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Ferulic acid modulates nitric oxide synthase expression in focal cerebral ischemia.

Laboratory Animal Research.2012;28(4):273-278. doi:10.5625/lar.2012.28.4.273

Nitric oxide (NO) is generated by three different NO synthase (NOS) isoforms, endothelial NOS (eNOS), inducible NOS (iNOS), and neuronal NOS (nNOS). It is known that eNOS produces NO, which exerts a protective effect, while iNOS produces NO with neurotoxic effects. Ferulic acid preserves neuronal cells against from cerebral ischemia and glutamate-induced excitotoxicity. This study confirmed the neuroprotective effect of ferulic acid and investigated the levels of three NOS isoforms in focal cerebral ischemia with or without ferulic acid. Rats were immediately treated with ferulic acid (100 mg/kg, i.v.) after middle cerebral artery occlusion (MCAO). Brains tissues were collected at 24 h after the onset of occlusion. The expressions of these three isoforms in cerebral ischemia with ferulic acid were analyzed using Western blot technique. Ferulic acid treatment significantly decreases the number of TUNEL-positive cells in the cerebral cortex against MCAO injury. The levels of eNOS decreased in MCAO-operated animals, while ferulic acid treatment attenuated the MCAO-induced decrease of eNOS. However, iNOS and nNOS expression levels increased during MCAO, and ferulic acid prevented injury-induced increase of these isoforms. Thus, these findings suggest that the up- and down modulation of three isoforms by ferulic acid is associated with a neuroprotective mechanism.
Animals ; Blotting, Western ; Brain ; Brain Ischemia ; Cerebral Cortex ; Coumaric Acids ; Infarction, Middle Cerebral Artery ; Neurons ; Neuroprotective Agents ; Nitric Oxide ; Nitric Oxide Synthase ; Protein Isoforms ; Rats

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Anti-obesity effects of Rapha diet(R) preparation in mice fed a high-fat diet.

Jihyun KIM ; Jangbeen KYUNG ; Dajeong KIM ; Ehn Kyoung CHOI ; Paul BANG ; Dongsun PARK ; Yun Bae KIM

Laboratory Animal Research.2012;28(4):265-271. doi:10.5625/lar.2012.28.4.265

The anti-obesity activities of Rapha diet(R) preparation containing silkworm pupa peptide, Garcinia cambogia, white bean extract, mango extract, raspberry extract, cocoa extract, and green tea extract were investigated in mice with dietary obesity. Male C57BL/6 mice were fed a high-fat diet (HFD) containing 3% Rapha diet(R) preparation for 8 weeks, and blood and tissue parameters of obesity were analyzed. The HFD markedly enhanced body weight gain by increasing the weights of epididymal, perirenal, and mesenteric adipose tissues. The increased body weight gain induced by HFD was significantly reduced by feeding Rapha diet(R) preparation, in which decreases in the weight of abdominal adipose tissue and the size of abdominal adipocytes were confirmed by microscopic examination. Long-term feeding of HFD increased blood triglycerides and cholesterol levels, leading to hepatic lipid accumulation. However, Rapha diet(R) preparation not only reversed the blood lipid levels, but also attenuated hepatic steatosis. The results indicate that Rapha diet(R) preparation could improve HFD-induced obesity by reducing both lipid accumulation and the size of adipocytes.
Abdominal Fat ; Adipocytes ; Animals ; Body Weight ; Bombyx ; Cacao ; Cholesterol ; Diet, High-Fat ; European Continental Ancestry Group ; Garcinia cambogia ; Humans ; Male ; Mangifera ; Mice ; Obesity ; Pupa ; Tea ; Triglycerides ; Weights and Measures

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Human leukocytes regulate ganglioside expression in cultured micro-pig aortic endothelial cells.

Jin Hyoung CHO ; Ji Su KIM ; Malg Um LIM ; Hyun Ki MIN ; Dong Hoon KWAK ; Jae Sung RYU ; Ju Taek LEE ; Sun Uk KIM ; Chang Hwan KIM ; Chang Hyun KIM ; Deog Bon KOO ; Kyu Tae CHANG ; Young Kug CHOO

Laboratory Animal Research.2012;28(4):255-263. doi:10.5625/lar.2012.28.4.255

Gangliosides are ubiquitous components of the membranes of mammalian cells that are thought to play important roles in various cell functions such as cell-cell interaction, cell adhesion, cell differentiation, growth control, and signaling. However, the role that gangliosides play in the immune rejection response after xenotransplantation is not yet clearly understood. In this study, the regulatory effects of human leukocytes on ganglioside expression in primary cultured micro-pig aortic endothelial cells (PAECs) were investigated. To determine the impact of human leukocytes on the expression of gangliosides in PAECs, we performed high-performance thin layer chromatography (HPTLC) in PAECs incubated with FBS, FBS containing human leukocytes, human serum containing human leukocytes, and FBS containing TNF-alpha. Both HPTLC and immunohistochemistry analyses revealed that PAECs incubated with FBS predominantly express the gangliosides GM3, GM1, and GD3. However, the expression of GM1 significantly decreased in PAECs incubated for 5 h with TNF-alpha (10 ng/mL), 10% human serum containing human leukocytes, and 10% FBS containing human leukocytes. Taken together, these results suggest that human leukocytes induced changes in the expression profile of ganglioside GM1 similar to those seen upon treatment of PAECs with TNF-alpha. This finding may be relevant for designing future therapeutic strategies intended to prolong xenograft survival.
Cell Adhesion ; Cell Communication ; Chromatography, Thin Layer ; Endothelial Cells ; Gangliosides ; Humans ; Immunohistochemistry ; Leukocytes ; Membranes ; Rejection (Psychology) ; Transplantation, Heterologous ; Tumor Necrosis Factor-alpha

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Analysis of reference interval and age-related changes in serum biochemistry and hematology in the specific pathogen free miniature pig.

Su Cheong YEOM ; Seong Yong CHO ; Chung Gyu PARK ; Wang Jae LEE

Laboratory Animal Research.2012;28(4):245-253. doi:10.5625/lar.2012.28.4.245

We analyzed 72 items related to serum biochemistry and hematology in 85 specific pathogen-free (SPF) Seoul National University (SNU) miniature pigs aged 1- to 36-months which originated from a Minnesota miniature pig. Almost all examined items were similar between male and female pigs. However, some items such as Cr level, B/C ratio, C.R.F, LDH, LAP and T4 were significantly different between male and female pigs (P<0.05). Thirty four examined items showed age-related changes, and the significant changes were observed in animals less than six months old. The values for BUN, K, uric acid, Ca, Ca++, and Pi were significantly higher in pigs younger than six months of age, which might reflect poor kidney function in young pigs. Additionally, TIBC, UIBC and RDW were significantly higher in young pigs, and RBC, Hb, HCT, MCHC and MCV were significantly lower in young pigs, thus indicating a similar physiology of iron deficiency anemia. These age-related specific phenotypes seemed to be normal, but it should be considered in the long-term experiment using the young pigs. In conclusion, in this study, we defined the normal reference intervals for SPF SNU miniature pigs, and we also determined that there are some physiological differences between the pig genders and ages. This study provides fundamental data for use in experiments involving SPF SNU miniature pigs.
Aged ; Anemia, Iron-Deficiency ; Animals ; Biochemistry ; Female ; Hematology ; Humans ; Kidney ; Male ; Minnesota ; Phenotype ; Specific Pathogen-Free Organisms ; Swine ; Uric Acid

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Early diagnosis of radiodermatitis using lactate dehydrogenase isozymes in hairless mice (SKH1-hr).

Sung Kyu CHO ; Won Dong KIM

Laboratory Animal Research.2012;28(4):239-244. doi:10.5625/lar.2012.28.4.239

In this study, we evaluate a method for the early diagnosis of radiodermatitis for use in the prevention and therapy of this condition. Hairless mice (SKH1-hr) were used to study the early diagnosis of radiodermatitis. Lactate dehydrogenase (LDH, EC 1.1.1.27) isozymes were analyzed using native-polyacrylamide gel electrophoresis and western blotting of blood serum and tissues collected from SKH1-hr mice. Radiodermatitis developed 24 days after the first X-irradiation. Reduced spleen weight was observed after the last X-irradiation (P<0.05). Thereafter the weight increased until 24 days after the first irradiation, finally reaching levels comparable to those in the sham-irradiated control group. LDH activity was the highest in skeletal muscle and lowest in blood serum. LDH C4, A4, A3B, A2B2, AB3, and B4 isozymes were detected, in the mentioned order, from the cathode. This result was similar in other mouse strains. In the irradiated group, LDH A4 isozyme levels were reduced in the serum until inflammation occurred, whereas those of B4 isozyme were elevated. The subunits A and B followed a similar trend to that of LDH A4 and B4 isozyme, respectively. Importantly, antibodies against LDH B4 isozyme could prove useful in the early diagnosis of radiodermatitis.
Animals ; Antibodies ; Blotting, Western ; Early Diagnosis ; Electrodes ; Electrophoresis ; Inflammation ; Isoenzymes ; L-Lactate Dehydrogenase ; Lactic Acid ; Mice ; Mice, Hairless ; Muscle, Skeletal ; Radiodermatitis ; Serum ; Spleen

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Toxicity of red Liriope platyphylla manufactured by steaming process on liver and kidney organs of ICR mice.

Sun Il CHOI ; Ji Eun KIM ; In Sik HWANG ; Hye Ryun LEE ; Young Ju LEE ; Moon Hwa KWAK ; Hong Joo SON ; Hee Seob LEE ; Jong Sub LEE ; Byeong Cheol KANG ; Dae Youn HWANG

Laboratory Animal Research.2012;28(4):229-238. doi:10.5625/lar.2012.28.4.229

Red Liriope platyphylla (RLP) produced by steaming process has been reported to enhance the secretion of insulin and nerve growth factor (NGF). However, there has been no report on the toxicity of RLP in the specific organs of mice. To investigate the toxic effect of RLP, we tried to observe a significant alteration on body weight, food/water intake, organ weight, liver pathology and kidney pathology in female ICR mice received 12.5, 25.0 and 50.0 mg/kg body weight/day of RLP via gavage for 10 days. Out of seven organs including brain, heart, lung, liver, kidney, spleen and ovary, two organs (heart and lung) showed significantly decreased weights in the medium dosage RLP-treated group, whereas weights of other organs were maintained at constant levels in all dosage groups. In the liver toxicity analysis, no significant increase of alkaline phosphatase (ALP), alanine aminotransferase (ALT), and aspartate amino-transferase (AST) were detected in any RLP-treated group compared to vehicle-treated group. The specific pathological changes induced by most of toxic compounds were not observed in the liver in microscopic examination. Furthermore, in the kidney toxicological analysis, a significant enhancement of the blood urea nitrogen (BUN) concentration was detected in the high dosage RLP-treated group compared to the vehicle-treated group. However, the serum creatinine (CA) concentration on the serum biochemistry as well as the pathological changes in microscopic examination were not significantly different between the vehicle- and RLP-treated groups. Therefore, these results suggest that RLP does not induce any specific toxicity in liver or kidney tissues of mice, although the BUN level slightly increased in 50.0 mg/kg of RLP-treated group.
Alanine Transaminase ; Alkaline Phosphatase ; Animals ; Aspartic Acid ; Biochemistry ; Blood Urea Nitrogen ; Body Weight ; Brain ; Creatinine ; Female ; Heart ; Humans ; Insulin ; Kidney ; Liver ; Lung ; Mice ; Mice, Inbred ICR ; Nerve Growth Factor ; Organ Size ; Ovary ; Spleen ; Steam ; Weights and Measures

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Development of a canine model for recurrent laryngeal injury by harmonic scalpel.

Kyu Eun LEE ; Hyeon Gun JEE ; Hoon Yub KIM ; Won Seo PARK ; Sung Hye PARK ; Yeo Kyu YOUN

Laboratory Animal Research.2012;28(4):223-228. doi:10.5625/lar.2012.28.4.223

Various energy devices had been used in thyroid surgery. Aim of study is to develop canine model for recurrent laryngeal nerve injury by harmonic scalpel and to evaluate feasibility of using this model for evaluating the safety use of harmonic scalpel during thyroid surgery. Nine dogs were divided into 3 groups according to distance between harmonic scalpel application and recurrent laryngeal nerve; group 1 (1 mm), 2 (2 mm), and 3 (3 mm). Vocal cord function was assessed pre- and postoperatively using video laryngoscopy. Harmonic scalpel was applied adjacent to left recurrent laryngeal nerve and, two weeks later, right recurrent laryngeal nerve at assigned distances. Recurrent laryngeal nerves were evaluated for subacute and acute morphologic changes. Laryngoscopy demonstrated 3 abnormal vocal cords in group 1, 1 in group 2, and no in group 3 (P=0.020). Subacute histologic changes were observed in nerves with abnormal function. Acute histologic changes were observed 5/8 (62.5%) in group 1, 1/7 (14.3%) in group 2, and not in group 3. We developed canine model for recurrent laryngeal injury. The functional outcomes matched with the histologic changes. These warrant further study to determine the safety margin for energy device in vicinity of recurrent laryngeal nerve.
Animals ; Dogs ; Laryngoscopy ; Recurrent Laryngeal Nerve ; Recurrent Laryngeal Nerve Injuries ; Thyroid Gland ; Vocal Cords

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Radiation-induced eosinophilic, polymorphic, and pruritic eruption in a pig skin model.

Joong Sun KIM ; Won Seok JANG ; Min Ji BAE ; Sunhoo PARK ; Seung Sook LEE

Laboratory Animal Research.2015;31(4):204-208. doi:10.5625/lar.2015.31.4.204

Eosinophilic, polymorphic and pruritic eruption associated with radiotherapy (EPPER) can occur in cancer patients after irradiation. In this study, we characterized the clinical and histopathological features of pig skin that developed widespread polymorphic and pruritic skin lesions following localized 50 Gy gamma-irradiation. The pigs developed pruritus 5-7 weeks after irradiation, and infiltration of the dermis by eosinophils was detected 4-7 weeks after irradiation. The irradiated animals also showed transiently increased numbers of peripheral eosinophils 5-7 weeks after treatment. Irradiation induced desquamation after 2-4 weeks, which and the desquamation gradually resolved after 7 weeks. These pathological changes correspond to those seen in irradiated human skin, indicating that this model could be useful for elucidating the pathogenesis of EPPER and for developing therapeutic and prophylactic methods.
Animals ; Dermis ; Eosinophils* ; Humans ; Pruritus ; Radiotherapy ; Skin* ; Swine

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Curcumin attenuates the middle cerebral artery occlusion-induced reduction in gamma-enolase expression in an animal model.

Sang Ah GIM ; So Ra LEE ; Fawad Ali SHAH ; Phil Ok KOH

Laboratory Animal Research.2015;31(4):198-203. doi:10.5625/lar.2015.31.4.198

Curcumin exerts a protective effect in cerebral ischemia through its anti-oxidant and anti-inflammatory activities. gamma-enolase is a glycolytic enzyme expressed in neurons that is known to exerts a neuroprotective effect. We investigated whether curcumin regulates gamma-enolase expression in focal cerebral ischemic injury in rats. Middle cerebral artery occlusion (MCAO) was performed to induce focal cerebral ischemia. Adult male rats were injected intraperitoneally with either vehicle or curcumin (50 mg/kg) 1 h after MCAO and cerebral cortex tissues were isolated 24 h after MCAO. We found that MCAO-induced injury resulted in a reduction in gamma-enolase expression in vehicle-treated animals using a proteomics approach. However, this reduction was attenuated in animals with MCAO treated with curcumin. Reverse-transcription PCR and Western blot analyses also showed that curcumin treatment prevented the MCAO injury-induced reduction in gamma-enolase expression. The results of this study suggest that curcumin exerts its neuroprotective function in focal cerebral ischemia by regulating the expression of gamma-enolase.
Adult ; Animals* ; Blotting, Western ; Brain Ischemia ; Cerebral Cortex ; Curcumin* ; Humans ; Infarction, Middle Cerebral Artery ; Male ; Middle Cerebral Artery* ; Models, Animal* ; Neurons ; Neuroprotective Agents ; Phosphopyruvate Hydratase* ; Polymerase Chain Reaction ; Proteomics ; Rats

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Tissue transglutaminase-interleukin-6 axis facilitates peritoneal tumor spreading and metastasis of human ovarian cancer cells.

Keunhee OH ; Hyeong Gon MOON ; Dong Sup LEE ; Young Bok YOO

Laboratory Animal Research.2015;31(4):188-197. doi:10.5625/lar.2015.31.4.188

Inflammation has recently been implicated in cancer formation and progression. As tissue transglutaminase (TG2) has been associated with both inflammatory signaling and tumor cell behavior, we propose that TG2 may be an important link inducing interleukin-6 (IL-6)-mediated cancer cell aggressiveness, including cancer stem cell-like characteristics and distant hematogenous metastasis. We evaluated the effect of differential TG2 and IL-6 expression on in vivo distant metastasis of human ovarian cancer cells. IL-6 production in human ovarian cancer cells was dependent on their TG2 expression levels. The size and efficiency of tumor sphere formation were correlated with TG2 expression levels and were dependent on TG2-mediated IL-6 secretion in human ovarian cancer cells. Primary tumor growth and propagation in the peritoneum and distant hematogenous metastasis into the liver and lung were also dependent on TG2 and downstream IL-6 expression levels in human ovarian cancer cells. In this report, we provide evidence that TG2 is an important link in IL-6-mediated tumor cell aggressiveness, and that TG2 and downstream IL-6 could be important mediators of distant hematogenous metastasis of human ovarian cancer cells. Intervention specific to TG2 and/or downstream IL-6 in ovarian cancer cells could provide a promising means to control tumor metastasis.
Axis, Cervical Vertebra* ; Humans* ; Inflammation ; Interleukin-6 ; Liver ; Lung ; Neoplasm Metastasis* ; Ovarian Neoplasms* ; Peritoneum

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