Aspergillus awamori-fermented mung bean seed coats enhance the antioxidant and immune responses of weaned pigs.

The potential benefits of Aspergillus-fermented mung bean seed coats (FMSC) for weaned pigs remain unexplored. Both in vitro and in vivo experiments were employed to evaluate the potential of FMSC supplement on the growth, antioxidant and immune responses of weaned pigs. The total polyphenols and DPPH scavenging capability of ethanol extract of FMSC exhibited a greater (p < 0.01) increase than those of pre-fermentation. With the addition of the polyphenol of FMSC extract, an increase in phagocytosis by neutrophils and proliferation of peripheral blood mononuclear cells (PBMC) were found. However, these observations were significantly inhibited (p < 0.05) in those activated cells. Next, 96 weaned pigs were allotted with a randomized complete block design into four dietary treatments, including 0 (control), 600, 1200 or 1800 mg/kg FMSC in a corn-soya bean meal basal diet for a 35-day trial. The pigs were injected with swine enzootic pneumonia (SEP) vaccines at day 3 and day 21 respectively. The results showed that dietary treatment failed to affect growth performance or serum SEP titre. The diet supplemented with 600-1800 mg/kg FMSC decreased faecal lactoferrin on day 21 and increased plasma trolox equivalent antioxidant capacity (TEAC) and erythrocytes catalase activity, as well as decreased (p < 0.01) plasma malondialdehyde (MDA) concentration on day 35. Diet supplementation of 1800 mg/kg FMSC increased phagocytosis by neutrophils and PBMC proliferation induced by pokeweed mitogen (PWM). However, the polymorphonuclear leucocytes (PMN)-positive respiratory burst cells were decreased in the supplementation of 1200 or 1800 mg/kg FMSC respectively. In addition, the serum haptoglobin concentration was decreased in the supplementation with 1200 mg/kg FMSC. Taken together, FMSC enriches polyphenols with antioxidative and immune modulated properties. After feeding FMSC, an improvement in antioxidative capability and immunocompetence was found, implying that FMSC could provide as a feed additive at optimal level 1200 mg/kg for weaned pigs.

The oncogenic role of androgen receptors in promoting the growth of oral squamous cell carcinoma cells.

OBJECTIVES: The aims of this study were to examine the expression of androgen receptors (AR) in oral squamous cell carcinoma (OSCC) cells and tumors and to determine the role of AR in regulating OSCC cell growth. MATERIALS AND METHODS: Four OSCC cell lines were used for analyzing AR expression and transcriptional activity. The effects of AR knockdown on the growth and tumorigenicity of OSCC cells were examined. A series of 11 benign, 22 premalignant, and 21 malignant lesions of the oral cavity were used for analyzing AR expression. RESULTS: OSCC cells expressed AR proteins with differential activities. Stimulation of AR by dihydrotestosterone in OSCC cells caused an increase in cyclin D1 expression and promoted cell growth, whereas treatment with bicalutamide led to decreased cyclin D1 expression and inhibited cell growth. Knockdown of AR expression in OSCC cells resulted in decreased proliferation, increased apoptosis, and inhibited tumorigenicity. Results from immunohistochemical studies showed that AR immunoreactivity was found in 27% (3/11) of benign lesions, while 68% (15/22) of premalignant and 67% (14/21) of malignant lesions showed positive AR staining. CONCLUSION: Our data suggest that OSCC cells express functional AR proteins which are critical for promoting cell growth and causing malignant disease.

Adenoviral gene transfer of bone morphogenetic protein-7 enhances functional recovery after sciatic nerve injury in rats.

Bone morphogenetic proteins (BMPs), members of the transforming growth factor-ß subfamily, function as instructive signals for neuronal lineage commitment and promote neuronal differentiation. However, the mechanism of BMP7 action in vivo after peripheral nerve injury is poorly understood. This study examines the efficacy of gene transfer of adenoviral (Ad) BMP7 on peripheral neuropathy. Transgene expression was found in both Ad-infected sciatic nerves and their respective remote neurons, indicating Ad transduction by a retrograde transport. After AdBMP7 infection to nerves, the sciatic nerves were crushed or transected. Hind limb functional behavior, including rotarod test and sciatic functional index, were conducted in rats weekly after nerve injury. Interestingly, enhanced BMP7 expression significantly improved hind limb functional recovery in AdBMP7-transduced rats when compared with AdGFP-transduced nerve-crushed or transected rats. Furthermore, AdBMP7 transduction reduced injury-induced macrophage activation, nerve demyelination and axonal degeneration. By contrast, AdBMP7 infection did not affect the hyperalgesia paw-withdrawal latency after nerve injury. We further examined the effect of AdBMP7 infection on sciatic nerve explant and Schwann cell cultures. Enhanced cell proliferation was significantly increased by AdBMP7 transduction in both cultures. Taken together, BMP7 overexpression by Ad gene transfer was beneficial in both nerves and Schwann cells on functional recovery after sciatic nerve injury in rats.

Roles of hepatocyte nuclear factors (HNF) in the regulation of reproduction in teleosts.

Hepatocyte nuclear factor (HNF) families are composed of liver-enriched transcription factors and upstream regulators of many liver-specific genes. HNF are involved in liver-specific gene expression, metabolism, development, cell growth and many cellular functions in the body. HNF genes can be activated or influenced by several hormones and insulin-like growth factors (IGF), and different combinations of the four HNF factors form a network in controlling the expression of liver-specific or liver-enriched genes. The functions of these factors and their interactions within the gonads of bony fishes, however, are not well understood, and the related literature is scant. Recently, several members of the HNF families have been detected in teleost gonads together with their downstream genes (IGF-I and IGF-II), suggesting that these HNF could be upregulated in vitro by steroid hormones. Thus, the hormone-HNF-IGF-gonad interaction may be an alternative axis in the reproductive mechanism that acts in concert with the conventional hypothalamus-pituitary-gonad pathway. This may help the early development and maturation of the gonad or gamete, sexual maturity or reversion and spawning-regulating mechanisms among fishes to be understood.

Extending donor source with bench coronary angiography: a case report.

This report proposes a safer, economical method to examine the marginal donor heart for a better chance of use, which also delivers comparable image quality of catheterization (CathLab) without creating potential damage to the kidney. Currently the examination of the coronary system mainly relies on the CathLab, which is not commonly accessible, and also results in nephrotoxic effects. Therefore, bench coronary angiography is hereby proposed because it is commonly available and economical as well as able to indicate coronary lesions for surgeons as well as the CathLab study. These benefits altogether provide a better chance to select usable hearts from older donors to help relieve the organ shortage.

Oral administration recombinant porcine epidermal growth factor enhances the jejunal digestive enzyme genes expression and activity of early-weaned piglets.

This study attempted to determine ingested porcine epidermal growth factor (pEGF) on the gastrointestinal tract development of early-weaned piglets. Thirty-two piglets (14-day weaned) were randomly allotted to supplemented with 0 (control), 0.5, 1.0, or 1.5 mg pEGF/kg diet. Each treatment consisted of four replicates with two pigs per pen for a 14 days experimental period. Piglets were sacrificed and gastrointestinal tract samples were collected to measure mucosa morphology, mRNA expression and activities of digestive enzymes in the gastrointestinal tract of piglets at the end of the experiment. Diets supplemented with pEGF failed to influence growth performance but tended to increase jejunal mucosa weight (p < 0.09) and protein content (p < 0.07). Piglets supplemental pEGF induced incrementally the gastric pepsin activity (p < 0.05) and stimulated jejunal alkaline phosphatase (ALP) and lactase activities accompanied with the increase of jejunal ALP and maltase mRNA expression. No effect of pEGF on the activities of all enzymes in ileum except the stimulation of ileal aminopeptide N mRNA expression. These results reveal that dietary pEGF supplementation might enhance gene expression and activities of digestive enzymes in the stomach and jejunum of piglets.

Synthetic (+)-antroquinonol exhibits dual actions against insulin resistance by triggering AMP kinase and inhibiting dipeptidyl peptidase IV activities.

BACKGROUND AND PURPOSE: The fungal product (+)-antroquinonol activates AMP kinase (AMPK) activity in cancer cell lines. The present study was conducted to examine whether chemically synthesized (+)-antroquinonol exhibited beneficial metabolic effects in insulin-resistant states by activating AMPK and inhibiting dipeptidyl peptidase IV (DPP IV) activity. EXPERIMENTAL APPROACH: Effects of (+)-antroquinonol on DPP IV activity were measured with a DPPIV Assay Kit and effects on GLP-1-induced PKA were measured in AR42J cells. Translocation of the glucose transporter 4, GLUT4, induced either by insulin-dependent PI3K/AKT signalling or by insulin-independent AMPK activation, was assayed in differentiated myotubes. Glucose uptake and GLUT4 translocation were assayed in L6 myocytes. Mice with diet-induced obesity were used to assess effects of acute and chronic treatment with (+)-antroquinonol on glycaemic control in vivo. KEY RESULTS: The results showed that of (+)-antroquinonol (100 µM ) inhibited the DPP IV activity as effectively as the clinically used inhibitor, sitagliptin. The phosphorylation of AMPK Thr(172) in differentiated myotubes was significantly increased by (+)-antroquinonol. In cells simultaneously treated with S961 (insulin receptor antagonist), insulin and (+)-antroquinonol, the combination of (+)-antroquinonol plus insulin still increased both GLUT4 translocation and glucose uptake. Further, (+)-antroquinonol and sitagliptin reduced blood glucose, when given acutely or chronically to DIO mice. CONCLUSIONS AND IMPLICATIONS: Chemically synthesized (+)-antroquinonol exhibits dual effects to ameliorate insulin resistance, by increasing AMPK activity and GLUT4 translocation, along with inhibiting DPP IV activity.

Immune localization of prolactin receptor in the mitochondria-rich cells of the euryhaline teleost (Oreochromis mossambicus) gill.

The present work demonstrates, by Western blotting and immunofluorescent staining, the presence and localization of prolactin (PRL) receptor in tilapia (ti) Oreochromis mossambicus gills. Gill epithelial cells that reacted with PRL receptor antibody were found to be labelled concomitantly with Con A, a marker of the apical crypts in mitochondria-rich (MR) cells. No positive staining was observed in pavement cells or mucus cells with PRL receptor antibody. This indicates that PRL receptors are located specifically in the gill MR cells. Further, the tiPRL receptors were found only in the MR cells of seawater-adapted tilapia gills. The effects of salinity and ions on the expression of tiPRL receptors are discussed.

Modulation of cell cycle regulatory protein expression and suppression of tumor growth by mimosine in nude mice.

Our previous results demonstrated that the plant amino acid mimosine blocked cell cycle progression and suppressed proliferation of human lung cancer cells in vitro by multiple mechanisms. Inhibition of cyclin D1 expression or induction of cyclin-dependent kinase inhibitor p21WAF1 expression was found in mimosine-treated lung cancer cells. However, whether mimosine may modulate the expression of these cell cycle regulatory proteins and suppress tumor growth in vivo is unknown. In this study, we examined the anti-cancer effect of mimosine on human H226 lung cancer cells grown in nude mice. Our results demonstrated that mimosine inhibits cyclin D1 and induces p21WAF1 expression in vivo. Furthermore, results of TUNEL analysis indicated that mimosine may induce apoptosis to suppress tumor growth in nude mice. Collectively, these results suggest that mimosine exerts anti-cancer effect in vivo and might be useful in the therapy of lung cancer.

In vitro regulation of immunoreactive beta-endorphin secretion from adult and fetal hypothalamus by sequential stimulation with corticotropin-releasing hormone.

Previous work has shown corticotropin-releasing hormone (CRH) stimulation of beta-endorphin (END) secretion from hypothalamus. We tested the hypothesis that CRH stimulation of beta-END (measured by radioimmunoassay) from hypothalamic explants is dependent on: (1) ovine CRH dose, (2) pattern and sequence of CRH stimulation, (3) androgen status, and (4) hypothalamic age. Hypothalami from adult male rats and day 17 fetal rats were studied. In adult hypothalami, CRH-stimulated immunoreactive (IR)-beta-END secretion with 10(-7) M was greater than that with 10(-8) M CRH and showed dose-dependent stimulation. Serial stimulation for 20 min by 10(-8) M CRH followed by a 40 min interval without CRH stimulation resulted in a brief stimulation of secretion of IR-beta-END and also secretion of IR-alpha-melanocyte-stimulating hormone (MSH), another peptide derived from pro-opiomelanocortin, the precursor of beta-END. Subsequent stimulation with 10(-6) M CRH showed a desensitization to stimulation despite readily releasable pools of IR-beta-END shown by potassium-induced depolarization. In addition, prolonged stimulation for 1 h with 10(-7) M CRH or increasing concentrations of CRH produced a sustained increase in IR-beta-END release as long as CRH was present. Dihydrotestosterone treatment had no effect on basal nor CRH-stimulated IR-beta-END release in orchiectomized rats. The pattern of IR-beta-END secretion from fetal hypothalamic explants exposed briefly (20 min) or for a prolonged period (1 h) to CRH was similar to that from adult explants. These results demonstrate that: (1) CRH-stimulated IR-beta-END secretion from hypothalamus is dose-dependent.(ABSTRACT TRUNCATED AT 250 WORDS)

Corticotropin-releasing hormone and dexamethasone do not alter secretion of immunoreactive beta-endorphin from dissociated fetal hypothalamic cell cultures.

Corticotropin-releasing hormone (CRH) and glucocorticoids are major regulators of the hypothalamic-pituitary-adrenal (HPA) axis controlling secretion of beta-endorphin and other pro-opiomelanocortin (POMC)-derived peptides from pituitary. Although previous work has shown that CRH stimulates secretion of beta-endorphin from adult hypothalamic explants, and that glucocorticoids can inhibit basal and stimulated secretion of POMC-derived peptides from pituitary, the role of glucocorticoids on hypothalamic beta-endorphin secretion is not known. Studies were performed to assess the effects of CRH and dexamethasone, a potent glucocorticoid, on secretion of immunoreactive (IR) beta-endorphin from dissociated fetal hypothalamic cell cultures. CRH (10(-9)-10(-6) M) did not stimulate secretion of IR-beta-endorphin from hypothalamic cells which did release IR-beta-endorphin upon potassium-induced depolarization. However, CRH did stimulate IR-beta-endorphin secretion from fetal hypothalamic explants which were similar to hypothalamic tissue from which dissociated hypothalamic cell cultures were derived. Exposure of cells to dexamethasone (10(-6) M) did not inhibit basal or potassium-stimulated release of IR-beta-endorphin. These results indicate that: (1) dissociated fetal hypothalamic cells in culture do not exhibit a functional CRH receptor coupled to stimulation of IR-beta-endorphin secretion; (2) exposure of hypothalamic cells to dexamethasone does not inhibit basal nor depolarization-induced release of IR-beta-endorphin; and (3) dissociated fetal hypothalamic cells may have limited utility in elucidating specific regulatory relationships because of in vitro conditions and/or cytoarchitectural relationships.

Activation of cyclic AMP second messenger system stimulates secretion of beta-endorphin from fetal hypothalamic cells.

Relatively little is known about physiological regulators of hypothalamic beta-endorphin (END) secretion and mechanisms by which they stimulate secretion. We sought to determine whether activation of the cyclic AMP (cAMP) second messenger pathway was involved in stimulating hypothalamic beta-END secretion from dissociated fetal hypothalamic cells in culture. Forskolin (FSK), a direct activator of adenylate cyclase which stimulates cAMP formation, stimulated immunoreactive (IR)-beta-END secretion. Because FSK can also stimulate independent of increased cAMP formation, we studied dibutyryl cAMP and 8-bromo-cAMP, analogues of cAMP, which also stimulated IR-beta-END secretion. From these studies we conclude: (1) activation of the cAMP second messenger system stimulates IR-beta-END secretion from hypothalamic cells and supports the rationale that endogenous regulators which stimulate this pathway could be involved in the physiological regulation of hypothalamic beta-END secretion; (2) coupling between the cAMP second messenger pathway and stimulation of hypothalamic beta-END secretion which is presumably present at maturity (adulthood) originates at early stages of development (fetal life).

Protein kinase C activators stimulate beta-endorphin secretion from hypothalamic cells.

Relatively little is known about the regulation of secretion of hypothalamic beta-endorphin, the potent opioid that is believed to play a variety of physiological roles in brain. Previous work has shown that arginine vasopressin (AVP), which acts in brain primarily via activation of the phosphoinositol (PI) second messenger system, stimulates secretion of hypothalamic beta-endorphin. To test the hypothesis that activators of protein kinase C (PKC), which is activated following PI hydrolysis, stimulates secretion of beta-endorphins from hypothalamus, we studied the separate effects of stimulators of PKC including phorbol ester 12-myristate-13-acetate (PMA) and 1-oleolyl-2-acetyl glycerol (OAG- a diacyl glycerol analogue) on secretion of immunoreactive (IR-) beta-endorphin (measured by RIA) from dissociated fetal rat hypothalamic cell cultures. We also studied AVP and angiotensin II (Ang II), hypothalamic peptides which activate the PI second messenger pathway, and interactions of PMA and forskolin (FSK), an activator of the cyclic AMP/protein kinase A (PKA) pathway. PMA, OAG, AVP, and Ang II stimulated IR-beta-endorphin secretion. The stimulatory effect of both PMA and FSK on IR-beta-endorphin secretion was greater than that of PMA or FSK alone and was essentially additive.(ABSTRACT TRUNCATED AT 250 WORDS)

Cyclooxygenase-2 level and culture conditions influence NS398-induced apoptosis and caspase activation in lung cancer cells.

Cyclooxygenases (COXs) catalyze the synthesis of prostaglandins (PGs) from arachidonic acid. Overexpression of COX-2 is frequently found in human cancers and is suggested to play an important role in tumorigenesis. Recent studies indicated that COX-2 inhibitors exert potent anti-cancer effects on a number of cancers. Interestingly, some COX-2 inhibitors potently induce apoptosis, while other COX-2 inhibitors primarily induce growth inhibition. Therefore, there is a variability in the effects that different COX-2 inhibitors have on cancer cells. In this study, we demonstrated that induction of apoptosis of high COX-2-expressing A549 lung cancer cells by a specific COX-2 inhibitor NS398 was observed in cells cultured under serum-free condition. However, this drug induced G1 growth arrest rather than apoptosis in A549 cells maintained in 10% serum medium. Conversely, low COX-2-expressing H226 lung cancer cells were resistant to NS398-induced apoptosis under both serum-free and serum-containing conditions. Moreover, our results showed that NS398-induced apoptosis is associated with activation of caspase-3, a cysteine protease that plays a crucial role in the execution phase of apoptosis. These results suggest that the cytotoxic effect of COX-2 inhibitors on cancer cells may be influenced by extracellular environments and the anti-cancer action of these inhibitors in vivo needs careful evaluation. Additionally, a correlation between the level of COX-2 expression and the extent of apoptosis induced by COX-2 inhibitors was found.

Effects of cortisol and salinity challenge on water balance in developing larvae of tilapia (Oreochromis mossambicus).

Effects of exogenous cortisol on drinking rate and water content in developing larvae of tilapia (Oreochromis mossambicus) were examined. Both freshwater- and seawater-adapted larvae showed increases in drinking rates with development. Drinking rates of seawater-adapted larvae were about four- to ninefold higher than those of freshwater-adapted larvae from day 2 to day 5 after hatching. Seawater-adapted larvae showed declines in drinking rate and water content at 4 and 14 h, respectively, after immersion in 10 mg L(-1) cortisol. In the case of freshwater-adapted larvae, the drinking rate decreased after 8 h of cortisol immersion, while the water content did not show a significant change even after 32 h of cortisol immersion. In a subsequent experiment of transfer from freshwater to 20 ppt (parts per thousand, salinity) seawater, immersion in 10 mg L(-1) cortisol for 8-24 h enhanced the drinking rate in larvae at 4 h after transfer, but no significant difference was found in water contents between cortisol-treated and control groups following transfer. These results suggest that cortisol is involved in the regulation of drinking activity in developing tilapia larvae.

Regulation of drinking rate in euryhaline tilapia larvae (Oreochromis mossambicus) during salinity challenges.

Euryhaline tilapia larvae are capable of adapting to environmental salinity changes even when transferred from freshwater (FW) to seawater (SW) or vice versa. In this study, the water balance of developing tilapia larvae (Oreochromis mossambicus) adapted to FW or SW was compared, and the short-term regulation of drinking rate of the larvae during salinity adaptation was also examined. Following development, wet weight and water content of both SW- and FW-adapted larvae increased gradually, while the dry weight of both group larvae showed a slow but significant decline. On the other hand, the drinking rate of SW-adapted larvae was four- to ninefold higher than that of FW-adapted larvae from day 2 to day 5 after hatching. During acute salinity challenges, tilapia larvae reacted profoundly in drinking rate, that is, increased or decreased drinking rate within several hours while facing hypertonic or hypotonic challenges, to maintain their constancy of body fluid. This rapid regulation in water balance upon salinity challenges may be critical for the development and survival of developing larvae.

Metallothionein induction in early larval stages of tilapia (Oreochromis mossambicus).

Amounts of whole-body metallothionein (MT) in tilapia (Oreochromis mossambicus) larvae increased to a peak (1,500 ng mg(-1) protein) 1 d after hatching (H1), decreased rapidly thereafter, and was maintained at a constant level (700 ng mg(-1)) 3 d after hatching (H3). Waterborne Cd(2+) could stimulate MT expression in newly hatched (H0) larvae in dose-dependent and time-dependent patterns. H0 larvae, which were treated with 35 microg L(-1) Cd(2+) for 24 h, showed a 1.7-fold increase in the MT amount (174.0+/-64.7) and a 6. 5-fold increase in accumulated Cd(2+) but no significant change in Ca(2+) content, compared with the H0 control (MT, 102.6+/-48.1). H3 larvae with the same treatment revealed about a 10-fold increase in accumulated Cd(2+), a 10% decrease in Ca(2+) content, but no change in MT (261.2+/-120.0), compared with the H3 control (MT, 330+/-74.0). H0 larvae could synthesize more MT to bind Cd(2+) for detoxification in 35 microg L(-1) Cd(2+), a dose that would not affect normal physiology or survival of H0 larvae. On the other hand, 35 microg L(-1) Cd(2+) caused H3 larvae to experience hypocalcemia, an abnormal physiological condition, in which H3 larvae could not synthesize sufficient MT, thus causing greater than 25% mortality. These results indicate for the first time that the inducibility of MT by waterborne Cd(2+) is development dependent, being correlated with inconsistent sensitivities to Cd(2+) during larval development.

Effects of cortisol on ion regulation in developing tilapia (Oreochromis mossambicus) larvae on seawater adaptation.

The yolk diameter of cortisol-treated tilapia (Oreochromis mossambicus) larvae, immersed in freshwater (FW) containing 10 mg L-1 cortisol from 48 h postfertilization to 12 d posthatching, was significantly larger than that of control larvae after 8 d of treatment, suggesting that inhibition on larval growth occurred only after a long-term treatment with cortisol. Tilapia embryos or larvae treated with 1-10 mg L-1 cortisol for 1-2 d and then transferred to 20-30 g L-1 seawater (SW) showed reduced cumulative larval mortality in SW compared with controls. Moreover, 4-5 d of cortisol treatments significantly diminished the degree of increase in larval body Na content after the transfer to SW. Significant effect of cortisol on body Na content of larvae occurred as early as 4-8 h after the transfer to SW, while no significant difference was found in the ouabain binding of yolk-sac epithelia between control and cortisol-treated larvae even 12 h after the transfer. Cortisol may be involved in the early phase of SW adaptation in developing larvae, and this mechanism may be achieved by other means than increasing the Na-K-ATPase of yolk-sac epithelia.

Presence of Na-K-ATPase in mitochondria-rich cells in the yolk-sac epithelium of larvae of the teleost Oreochromis mossambicus.

The purpose of this study is to provide biochemical evidence for the functions of the mitochondria-rich cell (MR cell) in the yolk-sac epithelium of the developing larvae of tilapia Oreochromis mossambicus. Western blotting with the antibody (6F) raised against avian Na-K-ATPase alpha1 subunit demonstrated the presence of Na-K-ATPase in yolk-sac epithelium of tilapia larvae and about 1. 46-fold more of the enzyme in seawater larvae than in freshwater ones. The yolk-sac MR cells were immunoreacted to the antibody (alpha5) against the alpha subunit of avian Na-K-ATPase and were double-labeled with anthroylouabain and dimethylaminostyrylethyl-pyridiniumiodine, suggesting the existence and activity of Na-K-ATPase in these cells. Binding of 3H-ouabain in the yolk sac of seawater larvae was much higher than in that of freshwater larvae (4.183+/-0.143 pmol/mg protein versus 1.610+/-0. 060 pmol/mg protein or 0.0508+/-0.0053 pmol/yolk sac versus 0. 0188+/-0.0073 pmol/yolk sac). These biochemical results are further evidence that yolk-sac MR cells are responsible for a major role in the osmoregulatory mechanism of early developmental stages before the function of gills is fully developed.

Effects of Chinese herbal prescriptions on copulatory activity in aged male rats: a preliminary study.

Four herbal prescription medicines, Chi-Pao-Mei-Jan-Tan, Gui-Fu-Ba-Wei-Wan, Huan-Shao-Tan; and San Tsai-Feng-Sui-Tan, were tested for their effects on sexual behavior in aged rats. Crude liquid extracts of these herbs were administered to the rats daily through oral tubing for 14 days. All four herbal prescriptions showed some effects in restoration of mount and intromission behaviors, but there was no effect on restoration of ejaculation in 26 month old rats that had exhibited no copulatory activity (no mount, intromission and ejaculation) previously. The effects of Chi-Pao-Mei-Jan-Tan were further tested in 26 month old rats with low mount and intromission activities but without ejaculation behavior, and in 15 month old rats (middle-age group) that showed normal mount and intromission behavior but no ejaculation activity. Chi-Pao-Mei-Jan-Tan was effective in improving the frequency of both mount and intromission, but failed to restore the ejaculation activity of the old rats with low mount and intromission behaviors. It was, however, very effective in restoration of ejaculation activity in middle-aged rats that exhibited normal mount and intromission behaviors. Serum testosterone (T) levels of Chi-Pao-Mei-Jan-Tan in tested old and middle-aged rats were determined by radioimmunoassay, and showed no difference before and after treatment. Our findings demonstrated that the four herbal prescriptions had some effects in restoration of mount and intromission behaviors, but not ejaculation activity in old rats, and that Chi-Pao-Mei-Jan-Tan was very effective in restoration of ejaculation activity in middle-aged rats. The promotional effect of Chi-Pao-Mei-Jan-Tan on copulatory behavior was not correlated with serum T levels.