Comparative studies of skeletal muscle proteome and transcriptome profilings between pig breeds.

Two genetically different pig breeds, the Korean native pig (KNP) and the Western meat-producing Landrace, show breed-specific traits in stress responsiveness (stress hormone levels), growth performance (live weight), and meat quality (intramuscular fat content). We analyzed expression levels within the proteome and transcriptome of the longissimus muscles of both breeds using two-dimensional electrophoresis (2-DE) and microarray analysis. We constructed a porcine proteome database focused mainly on mitochondrial proteins. In total, 101 proteins were identified, of which approximately 60% were metabolic enzymes and mitochondrial proteins. We screened several proteins and genes related to stress and metabolism in skeletal muscles using comparative analysis. In particular, three stress-related genes (heat shock protein beta-1, stress-70 protein, and heat shock 70 kDa protein) were more highly expressed in the Landrace than in the KNP breed. Six metabolism-related genes (peroxisome proliferative activated receptor alpha, short-chain acyl-CoA dehydrogenase, succinate dehydrogenase, NADH-ubiquinone oxidoreductase, glycerol-3-phosphate dehydrogenase, and sterol regulatory element binding protein-1c), all of which are involved in energy and lipid metabolism, were more highly expressed at the protein or mRNA level in the KNP breed. These data may reflect the breed dependence of traits such as stress responsiveness, growth performance, and meat quality.

The micronucleus frequency in cytokinesis-blocked lymphocytes of cattle in the vicinity of a nuclear power plant.

Cytogenetic and hematological analyses were performed on the peripheral blood lymphocytes (PBLs) obtained from Korean native cattle bred in the vicinity of three nuclear power plants (Wolsong, Uljin and Yeonggwang) and in a control area. The micronucleus (MN) rates for the cattle from the Wolsong, Uljin and Yeonggwang nuclear power plants and for the control area were 9.87 +/- 2.64, 8.90 +/- 3.84, 9.20 +/- 3.68 and 9.60 +/- 3.91 per 1,000 cytokinesis-blocked lymphocytes, respectively. The apparent difference is not statistically significant. The MN frequencies of PBLs from cattle bred in the four areas are within the background variation for this study. The MN frequencies and hematological values were similar regardless of whether the cattle were bred near a nuclear power plant or in the control area.

Somatostatin down-regulates LFA-1 activation by modulating Rap1 expression in CD4+ and CD8+ T cells.

Leukocyte function-associated antigen-1 (LFA-1) is one of the integrins that are expressed on the leukocytes, and has been shown to play an important role in leukocyte trafficking. The adhesive activity of LFA-1 is governed partially by the Rap1. This study examined that the relationship between LFA-1 and Rap1 mRNA expressions by anti-CD3 and anti-CD3+SOM treatment in the CD4+ and CD8+ T cells. The LFA-1 mRNA expression levels following the anti-CD3 and anti-CD3+SOM treatment for 30 min was greater on the CD8+ T cells, and the LFA-1 expression of the CD8+ T cells with anti-CD+SOM treatment was affected more severely than that of the CD4+ T cells. The Rap1 mRNA expression patterns following anti-CD3 and anti-CD3+SOM stimulation in the CD4+ and CD8+ T cells were similar to the LFA-1 expression patterns, and the expression level following anti-CD3+SOM treatment was suppressed more significantly in the CD8+ T cells. These results suggest that the difference in the Rap1 expression level after stimulation might explain the differences in the LFA-1 expression level on the T cell subsets, and that the down-regulation of Rap1 expression following SOM treatment is closely related to the diminished LFA-1 expression.

2,3,7,8-Tetrachlorodibenzo-p-dioxin activates ERK and p38 mitogen-activated protein kinases in RAW 264.7 cells.

2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) is a widespread environmental contaminant, exposure to it eliciting a broad spectrum of deleterious pathophysiological effects. Since mitogen-activated protein kinase (MAPK) pathways appear to play an important role in both cell survival and the apoptotic process, we assessed the effects of TCDD on the activation of extracellular signal-regulated kinase (ERK), Jun-N-terminal kinase (JNK), p38 MAPKs and caspase-3 in RAW 264.7 cells. TCDD treatment induced a transient upshift in ERK activity, followed by a decline, but a concomitant dramatic activation of p38. However, TCDD did not cause any apparent change in the activity of JNK, though it induced an up-regulation in caspase-3 activity. These results demonstrate that the equilibrium between the ERK and p38 pathways is critical to the fate of the cells, and that the activation of p38, upstream of caspase, plays an important role in the apoptotic process. The data obtained in this study also suggests that TCDD activates the MAPK pathway via an arylhydrocarbon receptor (AhR)-independent mechanism in RAW 264.7 murine macrophages.

A cytogenetic study of Korean native goat bred in the nuclear power plant using the micronucleus assay.

Cytogenetic and hematological analysis was performed on the peripheral blood lymphocytes (PBLs) obtained from Korean native goats bred in two nuclear power plants (Wolsong and Uljin) and a control area. The frequencies of gamma-ray-induced micronuclei (MN) in the cytokinesis-blocked (CB) lymphocytes at several doses were measured in three Korean native goats. The measurements performed after irradiation showed dose-related increases in the MN frequency in each of the donors. The results were analyzed using a linear-quadratic model with a line of best fit of y=0.1019D+0.0045D2+0.0093 (y=number of MN/CB cells and D=irradiation dose in Gy). The MN rates in the goats from the Wolsong and Uljin nuclear power plant, and the control area were 9.60+/-2.88, 6.83+/-1.47 and 9.88+/-4.32 per 1,000 CB lymphocytes, respectively. The apparent difference is not statistically significant. The MN frequencies of PBLs from goats bred in three areas means that the values are within the background variation in this experiment. The MN frequencies and hematological values were similar regardless of whether the goats were bred in the nuclear power plant or the control area.

Protective effect of ginsenosides, active ingredients of Panax ginseng, on kainic acid-induced neurotoxicity in rat hippocampus.

Ginsenosides are known to attenuate glutamate-induced cell injuries in vitro. We investigated the in vivo effect of ginsenosides on kainic acid (KA)-induced neurotoxicity in rat hippocampus using the methods of acid fuchsin (AF) staining and heat-shock protein-70 (HSP-70) immunoreactivity to detect neuronal death and stress, respectively. Pretreatment of ginsenosides (50 or 100 mg/kg for 7 days) via intraperitoneal (i.p.) administration significantly attenuated KA (10 mg/kg i.p.)-induced cell death by decreasing AF-positive neurons in both CA1 and CA3 regions of rat hippocampus compared with KA treatment alone. Pretreatment of ginsenosides (50 or 100 mg/kg for 7 days) via i.p. administration also significantly suppressed KA-induced induction of HSP-70 in both regions of rat hippocampus. These results show that ginsenosides are effective in protecting hippocampal CA1 and CA3 cells against KA-induced neurotoxicity.

Somatostatin controls LFA-1 gene expression by altering neuraminidase expression in spleen cells.

This study investigated whether neuraminidase (Neu) affects LFA-1 mRNA expression in spleen cells and whether somatostatin (SOM) and substance P (SP) treatment induce changes in the Neu mRNA expression level in spleen cells. Neu treatments down-regulated the LFA-1 mRNA levels after culturing for 2 h. SOM increased the Neu mRNA level slightly after 24-h culture and strongly after 48-h culture. These results suggest that prolonged exposure to SOM may regulate the Neu activation pathway, which in turn impairs the regulation of LFA-1 expression.

Modification of radiation response in mice by ginsenosides, active components of Panax ginseng.

We performed this study to determine the effect of extract of whole ginseng and ginsenosides (total saponin, panaxadiol and panaxatriol) on jejunal crypt survival, endogenous spleen colony formation, and apoptosis in jejunal crypt cells of mice irradiated with high- and low-doses of gamma-radiation. The radioprotective effect of ginseng was compared with the effect of diethyldithiocarbamate (DDC). The jejunal crypts were protected by pretreatment with extract of whole ginseng (i.p.: 50 mg/kg of body weight at 12 and 36 hours before irradiation, p < 0.005). Extract of whole ginseng (p < 0.005), total saponin (p < 0.01) or panaxadiol (p < 0.05) administration before irradiation (i.p.: 50 mg/kg of body weight at 12 and 36 hours before irradiation) resulted in an increase in the formation of the endogenous spleen colony. The frequency of radiation-induced apoptosis in the intestinal crypt cells was also reduced by pretreatment with extract of whole ginseng (p < 0.05), total saponin (p < 0.005) or panaxadiol (p < 0.05) (i.p. at 12 and 36 hours before irradiation). The radioprotective effect on the jejunal crypts and apoptosis in the DDC-treated group appeared similar to that in the ginseng-treated groups. Treatment with DDC showed no significant modifying effects on the formation of the endogenous spleen colony. In the experiment on the effect of ginsenosides, the result indicated that panaxadiol might have a major radioprotective effect. Although the mechanisms of this inhibitory effect remain to be elucidated, these results indicated that ginseng might be a useful radioprotector, especially since it is a relatively nontoxic natural product. Further studies are needed to better characterize the protective nature of ginseng extract and each ginsenoside.

Apoptosis in growing hair follicles following gamma-irradiation and application for the evaluation of radioprotective agents.

The usefulness of apoptotic fragments assay for investigating the radiation response of hair follicles and evaluation of radioprotective agents was examined in ICR mice. The extent of changes following 100 cGy (1000 cGy/min) was studied at 0, 2, 4, 8, 12, 16 or 20 hours after exposure. The maximal frequency was found 12 hours after exposure. The mice that received 50, 100, 200, 400 or 800 cGy of gamma-rays were examined 12 hours after irradiation. Measurements performed after gamma-ray irradiation showed a dose-related increase in apoptotic cells in each mouse studied. The dose-response curves were analyzed with a linear-quadratic model: the frequency (number per follicle) of apoptotic cells in the hair follicle was y = (0.05527 +/- 0.009574) D + (-0.00001988 +/- 0.00001337) D2 + 0.227 (r2 = 0.964, D = 100 cGy). The frequency of radiation (100 cGy)-induced apoptosis in hair follicles was reduced by pretreatment of diethyldithiocarbamate (DDC, i.p. at 30 minutes before irradiation, p < 0.05) or green tea (i.p. at 12 and 36 hours before irradiation, p < 0.01). From these results, it is thought that this model will be useful in the detection of radiation response and radioprotective agents.

Measurement of micronuclei by cytokinesis-block method in human, cattle, goat, pig, rabbit, chicken and fish peripheral blood lymphocytes irradiated in vitro with gamma radiation.

The frequencies of gamma-ray-induced micronuclei (MN) in cytokinesis-blocked (CB) lymphocytes at several doses were measured in three donors of seven species (human, cattle, goat, pig, rabbit, chicken, fish). Measurements performed after irradiation showed a dose-related increases in MN frequency in each of the donors of human, cattle, goat, pig and rabbit. The relative sensitivity of cattle, goat, pig and rabbit in peripheral blood lymphocytes (PBLs) compared with human PBLs was estimated by best fitting linear-quadratic model based on the radiation-induced MN data over the range from 0 cGy to 400 cGy. In the case of MN frequency with 0.2, the relative sensitivities of cattle, goat, pig and rabbit PBLs were 0.86, 0.98, 0.41 and 0.39, respectively. These data indicate that the induction of MN in CB cells following irradiation is similar in human, cattle and goat PBLs, while PBLs from pig and rabbit were much less sensitive to the MN induction effects of gamma-radiation than those from human. The micronucleus counts failed to show any evidence of radiation damage in the cells from chicken and fish. Measurements performed after irradiation showed a dose-related decrease in the formation of binucleated cells. We concluded that the use of CB cell from fish and chicken for detecting the results of radiation exposure was highly questionable. Our in vitro radiobiological study confirmed that the cytogenetic response obtained in blood from selected breeds of mammalian species can be utilized for application in environmental studies.

Effects of benzo[alpha]pyrene, 2-bromopropane, phenol and 2,3,7,8-tetrachlorodibenzo-p-dioxin on IL-6 production in mice after single or repeated exposure.

Altered IL-6 production regulation is associated with the pathogenesis of chronic inflammatory diseases, lymphoid malignancies, chronic infectious processes and certain types of autoimmune conditions. Here, we examine the effects of pollutants on IL-6 levels in mice serum and in culture supernatants of spleen cells. Mice were treated with vehicles (PBS or olive oil), benzo[alpha]pyrene (B[alpha]P, 100 mg/kg body weight), 2-bromopropane (2-BP, 3.5 g/kg), phenol (21.2 mg/kg), or 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD, 15 mg/kg). Serum IL-6 levels were significantly increased in the TCDD-treated group at 24 hours and 48 hours after a single exposure, whereas exposure to phenol, B[alpha]P or 2-BP did not cause a significant difference. IL-6 levels in culture supernatants of splenocytes were not affected at 24 hours and 48 hours after a single pollutant treatment. A repeated dose of TCDD (once/week for 4 weeks) resulted in a significant elevation of IL-6 levels in serum and its spontaneous production in culture supernatants of splenocytes. Repeatedly TCDD-treated mice contained more CD11b (Mac-1)-positive cells in the spleen and higher titers of tissue-specific autoantibodies than the vehicle-treated group. These results suggest that repeated exposure to TCDD might impair the regulation of immune response by deregulating the production of IL-6.

Dose-incidence relationships on the prenatal effects of gamma-radiation in mice.

The objective of this investigation was to evaluate dose-incidence relationships on the prenatal effects of gamma-radiation. Pregnant ICR mice were exposed on day 11.5 after conception, coincident with the most sensitive stage for the induction of major congenital malformations, with 0.5-4.0 Gy of gamma-radiations. The animals were sacrificed on day 18 of gestation and the fetuses were examined for mortality, growth retardation, change in head size and any other morphological abnormalities. With increasing radiation dose, incidence of small head, growth retarded fetuses, cleft palate, dilatation of cerebral ventricle and abnormalities of the extremities in live fetuses rose. The threshold doses of radiation that induced cleft palate and dilatation of cerebral ventricle, and abnormal extremities were between 1.0 and 2.0 Gy, and between 0.5 and 1.0 Gy, respectively.

Effects of benzo[a]pyrene, 2-bromopropane, phenol and 2,3,7,8-tetrachlorodibenzo-p-dioxin on proinflammatory cytokines gene expression by mice spleen cells.

The detrimental effects of environmental pollutants on the health of the individual are generally accepted, although the mechanisms of these effects remain to be incompletely understood. In the present study, we examined the effects of B[a]P, 2-BP, phenol and TCDD on proinflammatory cytokine gene expression in mice spleen cells which were stimulated with anti-CD3. 10(-9)M TCDD increased IFNgamma and TNFalpha gene expression, but suppressed IL-1 gene expression. 10(-6)M phenol inhibited IL-1, IL-6 and TNFalpha gene expression, and 10(-6)M of 2-BP downregulated TNFalpha gene expression. However, 10(-6)M of B[a]P did not influence on IL-1, IL-6, IFNgamma and TNFalpha gene expression. These findings suggest that TCDD may impair the immune functions of mice by enhancing proinflammatory cytokines production, whereas phenol and 2-BP may impair the functions by inhibiting the production of these cytokines.

The radioprotective effects of bu-zhong-yi-qi-tang: a prescription of traditional Chinese medicine.

We evaluated the effect of bu-zhong-yi-qi-tang, a prescription of traditional Oriental medicine, and its major ingredients on protection of the intestine and hematopoietic organs against radiation damage in this study. The jejunal crypt survival, endogenous spleen colony formation, and apoptosis in jejunal crypt cells were investigated in mice irradiated with high and low doses of gamma-rays. bu-zhong-yi-qi-tang administration before irradiation protected the jejunal crypts (p < 0.0001), increased the formation of the endogenous spleen colony (p < 0.05) and reduced the frequency of radiation-induced apoptosis (p < 0.05). In experiments on the effects of the individual ingredient of bu-zhong-yi-qi-tang, Rensan (Radix Ginseng), Danggui (Radix Angelicae gigantis), Shengma (Rhizoma Cimicifugae) and Chaihu (Radix Bupleuri) might have major radioprotective effects, and each might have different degrees of effect on these three endpoints. These results indicated that bu-zhong-yi-qi-tang might be a better agent than any one of its ingredients to satisfy all three endpoints. Although the mechanisms of this inhibitory effect remain to be elucidated, these results indicated that bu-zhong-yi-qi-tang might be a useful radioprotector, especially since it is a relatively non-toxic natural product. Further studies are needed to better characterize the protective nature of bu-zhong-yi-qi-tang extract and its ingredients.

Antiinflammatory activity of an herbal preparation (HemoHIM) in rats.

This study evaluated a new herbal preparation, HemoHIM, for its antiinflammatory activity against carrageenan-induced edema, the formation of granulation tissues by cotton pellet and experimental colitis by 2,4,6-trinitrobenzene sulfonic acid (TNBS). The HemoHIM was prepared by adding its ethanol-insoluble polysaccharide fraction to the total water extract of Angelica Radix, Cnidii Rhizoma and Paeonia Radix. The preparation (4 mg of solids/mL of drinking water, p.o., 50-100 mg/kg of body weight, i.p.) produced a dose-related inhibition of carrageenan-induced paw edema and cotton pellet-induced granuloma in rats. In addition, HemoHIM also reduced the degree of TNBS-induced colitis and improved the gross and histological changes such as thickening, dilatation, ulceration, and infiltration by polymorphonuclear leukocytes and multiple erosive lesions. These results demonstrate that the HemoHIM has a potent antiinflammatory effect.

In Vivo radioprotective effect of Panax ginseng C.A. Meyer and identification of active ginsenosides.

This study evaluated the effect of water extracts of Panax ginseng C.A. Meyer (PG), panaxadiol (PD), panaxatriol (PT), ginsenoside Rb(1), Rb(2), Rc, Rd, Re and Rg(1) on jejunal crypt survival, endogenous spleen colony formation and apoptosis in jejunal crypt cells in gamma-irradiated mice. Jejunal crypts were protected by pretreatment with PG, Rc and Rd. Administration of PG, PD, Rd and Re prior to irradiation resulted in an increase in the formation of endogenous spleen colonies. The frequency of radiation-induced apoptosis in intestinal crypt cells was also reduced by pretreatment with PG, PD, Rb(2), Rc, Rd, Re and Rg(1). In experiments on the effects of the individual ginsenosides, the rank order of activity was Rc > Rd > Rg(1) > Rb(2) > Re > Rb(1) on intestinal crypt survival assay, Re > Rb(2) > Rd > Rg(1) > Rb(1) > Rc on the spleen colony formation assay, and Rg(1) > Re > Rd > Rc > Rb(2) > Rb(1) on inhibiting the death of cells caused by apoptosis. The results indicated that Rc, Rd and Re may have a major radioprotective effect in mice irradiated with high and low doses of radiation. When the same experiments were performed using PD and PT, it was observed that most of the inhibitory effects came from PD rather than PT.

Teratological studies of prenatal exposure of mice to a 20 kHz sawtooth magnetic field.

In order to evaluate the importance of gestational age in possible effects due to exposure to a 20 kHz sawtooth magnetic field, pregnant ICR mice at gestational 2.5-15.5 days post-coitus, which is the most sensitive stage for the induction of major congenital malformations, were exposed in a carrousel irradiator. The mice were exposed to a 20 kHz intermediate frequency (IF) sawtooth magnetic field had a 6.5 microT peak intensity for 8 h/day. The animals were sacrificed on the 18th day of gestation; and the fetuses were examined for mortality, growth retardation, changes in head size, and other morphological abnormalities. From the above conditions, it is concluded that the exposure to a 20 kHz sawtooth magnetic field with 6.5 microT peak intensity does not inflict any adverse effect on fetuses of pregnant mice.

Regulation of leukocyte function-associated antigen 1-mediated adhesion by somatostatin and substance P in mouse spleen cells.

BACKGROUND: Interaction of the integrin leukocyte function-associated antigen (LFA)-1 (CD11a/CD18) with its ligands, the intercellular adhesion molecules (ICAM)-1, -2, and -3 (CD54, CD102, and CD50), is pivotal to many leukocyte adhesion events. METHOD: To define the mechanism of the movement of leukocytes to the inflammatory site by somatostatin (SOM) and substance P (SP), we examined the expression of the adhesion molecule LFA-1 and inside-out signals for integrins, protein kinase C (PKC), Ras, Rap1, and phosphoinositide (PI) 3-kinase, in anti-CD3-, anti-CD3+SOM-, anti-CD3+SP-stimulated or unstimulated spleen cells. RESULTS: SOM caused down-regulation of LFA-1 mRNA translation as well as of adhesion-stimulating molecules such as Rap1, Ras, and PI 3-kinase. On the other hand, SP slightly induced LFA-1 mRNA translation and activation signals for integrins. The early-phase alteration of LFA-1 mRNA translation after 3 h of culture may be due to the changes of CD8+ T cells rather than changes of CD4+ cells. In adhesion assays, SOM significantly decreased cell adhesion (p < 0.05). CONCLUSION: These data suggest that SOM treatment of spleen cells, especially in CD8+ T cells, leads to downregulation of LFA-1 mRNA translation, inside-out signaling molecules for integrins (Ras, Rap1 and PI 3-kinase, but not PKC), and consequently to a decrease in the LFA-1-mediated adhesion to ICAM-1.