Gender difference in clinical and genetic characteristics of Brugada syndrome: SADS-TW BrS registry.

BACKGROUND: Brugada syndrome (BrS) is a heritable sudden cardiac death (SCD) disease with male predominance. Information on gender difference of BrS remains scarce. AIM: To investigate the gender difference of BrS in Han Chinese. DESIGN: We consecutively enrolled 169 BrS patients (153 males and 16 females) from Han Chinese in Taiwan from 1998 to 2017. METHODS: Clinical characteristics, electrocardiographic parameters and SCN5A mutation status were compared between genders. RESULTS: The percentage of family history of SCD in females was slightly higher (31.3% vs. 15%, P = 0.15). Females exhibited longer QTc (457.8 ± 33.0 vs. 429.5 ± 42.1 ms, P < 0.01). Regarding cumulative event occurrence by age, Mantel-Cox test showed females had earlier age of onset of first cardiac events (SCD or syncope) than males (P = 0.049), which was mainly attributed to syncope (P < 0.01). Males with SCD exhibited longer QRS duration (114.2 ± 26.8 vs. 104.8 ± 15.3 ms, P = 0.02) and QTc (442.5 ± 57.4 vs. 422.9 ± 28.8 ms, P = 0.02). Males with syncope exhibited longer PR interval (181.2 ± 33.7 vs. 165.7 ± 27.1 ms, P = 0.01), whereas females with SCD or syncope had a trend towards slower heart rates (69.1 ± 9.6 vs. 82.2 ± 16.3 bpm, P = 0.10) than female with no or mild symptoms. There was no difference in the percentage of SCN5A mutation between genders. CONCLUSION: Gender difference is present in BrS. Females have longer QTc and suffer from syncope earlier than males. Risk of SCD in males is associated with boarder QRS complex and longer QTc, whereas risk of syncope is associated with longer PR interval in males and slower heart rate in females.

Size-effect induced short-range magnetic ordering in germanium nanostructures.

Formation of ordered magnetic states in germanium nanostructures embedded in SiO2 has been investigated. Samples with the nanostructures were prepared by sputtering deposition on Si(100) substrates, followed by thermal annealing in vacuum. Transmission electron microscopy, energy dispersive X-ray spectrometry, and Raman spectroscopy have been used to characterize the samples. Magnetic measurements were performed using a superconducting quantum interference device. Size-effect induced magnetic orderings in the germanium nanostructures were found to be present at room temperatures and below. Superparamagnetic behavior was observed at temperatures above 230 K, whereas thermal excitation of spin reorientation and magnetic coupling has been revealed at temperatures below 60 K. Inverted hysteresis loops with negative remanences and multiple plateaus revealed the ferri- or antiferromagnetic nature of the coupling. Inter-domain coupling and effect of magnetic anisotropy will be discussed based on the experimental results and simulations with a spin reorientation model.

Downregulation of hepatitis B surface antigen expression in human hepatocellular carcinoma cell lines by HD-03, a polyherbal formulation.

Hepatitis B virus is associated with chronic or acute liver diseases and with hepatocellular carcinoma. In the present study, we examined the activity of HD-03, a polyherbal formulation, on two hepatitis B surface antigen (HBsAg) expressing human hepatocellular carcinoma cell lines, PLC/PRF/5 and HepG2/A2. We observed that HD-03 downregulates HBsAg expression from these cell lines. Our studies have also shown that this effect is neither due to cytotoxicity on the cell lines nor due to blockade of the release of the antigen from the cells nor due to binding of the substance with the antigen. The possible mode of its antiviral activity is explained.

Protein kinase C-mediated tyrosine phosphorylation of paxillin and focal adhesion kinase requires cytoskeletal integrity and is uncoupled to mitogen-activated protein kinase activation in human hepatoma cells.

Treatment of cultured human hepatoma HepG2 cells with the protein kinase C (PKC) activator, 12-O-tetradecanoylphorbol-13-acetate (TPA), results in an increase in tyrosine phosphorylation of several proteins, including the focal adhesion kinase (FAK) and paxillin using anti-phosphotyrosine Western blotting and immunoprecipitation. However, when cells are in suspension or in the presence of cytochalasin D which disrupts the intracellular network of actin microfilaments, TPA loses its ability to stimulate tyrosine phosphorylation of FAK and paxillin but it still activates mitogen-activated protein kinase (MAPK) and induces PKC translocation from cytosol to the membrane in HepG2 cells. On the other hand, PD98059, a specific inhibitor of mitogen-activated protein kinase kinase, blocks TPA-induced MAPK activation but has no effect on TPA-induced tyrosine phosphorylation. Our findings suggest that TPA-induced tyrosine phosphorylation of FAK and paxillin in human hepatoma cells is PKC dependent and requires the integrity of the cell cytoskeleton but is uncoupled to the signal transduction pathway of PKC leading to the translocation of PKC and MAPK activation.

Suppressive effects of destruxin B on hepatitis B virus surface antigen gene expression in human hepatoma cells.

Destruxin B, a cyclodepsipeptide was originally identified as a plant pathogen from the fungus, Alternaria brassicae. We examined the antiviral activity of destruxin B and found that it suppresses the expression of the hepatitis B viral surface antigen (HBsAg) gene in human hepatoma Hep3B cells which carry an integrated viral gene in its chromosome. In contrast, destruxin B shows no cytotoxic effect on the viability of the cells. Furthermore, it can be shown that destruxin B can reversibly suppress HBsAg production by Hep3B cells in a concentration-dependent manner with EC50 of 0.5 microM. Northern blot analysis indicates that the suppression of HBsAg gene expression by destruxin B is mainly at the mRNA level. Destruxin B not only suppresses the endogenously expressed HBsAg in the Hep3B cells but also suppresses the HBsAg produced either from the stable transfected HBV DNA in another human hepatoma HuH-7 cell line which carry no endogenous HBV genome. These results suggest that destruxin B may have future potential for development as a specific anti-HBV drug.

Study of structure-activity correlation in destruxins, a class of cyclodepsipeptides possessing suppressive effect on the generation of hepatitis B virus surface antigen in human hepatoma cells.

A new destruxin [destruxin E2 chlorohydrin] was isolated from the culture medium of Metarrhizium anisopliae and its structure was determined by NMR spectroscopy and mass spectrometry. As compared with other destruxins, the new destruxin showed a lower suppressive activity on the production of hepatitis B virus surface antigen in human hepatoma Hep3B cells. NMR study coupled with molecular modeling by computer graphics has revealed that the hydrophobicity nature of the convex surface characteristic of all destruxin molecules plays an important role in their biological activity.

Identification of a protein kinase C (PKC) activator, daphnoretin, that suppresses hepatitis B virus gene expression in human hepatoma cells.

We examined the antiviral activity of a crude extract prepared from a Chinese medicinal herb Wikstroemia indica C.A. Mey. One active component, daphnoretin (7-hydroxyl-6-methoxy-3,7'-dicoumarylether), was identified, which showed strong suppressive effects on the expression of the hepatitis B surface antigen (HBsAg) in human hepatoma Hep3B cells. To examine the signaling pathway of daphnoretin on the Hep3B cells, we pretreated Hep3B cells with 12-O-tetradecanoylphorbol-13-O-acetate (200 nM) for 24 hr to down-regulate intracellular protein kinase C (PKC) levels and found that the PKC-down-regulated Hep3B cells did not respond at all to daphnoretin. Furthermore, daphnoretin induced translocation of PKC from the cytosol to the membrane and down-regulated intracellular PKC levels in the Hep3B cells, indicating that it may directly activate PKC. This hypothesis was supported by the observation that daphnoretin directly competed with [3H]phorbol dibutyrate for the binding of PKC in the whole cell and activated purified PKC activity in vitro. Our results demonstrated that daphnoretin, with a structure distinct from phorbol ester, is a PKC activator and has suppressive effects on HBsAg gene expression in human hepatoma cells.

Inhibition of hepatitis B surface antigen secretion on human hepatoma cells. Components from Rubia cordifolia.

The antiviral activity in the roots of Rubia cordifolia was examined, and three naphthohydroquinones, furomollugin (1), mollugin (2), and rubilactone (3), were isolated from it. Compounds 1 and 2 strongly suppressed the secretion of hepatitis B surface antigen (HBsAg), both with IC50 = 2.0 micrograms/mL, in human hepatoma Hep3B cells while having little effect on the viability of the cells. Evaluation of structurally related derivatives of 1 and 2 revealed that a 6-hydroxy group and a pyran or furan ring contribute to this suppressive effect.

A novel secondary metabolite relative to the degradation of PR toxin by Penicillium roqueforti.

A secondary metabolite different from PR-imine and PR-amide was produced in the liquid (YESC) and solid (buckwheat) culture medium of Penicillium roqueforti. We isolated and purified the compound in pure and colorless crystalline form. On the basis of elemental analysis, mass, 1H and 13C nuclear magnetic resonance, infrared, and UV spectroscopy, the compound was identified as PR-acid (C17H20O7). The structures of PR-acid and PR toxin (C17H20O6) are closely related. Moreover, we discovered that PR-acid disappeared concurrently with the PR toxin in the culture medium. Thus, we postulate that PR toxin is degraded to PR-acid in the culture of P. roqueforti.

Effect of anthraquinone derivatives on lipid peroxidation in rat heart mitochondria: structure-activity relationship.

Lipid peroxidation was induced in rat heart mitochondria with FeSO4 and the inhibitory effects of various anthraquinone derivatives were compared. Oxygen consumption and malondialdehyde formation were used to quantitate the amount of lipid peroxidation. Emodin [2], alizarin [13], and alizarin complexone [14] significantly inhibited lipid peroxidation; their potency as inhibitors of lipid peroxidation was higher than that of alpha-tocopherol. Structure-activity analysis showed that two hydroxyl groups arranged in either the ortho- or meta-position in the C ring of the anthraquinone nucleus are required for such derivatives to inhibit lipid peroxidation. The diphenyl-p-picrylhydrazyl test showed that alizarin [13] and alizarin complexone [14] are free-radical scavengers while emodin [2] is not. The mechanism for emodin [2] to inhibit lipid peroxidation is most likely due to inhibition on the propagation of lipid peroxyl radicals in the mitochondrial membrane.

Differential pathways of insulin action upon the hepatitis B surface antigen gene expression and cell proliferation in human hepatoma cells.

We have shown previously that insulin at the physiological concentration suppresses hepatitis B surface antigen (HBsAg) gene expression in cultured human hepatoma Hep3B cells, and this suppression phenomenon is concomitant with the stimulation of cell proliferation. We have now examined whether these two distinct insulin actions on the Hep3B cells are mediated through the same or different signaling pathways. After prolonged treatment with high concentration of tumor promoter, 12-O-tetradecanoyl phorbol-13-acetate (TPA), the protein kinase C-alpha (PKC-alpha) level in the Hep3B cells was diminished and could not be detected by Western blot analysis. Under this condition, TPA treatment has no effect on the number or affinity of the insulin receptor on Hep3B cells. However, insulin-stimulated cell proliferation was completely abolished in the PKC-alpha depleted cells. In contrast, insulin still suppressed HBsAg gene expression with the same ED50 (approximately 0.5 nM) as the control cell. The induction of proto-oncogene egr-1 (early-growth-regulatory-1) by insulin and TPA under similar conditions were also examined. Both insulin and TPA stimulated egr-1 gene expression up to 10-fold in the control cell, but neither of these two agents showed any effect on egr-1 gene expression in the PKC-alpha down-regulated Hep3B cells. These observations indicate that the PKC-alpha may be involved in the insulin induced egr-1 expression and cell proliferation but not in the insulin suppressed HBsAg gene expression in human hepatoma cells.

Clinical application of simultaneous pattern VEP and pattern ERG recording.

Thirteen eyes of optic neuropathy and 13 eyes of maculopathy during Sep. 1990-May 1992 were studied by simultaneous pattern VEP (PVEP) and pattern ERG (PERG) recording. In optic neuropathy, the P100 of PVEP was delayed in 7 eyes (ave. 108.87 msec; normal 94.41 +/- 3.51 msec), absent in 5 eyes, normal in 1 eye. Their PERG showed normal P50 amplitude (either normal or delayed latency) but small or absent N95 in 8 eyes. In maculopathy, the P100 of PVEP was delayed in 2 eyes (ave. 103.59msec), absent in 7 eyes, normal in 4 eyes. However, their PERG showed 12 eyes had small or absent P50. Combined PVEP & PERG test, when compared to PVEP alone, increased the diagnostic value from 57.1% to 100% for optic neuropathy and from 42.9% to 80% for maculopathy in this study. Thus, simultaneous PVEP & PERG recording is highly recommended in evaluation of anterior visual pathway disorders.

Active compounds from Saussurea lappa Clarks that suppress hepatitis B virus surface antigen gene expression in human hepatoma cells.

We have examined the antiviral activity of the crude extract prepared from the root of Saussurea lappa Clarks, a Chinese medicinal herb which is widely used for many illnesses including cancer. Two active components, costunolide and dehydrocostus lactone, were identified which show strong suppressive effect on the expression of the hepatitis B surface antigen (HBsAg) in human hepatoma Hep3B cells, but have little effect on the viability of the cells. Both costunolide and dehydrocostus lactone suppress the HBsAg production by Hep3B cells in a dose-dependent manner with IC50s of 1.0 and 2.0 microM, respectively. Northern blotting analysis shows that the suppression of HBsAg gene expression by both costunolide and dehydrocostus lactone were mainly at the mRNA level. Furthermore, the suppressive effect of costunolide and dehydrocostus lactone on HBsAg and hepatitis B e antigen (HBeAg), a marker for hepatitis B viral genome replication in human liver cells, was also observed in another human hepatoma cell line HepA2 which was derived from HepG2 cells by transfecting a tandemly repeat hepatitis B virus (HBV) DNA. Similarly, the mRNA of HBsAg in HepA2 cells was also suppressed by these two compounds. Our findings suggest that costunolide and dehydrocostus lactone may have potential to develop as specific anti-HBV drugs in the future.

The novel desmethyldestruxin B2, from Metarhizium anisopliae, that suppresses hepatitis B virus surface antigen production in human hepatoma cells.

We have examined the antiviral activity of a crude extract prepared from the culture medium of the fungus Metarhizium anisopliae. Eight active destruxins were identified which showed strong suppressive effect on the production of the hepatitis B surface antigen (HBsAg) in human hepatoma Hep3B cells. One new compound, desmethyldestruxin B2 [1], was isolated from M. anisopliae. This structure was determined based on its nmr and mass spectral data.

An immobilized-enzyme system for the determination of sialic acid using cloned neuraminidase from Clostridium perfringens A.T.C.C. 10543.

Neuraminidase expressed by cloned nanH of Clostridium perfringens has been immobilized and employed to determine the concentration of sialic acid in human serum. Two enzyme pairs, cloned neuraminidase-N-acetylneuraminate (NANA) lyase and pyruvate oxidase-peroxidase, have been respectively co-immobilized on to 1,12-aminododecane-agarose with glutaraldehyde. The relative specific activities of the co-immobilized neuraminidase and NANA lyase were 61 and 77%, and those of pyruvate oxidase and peroxidase were 51 and 96% of the corresponding soluble enzymes respectively. The optimal reaction pH at 37% C for each of the co-immobilized enzymes was about 1 pH unit higher than that of the corresponding soluble enzyme. The optimal reaction temperature of peroxidase was increased as a result of immobilization. The thermostability of the immobilized cloned neuraminidase, NANA lyase, pyruvate oxidase and peroxidase were increased 80-, 83-, 115- and 147-fold at 45 degrees C over the soluble forms respectively. The results correlated satisfactorily with those obtained by using a soluble enzyme system. The system is thus a reliable assay method for sialic acid in serum.

Linoleate-rich triacylglycerol in Panax pseudo-ginseng improves erythrocyte deformability in vitro.

With Reid's filtration techniques as a bioassay for evaluating red blood cell (RBC) deformability, we purified from Panax pseudo-ginseng an active component that improved the deformability of calcium-loaded RBC. NMR and mass spectrometric studies showed that the purified substance was a triacylglycerol (TG) with linoleic acid as the major fatty acid residue in the esterified positions of glycerol. The mechanism for this TG to improve RBC deformability could be a modification of membrane fluidity rather than a competitive antagonism with calcium ion.

Secondary metabolites resulting from degradation of PR toxin by Penicillium roqueforti.

PR toxin is a secondary metabolite of the fungus Penicillium roqueforti. It is lethal to rats, mice, and cats. Usually, the amount of PR toxin in the culture medium decreases from its maximum on day 15 to zero within 3 to 4 days. We found that two were secondary metabolites produced in the culture medium of this fungus while the production of PR toxin was decreasing. We isolated and purified the two compounds in pure and colorless crystalline form. On the basis of elemental analysis and mass, 1H and 13C nuclear magnetic resonance, infrared, and UV spectroscopies, the two compounds were identified as PR-imine (C17H21O5N) and PR-amide (C17H21O6N). The structures of both compounds and of PR toxin (C17H20O6) were closely related, and the peak production of PR toxin appeared earlier than those of PR-imine and PR-amide. Moreover, PR toxin was transformed to PR-imine when PR toxin was incubated with the culture medium on a given culture day. Thus, we propose that PR toxin is degraded into PR-imine and PR-amide in the culture medium of P. roqueforti.

Effect of an extract from Phyllanthus amarus on hepatitis B surface antigen gene expression in human hepatoma cells.

It has been suggested that Phyllanthus amarus may be helpful in the treatment of hepatitis B virus infection. We studied the effect of an aqueous extract of P. amarus on the cultured hepatoma cell line HepA2. This cell line had been transfected with tandemly arranged HBV DNA and continued to synthesize and secrete both HBsAg and HBeAg. Extract of P. amarus reversibly inhibited cellular proliferation and suppressed HBsAg production but not HBeAg production in HepA2 cells. We also found that P. amarus suppressed HBsAg gene expression at mRNA level in a time-dependent manner, and selectively abolished the HBsAg gene promoter driven CAT activity. Our results demonstrate that P. amarus contains some active components which can suppress the HBsAg gene expression in human hepatoma cells. Such suppression may contribute the antiviral activity of P. amarus in vivo.

Inhibitory effect of curcumin, an anti-inflammatory agent, on vascular smooth muscle cell proliferation.

The effects of curcumin, an anti-inflammatory agent from Curcuma longa, on the proliferation of blood mononuclear cells and vascular smooth muscle cells were studied. Proliferative responses were determined from the uptake of tritiated thymidine. In human peripheral blood mononuclear cells, curcumin dose dependently inhibited the responses to phytohemagglutinin and mixed lymphocyte reaction at the dose ranges of 10(-6) to 3 x 10(-5) and 3 x 10(-6) to 3 x 10(-5) M, respectively. Curcumin (10(-6) to 10(-4) M) dose dependently inhibited the proliferation of rabbit vascular smooth muscle cells stimulated by fetal calf serum. Curcumin had a greater inhibitory effect on platelet-derived growth factor-stimulated proliferation than on serum-stimulated proliferation. Cinnamic acid, coumaric acid and ferulic acid were much less effective than curcumin as inhibitors of serum-induced smooth muscle cell proliferation, suggesting that the cinnamic acid and ferulic acid moieties alone are not sufficient for activity, and that the characteristics of the diferuloylmethane molecule itself are necessary for activity. Curcumin may be useful as a new template for the development of better remedies for the prevention of the pathological changes of atherosclerosis and restenosis.