Shinji Ogura

Release of Phosphorylated HSP27 (HSPB1) from Platelets Is Accompanied with the Acceleration of Aggregation in Diabetic Patients

We investigated the relationship between HSP27 phosphorylation and collagen-stimulated activation of platelets in patients with diabetes mellitus (DM). Platelet-rich plasma was prepared from blood of type 2 DM patients. The platelet aggregation was analyzed in size of aggregates by an aggregometer using a laser scattering method. The protein phosphorylation was analyzed by Western blotting. Phosphorylated-HSP27 and PDGF-AB released from platelets were measured by ELISA. The phosphorylated-HSP27 levels at Ser-78 and Ser-82 induced by collagen were directly proportional to the platelet aggregation. Total HSP27 levels in platelets were decreased concomitantly with the phosphorylation. The released HSP27 levels were significantly correlated with the phosphorylated levels of HSP27 in the platelets stimulated by 0.3 μg/ml collagen. The low dose collagen-stimulated release of HSP27 was detected but relatively small in healthy donors. The released levels of PDGF-AB were in parallel with the levels of released HSP27. Area under the curve (AUC) of small aggregation (9-25 μm) induced by 0.3 μg/ml collagen was inversely proportional to the levels of released HSP27. AUC of large aggregation (50-70 μm) was directly proportional to the levels of released HSP27. Exogenous recombinant phosphorylated- HSP27 hardly affected the aggregation or the released levels of PDGF-AB induced by collagen. These results strongly suggest that HSP27 is released from human platelets accompanied with its phosphorylation induced by collagen, which is correlated with the acceleration of platelet aggregation in type 2 DM patients.

Effect of enteral diet enriched with eicosapentaenoic acid, gamma-linolenic acid, and antioxidants in patients with sepsis-induced acute respiratory distress syndrome

BackgroundIn this study, the effects of an enteral diet enriched with eicosapentaenoic acid (EPA), γ-linolenic acid (GLA), and antioxidants were compared with a standard enteral diet in critically ill patients with sepsis-induced acute respiratory distress syndrome (ARDS).MethodsThis study was a single-center, prospective, randomized, single-blind, controlled trial in our Advanced Critical Care Center. Patients were randomized to receive a continuous EPA, GLA, and antioxidant-enriched diet (study group), or an isocaloric standard diet (control group).ResultsTwenty-three of 46 patients were in the study group, and the other 23 were in the control group. Duration of mechanical ventilation, incidence of new nosocomial infections, changes over time in Sequential Organ Failure Assessment (SOFA) scores, and 60-day mortality were not significantly different between the two groups. The ratio of partial pressure of oxygen to fraction of inspired oxygen on day 7 was significantly higher in the study group (233.0 [185.5–282.8] vs. 274.0 [225.5–310.8], p = 0.021). Duration of ICU stay was significantly shorter in the study group than in the control group (24.0 [20.0–30.0] vs. 15.0 [11.0–24.0], p = 0.008).ConclusionsAn enteral diet enriched with EPA, GLA, and antioxidants did not improve duration of mechanical ventilation, SOFA score, incidence of new nosocomial infections, or mortality but did favorably influence duration of ICU stay in critically ill patients with sepsis-induced ARDS.

Granulocyte and Monocyte Adsorption Apheresis for Generalized Pustular Psoriasis: Therapeutic Outcomes in Three Refractory Patients

Generalized pustular psoriasis (GPP) is a type of neutrophilic dermatosis that is sometimes resistant to medications. In patients with neutrophilic skin diseases, granulocyte and monocyte adsorption apheresis (GMA) has been demonstrated to selectively and efficiently eliminate myeloid‐lineage leukocytes from the peripheral blood. We evaluated the efficacy and safety of repeated GMA therapy in three refractory GPP patients. Three GPP patients refractory to previous therapies received weekly GMA with five sessions per course, which was repeated when the symptoms reappeared. The efficacy was assessed by the disease severity scores 2 weeks after each course of GMA. The GPP severity scores of all three patients were reduced in all courses (N = 9); they were reduced by more than 3 points in six courses and by 2 points in three courses. After the first GMA course, the GPP severity scores were reduced by more than 3 points in all three patients. On average, the GPP severity scores were reduced by 4.67 and 3.67 points after the first course and repeated courses, respectively. The severity of edema and pustules were particularly improved in all patients and no adverse effects were observed. GMA showed efficacy for the treatment of refractory GPP patients as a non‐pharmacologic intervention without any associated adverse effects, and was particularly effective in the first course, but also effective in the subsequent courses.

Comparison of two pediatric flail chest cases

Flail chest is a rare complication in pediatric patients with blunt chest trauma. There is no general consensus on which treatment is most appropriate for flail chest in pediatric patients, although it has been reported that surgical fixation is associated with beneficial outcomes for flail chest in adults.The present report described two pediatric cases of flail chest, which was rare in pediatric blunt trauma. In small children, functional residual capacity is smaller, and the thorax is pliable due to high thoracic compliance. Therefore, it is only advisable to select intubation and mechanical ventilation treatment. Likewise, in pediatric flail chest, the available evidence does not suggest that ventilator management protocols should be adopted routinely, and the treatment for pediatric flail chest was not established completely.There were not huge different between the described patients, including injury severity and ventilation setting. However, one had a relapse of flail chest after extubation and chest taping was required, while the other patient’s condition was stable after decannulation.As described above, it is difficult to predict a recurrence of flail chest in pediatric patients even if treatment goes well. Therefore, T-piece trial should be considered prior to extubation.

Simple pneumopericardium due to blunt trauma progressing to tension pneumopericardium during transportation

Patients with simple pneumopericardium due to blunt thoracic trauma occasionally progressed to tension pneumopericardium, although pneumopericardium is believed to be benign in general. A 65-year-old man had both arms caught in a grinding machine and his face struck hard at work. He was diagnosed with bilateral degloving injuries of both arms and mediastinal emphysema on computed tomography. He required transfer to an advanced emergency medical service center for treatment. Although he was hemodynamically stable then, the patients condition deteriorated during transportation. The patient returned to the local hospital as cardiopulmonary resuscitation continued, repeat computed tomography was performed, which showed a substantial pneumopericardium and exacerbation of mediastinal and subcutaneous emphysema. After then, cardiopulmonary resuscitation was discontinued because there was no response. For the patient to be rescued in this situation, thoracotomy is required, although it should be reserved for patients with evidence of hemodynamic compromise attributable to cardiac tamponade.

Rho-kinase regulates human platelet activation induced by thromboxane A2 independently of p38 MAP kinase

We have previously demonstrated that ristocetin, an activator of GPIb/IX/V, induces the release of soluble CD40 ligand (sCD40L) via thromboxane A2 production in human platelets. It has been shown that thromboxane A2 induces the activation of Rho-kinase, a downstream effector of Rho, in human platelets. In the present study, we investigated the exact roles of Rho-kinase in thromboxane A2-induced platelet activation. We found that U46619, a thromboxane receptor (TP) agonist, induced the phosphorylation of cofilin, a target of Rho-kinase signaling, and that the cofilin phosphorylation by U46619 was suppressed by Y27632 or fasudil, specific inhibitors of Rho-kinase. Y27632 and fasudil markedly decreased large platelet aggregate formation by U46619. The release of sCD40L and secretion of platelet-derived growth factor (PDGF)-AB stimulated by U46619 were inhibited by Y27632 and fasudil. SB203580, a specific inhibitor of p38 mitogen-activated protein (MAP) kinase, reduced the sCD40L release and PDGF-AB secretion. Y27632 and fasudil failed to affect the phosphorylation of p38 MAP kinase whereas SB203580 had little effect on the phosphorylation of cofilin induced by U46619. In conclusion, our results strongly suggest that Rho-kinase regulates thromboxane A2-induced human platelet activation independently of p38 MAP kinase.

αB-crystallin reduces ristocetin‑induced soluble CD40 ligand release in human platelets: Suppression of thromboxane A2 generation

Our group has previously shown that αB-crystallin (HSPB5), a small heat shock protein, inhibits human platelet aggregation by ristocetin, an activator of glycoprotein Ib/IX/V. In addition, it was demonstrated that glycoprotein Ib/IX/V activation induces soluble CD40 (sCD40) ligand release via thromboxane (TX) A2. In the present study, the effect of αB-crystallin on the ristocetin-induced sCD40 ligand release in human platelets was investigated. The ristocetin-induced release of sCD40 ligand was suppressed by αB-crystallin. In addition, αB-crystallin reduced the ristocetin-stimulated production of 11-dehydro-TX B2, a stable metabolite of TXA2. αB-crystallin did not suppress the platelet aggregation induced by U46619, a TXA2 receptor agonist. αB-crystallin had little effect on the U46619-induced phosphorylation of p38 mitogen-activated protein kinase or sCD40 ligand release. In addition, αB-crystallin failed to reduce the binding of SZ2, a monoclonal antibody against the sulfated sequence in the α-chain of glycoprotein Ib, to the ristocetin-stimulated platelets. These results strongly suggest that αB-crystallin extracellularly suppresses ristocetin-stimulated release of sCD40 ligand by inhibiting the TXA2 production in human platelets.

Postinfarction Cardiac Remodeling Proceeds Normally in Granulocyte Colony-Stimulating Factor Knockout Mice.

Treatment with granulocyte colony-stimulating factor (G-CSF) reportedly mitigates postinfarction cardiac remodeling and dysfunction. We herein examined the effects of G-CSF knockout (G-CSF-KO) on the postinfarction remodeling process in the hearts of mice. Unexpectedly, the acute infarct size 24 hours after ligation was similar in the two groups. At the chronic stage (4 weeks later), there was no difference in the left ventricular dimension, left ventricular function, or histological findings, including vascular density, between the two groups. In addition, expression of vascular endothelial growth factor (VEGF) was markedly up-regulated in hearts from G-CSF-KO mice, compared with wild-type mice. Microarray failed in detecting up-regulation of VEGF mRNA, whereas G-CSF administration significantly decreased myocardial VEGF expression in mice, indicating that G-CSF post-transcriptionally down-regulates VEGF expression. When G-CSF-KO mice were treated with an anti-VEGF antibody (bevacizumab), cardiac remodeling was significantly aggravated, with thinning of the infarct wall and reduction of the cellular component, including blood vessels. In the granulation tissue of bevacizumab-treated hearts 4 days after infarction, vascular development was scarce, with reduced cell proliferation and increased apoptosis, which likely contributed to the infarct wall thinning and the resultant increase in wall stress and cardiac remodeling at the chronic stage. In conclusion, overexpression of VEGF may compensate for the G-CSF deficit through preservation of cellular components, including blood vessels, in the postinfarction heart.

Preliminary Experience with Air Transfer of Patients for Rescue Endovascular Therapy after Failure of Intravenous Tissue Plasminogen Activator

The present report describes our experience with air transfer of patients with acute ischemic stroke in whom intravenous tissue plasminogen activator (IV t-PA) failed for rescue endovascular therapy (EVT). Twenty-three consecutive patients in whom IV t-PA failed were transferred to our hospital for rescue EVT between February 2011 and April 2013. The amount of time required for transfer, distance, clinical outcomes, and complications were compared between patients transferred by ground (TG group; n = 17) and by air (TA group; n = 6). Computed tomography imaging on arrival revealed hemorrhagic transformation in 1 (5.9%) patient in the TG group, whereas none of the patients in the TA group developed any type of complication. The remaining 22 patients received rescue EVT. The elapsed time from the request call to arrival at our hospital did not significantly differ between the TG and TA groups (45.8 ± 4.9 min vs. 41.6 ± 2.3 min). However, the distance from the primary hospital to our institution was significantly longer for the TA group than for the TG group (38.8 ± 10.4 km vs. 13.5 ± 1.2 km, p = 0.001). The frequency of favorable outcomes (modified Rankin Scale 0–1 at 90 days after onset) in the TG and TA groups were 25.0% and 50.0%, respectively (p = 0.267). Air transfer for patients after IV t-PA failure allowed for more rapid delivery of patients over longer distances than ground transfer.

Phenotype and Physiological Significance of the Endocardial Smooth Muscle Cells in Human Failing Hearts

Background—Extravascular smooth muscle cells are often observed in the endocardium of human failing hearts. Here, we characterized the phenotype of those cells and investigated their physiological significance. Methods and Results—We examined left ventricular biopsy specimens obtained from 44 patients with dilated cardiomyopathy and 6 nonfailing hearts. In Masson trichrome–stained histological preparations, bundles of smooth muscle cells were seen localized in the endocardium in 23 of the 44 specimens (none of the 6 controls). These cells were immunopositive for &agr;-smooth muscle actin, type 2 smooth muscle myosin, desmin, and calponin, but were negative for embryonic smooth muscle myosin, vimentin, fibronectin, and periostin. This profile is indicative of a late differentiation (contractile) smooth muscle phenotype. Electron microscopy confirmed that phenotype, revealing the cells to contain abundant myofilaments with dense bodies but little rough endoplasmic reticulum or Golgi apparatus. In the endocardial smooth muscle–positive group, the left ventricular end-systolic volume index (73±34 versus 105±50 mL/m2; P=0.021), left ventricular peak wall stress (164±47 versus 196±43 dynes 103/cm2; P=0.023), and left ventricular end-systolic meridional wall stress (97±38 versus 121±37 dynes 103/cm2; P=0.036) were all significantly smaller, and the ejection fraction was larger (41±8.8 versus 33±9.3%; P=0.005) than in the endocardial smooth muscle–negative group. However, no histological parameters differed between the 2 groups. Conclusions—Endocardial smooth muscle cell bundles in hearts with dilated cardiomyopathy exhibit a mature contractile phenotype and may play a compensatory role mitigating heart failure by reducing left ventricular wall stress and systolic dysfunction.