Crystallization of Human Erythrocyte Band 3, the anion exchanger, at the International Space Station "KIBO".

Band 3 mediates the Cl- and HCO3- exchange across the red blood cell membrane and plays a pivotal role for delivering oxygen appropriately to metabolically active tissues. For understanding molecular mechanisms, it is essential to know the structure and function relationship. In terrestrial environments, however, nobody could make good quality crystals of Band 3 for the X-ray crystallographic study. In this study, we purified the transmembrane domain of Band 3 from human red blood cells and crystallized the purified Band 3 without the Fab fragment at the International Space Station "KIBO" under microgravity environments.

Therapeutic potential of recombinant thrombomodulin for lung injury after pneumonectomy via inhibition of high-mobility group box 1 in mice.

BACKGROUND: Surgical acute respiratory distress syndrome (ARDS) is an extremely critical condition which may occur after major lung resection. Despite advances in minimally invasive surgical procedures and progress in the therapeutic management of this disease, prognosis remains poor. In this study, we investigated the contribution of high-mobility group box 1 (HMGB1) in a surgical ARDS model and evaluated the possible therapeutic effect of recombinant thrombomodulin (rTM) for the treatment of surgical ARDS. METHODS: C57BL/6J mice underwent left pneumonectomy. rTM was injected at 12 hours before surgery, followed by 12 hours for 3 days after surgery. Lipopolysaccharide (LPS) was administered at 2 hours after surgery. We conducted a histologic analysis and measured HMGB1, IL-6, IL-1ß, and TNF-α in bronchoalveolar lavage fluid on day 3 after pneumonectomy. Data were compared between the treatment groups. RESULTS: On histologic analysis, left pneumonectomy followed by LPS administration induced both severe inflammatory cellular infiltration and alveolar wall congestion with hemorrhage. rTM administration rescued these histologic changes. The level of HMGB1, IL-6, IL-1ß, and TNF-α in bronchoalveolar lavage fluid was significantly increased by LPS administration after pneumonectomy and significantly decreased by rTM administration with LPS and pneumonectomy (p < 0.001). Also, LPS alone showed no statistical differences in HMGB1 or proinflammatory cytokine level compared with pneumonectomy (PNX) group. In addition, the survival outcome was also improved by rTM administration. CONCLUSIONS: LPS administration after left pneumonectomy could induce the severe lung injury. PNX and LPS have similar contribution to this model and may play a synergistic role in this process. rTM may have the potential therapeutic effect for surgical ARDS via suppression of HMGB1 and the secretion of proinflammatory cytokines induced by the administration of LPS after left pneumonectomy.

Optimized magnitude of cryosurgery facilitating anti-tumor immunoreaction in a mouse model of Lewis lung cancer.

BACKGROUND: Cryosurgery has reemerged as a less invasive local treatment with possible immune-regulatory effects. However, the optimal magnitude of cryosurgery for achieving immune-regulatory responses at abscopal tumor sites remains unclear. We aimed to investigate appropriate magnitude of cryosurgery for this goal using a mouse model. METHODS: C57BL/6J mice were inoculated with Lewis lung carcinoma cells or B16 melanoma cells in bilateral flanks. The left-sided tumor was cryoablated with repeated freeze/thaw cycles either once, twice, or thrice. The peritumoral injections of LPS were performed. Abscopal tumor volumes were measured, immunohistochemistry was performed for CD4, CD8, Foxp3, and Ki-67, and proinflammatory cytokines were measured in lavage fluid of cryoablated tumor. RESULTS: The growth rate of the abscopal tumor was slowest in the Cryosurgery ×2 group among the five experimental groups. The proportions of CD4(+) T cells and CD8(+) T cells in the abscopal tumor were also significantly higher in the Cryosurgery ×2 group. The levels of IL-1ß, IL-2, IL-6, IL-12ß, IFN-γ, and TNF-α in the peritumoral lavage fluid in Cryosurgery ×2 + LPS group were significantly increased compared with the other groups. CONCLUSIONS: This study suggested that achievement of approximately 73 % damaged area in the cryoablated tumor by two cycles of cryosurgery generates the most favorable immune-regulatory response for abscopal tumors via activation of anti-tumor immune cells as well as increased secretion of proinflammatory cytokines.

Efficacy and hemodynamic response of pleural carbon dioxide insufflation during thoracoscopic surgery in a swine vessel injury model.

PURPOSES: Thoracoscopic anatomical lung resection is a minimally invasive technique, but intraoperative massive bleeding is a critical complication. We investigated the hemostatic efficacy and safety of intrapleural carbon dioxide (CO2) insufflation in thoracoscopic surgery in a swine vessel injury model. METHODS: Swines were assigned to one of four groups subjected to thoracoscopic surgery under target intrathoracic pressures of 0, 5, 10, or 15 mmHg CO2 insufflation, respectively. A pin-hole injury of the right cranial lobe pulmonary vein was inflicted thoracoscopically and we compared the blood loss and hemodynamic changes in each group. RESULTS: There were no signs or echographic findings of air embolus. Both the blood loss per minute and total blood loss during the experiment were significantly lower in the 10 and 15 mmHg groups than in the 0 mmHg group (p > 0.05, respectively). The hemodynamic signs, including heart rate, mean arterial pressure, and peripheral oxygen saturation, were not significantly different in the 0 and 10 mmHg groups at most times, although they were significantly correlated with the insufflation pressure during the experiments (p < 0.05). CONCLUSIONS: CO2 insufflation in thoracoscopic major lung resection appears to be safe, even in the short term, and can help to control vessel injury.

Influence of activity level on the outcome of treatment of lateral ankle ligament rupture.

BACKGROUND: Few studies of treatment of lateral ankle ligament rupture have given careful consideration to the activity levels of patients. In the present study, we investigated whether a patient's activity level influences the outcome of the treatment of lateral ankle ligament rupture and whether treatment should be determined according to activity level. METHODS: A total of 115 consecutive patients with lateral ankle ligament rupture were treated with short-term cast immobilization followed by functional bracing. We divided preinjury activity levels and activity levels at follow-up into four classes according to the International Knee Documentation Committee standards. Follow-up examination 1 year after injury included clinical assessment (anterior drawer test, range of motion, maximum circumference of the calf, tenderness, swelling), reinjury assessment, and functional evaluation by means of the Karlsson scoring system. RESULTS: We found significant differences in ankle stability and reinjuries between the four groups classified by their preinjury activity level. The number of patients with ankle instability and reinjury was significantly higher in the high-activity group than in the low-activity groups. We also found significant difference between groups classified by activity level at follow-up with regard to the Karlsson score. CONCLUSIONS: The outcome of treatment of lateral ankle ligament rupture is significantly influenced by the patient activity level, and treatment may need to be determined according to this activity level.