First isolation of tryptophan from edible lotus (Nelumbo nucifera Gaertn) rhizomes and demonstration of its antioxidant effects.

Various vegetables were investigated for antioxidant activities in two assays, namely, inhibition of lysis of erythrocytes induced by peroxyl radicals and inhibition of lipid peroxidation. The lotus (Nelumbo nucifera Gaertn) rhizome showed the strongest antioxidant activity in both assays. The crude extract (L) of lotus rhizome was chromatographed on a macroporous adsorption resin named NKA. The resulting three fractions were designated L1, L2 and L3, respectively. L2 showed the highest antioxidant activity and was further fractionated by Sephadex LH-20 chromatography. Eight fractions were obtained and named from L2a to L2h, respectively. L2c showed the strongest activity in inhibiting hemolysis of erythrocytes and was further purified by high-performance liquid chromatography. L2c-3 was identified as tryptophan. Its inhibitory concentration of 50% (IC(50)) value in inhibiting hemolysis of erythrocytes was 156.3 microg/ml (i.e. 765.4 microM). This is the first report on isolation of tryptophan from the aqueous extract of lotus rhizome and demonstration of their antioxidant activities.

Isolation of adenosine, iso-sinensetin and dimethylguanosine with antioxidant and HIV-1 protease inhibiting activities from fruiting bodies of Cordyceps militaris.

According to previous studies, a close relationship between oxidative stress and AIDS suggests that antioxidants might play an important role in the treatment of AIDS. Cordyceps militaris was selected from nine edible mushrooms by assay of inhibition of erythrocyte hemolysis. Macroporous adsorption resin and HPLC were used to purify three micromolecular compounds named L3a, L3b and L3c. L3a was identified to be adenosine with the molecular formula C(10)H(13)N(5)O(4); L3b was 6,7,2',4',5'-pentamethoxyflavone with the molecular formula C(20)H(20)O(7), and L3c was dimethylguanosine with the molecular formula C(12)H(17)N(5)O(5). The compound 6,7,2',4',5'-pentamethoxyflavone was first isolated from C. militaris. The assay of inhibition of HIV-1 protease (HIV-1 PR) was based on the fact that the expression of this enzyme can inhibit the growth of E. coli. This is a new screening system for HIV-1 PR inhibitors. Both L3a and L3b showed high inhibition to HIV-1 PR. These compounds could be new anti-HIV-1 PR drugs.

Inhibitors from natural products to HIV-1 reverse transcriptase, protease and integrase.

Acquired immunodeficiency syndrome (AIDS), caused by human immunodeficiency virus type 1 (HIV-1) infection, is still one of the most challenging diseases of the early 21st century. Reverse transcriptase (RT), protease (PR) and integrase (IN) are three key enzymes of HIV-1. Despite the shortcomings of chemical drugs such as toxicity, lack of curative and multiple effects, the search for more and better anti-HIV agents has been focused on natural products. Many natural products have been shown to possess promising activities that could assist in the prevention and amelioration of the disease. Most of these natural anti-HIV agents have other medicinal values as well, which afford them further prospective as novel lead compounds for the development of new drugs. These natural products can deal with both the virus and the various disorders that are caused by HIV. In this review, natural inhibitors of RT, PR and IN have been found to be classified and the relationship between structure and inhibitory activity is discussed.

ISG15 expression in response to double-stranded RNA or LPS in cultured Fetal bovine lung (FBL) cells.

Fetal bovine lung (FBL) cells are used in the culture of viruses which infect cattle and ISG15 plays a role in innate immunity against viral infections. However, whether the expression of ISG15 gene can be induced in FBL cells is still unknown. In this work, the expression of ISG15 in cultured FBL cells was detected after stimulated with poly I:C or LPS. Real-time PCR analyses revealed that the transcript of ISG15 can be induced by poly I:C or LPS. The increased expression of free ISG15 was confirmed via Western blotting. Furthermore, immunofluorescence assays demonstrated that IRF-3 was translocated from the cytoplasm to the nucleus in the FBL cells treated with poly I:C. Chromatin immunoprecipitation assays showed that IRF-3 can bind to the promoter of the bISG15 gene. To demonstrate IRF-3 can promote the expression of bISG15, we establish a luciferase-reporter system of bovine ISG15 gene in 293 T cells. The luciferase assay showed that the over-expression of bovine IRF-3 could activate the promoter of bISG15 gene. Taken together, these results suggest that the expression of bISG15 can be induced in FBL cells stimulated with poly I:C or LPS, and IRF-3 may play a role in inducing the expression of ISG15 in FBL cells.