embryos not undergoing vitrification. In conclusion, supplementation with either CLA isomer did not improve embryo production, but inclusion of CLA cis-9,trans-11 before vitrification improved the quality of bovine IVF embryos after rewarming and culture.
- The effect of ethnicity on the vascular responses to cold exposure of the extremities. [JOURNAL ARTICLE]
- Eur J Appl Physiol 2014 Aug 1.
Cold injuries are more prevalent in individuals of African descent (AFD). Therefore, we investigated the effect of extremity cooling on skin blood flow (SkBF) and temperature (T sk) between ethnic groups.Thirty males [10 Caucasian (CAU), 10 Asian (ASN), 10 AFD] undertook three tests in 30 °C air whilst digit T sk and SkBF were measured: (i) vasomotor threshold (VT) test-arm immersed in 35 °C water progressively cooled to 10 °C and rewarmed to 35 °C to identify vasoconstriction and vasodilatation; (ii) cold-induced vasodilatation (CIVD) test-hand immersed in 8 °C water for 30 min followed by spontaneous warming; (iii) cold sensitivity (CS) test-foot immersed in 15 °C water for 2 min followed by spontaneous warming. Cold sensory thresholds of the forearm and finger were also assessed.In the VT test, vasoconstriction and vasodilatation occurred at a warmer finger T sk in AFD during cooling [21.2 (4.4) vs. 17.0 (3.1) °C, P = 0.034] and warming [22.0 (7.9) vs. 12.1 (4.1) °C, P = 0.002] compared with CAU. In the CIVD test, average SkBF during immersion was greater in CAU [42 (24) %] than ASN [25 (8) %, P = 0.036] and AFD [24 (13) %, P = 0.023]. Following immersion, SkBF was higher and rewarming faster in CAU [3.2 (0.4) °C min(-1)] compared with AFD [2.5 (0.7) °C min(-1), P = 0.037], but neither group differed from ASN [3.0 (0.6) °C min(-1)]. Responses to the CS test and cold sensory thresholds were similar between groups.AFD experienced a more intense protracted finger vasoconstriction than CAU during hand immersion, whilst ASN experienced an intermediate response. This greater sensitivity to cold may explain why AFD are more susceptible to cold injuries.
- Cardiac arrest patients have an impaired immune response, which is not influenced by induced hypothermia. [Journal Article]
- Crit Care 2014; 18(4):R162.
Induced hypothermia is increasingly applied as a therapeutic intervention in ICUs. One of the underlying mechanisms of the beneficial effects of hypothermia is proposed to be reduction of the inflammatory response. However, a fear of reducing the inflammatory response is an increased infection risk. Therefore, we studied the effect of induced hypothermia on immune response after cardiac arrest.A prospective observational cohort study in a mixed surgical-medical ICU. Patients admitted at the ICU after surviving cardiac arrest were included and during 24 hours body temperature was strictly regulated at 33°C or 36°C. Blood was drawn at three time points: after reaching target temperature, at the end of the target temperature protocol and after rewarming to 37°C. Plasma cytokine levels and response of blood leucocytes to stimulation with toll-like receptor (TLR) ligands lipopolysaccharide (LPS) from Gram-negative bacteria and lipoteicoic acid (LTA) from Gram-positive bacteria were measured. Also, monocyte HLA-DR expression was determined.In total, 20 patients were enrolled in the study. Compared to healthy controls, cardiac arrest patients kept at 36°C (n = 9) had increased plasma cytokines levels, which was not apparent in patients kept at 33°C (n = 11). Immune response to TLR ligands in patients after cardiac arrest was generally reduced and associated with lower HLA-DR expression. Patients kept at 33°C had preserved ability of immune cells to respond to LPS and LTA compared to patients kept at 36°C. These differences disappeared over time. HLA-DR expression did not differ between 33°C and 36°C.Patients after cardiac arrest have a modest systemic inflammatory response compared to healthy controls, associated with lower HLA-DR expression and attenuated immune response to Gram-negative and Gram-positive antigens, the latter indicative of an impaired immune response to bacteria. Patients with a body temperature of 33°C did not differ from patients with a body temperature of 36°C, suggesting induced hypothermia does not affect immune response in patients with cardiac arrest.ClinicalTrials.gov NCT01020916, registered 25 November 2009.
- The effect of adjusting tracheal tube cuff pressure during deep hypothermic circulatory arrest: A randomised trial. [Journal Article]
- Eur J Anaesthesiol 2014 Sep; 31(9):452-6.
Regular endotracheal tube cuff monitoring may prevent silent aspiration.We hypothesised that active management of the cuff of the tracheal tube during deep hypothermic cardiac arrest would reduce silent subglottic aspiration. We also determined to study its effect on postoperative mechanical ventilation and the incidence of postoperative positive tracheal cultures.A randomised clinical trial.The study was conducted in a University Teaching Hospital from September 2008 to November 2009.Twenty-four patients undergoing elective pulmonary endarterectomy were included in the study.After induction of general anaesthesia and tracheal intubation, the cuff of the tracheal tube was inflated to 25 cmH2O. Following this, 1 ml of methylene blue dye diluted in 2 ml of physiological saline was injected into the hypopharynx. Patients were randomly assigned to active cuff management during cooling and warming (where cuff pressure was monitored and the cuff was reinflated if it dropped below 20 cmH2O, or deflated if pressure exceeded 30 cmH2O) or passive monitoring (where cuff pressure was monitored but volume was not altered). Before weaning from cardiopulmonary bypass, fibreoptic bronchoscopy was performed. Silent aspiration was then diagnosed if blue dye was seen in the trachea below the cuff of the tube.The primary aim of this study was to determine the incidence of silent aspiration. Secondary outcomes included duration of postoperative mechanical ventilation of the lungs and incidence of positive culture of tracheal aspirate.Active cuff management patients were younger than controls (51.2 ± 11.6 vs. 63.2 ± 9 years, P = 0.028), but otherwise the two groups were similar. The primary endpoint was reached because we showed that silent aspiration was significantly less frequent in the study group (0/12 vs. 8/12 patients, P = 0.001). Significantly lower intracuff pressures were measured in the control group patients at several timepoints during cooling, just before hypothermic arrest and at all timepoints during rewarming.We recommend that the cuff of the tracheal tube should be checked regularly during surgery under deep hypothermia, and the cuff pressure adjusted as required.
- [Immunogenicity of allogeneic freezing periosteum and bone marrow]. [English Abstract, Journal Article]
- Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi 2014 May; 28(5):649-53.
To investigate the immunogenicity of freezing periosteum and bone marrow during allogeneic joint transplantation, and to explore proper pretreatment of allogeneic joint.The allogeneic periosteum and bone marrow were harvested from knee joints of 5 New Zealand white rabbits (aged, 6 months; weighing, 2.6-3.0 kg). After gradient cooling, the tissue was cryopreserved for 1 month. The freezing periosteum and bone marrow were grinded to pieces after rewarming to prepare the suspension of periosteum and bone marrow. Eighteen Chinchilla rabbits (aged, 6 months; weighing, 2.1-2.8 kg) were divided into 3 groups randomly: normal saline injection group (group A, n = 6), periosteum injection group (group B, n = 6), and bone marrow injection group (group C, n=6). The normal saline, periosteum suspension, and bone marrow suspension were injected into the peritoneal cavity in groups A, B, and C, respectively. The concentrations of interleukin 2 (IL-2), IL-6, and tumor necrosis factor alpha (TNF-alpha) in serum and the ratio of CD4' T cell/CD8+ T cell in venous blood were measured before injection, at 1 week and 2 weeks after injection.There was no significant difference in the concentration of IL-2 between before and after injection in the same group (P=0.241), and between groups (P = 0.055). The concentration of IL-6 after injection was significantly lower than that before injection in the same group (P = 0.040), but no significant difference was found between groups (P = 0.357). The concentration of TNF-a showed no significant difference between before and after injection in the same group (P = 0.925), but the concentration of TNF-a in group B was significantly higher than that in groups A and C (P < 0.05). The ratio of CD4+ T cell/CD8+ T cell of venous blood had no significant difference between before and after operation in the same group (P = 0.248), and between groups (P=0.646).The freezing periosteum and bone marrow are lowly immunogenic. In order to decrease the immunogenicity of the joint, preserving the periosteum and removing the marrow cavity are recommended.
- The "neurovascular unit approach" to evaluate mechanisms of dysfunctional autoregulation in asphyxiated newborns in the era of hypothermia therapy. [REVIEW]
- Early Hum Dev 2014 Jul 22.
Despite improvements in obstetrical and neonatal care, and introduction of hypothermia as a neuroprotective therapy, perinatal brain injury remains a frequent cause of cerebral palsy, mental retardation and epilepsy. The recognition of dysfunction of cerebral autoregulation is essential for a real time measure of efficacy to identify those who are at highest risk for brain injury. This article will focus on the "neurovascular unit" approach to the care of asphyxiated neonates and will address 1) potential mechanisms of dysfunctional cerebral blood flow (CBF) regulation, 2) optimal monitoring methodology such as NIRS (near infrared spectroscopy), and TCD (transcutaneous Doppler), and 3) clinical implications of monitoring in the neonatal intensive care setting in asphyxiated newborns undergoing hypothermia and rewarming. Critical knowledge of the functional regulation of the neurovascular unit may lead to improved ability to predict outcomes in real time during hypothermia, as well as differentiate non-responders who might benefit from additional therapies.
- Two case studies and a review of paroxysmal cold hemoglobinuria. [Journal Article]
- Lab Med 2014; 45(3):253-8.
Paroxysmal cold hemoglobinuria (PCH) is an acquired hemolytic anemia caused by immunoglobulin G (IgG) antibodies that sensitize red blood cells (RBCs) at cold temperatures by fixing complement to the RBCs causing intravascular hemolysis on rewarming. PCH usually appears in young children as recurrent high fevers, chills, and passage of red-brown urine. The diagnostic test for PCH is the Donath-Landsteiner test, an in vitro assay for biphasic hemolysis. Herein, we present 2 cases of PCH that occurred within 12 months of each other. We quickly diagnosed the second case and treated the patient successfully, in part due to our recognition of its characteristics based on the first case. PCH is a hemolytic anemia for which there is a specific diagnostic test; the timely recognition of this entity by physicians and laboratory staff will allow prompt, supportive therapy and will raise the odds of quick resolution of hemolysis.
- Plasma proteomic changes during hypothermic and normothermic cardiopulmonary bypass in aortic surgeries. [Journal Article]
- Int J Mol Med 2014 Oct; 34(4):947-56.
Deep hypothermic circulatory arrest (DHCA) is a protective method against brain ischemia in aortic surgery. However, the possible effects of DHCA on the plasma proteins remain to be determined. In the present study, we used novel high‑throughput technology to compare the plasma proteomes during DHCA (22˚C) with selective cerebral perfusion (SCP, n=7) to those during normothermic cardiopulmonary bypass (CPB, n=7). Three plasma samples per patient were obtained during CPB: T1, prior to cooling; T2, during hypothermia; T3, after rewarming for the DHCA group and three corresponding points for the normothermic group. A proteomic analysis was performed using isobaric tag for relative and absolute quantification (iTRAQ) labeling tandem mass spectrometry to assess quantitative protein changes. In total, the analysis identified 262 proteins. The bioinformatics analysis revealed a significant upregulation of complement activation at T2 in normothermic CPB, which was suppressed in DHCA. These findings were confirmed by the changes of the terminal complement complex (SC5b‑9) levels. At T3, however, the level of SC5b‑9 showed a greater increase in DHCA compared to normothermic CPB, while 48 proteins were significantly downregulated in DHCA. The results demonstrated that DHCA and rewarming potentially exert a significant effect on the plasma proteome in patients undergoing aortic surgery.
- Induced Hypothermia During Resuscitation From Hemorrhagic Shock Attenuates Microvascular Inflammation In The Rat Mesenteric Microcirculation. [JOURNAL ARTICLE]
- Shock 2014 Jul 18.
Microvascular inflammation occurs during resuscitation following hemorrhagic shock, causing multiple organ dysfunction and mortality. Pre-clinical evidence suggests that hypothermia may have some benefit in selected patients by decreasing this inflammation, but this effect has not been extensively studied.Intravital microscopy was used to visualize mesenteric venules of anesthetized rats in real time to evaluate leukocyte adherence and mast cell degranulation. Animals were randomly allocated to normotensive or hypotensive groups, and further subdivided into hypothermic and normothermic resuscitation (N=6 per group). Animals in the shock groups underwent mean arterial blood pressure reduction to 40-45 mmHg for 1 hour via blood withdrawal. During the first two hours following resuscitation by infusion of shed blood plus double that volume of normal saline, rectal temperature of the hypothermic groups were maintained at 32-34°C, while the normothermic groups were maintained between 36-38°C. The hypothermic group was then rewarmed for the final two hours of resuscitation.Leukocyte adherence was significantly lower after 2 hours of hypothermic resuscitation compared with normothermic resuscitation: (2.8±0.8 vs 8.3±1.3 adherent leukocytes, p=0.004). Following rewarming, leukocyte adherence remained significantly different between hypothermic and normothermic shock groups: (4.7±1.2 vs 9.5±1.6 adherent leukocytes, p=0.038). Mast cell degranulation index (MDI) was significantly decreased in the hypothermic (1.02±0.04 MDI) vs normothermic (1.22±0.07 MDI) shock groups (p=0.038) after the experiment.Induced hypothermia during resuscitation following hemorrhagic shock attenuates microvascular inflammation in rat mesentery. Furthermore, this decrease in inflammation is carried over after rewarming takes place.
- Extracorporeal-Assisted Rewarming in the Management of Accidental Deep Hypothermic Cardiac Arrest: A Systematic Review of the Literature. [REVIEW]
- Heart Lung Circ 2014 Jun 27.
A systematic review of the literature surrounding the use of Extra-Corporeal Assisted Rewarming (ECAR) in patients presenting with deep hypothermia or hypothermic cardiac arrest was undertaken using a structured protocol. Thirty-one papers were deemed suitable for review, 13 of these were of sufficient quality to permit systematic data analysis. The primary outcome measure was survival to hospital discharge. The secondary outcome measure was functional neurological status at last follow-up. Analysis revealed a 67.7% survival to discharge and a 61.5% rate of good neurological recovery for patients presenting with pure hypothermic cardiac arrest. This was in marked contrast to a 23.4% survival and a 9.4% rate of good neurological outcome in those presenting with a mixed hypoxic/hypothermic arrest. Other data revealed a survival benefit for patients presenting with deep hypothermia without cardiac arrest treated with ECAR compared to those treated with conventional rewarming techniques. Hypoxic arrest, serum potassium >10mmol/L and presenting rhythm of asystole were found likely be significant predictors of poor outcome. Innovative reperfusion and rewarming strategies are also reviewed.