Inhibitory Effect of Imiquimod-Induced Psoriasis-Like Skin Inflammation in Mice by Histamine H4 Receptor Agonist 4-Methylhistamine.

Psoriasis is a chronic inflammatory immune-mediated autoimmune skin disorder. The histamine H4 receptor (H4R) agonist 4-methylhistamine (4-MH) plays an important role in immunomodulation of inflammatory responses associated with allergic inflammatory diseases. In this study, we investigated the effects of H4R agonist 4-MH on the development of imiquimod (IMQ)-induced psoriasis-like skin inflammation in mice and explored the immunoregulatory mechanism involved. The total clinical severity scores were significantly ameliorated by treatment with 4-MH (20 mg/kg) and 4-MH (40 mg/kg). Histological analysis of the skin revealed that 4-MH (20 mg/kg) and 4-MH (40 mg/kg) significantly attenuated the psoriatic phenotypes, including epidermal hyperplasis, hyperkeratosis and lymphocytes infiltration. Treatment with 4-MH (20 mg/kg) and 4-MH (40 mg/kg) led to reductions in the levels of Th1 cytokines (TNF-α, IFN-α, and IL-27) in the serum and dorsal skin, whereas Th17 cytokines levels (IL-17A and IL-23) did not change in response to treatment with 4-MH (20 mg/kg) and 4-MH (40 mg/kg). Furthermore, the number of CD4(+) CD25(+) FoxP3(+) regulatory T (Treg) cells was significantly increased by treatment with 4-MH (40 mg/kg). Taken together, these results imply that H4R agonist 4-MH might be an effective immunomodulatory approach for treatment of patients with psoriasis and the effects may be related to inhibited epidermal alteration, selectively reduced Th1 pro-inflammatory cytokines, and recruited CD4(+) CD25(+) FoxP3(+) Treg cells.

Effects of Dietary Fermented Chlorella vulgaris (CBT(®)) on Growth Performance, Relative Organ Weights, Cecal Microflora, Tibia Bone Characteristics, and Meat Qualities in Pekin Ducks.

Fermented Chlorella vulgaris was examined for its effects on growth performance, cecal microflora, tibia bone strength, and meat qualities in commercial Pekin ducks. A total of three hundred, day-old male Pekin ducks were divided into three groups with five replicates (n = 20 ducklings per replicate) and offered diets supplemented with commercial fermented C. vulgaris (CBT(®)) at the level of 0, 1,000 or 2,000 mg/kg, respectively for 6 wks. The final body weight was linearly (p = 0.001) increased as the addition of fermented C. vulgaris into diets increased. Similarly, dietary C. vulgaris linearly increased body weight gain (p = 0.001) and feed intake (p = 0.001) especially at the later days of the feeding trial. However, there was no C. vulgaris effect on feed efficiency. Relative weights of liver were significantly lowered by dietary fermented C. vulgaris (linear effect at p = 0.044). Dietary fermented C. vulgaris did not affect total microbes, lactic acid bacteria, and coliforms in cecal contents. Finally, meat quality parameters such as meat color (i.e., yellowness), shear force, pH, or water holding capacity were altered by adding fermented C. vulgaris into the diet. In our knowledge, this is the first report to show that dietary fermented C. vulgaris enhanced meat qualities of duck meats. In conclusion, our study indicates that dietary fermented C. vulgaris exerted benefits on productivity and can be employed as a novel, nutrition-based strategy to produce value-added duck meats.

Carcass Characteristics and Meat Quality of Korean Native Ducks and Commercial Meat-type Ducks Raised under Same Feeding and Rearing Conditions.

This study was conducted to compare carcass characteristics and physico-chemical meat quality in two different genotype ducks raised under identical feeding and rearing conditions. A total of ninety 1-d-old Korean native ducks (KND, n = 45) and commercial meat-type ducks (Grimaud, n = 45) were fed same experimental diets during 56 d and 42 d, respectively to obtain similar slaughter weights. The experimental diet for starter period contained 20% crude protein (CP) and 2,900 kcal nitrogen corrected true metabolizable energy (TMEn)/kg of diet and that for grower period contained 17% CP and 3,050 TMEn/kg of diet. Average daily gain and feed efficiency of KND were inferior to those of commercial meat-type ducks (p<0.05). Carcass weight was not different between two genetically different ducks, but carcass yield of KND was significantly higher (p<0.05) than that of commercial meat-type ducks. There were no significant differences in cooking loss and pH of breast meat between two genetically different ducks, but water holding capacity of KND was significantly higher than that of commercial meat-type ducks. The linoleic acid and total polyunsaturated fatty acid of breast meat from KND were significantly higher (p<0.05) than the corresponding part from commercial meat-type ducks. Significant differences were detected in water holding capacity and the content of linoleic acid and polyunsaturated fatty acid, which were significantly higher in KND, whereas growth performance tended to be superior in commercial ducks. At the market weight, the meat from KND was judged to have better qualities with regard to higher water holding capacity and greater content of polyunsaturated fatty acid compare with meat from commercial meat-type duck.

Comparison of growth performance, carcass characteristics and meat quality of korean local chickens and silky fowl.

This study was conducted to compare growth performance, carcass characteristics and meat quality of 4 breeds of local chicken. A total of 480 1-d-old chicks were distributed to 16 pens, with 4 treatments of breed, 4 replicates and 30 chicks per pen. Three Korean local breeds of white-mini broiler, Hanhyup-3-ho, and Woorimatdag, and a breed of silky fowl were raised under identical rearing and feeding conditions for 31-d, 37-d, 36-d, and 59-d, respectively. The BW and feed consumption on a pen basis were weekly measured for all pens, and ADFI, ADG and gain:feed were calculated for each pen. The ADFI and ADG of 3 breeds of Korean local chicken were greater than those of silky fowl (p<0.05). Within the Korean local breeds, ADFI of white-mini broiler was the highest (p<0.05), and ADG of Hanhyup-3-ho and white-mini broiler was the highest (p<0.05). Gain:feed of silky fowl was less than that of the 3 breeds of Korean local chicken. The carcass and breast yield of white-mini broiler were the greater than those of other breeds (p<0.05). The breast meat color (CIE L*, a*, and b*) of 3 breeds of Korean local chicken were higher than that of silky fowl (p<0.05). The breast meat of Hanhyup-3-ho had greater cooking loss (p<0.05), whereas water holding capacity and pH were less than those of other breeds (p<0.05). The color score of 3 breeds of Korean local chicken was higher than that of silky fowl (p<0.05). Woorimatdag had a higher score on tenderness (p<0.05), whereas flavor score was less than that of other breeds (p<0.05). In conclusion, 4 local breeds of chicken have some unique features and seem to have more advantages, and this information can help consumers who prefer healthy and premium chicken meat.

Growth performance and carcass characteristics of korean native ducks fed diets with varying levels of limiting amino acids.

There are multiple experiments conducted with male Korean native ducks (KND) to evaluate the optimal levels of limiting amino acids (AA). In Exp. 1, a total of 450 one-d-old male KNDs were divided into five groups with six replicates and fed experimental diets with varying levels of lysine, total sulfur amino acids (TSAA) and threonine (T1, 0.90/0.74/0.70%; T2, 1.00/0.82/0.77%; T3, 1.10/0.90/0.85%; T4, 1.20/0.98/0.93%; T5, 1.30/1.07/1.01%) to 21 d of age. In Exp. 2, one-d-old male KND were received and fed commercial starter diet from hatching to 21 d of age, and then divided into five groups with six replicates and fed one of five diets varying levels of lysine, TSAA, and threonine (T1, 0.73/0.62/0.54%; T2, 0.80/0.68/0.60%; T3, 0.87/0.74/0.65%; T4, 0.94/0.80/0.70%; T5, 1.01/0.86/0.75%) during 22 to 56 d of age, respectively. The BW gain was linearly increased as dietary limiting AA levels increased to 1.20% lysine, 0.98% TSAA and 0.93% threonine. There were no significant differences in feed intake, gain:feed and uniformity among groups. In Exp. 2, the BW gain and gain:feed were not affected by dietary limiting AA levels. There were no significant differences in carcass characteristics and meat quality among groups. The growth performance and carcass characteristics did not show the significant response to increasing dietary limiting AA levels in KND during 22 to 56 d of age. In conclusion, the levels of lysine, TSAA and threonine necessary to maximize growth for starter phase were at least 1.20%, 0.98%, and 0.93%, respectively. On the other hands, KND require relatively low levels of limiting AA for late growth and carcass yield. The dietary levels of 0.73% lysine, 0.62% TSAA and 0.54% threonine appear to be adequate during growing phase.

Tauroursodeoxycholic acid enhances the pre-implantation embryo development by reducing apoptosis in pigs.

Apoptosis is an important determinant of the normal development of pre-implantation embryos in vitro. Recently, endoplasmic reticulum (ER) stress-mediated apoptosis has been extensively investigated in a wide variety of diseases. Efficient functioning of the ER is essential for most cellular activities and survival. Tauroursodeoxycholic acid (TUDCA), an endogenous bile acid, has been reported to attenuate ER stress-mediated cell death by interrupting the classic pathways of apoptosis. Therefore, in this study, the anti-apoptotic effect of TUDCA on ER stress-induced apoptosis was examined in pre-implantation pig embryos. Also, tunicamycin was used to investigate the effects of ER stress on pig embryo development. After in vitro maturation and fertilization, presumptive pig embryos were cultured in NCSU-23 medium supplemented with TUDCA or TM for 6 days at 39 °C, 5% CO(2) in air. All data were analysed using one-way anova and Duncan's multiple range test in the statistical analysis system (SAS). In addition, we also determined the optimal TM and TUDCA concentrations. Samples were treated with TM at concentrations of 0, 1, 2 or 5 µm and with TUDCA at concentrations of 0, 100, 200 or 300 µm. When TM was used during in vitro culture, only 8.2% (8/97) of the embryos developed to the blastocyst stage when the treatment concentration was 1 µm compared with 27.4% (28/102) of the embryos in the control group (p < 0.05). In contrast, the frequency of blastocyst formation and the number of cells were higher when treated with 200 µm TUDCA compared with the control group (32.8% and 39.5 vs 22.2% and 35.6, p < 0.05). Moreover, the developmental rate to the blastocyst stage embryo in the group treated with TM and TUDCA was not significantly different from that of the control group (17.8%, 26/142 vs 24.9%, 36/145). Furthermore, the blastocyst cell number was enhanced (31.9 vs 36.9) and apoptosis reduced (TUNEL-positive nuclei number, 6.0 vs 3.2) by TUDCA treatment in pig embryos. In the real-time quantitative RT-PCR analysis, the expression of anti-apoptotic Bcl-XL gene was shown to be increased in the blastocyst stage because of TUDCA treatment, whereas expression of pro-apoptotic Bax was decreased. In addition, we also found that TUDCA decreased the rate of TM-induced apoptosis in the pre-implantation stage. Taken together, our results indicate that TUDCA improves the developmental competence of pig embryos by modulating ER stress-induced apoptosis during the pre-implantation stage.

Exogenous dibutyryl cAMP affects meiotic maturation via protein kinase A activation; it stimulates further embryonic development including blastocyst quality in pigs.

High concentrations of cyclic AMP in germinal vesicle oocytes generally inhibit GVBD. Thus, maintaining the GV stage in growing oocytes is essential for the developmental competence of the eggs. In this study, we traced the effects of dibutyryl cyclic AMP on meiotic maturation and early embryonic development in pigs. We also investigated several blastocyst qualities, including structural integrity, mitochondrial membrane potential, and apoptosis, which are affected by dbcAMP. To determine whether increased concentrations of cAMP inhibit GVBD, we explored the meiotic patterns and during maturation of pig oocytes. When treated with dbcAMP for 22h, 91.1% of the oocytes were arrested in the GV stage compared to only 38.8% of the oocytes in the control group (P<0.05). After completion of IVM, a higher proportion of the dbcAMP-treated oocytes were in metaphase II than the untreated ones (91.3% vs. 72.8%, P<0.05). Western blot analysis showed a reduction (at 22h) and/or increase (at 44h) in MPF and MAP kinase activities in porcine oocytes treated with dbcAMP for the first 22h of IVM compared to the untreated control. We also confirmed that protein kinase A activity increased in dbcAMP-treated oocytes, indicating an elevated intracellular concentration of cAMP. After IVF, the frequency of polyspermy in the dbcAMP-treated group decreased compared to that in the control group (22.4% vs. 47.4%, P<0.05). Furthermore, blastocyst formation, the blastocyst cell number, mitochondrial membrane potential, and apoptosis were enhanced and/or reduced by dbcAMP in both IVF and SCNT embryos. We concluded that synchronizing meiotic resumption by dbcAMP treatment improved the developmental capacity and embryonic qualities of IVF and SCNT embryos by increasing the mitochondrial membrane potential and decreasing the incidence of apoptosis in preimplantation-stage porcine embryos.

Inhibitory effects of Cnidium officinale Makino and Tabanus fulvus Meigan on the high glucose-induced proliferation of glomerular mesangial cells.

This study describes a potent activity of Cnidium officinale Makino (Cnidii rhizoma) and Tabanus fulvus Meigan (Tabanus) as an inhibitor of high glucose-induced proliferation of glomerular mesangial cells (GMCs). Raising the ambient glucose concentration from 5.6 to 25 mM for 24 h caused a dramatic increase in [3H]thymidine incorporation, and these increases were attenuated by treatment of GMCs with the extracts of Cnidii rhizoma and Tabanus (2.5-20 microg/ml) in a dose-dependent manner. In contrast, extracts of Cnidii rhizoma or Tabanus (20 microg/ml) did not change the growth of GMCs cultured under normal glucose condition. To clarify the mechanism involved in anti-proliferative activity of these medicines, this study examined the effects of Cnidii rhizoma and Tabanus on high glucose-stimulated extracellular matrix (ECM) protein accumulation and transforming growth factor-beta1 (TGF-beta1) production. Exposure of GMCs to high glucose significantly stimulated the ECM protein, collagen and fibronectin, accumulation and TGF-beta1 secretion, and these changes were dramatically diminished by treatment of GMCs with extracts of Cnidii rhizoma or Tabanus (10 microg/ml). Taken together, these results indicate that Cnidii rhizoma and Tabanus inhibit the high glucose-induced GMC proliferation partially through suppressing the ECM accumulation and TGF-beta1 production, suggesting that these medicines may be a promising agent for treating the development and progression of diabetic glomerulopathy.

Seungnoidan increases cerebrocortical ATP and acetylcholine contents in ovariectomized rats.

This study investigated the effects of Seungnoidan (SND), which has been widely used as a remedy for cerebroneuronal diseases in Korean folk medicine, on the cerebrocortical adenosine triphosphate (ATP) and acetylcholine (ACh) contents in ovariectomized (OVX) rats. Female Sprague-Dawley rats were ovariectomized and maintained for 12 weeks to deplete ovarian steroid hormones, followed by oral administration of SND at 500 mg/kg/day for 14 weeks. SND markedly attenuated the high rate of body weight increase in OVX rats, and also reduced the decline of cerebral weight caused by ovariectomy (p < 0.05). Superfusion of SND at 50 mg/kg significantly increased the rate of cerebral blood fl ow, but did not change the mean arterial blood pressure. Deprivation of ovarian steroid hormones significantly decreased the cerebral ATP, choline and ACh contents, and these reductions were reduced by treatment of OVX rats with SND (p < 0.01). Additionally, SND also significantly elevated the cerebral choline acetyltransferase activities reduced by OVX (p < 0.01). Taken together, these results suggest that the pharmacological properties of SND may be implicated in the improvement of metabolic pathways of cerebral energy and cholinergic neurotransmitter function induced by deprivation of ovarian steroid hormones, and SND may be a promising herbal remedy for treatment of cerebral dysfunctions including dementia.

Effects of hwangryunjihwang-tang on in vitro ertilization and ovulation in mice fed a high-fat diet.

It has been generally accepted that hwangryunjihwang-tang (h-tang) is a useful prescription for treating polydipsia and to prevent obesity induced by a high-fat diet. The aim of this study was to clarify whether h-tang improved reproductive dysfunction caused by obesity in mice. Mice were fed a high density protein and lipid diet for 4 weeks, followed by administration of h-tang at 480 mg/kg body weight per day for 4 days. Thereafter, changes of body weight, ovulation rate, in vitro and in vivo fertilization, embryonic development and implantation rate were measured. H-tang markedly reduced the body weight of obese mice fed a high-fat diet, but not mice fed a normal diet. H-tang significantly improved ovulation rates, in vitro and in vivo fertilization rates and embryonic development. These results indicate pharmacological reversal of reproductive dysfunction caused by obesity, perhaps by adjusting internal secretions and metabolic functions.

Differential distribution of ganglioside GM3 in seminiferous tubule and epididymis of adult rats.

Gangliosides are ubiquitous membrane components in mammalian cells and are suggested to play important roles in various functions such as cell-cell interaction, adhesion, cell differentiation, growth control and signaling. Among all ganglio-series gangliosides, GM3 has the simplest carbohydrate structure, and has been shown as a major ganglioside in male reproductive system. To study GM3 distribution in the seminiferous tubule and epididymis, frozen sections were stained with specific monoclonal antibody (MAb) against ganglioside GM3. In the seminiferous tubule of testis, pachytene spermatocytes and spermatids expressed ganglioside GM3, but not in spermatogonia and sertoli cells. Spermatogonia and sertoli cells near the basement membrane were negatively reacted to anti-GM3. In the epididymis, GM3 was expressed only in some interstitial cells. Taken together, these results suggest that the expression of ganglioside GM3 in rat seminiferous tubule and epididymis is spatio-temporally regulated during spermatogenesis.

Developmental patterns of ceramide glucosyltransferase (GlcT-1) expression in the mouse: in situ hybridization using DIG-labeled RNA probes.

Glycosphingolipids (GSLs) play significant roles in a variety of cell membrane events, including cellular interactions, signaling, and trafficking. Ceramide glucosyltransferase (glucosylceramide synthase, GlcT-1, EC 2.4.1.80) catalyzes the initial step in GSL synthesis, the transfer of glucose from UDP-glucose to ceramide. The reaction product of glucosylceramide serves as a core structure for over 400 species of GSLs. The enzyme is a key regulatory factor controlling intracellular levels of ceramide and GSLs. Appearance and differential distribution of GlcT-1 mRNA during mice postimplantation embryogenesis [embryonic (E) days; E9, E11, E13, E15] were investigated by in situ hybridization with digoxigenin-labeled RNA probes, coupled with alkaline phosphatase detection. On E9, tissues of the mesencephalon, myelecephalon, diencephalons, and telencephalon expressed GlcT-1. On E11, it was expressed to a detectable extent in various tissues including mesencephalon, myelecephalon, diencephalon, telencephalon, nose, lung, liver, vertebra, tail, spinal cord, and tongue. The expression patterns of E13 were similar to those of E11, except that the heart and stomach became positive. On E15, a specific signal for GlcT-1 was detected in all organs of the embryo. These results provide the first evidence that GlcT-1 is differentially expressed during postimplantation embryogenesis.

Bombycis corpus extract prevents amyloid-beta-induced cytotoxicity and protects superoxide dismutase activity in cultured rat astrocytes.

Bombycis corpus(BC) or Bombyx Batryticatus, a batryticated silkworm and white-stiff silkworm, is a drug consisting of the dried larva of silkworm, Mobyz mori L., dead and stiffened due to the infection of Beauveria (Bals.) Vuill. In the present study, we have examined the protective effect of the water extracts against Amyloid- beta(A beta) 25-35 peptide-induced cytotoxicity by microscopic observation and LDH assay, and its action on antioxidative enzymes using cultured astrocyte cells. A beta 25-35-induced cell death was protected by the application of water extract of BC in a dose-dependent manner, and concentrations of 10(-6)to 10(-7)g ml(-1)showed a significant effect compared to exposure of A beta 25-35 alone. When antioxidative enzyme activities such as catalase, superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and glutathione-S transferase (GST) were assayed after A beta 25-35 treatment, most enzyme activities were decreased in a similar fashion. BC treatment of A beta 25-35-treated astrocytes did not affect the enzyme activities of catalase, GSH-Px and GST. However, only SOD activity was enhanced by BC treatment and this may result from the potentiation of the antioxidative ability of BC. The protective effect of BC against cytotoxicity induced by Abeta 25-35 strongly indicates that BC could be a protective agent for free radical generating compounds, and that Abeta 25-35 is not only a potent lipid peroxide inducer, but can also cause changes in antioxidative enzymes. From the results, it was concluded that BC has a protective effect against Abeta -induced cytotoxicity in cultured astrocyte cells through the inhibition of lipid peroxidation and protection of antioxidative enzymes.

Developmental patterns of mST3GalV mRNA expression in the mouse: in situ hybridization using DIG-labeled RNA probes.

mST3GalV synthesizes ganglioside GM3, the precursor for simple and complex a- and b- series gangliosides, and the expression and regulation of mST3GalV (CMP-NeuAc: lactosylceramide alpha2,3-sialyltransferase) activity is central to the production of almost all gangliosides, a class of glycosphingolipids implicated in variety of cellular processes such as transmembrane signaling, synaptic transmission, specialized membrane domain formation and cell-cell interactions. To understand the developmental expression of mST3GalV in mice, we investigated the spatial and temporal expression of mST3GalV mRNA during the mouse embryogenesis [embryonic (E) days; E9, E11, E13, E15] by in situ hybridization with digoxigenin-labeled RNA probes. All tissues from E9 and E11 were positive for mST3GalV mRNA. On E13, mST3GalV mRNA was expressed in various neural and non-neural tissues. In contrast to these, on E15, the telencephalon and liver produced a strong expression of mST3Gal V which was a quite similar to that of E13. In this stage, mST3GalV mRNA was also expressed in some non-neural tissues. These data indicate that mST3GalV is differently expressed at developmental stages of embryo, and this may be importantly related with regulation of organogenesis in mice.

Radish extract stimulates motility of the intestine via the muscarinic receptors.

The effects of radish (Brassica oleraceae, Cruciferae) on gastrointestinal motility were examined using rat intestinal segments with myenteric plexus in-vitro and measuring the intestinal transit of charcoal in-vivo. Radish extract (10 microg mL(-1) to 2 mg mL(-1)) caused a dose-dependent increase in contractions of the duodenum, jejunum and ileum, and 1 mg mL(-1) was the maximum effective dose. The largest contraction by the extract was found in ileal segments. The extract-induced (0.5 mg mL(-1)) ileal contraction was remarkably inhibited by pretreatment of segments with atropine (10(-7) M) for 10 min, but not by hexamethonium (0.5 mM). Moreover, antagonists of the muscarinic receptor reduced the radish-induced ileal contraction by a different ratio. The rank order of inhibitory effects was 4-diphenylacetoxy-N-methyl-(2-chloroethyl)-piperidine methiodide (90.5% of control) > tropicamide (67.4%) > pirenzepine (42.8%) > methoctramine (16.7%). Oral administration of radish extract (300-500 mg kg(-1) body weight) to mice remarkably improved the intestinal transit of charcoal, and this was significantly attenuated by co-administration of atropine (50 mg kg(-1)). Taken together, these results suggest that radish extract stimulates gastrointestinal motility through activation of muscarinic pathways.

Shibimijihwang-tang elevates intracellular ATP and choline content in the cerebral cortex of ovariectomized rats.

Shibimijihwang-tang (SJT) has been used traditionally to improve systemic blood circulation and biological energy production in patients with circulatory and neuronal diseases. The object of this study is to determine the effect of SJT extract on the intracellular adenosine triphosphate (ATP) and choline content in the cerebral cortex of ovariectomized (OVX) rats. Bilateral ovaries of 8-week-old rats were removed. Rats were maintained for 12 weeks to deplete ovarian steroid hormones, followed by treatment with SJT at 500 mg/kg body weight per day for 12 weeks. High rate of body weight increase in the OVX rats was markedly reduced by treatment with SJT, but the change in body weight of normal rats was not affected by it. SJT also significantly reduced the decline of cerebral weight in the OVX rats (P<0.05). Tissue glucose content in the cerebral cortex of OVX rats was significantly increased by SJT treatment (P<0.05). A decline in cerebral ATP content in OVX rats was dramatically restored by SJT administration (P<0.01), but SJT did not change the cerebral ATP content in normal rats. Cerebral choline content also declined following ovariectomy. This reduction was significantly elevated by SJT treatment (P<0.05), but SJT did not affect the change in cerebral choline in normal rats. Taken together, these results demonstrate that SJT can reduce the decrease in brain weight, cerebral ATP and choline content caused by deprivation of ovarian steroid hormones. This suggests that pharmacological properties of SJT may play a role in improvement of reduced cerebral energy production and cholinergic neurotransmitter synthesis caused by deficiency of ovarian steroid hormones in the cerebroneuronal cells of postmenopausal women.

Distribution of ganglioside GM3 in the rat ovary after gonadotropin stimulation.

Gangliosides are ubiquitous membrane components in mammalian cells and are suggested to play important roles in various cell functions, such as cell-cell recognition, differentiation and transmembrane signalling. Ovaries have been shown to contain GM3 as a major ganglioside. To study GM3 distribution during gonadotropin stimulation in the hypophysectomized rat ovary, ovarian sections and cultured granulosa cells were stained with specific monoclonal antibody against GM3. Interstitial cells of follicles of immature hypophysectomized rat ovary expressed ganglioside GM3. Theca cells of early antral follicles but not primary follicles expressed GM3. No granulosa cells of these follicles expressed GM3. When a surge dose of FSH/LH was injected, Graafian follicles were formed and GM3 expression was detected in granulosa cells of these follicles. After ovulation, cumulus cells kept expressing GM3 in the ampulla region of ovulated oviduct. The follicles did not show GM3 expression in their granulosa cells after an ovulatory dose of FSH/LH. At 48 h after in vitro culture with FSH/LH of granulosa cells from preantral follicles, GM3 was expressed to a detectable extent on the outer part of the granulosa layer. Finally, at 72 h after culture, all granulosa cells became positive to anti-GM3 antibody. These data suggest that the expression of ganglioside GM3 in the hypophysectomized rat ovary is spatiotemporally regulated by FSH/LH during follicular development and ovulation.

Rehmannia glutinosa inhibits tumour necrosis factor-alpha and interleukin-1 secretion from mouse astrocytes.

We investigated whether an aqueous extract of Rehmannia glutinosa steamed root (RGAE) inhibits secretion of tumour necrosis factor-alpha (TNF-alpha) and interleukin-1 (IL-1) from primary cultures of mouse astrocytes. RGAE dose-dependently inhibited the TNF-alpha secretion by astrocytes stimulated with substance P (SP) and lipopolysaccharide (LPS). IL-1 has been shown to elevate TNF-alpha secretion from LPS-stimulated astrocytes while having no effect on astrocytes in the absence of LPS. We therefore investigated whether IL-1 mediated inhibition of TNF-alpha secretion from astrocytes by RGAE. Treatment of RGAE to astrocytes stimulated with both LPS+SP decreased IL-1 secretion. Moreover, incubation of astrocytes with IL-1 antibody abolished the synergistic cooperative effect of LPS+SP. These results suggest that RGAE may inhibits TNF-alpha secretion by inhibiting IL-1 secretion and that RGAE has an anti-inflammatory activity in the central nervous system curing some pathological disease states.

Developmental patterns of GalBeta1,3(4)GlcNAc alpha2,3-sialyltransferase (ST3Gal III) expression in the mouse: in situ hybridization using DIG-labeled RNA probes.

Sialic acids are key determinants for biological processes, such as cell-cell interaction and differentiation. Sialyltransferases contribute to the diversity in carbohydrate structure through their attachment of sialic acid in various terminal positions on glycolipid and glycoprotein (N-linked and O-linked) carbohydrate groups. Galbeta 1,3(4)GlcNAc alpha2,3-sialyltransferase (ST3Gal III) is involved in the biosynthesis of sLe(x)and sLe(a) known as selectin ligands and tumor-associated carbohydrate structures. The appearance and differential distribution of ST3Gal III mRNA during mice embryogenesis [embryonic (E) days; E9, E11, E13, E15] were investigated by in situ hybridization with digoxigenin-labeled RNA probes coupled with alkaline phosphatase detection. On E9, all tissues were positive for ST3Gal III mRNA expression, whereas ST3Gal III mRNA on E11 was not detected throughout all tissues. On E13, ST3Gal III mRNA was expressed in different manner in various tissues. In this stage, ST3Gal III mRNA was positive only in the liver, pancreas and bladder. On E15, specific signal for ST3Gal III was detected in the liver, lung and forebrain. These results indicate that ST3GAI III is differently expressed at developmental stages of mice embryo, and this may be importantly related with regulation of organogenesis in mice.

Inhibition of mast cell-dependent anaphylaxis by sodium salicylate.

Sodium salicylate (NaSal) is a commonly used agent with a wide pharmacological spectrum. The objective of the present study was to investigate the effect of NaSal on anaphylaxis. NaSal (10-1 and 1 mm) significantly inhibited systemic anaphylaxis induced by compound 48/80 in rats. NaSal also significantly inhibited local anaphylaxis activated by anti-dinitrophenyl (DNP) immunoglobulin E (IgE). NaSal (10-1 and 1 mm) significantly inhibited histamine release from rat peritoneal mast cells (RPMC) activated by compound 48/80 or anti-DNP IgE. Northern-blot analysis demonstrated that a significantly reduced level of the mRNA of L-histidine decarboxylase was expressed in mast cells treated with NaSal, compared with that without NaSal. NaSal (10-2 and 10-1 mm) had a significant inhibitory effect on anti-DNP IgE-induced tumour necrosis factor-alpha secretion from RPMC. The level of cyclic AMP in RPMC, when NaSal (1 mm) was added, transiently and significantly increased about sixfold compared with that of basal cells. These results suggest a possible use of NaSal in managing mast cell-dependent anaphylaxis.