In skeletal muscles, oxygen free radicals generated during ischemia -reperfusion are known as inducers that cause cellular injury and apoptosis and contribute to the pathogenensis of reperfusion injury. Ischemia -reperfusion for 2 hours may cause reversible changes, while prolonged ischemia -reperfusion causes irreversible changes. Following ischemia -reperfusion, diverse signals are transduced to induce a variety of gene expression. Ischemic preconditioning, defined as brief episodes of ischemia and reperfusion, is known to provide protection from the consequences of prolonged ischemia followed by reperfusion. NF -kappa B is a transcription factor that activated during ischemic preconditioning and ischemia -reperfusion. It initiates inflammation through inducing transcription of proinflammatory, procoagulant and vasoactive gene, while mediates the expression of cytoprotective proteins that block apoptosis or inhibit inflammation. The present study was performed to study the change of NF -kappa B immunoreacitvity in rat anterior tibialis and soleus muscles in response to ischemia -reperfusion and preconditioning. Experimental animals, Sprague -Dawley rats (250 ~300 g), were divided into 6 groups; 1) control, 2) ischemic preconditioning, 3) 2 hours of ischemia, 4) 4 hours of ischemia, 5) 2 hours of ischemia after ischemic preconditioning, 6) 4 hours ischemia after ischemic preconditioning. For ischemic preconditioning, left common iliac artery was occluded three times for 5 minutes followed by 5 minutes of reperfusion using vascular clamp. Ischemia was done by occlusion of the same artery for 2 or 4 hours. The specimens of tibialis anterior and soleus muscles were obtained 0, 1, 3, 6, and 24 hours after onset of reperfusion. The specimens were paraffin sectioned at 6 micrometer and NF -kappa B expression was examined using immunohistochemical methods. The results obtained were as follows : 1. In normal control group, immunoreactivity of NF -kappa B was moderate to strong in tibialis anterior muscles and weak in soleus muscles. 2. In tibialis anterior, immunoreactivity of NF -kappa B was decreased in 2 and 4 hours of ischemia comparede with normal control group. In soleus muscle, immunoreactivity of NF -kappa B was decreased in 2 hours of ischemia but it was comparable to that of normal control group in 4 hours of ischemia. 3. Ischemia for 4 hours induced more remarkable change in NF -kappa B immunoreactivity than that for 2 hours. 4. After ischemic preconditioning, changes in NF -kappa B immunoreactivity after 2 and 4 hours of ischemia were decreased compared with normal control group. 5. In ischemia for 2 and 4 hours, changes in NF -kappa B immunoreactivity of tibialis anterior muscles were more severe than that of soleus muscles. These results suggest that in the skeletal muscle, changes in NF -kappa B immunoreactivity of 4 hours of ischemia were more remarkable than that of 2 hours ischemia, and changes in NF -kappa B of tibialis anterior muscles were more remarkable than that of soleus muscles. Ischemic preconditiong attenuated the alteration of the NF -kappa B immunoreactivity induced by ischemia -reperfusion in the muscles.
Animals ; Apoptosis ; Arteries ; Free Radicals ; Gene Expression ; Iliac Artery ; Inflammation ; Ischemia ; Ischemic Preconditioning ; Muscle, Skeletal ; Muscles* ; Oxygen ; Paraffin ; Rats* ; Reperfusion ; Reperfusion Injury ; Transcription Factors

Akt, heat shock protein (HSP72)72, and HSP90 induced by ischemic preconditioning protect cells from the ischemic injury. The purpose of this study was to examine the alterations of the level of phospho-Akt, HSP72, and HSP90 in the rat tibialis anterior and soleus muscles after cyclic episodes of ischemic preconditioning. Sprague-Dawley rats aged 35 weeks were divided into control and ischemic preconditioning (IP) groups. The IP group was divided into 3 subgroups based on cycles of IP. Left common iliac artery was occluded 3, 6, and 10 times for 5 minutes, followed by 5 minutes reperfusion. The experimental animals were sacrificed at 0, 3, 6, 24, and 72 hours after reperfusion, and left tibialis anterior and soleus muscles were removed. The expression of phospho-Akt, HSP72, and HSP90 were examined with immunohistochemical methods and Western blot analysis. The results were as follows; 1. In the 3 and 6 times of IP groups, the expression of phospho-Akt (p-Akt) was increased at 0 and 3 hours after reperfusion, compared with control group. The expression of p-Akt in the 10 times of IP group was lower than that in 3 and 6 times of IP groups. At 72 hours after reperfusion, the expression of p-Akt showed no difference among the IP groups. The expression of p-Akt was higher in Soleus than that in Tibialis anterior. 2. The expression of HSP72 in 3 times of IP group increased at 0 and 3 hours after reperfusion, compared with 6 and 10 times of IP groups. The expression of HSP72 in the 10 times of IP group was lower than that in 3 and 6 times of IP groups. At 72 hours after reperfusion, the expression of HSP72 showed no difference among the IP groups. The expression of HSP72 was higher in Soleus than that in Tibialis anterior. 3. In the 3 and 6 times of IP groups, the expression of HSP90 increased at 0 and 3 hours after reperfusion, compared with control group. The expression of HSP90 in the 10 times of IP group was lower than that in 3 and 6 times of IP groups. At 24 hours after reperfusion, the expression of HSP90 showed no difference with increasing episode of IP. The expression level of HSP90 was higher in Soleus than that in Tibialis anterior. These findings suggest that ischemic preconditioning increases the expression of p-Akt, HSP72 and HSP90 at early phase after reperfusion in the rat tibialis anterior and soleus muscles. However, increased cycles of ischemic preconditioning may not induce the expression of them.
Animals ; Blotting, Western ; Heat-Shock Proteins ; Iliac Artery ; Ischemic Preconditioning* ; Muscles* ; Phosphorylation* ; Rats* ; Rats, Sprague-Dawley ; Reperfusion

Ischemic preconditioning (IP), short pre-treatment sublethal ischemia, induces a state of protection against subsequent prolonged ischemia-reperfusion. The purpose of this study was to investigate the expression of HO-1, HSP70, and iNOS proteins in the liver subjected to the courses of reperfusion after repetitive cycles of remote IP in the rat. Using thirty five week-old rats, the remote preconditioning was undertaken by vascular clamp occlusion of blood flow to one hindlimb, with 3 and 10 cycles of 5 minutes occlusion followed by 5 minutes reperfusion. The liver was removed 0, 3, 6, 24, and 72 hours of reperfusion after remote IP and assayed by immunohistochemical staining and Western blotting analyses for anti-HO-1, anti-HSP70, and anti-iNOS antibodies. The expression of HO-1 in rat liver increased at 72 hours of reperfusion groups after 3 and 10 cycles of remote IP, compared with normal control groups. The expression of HSP70 in rat liver increased at 6 hours of reperfusion groups after 3 cycles of remote IP, compared with normal control groups. The expression of HSP70 in rat liver increased at 0 hour of reperfusion groups after 10 cycles of remote IP, compared with normal control groups and decreased at 24 and 72 hours of reperfusion groups. The expression of iNOS in rat liver increased at 24 hours of reperfusion groups, but decreased at 72 hours of reperfusion groups after 3 and 10 cycles of remote IP, compared with normal control groups. In summary, these results showed that at early phase of reperfusion after remote IP, HSP70 expression was increased in rat liver. However, at 72 hrs of reperfusion after remote IP, HO-1 expression was increased and iNOS expression was decreased in rat liver.
Animals ; Blotting, Western ; Hindlimb ; Ischemia ; Ischemic Preconditioning ; Liver ; Proteins ; Rats ; Reperfusion

No abstract available.
Dermatitis, Allergic Contact

Akt, a key protein of cell survival, can promote cell growth and survival by activations of various cellular protective factors. Ischemic preconditioning (IP) has been known to reduce ischemic injury through upregulation of phosphorylation of Akt (p-Akt). CuZn-superoxide dismutase (SOD-1), an antioxidant enzyme, scavenges reactive oxygen species and protects cell from oxidative stress by increasing the activaiton of Akt. The present study was performed to examine the effects of IP on the expression of p-Akt and SOD-1 in the ischemicreperfused rat skeletal muscles. Thirty weeks old male SD rats were divided into 4 groups, such as controls, IP, 4 hour ischemia and 4 hour ischemia with IP. For IP, commom iliac artery was occluded three times for 5 min ischemia followed by 5 min reperfusion using rodent vascular clamps. Ischemia was induced by occlusion on the same artery for 4 hours. The Tibialis anterior and Soleus were removed at 0, 1, 3, and 24 hours of reperfusion. The expressions of p-Akt (Ser 473) and SOD-1 were examined with immunohistochemistry and Western blot analysis.In the IP group, the p-Akt and SOD-1 were increased, compared to the control group. In the ischemia group, the p- Akt and SOD-1 were decreased, compared to the control group, and were more abundant when reperfusion time were increased. IP increased the p-Akt and SOD-1 after 4 hour ischemia, and the p-Akt and SOD-1 were higher in Soleus compared to Tibialis anterior. These findings suggest that IP increases p-Akt and expression of SOD-1 in the ischemic-reperfused rat skeletal muscles, and that upregulations of p-Akt and SOD-1 induced by IP were higher in the red muscle fiber, Soleus, than the white muscle fiber, Tibialis anterior.
Animals ; Arteries ; Blotting, Western ; Cell Survival ; Humans ; Iliac Artery ; Immunohistochemistry ; Ischemia ; Ischemic Preconditioning ; Male ; Muscle Fibers, Fast-Twitch ; Muscle Fibers, Slow-Twitch ; Muscle, Skeletal ; Oxidative Stress ; Phosphorylation ; Rats ; Reactive Oxygen Species ; Reperfusion ; Rodentia ; Up-Regulation

A balance between production and degradation of reactive oxygen species has an important role in the cardiovascular homeostasis, and is known to contribute to hypertension. Under oxidative stress, an upregulation of inducible NOS (iNOS) induces ischemic-reperfusion injury, and is involved in the pathophysiology of the hypertension. Ischemic-reperfusion injury of the skeletal muscle results from reactive oxygen species, and overexpression of iNOS in the skeletal muscle increases the ischemic injury. Superoxide dismutase (SOD), antioxidant, is a major enzyme for degradation of reactive oxygen species (ROS). The purpose of this study was to observe the effect ischemic preconditioning (IP) of the lower limb on the expression of iNOS, CuZnSOD and MnSOD in the white and red muscle of the spontaneously hypertensive rat (SHR). Nine weeks old male normotensive rat (Wistar-Kyoto rat, WKY) and SHR were divided into control and IP groups. The IP group was further divided into 3 (3IP) and 10 (10IP) times of IP. Left common iliac artery was occluded 3 and 10 times for 5 min of ischemia-5 min of reperfusion using rodent vascular clamp. The animals were sacrificed at 0, 0.5, 1, and 3 hours after reperfusion and the Tibialis anterior and Soleus were removed. The expressions of iNOS, CuZnSOD and MnSOD in the skeletal muscle were examined with immunohistochemical methods and Western blot analysis. iNOS was expressed in Tibialis anterior, but in Soleus after IP. The expression of iNOS was increased in both WKY and SHR, it was higher in SHR than WKY. CuZnSOD and MnSOD were expressed in Tibialis anterior and Soleus, higher in Soleus, after IP. The expression of CuZnSOD and MnSOD were increased in both WKY and SHR, higher in WKY than SHR. It is consequently suggested that hypertensive individual and white muscle are more sensitive to ischemic injury of the skeletal muscle as considering their high expression of iNOS and low expression of SODs.
Animals ; Blotting, Western ; European Continental Ancestry Group ; Homeostasis ; Humans ; Hypertension ; Iliac Artery ; Ischemic Preconditioning ; Lower Extremity ; Male ; Muscle, Skeletal ; Muscles ; Oxidative Stress ; Rats ; Rats, Inbred SHR ; Reactive Oxygen Species ; Reperfusion ; Rodentia ; Superoxide Dismutase ; Superoxides ; Up-Regulation

The Valeri method, introduced in 1972 with dimethyl sulfoxide (DMSO) as a cryoprotective agent, has evolved into a no-wash technique widely employed for cryopreservation of platelets. Cryopreserved platelets (CP) are a viable alternative to liquid platelets (LP) and address the limitations of LP. CP have an extended shelf file, are a reliable supply for rare blood types, and can be transported to challenging terrains for use in military settings and remote areas and places where blood supply is imbalanced. Despite CP exhibiting a lower recovery rate compared to LP, the superior hemostatic efficacy makes it advantageous for use in bleeding patients. Some countries have already implemented CP for civilian use in disasters caused by natural hazards or human-induced events, and clinical trials are underway to expand applications among civilians. The absence of national regulations and standardized guidelines for CP preparation and evaluation is a significant obstacle to the extensive use of CP. A consensus is needed among academic societies, blood centers, the military, and governments to lend support and interest in the development of CP as a viable alternative to LP. This review presents information on the initial attempts to produce CP, in vitro changes of thawed CP, utilization of CP, and the current usage status in various countries. The goal was to evaluate the potential need to introduce CP domestically and provide insights on the strengths and challenges.

Adriamycin, a member of anthracycline class isolated from the culture medium of Streptomyces peucetius var. caesius, is one of the most effective and useful anti-neoplastic agents for the treatment of hematological and solid malignancy. The efficacy of adriamycin has been shown to mainly rely on free radicals generated during its inhibitory activity of DNA synthesis and its metabolism. However, the production of free radicals elicited side effects including chronic cardiotoxicityits. Such aspect occasionally limits the therapeutic use of adriamycin. This study was aimed to investigate the dose effect of adriamycin on apoptosis induction of the myocardium of left ventricle along with the effect of vitamin C on adriamycin activity. SD rats weighing 250~300 g were injected intraperitoneally with either 20 mg/kg or 5 mg/kg of adriamycin and drunk vitamin C at 1000 mg/animal/day. Rats were sacrificed at days 1, 3, 5, and 14 after injection but rats injected with 20 mg adriamycin died at day 6. The heart were paraffin -sectioned at 6 micrometer thickness, stained following TUNEL methods. Stained cells undergoing apoptotic death in myocardium of left ventricle were counted and statistically analyzed. The results were obtained as follows; 1. The muscular layer of left ventricle from nomal rats contained 4.2+/-12.3 apoptotic cells. 2. Distributions of apoptotic cells from rats treated with 20 mg/kg adriamycin were 26.7+/-43.2, 17.7+/-41.4, and 17.8+/- 31.8 at days 1, 3, and 5, respectively. 3. Distributions of apoptotic cells from rats treated with 5 mg/kg adriamycin were 10.8+/-19.5, 4.1+/-12.4, 5.6+/-10.2, 10.8+/-17.3 at days 1, 3, 5, and 14, respectively. 4. Coadministration of 20 mg/kg adriamycin and vitamin C resulted in 8.8+/-19.5, 4.1+/-12.4, and 16.2+/-33.6 apop-totic cells at days 1, 3, and 5 post administration. 5. Coadministration of 5 mg/kg adriamycin and vitamin C resulted in 17.0+/-32.3, 12.2+/-19.7, 7.1+/-14.0, and 7.2+/-16.7 apoptotic cells at days 1, 3, 5, and 14 post administration. Taken together, these results demonstrated that treatment of rats with 20 mg/kg adriamycin induced more apoptotic cells than that with 5 mg/kg adriamycin and vitamin C reduced such cytotoxic effects of adriamycin.
Animals ; Apoptosis* ; Ascorbic Acid* ; DNA ; Doxorubicin ; Free Radicals ; Heart ; Heart Ventricles ; In Situ Nick-End Labeling ; Metabolism ; Myocardium ; Paraffin ; Rats* ; Streptomyces ; Vitamins*

As the interest in health is increasing and the population enjoyed the leisure sports is steadily increasing, the stress fracture, fracture or variant of metatarsal bone of foot has been shown frequently. The mistaken estimation about the length and rank of metatarsal bones during the osteotomy of the metatarsal bones of foot can be complicated. It is essential to have detailed knowledges about the anatomical structure of surgical region. This study aimed to investigate the metatarsal bones of foot and to develop a regression equation that can predict the length of metatarsal bones during the osteotomy. The subject of this study is fifty four feet (30M/224F). We measured the whole length and the article length of metatarsal bone. Also, we measured the whole width and the article width in the head, body, base of the metatarsal bone. The data was analyzed using SPSS win 13.0. The regression equation models of length of the metatarsal bones were developed by multiple regression analysis. The regression equation predicted first metatarsal length was second metatarsal articular length x0.770+7.780, second metatarsal length was third metatarsal length x0.976+6.050, third metatarsal length was fourth metatarsal length x1.000+0.922, fourth metatarsal length was third metatarsal length x0.917+4.167, fifth metatarsal length was fourth metatarsal length x0.901+7.972. The results of this study would be useful to clarify the characteristics of the metatarsal bone of the foot, to develop a regression equation for prediction of the length of the metatarsal bone.
Foot ; Fractures, Stress ; Head ; Leisure Activities ; Metatarsal Bones ; Osteotomy ; Sports

Remote ischemic preconditioning (IP), brief tolerating cycles of ischemia and reperfusion in remote non-vital organs, can reduce ischemic injury of the heart. IP induces cardiac protection by down-regulating iNOS or up-regulating eNOS. In addition, Akt has been known to protect myocardium against ischemia-reperfusion injury. This study was undertaken to observe the expression of iNOS, eNOS, Akt and phospho-Akt (p-Akt) in the rat myocardium after IP. Thirty-five weeks-old male Sprague-Dawley rats were divided into control and IP groups. The IP group was further subdivided into 3 groups based on the number of cycles of IP. For IP, left commom iliac artery was occluded 3, 6 and 10 cycles for 5 min of ischemia alternating with 5 min of reperfusion. The rat were sacrificed at 0, 3, 6, 24 and 72 hours of IP and the heart was removed. The expression of iNOS, eNOS, Akt and p-Akt in the rat myocardium was examined by immunohistochemical staining and Western blot analysis. The expression of iNOS was increased by IP and was higher in 10IP groups than 3IP and 6IP group. The expression of eNOS was increased or decreased by IP and was showed no difference with increasing episode of IP. The expression of Akt was decreased by IP at 24 and 72 hours after reperfusion, and showed no differences with increasing episode of IP. The expression of p-Akt was increased by IP and showed no difference with increasing episode of IP. These results suggest that hind limb ischemic preconditioning provides cardiac protection through up-regulation of eNOS and phosphorylation of Akt, however excessive episodes of remote preconditioning may induce the myocardial ischemic injury through overexpression of iNOS.
Animals ; Blotting, Western ; Extremities ; Heart ; Humans ; Iliac Artery ; Ischemia ; Ischemic Preconditioning ; Male ; Myocardium ; Phosphorylation ; Rats ; Rats, Sprague-Dawley ; Reperfusion ; Reperfusion Injury ; Up-Regulation