Antimelanogenic effects of luteolin 7-sulfate isolated from Phyllospadix iwatensis Makino.

BACKGROUND: Abnormal deposition of melanin may cause an aesthetic skin problem; therefore, the control of unwanted excessive melanin synthesis is the major goal of cosmetic research. OBJECTIVES: To identify novel tyrosinase (TYR) inhibitors from marine plants and examine their cellular antimelanogenic effects. METHODS: The extracts of 50 marine plants endemic to Korea were screened against human TYR. Active constituents were then isolated from the selected plant extracts that showed potential and their chemical structures elucidated. Furthermore, their antimelanogenic effects were examined using murine melanoma B16/F10 cells and human epidermal melanocytes (HEM). RESULTS: Among the tested extracts, that of Phyllospadix iwatensis Makino exhibited the strongest human TYR inhibitory activity. The active constituents were purified from the butanol fraction of the P. iwatensis extract and identified as hispidulin 7-sulfate and luteolin 7-sulfate. Luteolin 7-sulfate inhibited human TYR more strongly than hispidulin 7-sulfate, luteolin, hispidulin and arbutin. Furthermore, luteolin 7-sulfate showed lower cytotoxicity than luteolin in both B16/F10 cells and HEM. Luteolin 7-sulfate attenuated cellular melanin synthesis more effectively in B16/F10 cells and HEM stimulated by α-melanocyte-stimulating hormone and l-tyrosine than arbutin. CONCLUSIONS: This study demonstrates that luteolin 7-sulfate isolated from P. iwatensis is a human TYR inhibitor with advantageous antimelanogenic properties, and would be useful for development as a therapeutic agent for the control of unwanted skin pigmentation.

Treatment outcomes and moxifloxacin susceptibility in ofloxacin-resistant multidrug-resistant tuberculosis.

SETTING: A tertiary referral centre in Seoul, South Korea. OBJECTIVE: To investigate the effect of moxifloxacin (MFX) susceptibility and later-generation fluoroquinolone (FQ) use on the treatment outcomes of ofloxacin (OFX) resistant multidrug-resistant tuberculosis (MDR-TB). DESIGN: Of 223 patients diagnosed with MDR-TB between January 2006 and December 2012, 70 (31.4%) patients with OFX-resistant MDR-TB were enrolled in this retrospective cohort study. Their treatment outcomes were analysed. RESULTS: The mean age (standard deviation) of the 70 patients was 40.6 (12.9) years; 43 (61.4%) were male and 26 (37.1%) had extensively drug-resistant TB. Of the 70 patients, 22 (31.4%) had MFX-susceptible TB, while the remaining 48 (68.6%) were MFX-resistant. The MFX-susceptible and -resistant groups were comparable in terms of baseline characteristics (including age, sex and radiological severity), and respectively 90.9% (20/22) and 70.8% (34/48) were treated with later-generation FQ-containing regimens (P = 0.074; mainly MFX [40/54, 74.1%]). Treatment success was achieved in 72.7% (16/22) of the MFX-susceptible patients and in 41.7% (20/48) of the MFX-resistant patients (P = 0.021). Treatment failure was significantly higher in the MFX-resistant group (41.7% [20/48] vs. 9.1% [2/22]; P = 0.006). CONCLUSION: Patients with OFX-resistant MDR-TB had significantly better treatment outcomes when susceptible to MFX. This probably reflects the effect of later-generation FQ treatment.

Anti-glutamatergic effect of riluzole: comparison with valproic acid.

Riluzole, an anti-amyotrophic lateral sclerosis drug, known to decrease presynaptic glutamate release, is viewed as a candidate supplementary medication for epilepsy. In the present study, we compared the effects of riluzole and valproate (VPA) in the pilocarpine-induced limbic seizure model and in the gamma-hydroxybutyrate lactone (GBL)-induced absence seizure model. We applied immunohistochemical study for vesicular transporter 1 (VGLUT1) and extracellular recording in the rat dentate gyrus of both pilocarpine- and GBL-induced seizure models to measure effects of riluzole and VPA. Both VPA and riluzole treatments reduced VGLUT1 immunoreactivity. Riluzole treatment completely inhibited pre-ictal spikes and spike-wave discharges in the pilocarpine- and GBL-induced epilepsy models, whereas VPA partially inhibited these phenomena. In both seizure models, the anti-epileptic effects of VPA and riluzole are basically related to anti-glutamatergic (reducing field excitatory postsynaptic potential slope and excitability ratio), not GABAergic (paired-pulse inhibition) effect. Riluzole was more effective at reducing seizure activity in both epilepsy models than VPA. These results suggest that riluzole is a potential antiepileptic drug with activity against limbic seizure and absence seizure.

Vagus nerve stimulation in pediatric intractable epilepsy: a Korean bicentric study.

OBJECTIVE: To present our experience with vagus nerve stimulation (VNS) and to evaluate the long-term efficacy and safety of the procedure in pediatric intractable epilepsy. METHODS: This study included sixteen patients, who were implanted with a vagus nerve stimulator and could be followed up for at least more than 12 months in two epilepsy centers. Data including seizure frequency, EEG, quality of life measures and adverse events were prospectively filed over a 5-year period. RESULTS: VNS resulted in a > 50% reduction in seizure frequency in 50.0% (8/16) of children with 31.3% (5/16) of patients achieving a > 90% reduction. Additionally, enhancements in quality of life were as follows: memory in 50.0% (8/16), mood in 62.5% (10/16), behavior in 68.8% (11/16), alertness in 68.8% (11/16), achievement in 37.5% (6/16), and verbal skills in 43.8% (7/16) of the patients. Adverse events included hoarseness in two patients, dyspnea during sleep in two patients, and sialorrhea in one patient. However, these events were tolerable or could be controlled by the adjustment of output currents. In one patient, wound revision was required. CONCLUSION: Our data supports the role of VNS as an alternative therapy for pediatric intractable epilepsy.

A study of NH3-N and P refixation by struvite formation in hybrid anaerobic reactor.

This research is concerned with the removal of ammonia nitrogen and phosphorus in foodwaste by crystallization. Reductions have been achieved by struvite formation after the addition of magnesium ions (Mg2+). Magnesium ions used in this study were from magnesium salts of MgCl2. The results of our analysis using scanning electron microscopy and energy dispersive X-ray analysis showed that the amount of struvite in precipitated sludge grew enough to be seen with the naked eye (600-700 microm). EDX analysis also showed that the main components of the struvite were magnesium and phosphorus. NH3-N removal efficiency using MgCl2 was 67% while PO4-P removal efficiency was 73%. It was confirmed that nitrogen and phosphorus could be stabilized and removal simultaneously through anaerobic digestion by Mg, NH3 and PO4-P, which were necessary for struvite formation.

Molecular characterization of a cDNA encoding putative calcium binding protein, HvCaBP1, induced during kernel development in barley (Hordeum vulgare L.).

Seed development is known to involve complex physiological and molecular events. In order to gain information on the molecular events that occur in the grains of barley during kernel development, we conducted suppression subtractive hybridization (SSH) using grains of barley cv. Karl at 14 days after fertilization (DAF) as the tester and grains at 5 DAF as the driver. We isolated an SSH clone that showed homology with a specific calcium binding protein in rice called EFA27. Screening the cDNA library, we identified two clones as a calcium binding protein. These clones, each carrying one calcium-binding EF-hand motif, were designated HvCaBP1 (Hordeum vulgare Calcium Binding Protein 1). HvCaBP1 possesses an N-terminal region with a conserved single Ca(2+)-binding EF-hand motif and one transmembrane helix. Northern hybridization showed that the highest expression occurred in grains and that expression increased in kernels at 8 DAF. As shown in situ hybridization, the HvCaBP1 gene was highly expressed in the embryo and tissues of the endosperm near the embryo and was detected in the vascular tissues of the glume in the kernel at 8 DAF. Accumulation of HvCaBP1 mRNAs subsequently increased in vegetative tissues for up to 48 h after abscisic acid (ABA) treatment. Transcripts of HvCaBP1 mRNAs may be regulated by endogenous ABA in the grains during kernel development.

The induction of BDNF and c-fos mRNA in the hippocampal formation after febrile seizures.

In this study we investigated the expression of brain-derived neurotrophic factor (BDNF) and c-fos mRNA in the hippocampal formation after febrile seizures (FSs) with in situ hybridization histochemistry using riboprobes. The induction of BDNF mRNA was firstly observed in the dentate gyrus at 30 min after FSs. The expression in the dentate gyrus peaked at 3 h and returned to basal level at 24 h. It was also observed in the CA3 of hippocampus from 2 to 3 h. The induction of c-fos mRNA was observed in the dentate gyrus at 30 min and 1 h. These observations suggest that BDNF and c-fos are the genes whose expression can be altered by FSs and might be related to pathologic alterations after FSs.

Multijet and multistage aerosol concentrator: design and performance analysis.

A multijet and multistage aerosol concentrator was designed and fabricated with two virtual impactors in a series. Collection efficiency, internal loss, and concentration factors were calculated at ambient conditions for each stage. The total inlet flow rate of the aerosol concentrator was set at 1000 L/min(-1), while the minor flow rate for the first stage was at 6.0% of the total inlet flow and the minor flow rate of the second stage was at 6.7% of the first stage minor flow. The aerosol concentrator was calibrated using polystyrene latex particles in aerodynamic sizes ranging from 0.5 to 10 microm. Several configurations of the multijet acceleration nozzles and multitube receptors were designed in this study. The effects of the different designs were subsequently evaluated through experimentation. It was found that a properly designed multijet and multistage aerosol concentrator can significantly improve aerosol concentration performance. Results showed that the concentration factor increases from 1 to 240 over the particle size range studied. Applications of the multijet and multistage aerosol concentrator with high-volume flow rate can vary widely, from detection of biological aerosols at low concentration, laboratory aerosol sampling, clean room monitoring, and ambient aerosol measurements.

Effects of pH and trace minerals on long-term starvation of Leuconostoc mesenteroides.

Laboratory experiments have definitively shown that exopolymer-producing bacteria have the potential to modify the flow of fluids in oil reservoirs to enhance oil production. Once injected into the reservoir, they will be subjected to a wide range of pH values and to starvation resulting from nutrient depletion. For successful field implementation it is necessary to have a fundamental understanding of these effects on the viability of bacteria. This paper addresses the effects of pH and trace minerals on cell viability of Leuconostoc mesenteroides during carbon source depletion. Two different carbon sources were used to grow cells before transferring the cells to starvation conditions: sucrose and a combination of glucose and fructose. These substrates were chosen because L. mesenteroides produces a significant amount of water-insoluble exopolymers (dextran) under sucrose-fed conditions, which may enhance cell survival under harsh conditions. The effects of dextran on the cell viability were tested at different pH values with and without trace minerals. The rate of cell death followed an exponential-decay law for different values of the solution pH. The optimal solution pH for survival was pH 5, whereas cells died rapidly at pH 3 and below and at pH 13 and above. The sucrose-fed cells showed a greater viability than cells fed glucose and fructose for all pH ranges tested. The results indicated that water-insoluble exopolymers help cells survive for longer periods of time under starvation conditions. The effects of trace minerals on cell culturability were tested at two pH values, 4.5 and 7. For both cases, cells showed a greater culturability (smaller decay rate constant) in the presence of trace minerals than without trace minerals. It was also found that the effects of trace minerals on cell culturability were greater for glucose-fructose-fed cells than for sucrose-fed cells. The Michaelis pH function theory was used for comparing the relationships between the cell decay rate and pH.

Aromatase and sulfatase inhibitors from Lepiota americana.

From an edible mushroom Lepiota americana Pk., (Agaricaceae), 2-aminophenoxazin-3-one that inhibited aromatase at IC50 = 5.7 microM and 3 beta-hydroxy-5,8-epidioxyergosta-6,22-diene that inhibited sulfatase at IC50 = 0.9 microM were isolated. Neither 2-aminophenoxazin-3-one was active against sulfatase nor was 3 beta-hydroxy-5,8-epidioxyergosta-6,22-diene active against aromatase.

Anterior thalamoperforating artery aneurysm associated with internal carotid artery occlusion: case report.

OBJECTIVE AND IMPORTANCE: We describe a rare case of a ruptured distal anterior thalamoperforating artery aneurysm associated with right internal carotid artery occlusion. CLINICAL PRESENTATION: A 59-year-old woman experienced sudden occipital headache, vomiting, and subsequent coma as a result of massive intraventricular hemorrhage. An initial angiogram revealed only an occlusion of the right internal carotid artery just distal to the posterior communicating artery. Repeat angiography 1 month later, however, revealed a saccular aneurysm at a distal anterior thalamoperforating artery in addition to the occlusion of the internal carotid artery. INTERVENTION: We approached this aneurysm through the right temporal horn after opening the ambient cistern. The aneurysm, which was located in the brain parenchyma just medial to the temporal horn, was successfully resected. CONCLUSION: This rare aneurysm probably developed as a result of hemodynamic stress on the anterior thalamoperforating artery after occlusion of the internal carotid artery and/or secondary to chronic hypertension.

cDNA cloning of the CEPUS, a secreted type of neural glycoprotein belonging to the immunoglobulin-like opioid binding cell adhesion molecule (OBCAM) subfamily.

GP55 is a family of glycoproteins distributed predominantly in the nervous system, and its previously characterized members, including the GP55A (EMBL Y08170) and E19S (EMBL Y08171) reveal a typical glycosyl phosphatidyl inositol (GPI)-anchored pattern for membrane proteins. CEPUS identified in this study appeared to represent the third member of GP55. This 3.2 kb long complete cDNA clone from the chicken brain exhibited 3 Ig-like domains. The open reading frame of CEPUS contains 313 amino acids, which can encode a 31.7 kDa core protein (pI 5.75) for the mature form. The signal peptide cleavage site was predicted at Gln25. The structural features of the CEPUS cDNA sequence represented a soluble counterpart to the recently identified cerebellar Purkinje cell specific antigen, CEPU-1. The sequence difference between CEPU-1 and CEPUS was only found in the C-terminus in which the CEPUS lacked the GPI-anchored binding site. It displays significant sequence homology to GP55-related molecules, including OBCAM, GP55A, E19S/LAMP, neurotrimin, and CEPU-1, which are all membrane attached types. The absence of the hydrophobic tail sequence in CEPUS may, therefore, suggest that CEPUS would represent the first identified secreted member in this group of genes. We defined that this molecule forms the opioid-binding cell adhesion molecule (OBCAM) subfamily in the molecular phylogeny. Structurally, these molecules represent acidic proteins (pI 5.47-6.09). Six cysteins, as well as 5 Asn-linked potential glycosylation sites were evolutionary-conserved, suggesting that this OBCAM subfamily resembles immunoglobulin-like and highly glycosylated molecules. The presence of CEPUS would probably suggest to us that the spatial/local expression of the CEPU gene may provide a favorable route for migrating CEPU-positive population of neurons to generate a neuron-specific guidance in developing neurons in vivo.

Acyl-CoA: cholesterol acyltransferase inhibitory activity of ginseng sapogenins, produced from the ginseng saponins.

Ginseng sapogenins were produced from ginseng saponins, isolated from Korean ginseng roots. Ginseng saponins very mildly inhibited acyl-CoA:cholesterol acyltransferase (ACAT) in vitro, however, the sapogenins showed strong inhibitory activity on microsomal ACAT. Therefore, the sapogenins will be one of key ingredients of ginseng affected a lowering of the serum total cholesterol level.

Semisynthesis of abrusoside A methyl ester.

Abrusoside A methyl ester was prepared from abrusogenin through methylation (CH2N2) and a subsequent coupling reaction with 1-chloro-2,3,4,6-tetra-O-acethylglucopyranose in the presence of AgOTf and TMU in CH2Cl2, followed by deacetylation using K2CO3 in MeOH-H2O.

No effect of expression of neuron-restrictive silencer factor (NRSF) protein on N-type Ca2+ channel alpha 1B gene promoter activity in NS20Y cells.

The expression of the N-type voltage-sensitive calcium channel alpha1B gene is restricted to neurons by an 5'-upstream region (-3992 to -1788) containing negative regulation element(s) active in non-neuronal cells (Kim et al., 1997). The neuron-restrictive silencer factor (NRSF) represses the transcription of several neuronal genes in non-neuronal cells by binding to a 21 bp DNA element, termed the neuron-restrictive silencer element (NRSE). To analyze the involvement of NRSF in the neuron-specific expression of the alpha1B gene, the coding region of NRSF cDNA was cloned by PCR and the exogenous NRSF cDNA was transiently cotransfected with tester plasmids into NS20Y neuronal cells which do not contain the endogenous NRSF protein. The luciferase activity of a positive control plasmid NRSEL containing a single copy of the NRSE sequence of the SCG10 gene was repressed 5 fold in HeLa cells containing the endogenous NRSF protein, and its activity was repressed to 44-27% of the control with an increasing amount of exogenous DNA in NS20Y cells. Unlike NRSEL, the promoter activity of the alpha1B subunit-luciferase fusion construct (-3992L) was about 15 fold repressed in HeLa cells compared to NS20Y cells, while any remarkable changes was undetectable in the NRSF expressed NS20Y. These results suggest that the repression of the alpha1B gene in non-neuronal cells may not be mediated by the NRSF function.

Quantitative analysis of gene expression in organotypic slice-explant cultures by particle-mediated gene transfer.

Biolistics, also known as particle-mediated gene transfer, has been used as an effective, method to transfect primary neurons in cultured slices when all other methods have proven unsuccessful. Most of these uses have provided qualitative or semi-quantitative data based on visual assays such as immunohistochemistry. In this paper, we describe a quantitative method of biolistics to analyze gene expression in organotypic cultures of hippocampus and hypothalamus. The method involves co-transfection of the experimental promoters and standard (cytomegalovirus or Rous sarcoma virus) promoters coupled to different reporters (luciferase or beta-galactosidase), with the standard promoter-reporter construct used to 'normalize' the experimental data. Examples and validations of this technique with various cell specific promoters are given: for example, astrocyte-specific and neuron-specific (alpha-tubulin and N-type calcium channel alpha-1B gene) promoters and various tissues (Neuro 2A cells and hippocampal and hypothalamic organotypic slice-explants). An analysis of deletion constructs of the alpha 1B calcium channel subunit gene is described. This method should provide a new opportunity for the analysis of gene expression in diverse neuronal phenotypes.

Anticomplementary activity of constituents from the heartwood of Caesalpinia sappan.

The anticomplementary activity of compounds isolated from the heartwood of Caesalpinia sappan L. (Leguminosae) was investigated. The sterol mixture (campesterol 11.2%, stigmasterol 18.9% and beta-sitosterol 69.9%) was most potent and brazilin, brazilein, and protosappanin E showed a new anticomplementary activity on the complement system in vitro.

Magnone A and B, novel anti-PAF tetrahydrofuran lignans from the flower buds of Magnolia fargesii.

In a search for platelet-activating-factor (PAF) antagonists, two new lignan compounds were isolated from the Chinese crude drug shin-i, the flower buds of Magnolia fargesii. Their structures were elucidated as (2S,3R,4R)-tetrahydro-2-(3,4-dimethoxyphenyl)-4-(3, 4-dimethoxybenzoyl)-3-(hydroxymethyl)furan (magnone A, 1) and (2S,3R, 4R)-tetrahydro-2-(3,4,5-trimethoxyphenyl)-4-(3, 4-dimethoxybenzoyl)-3-(hydroxymethyl)furan (magnone B, 2). Magnones A and B showed antagonistic activity against PAF in the [3H]PAF receptor binding assay with the IC50 values of 3.8 x 10(-5) M and 2.7 x 10(-5) M, respectively.

Ginsenoside Rh2 and Rh3 induce differentiation of HL-60 cells into granulocytes: modulation of protein kinase C isoforms during differentiation by ginsenoside Rh2.

Ginsenoside Rh1 or Rh2 differentiated B16 melanoma or F9 teratocarnoma to phenotypic normal melanocyte-like cells or parietal endoderm-like cells. Ginsenoside Rh3 and Rh4 were recently isolated from Panax ginseng, but their biochemical and pharmacological effects remain unidentified. The present study investigated whether the ginsenoside Rh group (G-Rh1, -Rh2, -Rh3 and -Rh4) having similar structures induce differentiation of HL-60 cells and whether protein kinase C (PKC) is involved in differentiation by ginsenoside. Differentiation was assessed by Wright-Giemsa stain and nitroblue tetrazolium reduction. G-Rh2 and G-Rh3 induced differentiation of HL-60 cells into morphologically and functionally granulocytes but G-Rh1 and G-Rh4 did not. G-Rh2 and G-Rh3 arrested the cell cycle at the G1/S phase, consistent with the ability to induce differentiation in a decreasing order of retinoic acid > G-Rh2 > G-Rh3. During differentiation by G-Rh2, Ca2+/phospholipid-dependent PKC activity was increased in both the cytosol and total cell extract and Ca2+/phospholipid-dependent phosphorylation of 38 and 200 kDa endogenous proteins increased, while phosphorylation of 60, 64, 66 and 97 kDa proteins was Ca2+/phospholipid-independent. When cytosolic PKC isoforms were analyzed by immunoblotting, no significant change was observed in the alpha level, however, the immunoreactive 60 kDa band of a similar mass to the PKC catalytic fragment appeared following treatment with G-Rh2. The beta isoform was gradually increased with prolonged treatment. The gamma isoform was not detected in the cytosol of untreated cells, whereas a small amount was detected 5 days after treatment. It is concluded that G-Rh2 and G-Rh3 can induce differentiation of HL-60 cells into granulocytes and modulation of PKC isoform levels may contribute to differentiation of HL-60 cells by G-Rh2.

Can antioxidant supplementation slow the aging process?

The oxidative stress theory of aging is well supported by accumulated evidence from various aging intervention studies. Early antioxidant supplementation studies indicate life span extensions by antioxidant feeding in various experimental organisms. Data collected under tightly controlled conditions show that the feeding of 2-mercaptoethanol (0.25%) effectively prolonged both the median and maximum life spans of mice. Evidence has been obtained showing dietary vitamin E to protect against oxidative damage to DNA in human lymphocytes and white blood cells. Other clear evidence of vitamin E's protective effect has been seen in its suppressive action of LDL oxidation both in vitro and in vivo. New evidence on the physiological roles of antioxidants, in addition to their well-known role as free radical scavengers, is emerging from recent research. For instance, the beneficial effect of vitamin E in improving glucose transport and the insulin sensitivity and its putative role as a regulator of cell proliferation should open new research dimensions. This presentation will review some of the anti-aging aspects of dietary antioxidant supplementation, as well as the potential problems of its long-term administration that stem from our lack of knowledge about free radical metabolism and the regulation of endogenous defense mechanisms.