In vitro antiviral activities of Caesalpinia pulcherrima and its related flavonoids.

The aim of this study was to search for new antiviral agents from Chinese herbal medicine. Pure flavonoids and aqueous extracts of Caesalpinia pulcherrima Swartz were used in experiments to test their influence on a series of viruses, namely herpesviruses (HSV-1, HSV-2) and adenoviruses (ADV-3, ADV-8, ADV-11). The EC50 was defined as the concentration required to achieve 50% protection against virus-induced cytopathic effects, and the selectivity index (SI) was determined as the ratio of CC50 (concentration of 50% cellular cytotoxicity) to EC50. Results showed that aqueous extracts of C. pulcherrima and its related quercetin possessed a broad-spectrum antiviral activity. Among them, the strongest activities against ADV-8 were fruit and seed (EC50 = 41.2 mg/l, SI = 83.2), stem and leaf (EC50 = 61.8 mg/l, SI = 52.1) and flower (EC50 = 177.9 mg/l, SI = 15.5), whereas quercetin possessed the strongest anti-ADV-3 activity (EC50 = 24.3 mg/l, SI = 20.4). In conclusion, some compounds of C. pulcherrima which possess antiviral activities may be derived from the flavonoid of quercetin. The mode of action of quercetin against HSV-1 and ADV-3 was found to be at the early stage of multiplication and with SI values greater than 20, suggesting the potential use of this compound for treatment of the infection caused by these two viruses.

Antiviral activity of Plantago major extracts and related compounds in vitro.

Plantago major L., a popular traditional Chinese medicine, has long been used for treating various diseases varying from cold to viral hepatitis. The aim of present study was to examine the antiviral activity of aqueous extract and pure compounds of P. major. Studies were conducted on a series of viruses, namely herpesviruses (HSV-1, HSV-2) and adenoviruses (ADV-3, ADV-8, ADV-11). The antiviral activity of EC50 was defined as the concentration achieved 50% cyto-protection against virus infection and the selectivity index (SI) was determined by the ratio of CC50 (concentration of 50% cellular cytotoxicity) to EC50. Results showed that aqueous extract of P. major possessed only a slight anti-herpes virus activity. In contrast, certain pure compounds belonging to the five different classes of chemicals found in extracts of this plant exhibited potent antiviral activity. Among them, caffeic acid exhibited the strongest activity against HSV-1 (EC50=15.3 microg/ml, SI=671), HSV-2 (EC50=87.3 microg/ml, SI=118) and ADV-3 (EC50=14.2 microg/ml, SI=727), whereas chlorogenic acid possessed the strongest anti-ADV-11 (EC50=13.3 microg/ml, SI=301) activity. The present study concludes that pure compounds of P. major, which possess antiviral activities are mainly derived from the phenolic compounds, especially caffeic acid. Its mode of action against HSV-2 and ADV-3 was found to be at multiplication stages (postinfection of HSV-1: 0-12 h; ADV-3: 0-2 h), and with SI values greater than 400, suggesting the potential use of this compound for treatment of the infection by these two viruses.

Antileukemic activity of Bidens pilosa L. var. minor (Blume) Sherff and Houttuynia cordata Thunb.

To evaluate the anti-leukemic activity of Bidens pilosa L. var. minor (Blume) Sherff and Houttuynia cordata Thunb., cytotoxicity tests with an XTT-based colorimetric assay were used. Five leukemic cell lines, namely L1210, U937, K562, Raji and P3HR1, were cultured with hot water extracts of B. pilosa var. minor or H. cordata. Hot water extracts of B. pilosa var. minor inhibited these five leukemic cells with IC50s between 145 microg/ml and 586 microg/ml. The effect was greatest on four cell lines, namely L1210, P3MR1, Raji and K562, with IC50s below 200 microg/ml and a selective index of more than 5. Hot water extract of H. cordata inhibited these five leukemic cells with IC50s between 478 microg/ml and 662 microg/ml. The selective index was between 1.5 and 2.1. B. pilosa var. minor was more effective than H. cordata in inhibiting most of the leukemic cells in our study. We suggest that B. pilosa L. var. minor (Blume) Sherff may prove to be a useful medicinal plant for treating leukemia.

A highly selective beta1-adrenergic blocker with partial beta2-agonist activity derived from ferulic acid, an active component of Ligusticum wallichii Franch.

Short-term injection of ferulinolol (0.1, 0.5, and 1.0 mg/kg, i.v.) produced dose-dependent bradycardia responses in pentobarbital-anesthetized Wistar rats, whereas it had no significant effects on the blood pressure. Ferulinolol markedly inhibited the tachycardia effects induced by (-)isoproterenol but did not show any blocking effect on the arterial pressor responses induced by (-)phenylephrine. These findings clearly suggested that ferulinolol had a beta-adrenergic blocking activity; nevertheless, it did not involve an alpha-adrenergic blocking action. In isolated guinea pig tissues, ferulinolol competitively antagonized (-)isoproterenol-induced positive inotropic and chronotropic effects of the atria and tracheal relaxation responses. The parallel shift to the right of the concentration-response curve of (-)isoproterenol suggested that ferulinolol was a beta-adrenoceptor-competitive antagonist. The apparent pA2 values for ferulinolol on right atria, left atria, and trachea were 7.62 +/- 0.05, 7.54 +/- 0.01, and 6.28 +/- 0.11, respectively. Ferulinolol was more potent on the atria than on tracheal tissues, demonstrating that it possessed beta1-adrenoceptor selectivity. The intrinsic sympathomimetic activity (ISA) of ferulinolol and propranolol were determined on isolated atria and trachea from reserpine-treated guinea pig. Propranolol caused significantly negative inotropic and chronotropic effects at > or =1 microM, whereas ferulinolol possessed fewer cardiodepressant activities than propranolol. In reserpine-treated tracheal strips, ferulinolol produced dose-dependent relaxant responses, but propranolol was without effectiveness. Preincubating the preparations with ICI 118,551 (0.1, 1.0, and 10 nM), a beta2-adrenoceptor antagonist, significantly shifted the concentration-relaxation curves of ferulinolol to a region of higher concentrations. These results implied that ferulinolol had a partial beta2-agonist activity. Further, binding characteristics of ferulinolol and various beta-adrenoceptor antagonists were evaluated in [3H]CGP-12177 binding to rat ventricular or lung membranes. The Ki values of ferulinolol, atenolol, metoprolol, and (-)propranolol were 103, 262, 123, and 0.23 nM, respectively, in ventricular membranes, and 2,412, 7,539, 2,186, and 0.72 nM, respectively, in lung membranes. In conclusion, ferulinolol was found to be a highly selective beta1-adrenoceptor antagonist with partial beta2-agonist activity but was devoid of alpha-adrenoceptor blocking action.

Effects of suramin-related and other clinically therapeutic polyanions on protein kinase C activity.

The mechanism of the antineoplastic effects of suramin may involve interference with signal transduction, but in general is not well understood. We examined several polyanions to determine their effects on the kinase activity of the protein kinase C (PKC) beta1 and other PKC isoforms. Similar to suramin, a phosphorothioate oligodeoxynucleotide 28-mer homopolymer of cytidine (SdC28) inhibited the phosphatidylserine and Ca2+-dependent phosphorylation of an epidermal growth factor receptor octapeptide substrate. The inhibition by suramin was mixed competitive/noncompetitive with respect to ATP, but uncompetitive with respect to substrate. In contrast, the inhibition by SdC28 was competitive with respect to substrate (Ki = 5.4 microM) and not competitive with respect to ATP. The PKC alpha and beta1 isoforms were inhibited to the same extent with SdC28, while PKC epsilon was not inhibited. SdC28, in the absence of lipid cofactor, stimulated substrate phosphorylation, and in the absence of substrate induced PKC beta1 autophosphorylation. Similar behavior was seen with another polyanion, the polysulfated carbohydrate pentosan polysulfate (polyxylyl hydrogen sulfate). H4, a bis-naphthalene disulfonate tetraanion structurally related to suramin, also inhibited kinase activity but was not competitive with respect to ATP. Dianions closely related to H4 failed to inhibit PKC beta1, suggesting that multiple (>2) negative charges are required. The interactions of polyanions with PKC are complex, and are dependent on the molecular structure of the polyanion, the presence of cofactors, and the PKC isoform.

Carbon dioxide-enhanced ultrasonography of liver tumors.

CO2 gas-enhanced ultrasonography was performed in 37 patients (47 studies) for the purpose of detecting small tumors and evaluating differential diagnosis. With conventional ultrasonography, 62 lesions were identified in 25 patients with HCC, 13 tumors were identified in eight patients with hemangioma, and multiple tumors were found in four patients with metastatic adenocarcinoma. CO2-enhanced ultrasonography detected five additional hemangiomas, 12 additional nodules in HCC, and the same number of metastatic nodules. The patterns of CO2 enhancement were characterized as homogeneous, heterogeneous, rim, internal spotted, negative, and mixed (more than one pattern in one lesion). The rim enhancement pattern was found to be specific for hemangioma. The internal spotted enhancement pattern was found exclusively in HCC. All the lesions that demonstrated negative enhancement were treated HCC. All the metastatic tumors demonstrated the mixed rim and internal spotted enhancement pattern. We suggest that CO2-enhanced ultrasonography is a useful tool in detecting small liver tumors. It can also help in the differentiation among various hepatic tumors.

Inhibitory effect of garlic (Allium sativum) on sodium transport in isolated toad skin.

The effect of an aqueous fraction from the bulbs of Allium sativum (GE) was investigated in toad skin. When added to the inner (serosal) solution, GE caused a maximal reversible reduction of the transepithelial potential difference and short circuit current of 38% and 45%, respectively. When added to the outer (mucosal) solution, the effect was only partially reversible. Isaacson's amiloride test showed that GE decreased sodium potential (ENa.) and sodium conductance (GNa.). The net Na+ flux decreased due principally to a fall in Na+ flux in the active direction. GE decreased Na(+)-K+ ATPase activity in vitro. Partial replacement of sodium by choline in the outer solution reduced the effect of GE on the skin and substitution of normal Ringer's solution with isethionate Ringer's solution in the outer solution significantly enhanced the effect of GE on the skin. These results indicate that GE decreases active Na transport in the toad skin.

Diuretic, natriuretic and hypotensive effects produced by Allium sativum (garlic) in anaesthetized dogs.

Gastric administration of encapsuled garlic powder to anaesthetized dogs induced dose-dependent (2.5 to 15 mg/kg) natriuretic and diuretic responses which reached maximum 30-40 min after garlic administration and decreased to basal levels after 100-150 min. A simultaneous decrease in arterial blood pressure was observed which continued past the 250 min-mark. High garlic doses (15 and 20 mg/kg) provoked bradycardia and T-wave inversion during the first 10-15 min of the experiment with recordings returning to normal and staying normal throughout the remainder of the experiment.

[Purification and characterization of phytomitogens isolated from Phaseolus vulgaris bean].

The physical properties and biological activities of commercial PHAP (Difco) with eleven phytoproteins purified from red kidney beans (Phaseolus vulgaris) are compared. In gel filtration pattern, there seems no apparent difference among F2, F9 and F10. However, F1 component appears second sharp peak which given no biological activity. From results of cellulose acetate electrophoresis, it showed that majority of F1, F5, and F7 are anodal proteins, while only minor part of F2, F9, F10 and F12 possess this property. As to biological activities, higher potency for hemagglutination are obtained from F8(467%), F12(350%0, F9(280%) and F10(100%). On the other hand, fractions such as F10(90%), F2(87%), F8(86%) and F12(64%) reveal more capability of causing lymphocyte mitogenesis. For lymphocyte transformation, F10 lymphocyte transformation, F10(100%), F9(96%), F2(93%), F3(86%), F5(84%), F1(80%), F8(78%), and F12(70%) are predominance in their activity. In lymphocyte transformation test, F9 and F10 exhibit highest activity if their concentrations are lower than 20 micrograms/ml. But there have no significant difference among F1, F3 and F10 when the test concentrations are over 20 micrograms/ml. For patients of bladder carcinoma, the lymphocyte transformation pattern are resemblance to that of normal individuals. In summary, according to experimental data compared as above, the biological activities of F1, F2, F3, and F9 are similar with standard F10 (PHAP) and might applied in assay of cellular immunity and other cytogenetic studies.