Onion (Allium cepa L.) peel extract (OPE) regulates human sperm motility via protein kinase C-mediated activation of the human voltage-gated proton channel.

Onion (Allium cepa L.) and quercetin protect against oxidative damage and have positive effects on multiple functional parameters of spermatozoa, including viability and motility. However, the associated underlying mechanisms of action have not yet been identified. The aim of this study was to investigate the effect of onion peel extract (OPE) on voltage-gated proton (Hv1) channels, which play a critical role in rapid proton extrusion. This process underlies a wide range of physiological processes, particularly male fertility. The whole-cell patch-clamp technique was used to record the changes in Hv1 currents in HEK293 cells transiently transfected with human Hv1 (HVCN1). The effects of OPE on human sperm motility were also analyzed. OPE significantly activated the outward-rectifying proton currents in a concentration-dependent manner, with an EC50 value of 30 µg/mL. This effect was largely reversible upon washout. Moreover, OPE induced an increase in the proton current amplitude and decreased the time constant of activation at 0 mV from 4.9 ± 1.7 to 0.6 ± 0.1 sec (n = 6). In the presence of OPE, the half-activation voltage (V1/2 ) shifted in the negative direction, from 20.1 ± 5.8 to 5.2 ± 8.7 mV (n = 6), but the slope was not significantly altered. The OPE-induced current was profoundly inhibited by 10 µm Zn2+ , the most potent Hv1 channel inhibitor, and was also inhibited by treatment with GF109203X, a specific protein kinase C (PKC) inhibitor. Furthermore, sperm motility was significantly increased in the OPE-treated groups. OPE exhibits protective effects on sperm motility, at least partially via regulation of the proton channel. Moreover, similar effects were exerted by quercetin, the major flavonoid in OPE. These results suggest OPE, which is rich in the potent Hv1 channel activator quercetin, as a possible new candidate treatment for human infertility.

Effects of eupatilin on the contractility of corpus cavernosal smooth muscle through nitric oxide-independent pathways.

Eupatilin (5,7-dihydroxy-3,4,6-trimethoxyflavone) is one of the main compounds present in Artemisia species. Eupatilin has both antioxidative and anti-inflammatory properties and a relaxation effect on vascular contraction regardless of endothelial function. We evaluated the relaxant effects of eupatilin on the corpus cavernosum (CC) of rabbits and the underlying mechanisms of its activity in human corpus cavernosum smooth muscle (CCSM) cells. Isolated rabbit CC strips were mounted in an organ bath system. A conventional whole-cell patch clamp technique was used to measure activation of calcium-sensitive K+ -channel currents in human CCSM cells. The relaxation effect of eupatilin was evaluated by cumulative addition (10-5  m ~ 3 × 10-4  m) to CC strips precontracted with 10-5  m phenylephrine. Western blotting analysis was performed to measure myosin phosphatase targeting subunit 1 (MYPT1) and protein kinase C-potentiated inhibitory protein for heterotrimeric myosin light chain phosphatase of 17-kDa (CPI-17) expression and to evaluate the effect of eupatilin on the RhoA/Rho-kinase pathway. Eupatilin effectively relaxed the phenylephrine-induced tone in the rabbit CC strips in a concentration-dependent manner with an estimated EC50 value of 1.2 ± 1.6 × 10-4  m (n = 8, p < 0.05). Iberiotoxin and tetraethylammonium significantly reduced the relaxation effect (n = 8, p < 0.001 and p = 0.003, respectively). Removal of the endothelium or the presence of L-NAME or indomethacin did not affect the relaxation effect of eupatilin. In CCSM cells, the extracellular application of eupatilin 10-4  m significantly increased the outward currents, and the eupatilin-stimulated currents were significantly attenuated by treatment with 10-7  m iberiotoxin (n = 13, p < 0.05). Eupatilin reduced the phosphorylation level of MYPT1 at Thr853 of MLCP and CPI-17 at Thr38. Eupatilin-induced relaxation of the CCSM cells via NO-independent pathways. The relaxation effects of eupatilin on CCSM cells were partially due to activation of BKCa channels and inhibition of RhoA/Rho-kinase.

Additive effects of Artemisia capillaris extract and scopoletin on the relaxation of penile corpus cavernosum smooth muscle.

The objective was to investigate the cellular effect and action mechanism of Artemisia capillaris extract (ACE) and its component, scopoletin, on penile corpus cavernosum smooth muscle (PCCSM). In vitro study with PCCSM, the precontracted PCCSM with phenylephrine was treated with ACE or scopoletin. Cyclic nucleotides in the perfusate were measured by radioimmunoassay and expression of protein and mRNA of endothelial nitric oxide synthase (eNOS) and neuronal nitric oxide synthase in the perfused PCCSM were measured by western blot and real-time PCR, respectively. The interaction of ACE or scopoletin with udenafil was also evaluated. ACE and scopoletin exerted a significant and concentration-dependent relaxation in PCCSM. The perfusion with ACE or scopoletin significantly increased cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP) and the perfusion with ACE or scopoletin increased the expression of eNOS mRNA and protein. Furthermore, ACE or scopoletin enhanced udenafil-inducing relaxation in PCCSM. ACE and scopoletin relaxed the PCCSM mainly by activating nitric oxide-cGMP system and cAMP pathway and they may be additive therapeutic candidates for ED patients who do not completely respond to udenafil.

Effect of fulminant hepatic failure porcine plasma supplemented with essential components on encapsulated rat hepatocyte spheroids.

The development of bioartificial liver (BAL) systems has required detailed information about the functional capabilities of cultured hepatocytes during blood or plasma passage. In this study we investigated the effects of porcine plasma and various supplements on the viability and function of adult rat hepatocytes in vitro. Primary rat hepatocytes cultured in porcine plasma supplemented with various substances showed albumin synthesis rates and viability equal to or higher than those of controls. Supplementation with calcium chloride, magnesium sulfate, trace elements, amino acids, insulin, and epidermal growth factor were essential to maintain viability and high albumin synthesis. Especially, trace elements showed significantly higher and longer albumin secretion. Isolated rat hepatocytes were cultured in Spinner flasks for 24 hours to form spheroids that were harvested and encapsulated with chitosan-alginate solution before transfer to the bioreactor in the BAL system. Encapsulated rat hepatocyte spheroids cultured with porcine plasma including trace elements showed higher viability (57%) than controls (40%) after 24 hours, with ammonia removal values of 30.92 µg/10(6) cells versus the control 9.04 µg/10(6) cells. After 24 hours of operation the urea secretion value of encapsulated rat hepatocyte spheroids cultured in porcine plasma in the presence versus absence of trace elements was 76.73 µg/10(6) cells and 18.80 µg/10(6) cells, respectively. We concluded that encapsulated hepatocyte spheroids in a packed-bed bioreactor operated with human plasma including trace elements enhanced cell viability and liver function as a bases for an in vivo clinical trial of the BAL system.

Feasibility of nonthermal atmospheric pressure plasma for intracoronal bleaching.

AIM: To investigate the effect of nonthermal atmospheric pressure plasma on intracoronal tooth bleaching in blood stained human teeth. METHODOLOGY: Forty extracted single-root and blood stained human teeth were used. The teeth were randomly divided into two groups (n=20): group 1 received 30% HP activated by nonthermal atmospheric pressure plasma in the pulp chamber for 30 min, whilst group 2 received 30% HP alone in the pulp chamber for 30 min. The overall colour changes (ΔE) were assessed using the Commission Internationale de L'Eclairage (CIE) Lab Colour System. The data were analysed using Student's t-test to determine the significant differences. RESULTS: The temperature of all teeth was maintained at approximately 37 °C during plasma bleaching. The plasma treatment with 30% HP resulted in significantly higher bleaching efficacy compared to 30% HP alone in discoloured teeth (P<0.05). The average ΔE values of group 1 and group 2 were 9.24 (0.37) and 4.47 (1.62), respectively, at 30 min. CONCLUSIONS: The application of nonthermal atmospheric pressure plasma to intracoronal bleaching could be a novel and efficient therapy in the bleaching of haemorrhagically stained teeth.

Appearance of novel G-bacteria belonging to acidobacteria in a dairy wastewater treatment plant.

A dairy wastewater treatment plant operates a sequencing batch reactor (SBR) and stimulates enhanced biological phosphorus removal (EBPR) process with alternating anaerobic and aerobic cycles. Occasionally, the plant suffers from a high suspended solids problem in the supernatant. Interestingly, the occurrence of high suspended solids coincided with times when the EBPR process failed to remove phosphorus. To find out if there was a relationship between the EBPR failure and the high suspended solids problem, effluent samples were collected from the site during the period of poor phosphorus removal and examined microscopically. It was found that cocci-shaped bacteria (3-4 microm in diameter) were abundant in the effluent samples and they were clustered in tetrads. These were believed to be G-bacteria and results of both Gram and Neisser staining tests were negative, suggesting that they had few intracellular polyphosphate granules. Using polymerase chain reaction (PCR), cloning and deoxyribonucleic acid (DNA) sequence analysis, the phylogenetic information of in situ G-bacteria was obtained. It was found that all of the recovered clones were clustered in the phylum of Acidobacteria.

Surgical and pathological outcomes of laparoscopic surgery for transverse colon cancer.

PURPOSE: Several multi-institutional prospective randomized trials have demonstrated short-term benefits using laparoscopy. Now the laparoscopic approach is accepted as an alternative to open surgery for colon cancer. However, in prior trials, the transverse colon was excluded. Therefore, it has not been determined whether laparoscopy can be used in the setting of transverse colon cancer. This study evaluated the peri-operative clinical outcomes and oncological quality by pathologic outcomes of laparoscopic surgery for transverse colon cancer. MATERIALS AND METHODS: Analysis of the medical records of patients who underwent laparoscopic colorectal resection from August 2004 to November 2007 was made. Computed tomography, barium enema, and colonoscopy were performed to localize the tumor preoperatively. Extended right hemicolectomy, transverse colectomy, and extended left hemicolectomy were performed for transverse colon cancer. Surgical outcomes and pathologic outcomes were compared between transverse colon cancer (TCC) and other site colon cancer (OSCC). RESULTS: Of the 312 colorectal cancer patients, 94 patients underwent laparoscopic surgery for OSCC, and 34 patients underwent laparoscopic surgery for TCC. Patients with TCC were similar to patients with OSCC in age, gender, body mass index, operating time, blood loss, time to pass flatus, start of diet, hospital stay, tumor size, distal resection margin, proximal resection margin, number of lymph nodes, and radial margin. One case in TCC and three cases in OSCC were converted to open surgery. CONCLUSIONS: Laparoscopic surgery for transverse colon cancer and OSCC had similar peri-operative clinical and acceptable pathological outcomes.

High-dose prednisolone and psoralen ultraviolet A combination therapy in 36 patients with vitiligo.

It is well known that systemic corticosteroids arrest the progress of vitiligo and lead to repigmentation, but they may produce unacceptable side-effects. The use of high-dose prednisolone therapy to minimize the side-effects of systemic steroids has been reported, but there have been no reports on the effectiveness of such treatment combined with phototherapy. We evaluated the efficacy and safety of combination therapy with intravenous prednisolone and psoralen ultraviolet A (PUVA). In 36 patients with vitiligo, intravenous methylprednisolone for 3 days was followed by PUVA twice weekly. After 6 months, vitiligo lesions on the face were reduced in size by 57.5%, on the upper extremities by 34.5%, on the trunk by 30.4% and on the lower extremities by 26.3%. Overall, improvement was seen in 13 patients (36.1%), with >50% repigmentation. Side-effects were mild and transient. We conclude that combination treatment of high-dose prednisolone therapy and PUVA may represent a highly effective therapeutic option for generalized vitiligo.

Mitigating nutrient leaching with a sub-surface drainage layer of granulated tires.

Markets for scrap tires have expanded since the early 1990s with the development of value-added applications such as tire-derived fuel and crumb-rubber-amended asphalt. Granulated tires have also displayed the ability to adsorb volatile organic compounds, indicating that the rubber material can be a useful filter media. Sand-based root zones, typically used for golf course putting green and athletic field construction, lack sufficient cation exchange capacity to restrict nitrogen and phosphorus migration through the root zone and into sub-surface drainage systems. Therefore, the adsorptive properties of tire rubber for retaining nitrogen and phosphorus were studied when applied as a distinct sub-surface drainage or intermediate layer in golf course putting greens. A statistically significant reduction in the concentration of nitrate in leachate was achieved by replacing traditional pea gravel with equally sized granulated tires for the drainage layer media, although the mechanism of nitrate mitigation remains unclear. The results indicate that using granulated tires as a drainage layer or fill material beneath sand-based root zones does not compromise the function of the profile or quality of the vegetation while creating an environmentally beneficial and value-added option for scrap tire reuse.

Improvement of cadmium ion removal by base treatment of juniper fiber.

Juniper is a small-diameter underutilized lignocellulosic material. We evaluated the efficacy of base-treated juniper fiber (BTJF) for cadmium (Cd2+) sorption and the viability of juniper fiber as a sorbent for removing Cd2+ from water. Fourier transform infrared spectroscopy analysis indicated that carboxylate ion is a major functional group responsible for Cd2+ sorption. The apparent ideal sodium hydroxide concentration for base treatment is approximately 0.5M. A batch sorption isotherm test showed that equilibrium sorption data were better represented by the Langmuir model than the Freundlich model. After base treatment, the maximum Cd2+ sorption loading, Qmax, was greatly improved (9.18-29.54 mg/g), despite a decrease in specific surface area. A pseudo-second-order kinetic model fitted well for the sorption of Cd2+ onto BTJF. Initial metal ion concentration and treatment alkalinity were found to be major parameters influencing the kinetics of the sorption reaction. As a result of its strong ability to bind cadmium and its faster kinetics in low concentration, BTJF could be an inexpensive and efficient sorbent for removing heavy metals from stormwater runoff.

Nitric oxide synthase isoforms I, III and protein kinase-Ctheta in skeletal muscle fibres of normal and streptozotocin-induced diabetic rats with and without Ginkgo biloba extract treatment.

The expression of nitric oxide synthase (NOS) isoforms I, III and protein kinase-Ctheta (PKCtheta) in rat vastus lateralis muscle was demonstrated immunohistochemically and then correlated to the physiological metabolic fibre types: SO (slow-oxidative), FOGI, FOGII (fast-oxidative glycolytic; I more glycolytic, II more oxidative), and FG (fast-glycolytic). NOS expression in muscles from different experimental groups (normal and diabetic rats, with and without Ginkgo biloba extract treatment) was assayed by Western blotting. Generally, NOS I and PKCtheta were co-expressed in fibres with predominantly oxidative metabolism (SO, FOGII). This suggests an interplay of PKCtheta and NOS I in nitric oxide production by oxidative fibres. NOS III was more highly expressed in fibres with predominantly glycolytic metabolism (FOGI, FG). A somewhat lower NOS I immunoreactivity was also found in NOS III positive fibres suggesting that NOS III and NOS I are co-expressed in these fibres. Western blotting revealed that NOS I as well as NOS III expression in the vastus lateralis muscle was down-regulated in diabetes and increased after Ginkgo biloba extract treatment. These effects may be associated with a diminished glucose uptake by myocytes of diabetic musclesand with an improved muscle function after Ginkgo biloba treatment.

A biological phosphorus removal potential test for wastewaters.

A simple test was proposed to assess whether phosphorus in a wastewater can be removed using a biological phosphorus removal (BPR) process. The test includes the measurement of phosphorus release during 2 hours of the anaerobic stage in a batch reactor containing phosphorus-accumulating organisms (PAOs) and estimation of the effluent phosphorus concentration using biochemical relationships. The BPR potential test developed allowed for the successful evaluation of BPR feasibility for five wastewater samples. The BPR potential test was validated by comparing the test results with the effluent phosphorus concentrations measured in a sequencing batch reactor (SBR). An effluent phosphorus concentration of 1.9 mg P/L predicted for the BPR potential test performed was close to the effluent phosphorus concentration of 1.8 mg P/L obtained from an SBR on the same day. During the anaerobic stage, phosphorus release was significantly affected by the sludge concentration initially, but became insignificant after 2 hours. The initial sludge concentration affected the phosphorus release rate; thus, it is recommended that the BPR potential test be conducted at a target mixed liquor volatile suspended solids concentration. It is also recommended that the BPR potential test be conducted at the site where the PAO-containing sludge is available and the wastewater sample can be delivered at 4 degrees C in less than 24 hours. The PAOs in different sludges had almost identical phosphorus release after 2 hours; however, the characteristics of facultative bacteria in sludges affected the phosphorus release. If the wastewater is prefermented for at least 3 days before the BPR potential test, the amount of phosphorus released by various PAO-containing sludges is expected to be identical.

Comparison of fatty acid composition and kinetics of phosphorus-accumulating organisms and glycogen-accumulating organisms.

It was demonstrated that glycogen-accumulating organisms (GAOs) were able to compete with phosphorus-accumulating organisms (PAOs) for acetate in a biological phosphorus removal (BPR) process, leading to a loss of BPR capability. Cellular fatty acid composition, which serves as a fingerprint for microbial identification, was used to determine microbial population change and to investigate the competition mechanisms of PAOs and GAOs. Analysis of cellular fatty acid composition indicated that PAOs grown with acetate and glucose were different species and that GAOs and PAOs grown with the same substrate were also different species. Glycogen-accumulating organisms seemed to coexist with PAOs even in a well-developed BPR process. The GAOs were able to accumulate more poly-beta-hydroxybutyrate (PHB) and glycogen than PAOs during the anaerobic stage of the BPR process. The GAOs synthesized more in-cell glycogen than PAOs. The growth rate for PAOs was always greater than that for GAOs at various acetate or glucose concentrations, while GAOs had higher acetate uptake and PHB synthesis rates than PAOs. Therefore, GAOs are thought to compete with PAOs only at long solids retention times (> or = 20 days).

A prospective randomized trial comparing intravenous 5-fluorouracil and oral doxifluridine as postoperative adjuvant treatment for advanced rectal cancer.

BACKGROUND: Postoperative adjuvant chemoradiation treatment after curative resection for rectal cancer was needed to reduce recurrence and improve a survival rate. Intravenous 5-fluorouracil (5-FU) and leucovorin has been a mainstay of chemotherapy, but oral 5-FU derivatives have been shown a comparable antitumor activity. Intravenous 5-FU and oral doxifluridine were compared with respect to therapeutic efficacy, drug toxicity, and quality of life. METHODS: A total of 166 patients were randomized to receive intravenous 5-FU (450 mg/m2/day) or oral doxifluridine (900 mg/m2/day) in combination with leucovorin (20 mg/m2/day) for depth of invasion, nodal status, metastasis (TNM) stage II and III patients between October 1997 and February 1999. Consecutive daily intravenous infusion for 5 days per every month for a total of 12 cycles (IV arm, n = 74) and oral doxifluridine daily for 3 weeks and 1 week rest for a total of 12 cycles (oral arm, n = 92). Drug toxicity and quality of life were observed. Quality of life was scored according to 22 daily activity items (good, > or =71; fair, < 70; poor, < 52). RESULTS: There was no difference of sex between two groups (IV arm: male/female = 45/29, oral arm: male/female = 59/33). The mean age was 52.3 vs. 59.5, respectively. There was also no difference of TNM stage distribution and type of operation between groups (P>.05). Mean numbers of chemotherapy cycles were 6.5+/-3.7 (IV arm) vs. 7.2+/-4.3 (oral arm), respectively. The rate of recurrence was 9/74 (12.1%) in the IV arm and 6/92 (6.5 %) in the oral arm, respectively (P = .937). Local recurrence was 2/74 (stage III; 2.7%) in the IV arm and 1/92 (stage II; 1.1%) in the oral arm, respectively. Systemic recurrence was 7/74 (stage III; 9.4%) in the IV arm and 5/92 (stage III; 5.4%) in the oral arm, respectively. The most common site of systemic recurrence was the liver. Toxicity profile was as follows: leukopenia (30/74 vs. 17/92) and alopecia (21/74 vs. 13/92) were statistically more common in the IV arm. Diarrhea was more common in the oral arm. Poor quality of life score between two groups was observed at 1 month (23.9% vs. 13%) and 2 months (15.8% vs. 3.7%) after chemotherapy. Good quality of life score was observed at 1 month (19.5% vs. 49%) and 2 months (47% vs. 72%), respectively (P<.05). CONCLUSIONS: Oral doxifluridine with leucovorin shows a comparable therapeutic efficacy to intravenous 5-FU regimen with high quality of life as postoperative adjuvant therapy. The oral regimen also can be safely given with appropriate toxicity and tolerability.

Anticonvulsant compounds from the wood of Caesalpinia sappan L.

80% Aqueous MeOH extracts from the wood of Caesalpinia sappan, which showed remarkable anticonvulsant activity, were fractionated using EtOAc, n-BuOH, and H2O. Among them, the EtOAc fraction significantly inhibited the activities of two GABA degradative enzymes, succinic semialdehyde dehydrogenase (SSADH) and succinic semialdehyde reductase (SSAR). Repeated column chromatographies for the fraction guided by activity test led to the isolation of the two active principal components. Their chemical structures were determined to be sappanchalcone and brazilin based on spectral data. The pure compounds, sappanchalcone (1) and brazilin (2), inactivated the SSAR activities in a dose dependent manner, whereas SSADH was inhibited partially by sappanchalcone and not by brazilin.

Reduction of electrically evoked neural activity by ginseng saponin in rat hippocampal slices.

It is well established that ginseng saponin has positive influences on various neural diseases, but little is known about its electrophysiological effects in the central nervous system. In this study, we examined the electrophysiological effects of ginseng saponin in rat hippocampal slices. Total saponin from ginseng root reduced the slope of fEPSPs (field excitatory postsynaptic potentials) in the CA1 area in a dose-dependent manner (9.1 +/-5.4%, 48.4+/-12.1%, and 60.5+/-15.3% at 10, 50, and 100 microg/ml, respectively), which was reversed within 10 min of washout. Seven different ginsenosides resulted in varied degrees of fEPSPs reduction. The rank order of reduction was Rb1, Rg1 >Rg2, Rh1, Rc>Rd, Re within a range of 5-64% reduction. No difference in the suppressive action between protopanaxadiol (Rb1, Rc, Rd) and protopanaxatriol (Rg1, Rg2, Re, Rh1) saponins was shown; the slope of fEPSPs was reduced by 38% and 40% on average, respectively. The possible role of gamma-aminobutyric acid (GABA(A)) receptor in the suppressive action of ginseng saponins was tested using whole cell patch recording in acutely isolated hippocampal neurons. Ginsenosides did not induce chloride current nor modified GABA-induced current. Also, the suppressive effect of ginsenosides on fEPSPs was still observed in the presence of the GABA(A) receptor antagonist, bicuculline methiodide 50 microM. These results suggest that the suppressive effect is not attributable to regulation of GABA(A) receptor activation.

Korean red ginseng saponins with low ratios of protopanaxadiol and protopanaxatriol saponin improve scopolamine-induced learning disability and spatial working memory in mice.

The effects of two ginseng saponins having a different ratio of protopanaxadiol (PD) and protopanaxatriol saponins (PT) on the learning impairment induced by scopolamine, and learning and memory in mice were investigated in a passive avoidance task and a Morris water maze task. The ratio of PD and PT was 1.24 and 1.46, respectively. Before training, the ginseng saponins were administered intraperitoneally at doses of 50 and 100 mg/kg. The two saponins improved the scopolamine-induced learning impairment at different dosages in mice, 50 and 100 mg/kg, respectively. However, the two saponins did not show a favorable effect on learning and memory in normal mice. Korean red ginseng saponin with a low PD/PT ratio had an improving effect on spatial working memory, but the saponin with a high PD/PT ratio did not. This finding suggests that the PD/PT ratio of the ginseng saponins may be an important factor in the pharmacological role of red ginseng as a medicinal herb.

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.

Isolation and identification of succinic semialdehyde dehydrogenase inhibitory compound from the rhizome of Gastrodia elata Blume.

In our search for the anticonvulsant constituent of Gastrodia elata repeated column chromatographies guided by activity assay led to isolation of an active compound, which was identified as gastrodin on the basis of spectral data. Brain succinic semialdehyde dehydrogenase (SSADH) was inactivated by preincubation with gastrodin in a time-dependent manner and the reaction was monitored by absorption and fluorescence spectroscopic methods. The inactivation followed pseudo-first-order kinetics with the second-rate order constant of 1.2 x 10(3) M-1min-1. The time course of the reaction was significantly affected by the coenzyme NAD+, which affected complete protection against the loss of the catalytic activity, whereas substrate succinic semialdehyde failed to prevent the inactivation of the enzyme. It is postulated that the gastrodin is able to elevate the neurotransmitter GABA levels in central nervous system by inhibitory action on one of the GABA degradative enzymes, SSADH.