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A protein designated hypogin, with a prominent suppressive action on the growth of the fungi Mycosphaerella arachidicola, Fusarium oxysporum and Coprinus comatus, was isolated from seeds of the peanut Arachis hypogaea. The protein inhibited human immunodeficiency virus (HIV) reverse transcriptase and enzymes associated with HIV infection including alpha-glucosidase and beta-glucosidase. The proliferative response of mouse splenocytes was attenuated in the presence of the protein. The protein exhibited a molecular mass of 7.2 kDa in tricine gel electrophoresis and gel filtration on Superdex 75 and an N-terminal sequence resembling peanut allergen Ara H1. The isolation procedure involved affinity chromatography on Affi-gel blue gel and ion-exchange chromatography on CM-Sepharose. The protein was adsorbed in both chromatographic media.

An antifungal protein, with a molecular weight of 15 kDa and an N-terminal sequence analogous to those of chitinases, was first isolated from the Chinese medicinal material Panax notoginseng, using cation exchange chromatography and affinity chromatography. The protein was adsorbed on CM-cellulose, Affi-gel Blue Gel and Mono S. It exerted antifungal activity against Coprinus comatus, Fusarium oxysporum and Mycosphaerella arachidicola but not against Rhizoctonia solani. The protein was devoid of ribonuclease activity against yeast tRNA.

Ten polymorphic SSR markers of Flammulina velutipes were developed and characterized with FIASCO methods. The polymorphism information content (PIC) ranged from 0.13 to 0.69. This is the first report on development of SSR markers in F. velutipe.

A purification protocol that encompassed anion exchange chromatography (EC) on DEAE-cellulose, cation EC on CM-cellulose, anion EC on Q-Sepharose, and fast protein liquid chromatography-gel filtration of Superdex 75 was used to isolate a ribonuclease from dried fruiting bodies of Ramaria formosa. The ribonuclease was unadsorbed on CM-cellulose but adsorbed on both DEAE-cellulose and Q-Sepharose. It displayed a molecular mass of 29-kDa in both gel filtration and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The ranking of its ribonucleolytic activity toward polyhomoribonucleotides was poly(U) poly(C) poly(G) poly(A). It exhibited a pH optimum of pH 5 and a temperature optimum at 60 °C. The ribonuclease inhibited HIV-1 reverse transcriptase with an IC50 of 3 µM.

The antithrombotic and thrombolytic activities of Agkisacutacin (Agk), a component isolated from Agkistrodon acutus, were determined in vitro and in vivo. The models employed included Chandler's model, arterio-venous shunt model and pulmonary embolus model. The effects of Agkisacutacin on coagulation, plasma fibrinogen and platelet aggregation induced by collagen, adenosine diphosphate (ADP) and thrombin were also investigated. The results showed that Agkisacutacin can significantly inhibit thrombus formation in Chandler's and arterio-venous shunt models, and accelerate thrombolysis of pulmonary emboli in rats. The data suggested that Agkisacutacin possessed antithrombotic and thrombolytic activities. Agkisacutacin was also partial characterized.

The aqueous and methanol extracts of thirty-one herbs traditionally used as anti-fever remedies in China were screened for their in vitro inhibition on human immunodeficiency virus type-1 protease (HIV-1 PR). The activity of recombinant HIV-1 protease was determined by sequence-specific cleavage at the Tyr-Pro bond of the fluorogenic substrate (Arg-Glu(EDANS)-Ser-Gln-Asn-Tyr-Pro-Ile-Val-Gln-Lys(DABCYL)- Arg) or by HPLC anaylsis of the cleavage products after incubation of the enzyme with a synthetic peptide substrate (Acetyl-Ser-Gln-Asn-Tyr-Pro-Val-Val-amide). Among the herbal extracts examined, the aqueous extracts of Prunella vulgaris and Scutellaria baicalensis and the methanol extracts of Woodwardia unigemmata, Paeonica suffruticosa and Spatholobus suberectus elicited significant inhibition (>90%) at a concentration of 200 microg/ml.

Six strains of crown gall bacteria were isolated from flowering cherry. It was revealed by Otten paper electrophoresis that of the six strains, only BYH18-4 possessed the octopine type Ti plasmid, the remainder having nopaline type Ti plasmid. BYH5-1 was identified by physiological and biochemical tests to be Agrobacterium tumefaciens (originally biovar 1). The other five were A. rhizogenes (originally biovar 2). It was demonstrated with Stonier's method of double layer medium that flowering cherry crown gall bacteria exhibited different sensitivities to agrocin produced by biocontrol strain K1026. Strain K1026 on greenhouse-grown sunflower seedlings exerted a relatively potent inhibitory action on flowering cherry crown gall bacteria. Artificial inoculation showed that K1026 produced 67–99% inhibition of flowering cherry crown gall disease, compared with the treatment of inoculation with crown gall bacteria only.

This paper reports the isolation and characterization from bovine milk of two proteins: angiogenin-1, a recently discovered angiogenin, and lactogenin, a novel protein. Both proteins were adsorbed on and eluted closely from CM-Sepharose and Mono S. Lactogenin possessed a molecular weight (17 kDa) slightly higher than that of angiogenin-1 (15 kDa). Lactogenin had a higher ribonucleolytic (RNase) activity than angiogenin-1 towards yeast transfer RNA (tRNA). The Km values estimated for the RNase activities of angiogenin-1 and lactogenin were 51 microM and 40 microM respectively. Both were specific for poly C. The optimal pH for the RNase activities of angiogenin-1 and lactogenin was 7.75 and 7.5 respectively. Comparison of the amino acid sequences of cyanogen bromide fragments and the pyroglutaminase-treated N-terminal fragment of lactogenin with the sequence of bovine liver RNase (RNase BL4) revealed identity in residues 3-22, 24, 26-27, 37, 41-44, 46-50, 54, 56, 63, 72-80, and 83. Considerable similarity to the N-terminal sequence of angiogenin-2 was also noted. Both lactogenin and angiogenin-1 inhibited cell-free translation in a rabbit reticulocyte lysate system with an IC(50) below 100 nM.

A new ribosome-inactivating protein (RIP) (δ-momorcharin) and a candidate RIP (e-momorcharin) were isolated, respectively, from the seeds and fruits of the bitter gourd Momordica charantia, by affinity chromatography on Affi-gel blue gel and ion exchange chromatography on Mono S with a fast protein liquid chromatography (FPLC) system. Both δ- and e-momorcharins were adsorbed on Affi-gel blue gel and were eluted from the cation exchange Mono S column earlier than α- and β-momorcharins, the two previously reported RIPs, δ- and e-momorcharins possessed a molecular weight of 30 and 24 kDa respectively and inhibited cell-free translation in rabbit reticulocyte lysate with an IC50 of 0.15 and 170 nM. The sequence of the first seven N-terminal amino acids in δ-momorcharin was DVNFGLA, which was different from the N-terminal sequences DVSFRLS and DVNFDLS for α- and β-momorcharins respectively.

The antithrombotic activity of low-molecular-weight heparin calcium (LMWH-Ca) was studied in venous and arterial thrombosis models, arterial thrombosis model in rats, arterio-venous shunt model and venous thrombosis model in rabbits. The data showed that LMWH-Ca reduced thrombus formation in a dose-dependent manner. It suggests that LMWH-Ca is a potent antithrombotic agent for venous thrombosis, and also may be a beneficial therapeutic agent in arterial thrombosis.

A lectin, with a specificity for N-acetylgalactosamine and N-acetylglucosamine and a molecular weight of 43 kDa, was isolated from fruiting bodies of the edible shiitake mushroom Lentinus edodes. The purification procedure entailed extraction with aqueous buffer, ammonium sulfate precipitation, gel filtration on Sephadex G-100 and affinity chromatography on N-acetylgalactosamine-agarose. The lectin was unique in that it was tenaciously bound on anion exchangers including DEAE-cellulose, DEAE-Sepharose, DEAE-Sephadex, Q-Sepharose, Dowex and PEI-cellulose and also on hydroxyapatite and phenyl Sepharose. It was largely unadsorbed on cation exchangers including CM-cellulose, CM-Sepharose, SP-Sepharose and Amberlite, and also on protein G-Sepharose, Red Sepharose and Affi-gel Blue gel, wheat germ lectin-Sepharose and p-aminophenyl- d -glucopyranoside agarose.

DNA fingerprints distinctive among the samples from different localities in China were successfully reproduced for the Chinese herb Dangshen, the roots of CODONOPSIS PILOSULA, (Campanulaceae). Similarity index (S.I.) analysis revealed that C. PILOSULA samples from the same province generated similar DNA fingerprints, while samples of different provinces displayed different DNA fingerprints. This method may be a general and valuable tool for locality authentication of other Chinese herbal medicinal materials.

In a study on the embryotoxicity of pineal indoles on developing chick embryos in vivo, the pineal indoles—namely, melatonin (MEL), methoxytryptamine (MTA) and methoxytryptophol (MTP)—were injected into the yolk sacs of the chick embryos through the air chambers of the eggs on the 4th day of incubation. The eggs were opened and the embryos examined after 6, 10 or 14/15 days of incubation. Abnormalities were found to occur mainly in the 6- and 10-day-old embryos, which exhibited external malformations such as twisted vertebral column, abdominal hernia, exteriorization of heart and viscera, defects of eye, beak and limb. From the results obtained from embryos on the 14th or 15th day of incubation, MEL was found to be the most toxic indole in regard to the mortality induced, whereas MTA had the highest teratogenicity because of the frequent incidence of abnormal embryos. Effect of MTP treatment on the development of chick embryos varied greatly between doses, and there were no abnormal embryos found on the 14th or l5th day of incubation.

Bacillus licheniformis α-amylase (BLA) is routinely used as a model thermostable amylase in biochemical studies. Its starch hydrolysis activity has recently been studied in Tris buffer. Here, we address the question that whether the application of Tris buffer may influence the results of BLA activity analyses. Based on the inhibition studies and docking simulations, we suggest that Tris molecule is a competitive inhibitor of starch-hydrolyzing activity of BLA, and it has a high tendency to bind the enzyme active site. Hence, it is critically important to consider such effect when interpreting the results of activity studies of this enzyme in Tris buffer.

One of the ribosome inactivating proteins (RIPs), beta-momorcharin, and a lectin were isolated from seeds of the bitter melon Momordica charantia (Family Cucurbitaceae) in accordance with published procedures. Both beta-momorcharin and M. charantia lectin were then subjected to amino acid sequencing. beta-momorcharin exhibited considerable homology in sequence to other Cucurbitaceae RIPs, and also to the A chains of abrin and ricin which are type II (double-chained) RIPs. The resemblance between alpha-momorcharin and other RIPs is closer than that between beta-momorcharins and other RIPs. M. charantia lectin manifested a certain extent of sequence similarity to beta-momorcharin and other RIPs, and some degree of homology in sequence to lectins from Cucurbita maxima, Cucurbita argyrosperma, Sambucus nigra and Ricinus Communis.

Adolescent idiopathic scoliosis (AIS) is a common orthopedic disorder of unknown etiology and pathogenesis. Melatonin and melatonin pathway dysfunction has been widely suspected to play an important role in the pathogenesis. Many different types of animal models have been developed to induce experimental scoliosis mimicking the pathoanatomical features of idiopathic scoliosis in human. The scoliosis deformity was believed to be induced by pinealectomy and mediated through the resulting melatonin-deficiency. However, the lack of upright mechanical spinal loading and inherent rotational instability of the curvature render the similarity of these models to the human counterparts questionable. Different concerns have been raised challenging the scientific validity and limitations of each model. The objectives of this review follow the logical need to re-examine and compare the relevance and appropriateness of each of the animal models that have been used for studying the etiopathogenesis of adolescent idiopathic scoliosis in human in the past 15 to 20 years.

A novel lectin has been purified from the fruiting bodies as well as cultured mycelia of the edible mushroom Volvariella volvacea. The lectin, designated as VVL, was a homodimeric protein with a molecular weight of 32 kDa as demonstrated by gel filtration and SDS-PAGE. VVL had no carbohydrate moiety, and its hemagglutinating activity was inhibited by thyroglobulin but not by simple carbohydrates such as monomeric or dimeric sugars. The immunomodulatory activity of VVL was demonstrated by its potent stimulatory activity toward murine splenic lymphocytes. VVL was also found to markedly enhance the transcriptional expression of interleukin-2 and interferon-gamma by reverse transcriptase-polymerase chain reaction. As revealed by its N-terminal amino acid sequence, VVL possessed a molecular structure distinct from other immunomodulatory proteins previously reported in the same fungus.

A novel single-chained ribosome-inactivating protein (RIP) with a molecular weight of approximately 29 000 was purified from fruiting bodies of the edible mushroom Volvariellavolvacea with a procedure involving ammonium sulfate precipitation, ion-exchange chromatography on DEAE-cellulose, and gel filtration on Superdex 75. The mushroom RIP, designated volvarin, exhibited a potent inhibitory action on protein synthesis in the rabbit reticulocyte lysate system with an IC(50) value of 0.5 nM. Like most plant RIPs, volvarin acted as an N-glycosidase that depurinated rRNA from rabbit reticulocyte lysate, releasing a characteristic RNA fragment after treatment with aniline. It also exerted a deoxyribonuclease activity on supercoiled SV-40 DNA and demonstrated a strong abortifacient effect in mice.

A microplate assay, for use with a variety of glycohydrolase enzymes, was developed to aid the screening of Chinese medicinal herb extracts for the presence of potential anti-viral and anti-lymphoma compounds. The microplate assay method described offers greater convenience, speed and reproducibility over existing methods. The enzymes tested were alpha-glucosidase, beta-glucosidase and beta-glucuronidase. The assay can be easily adapted for use with other glycohydrolase enzymes. Of the 12 herb extracts examined four did not inhibit any of the enzymes (50% inhibition), one inhibited alpha-glucosidase only (50% inhibition), six inhibited beta-glucuronidase only, and one inhibited both alpha-glucosidase and beta-glucuronidase. None of the extracts were capable of inhibiting beta-glucosidase to any significant extent.

1. Coriolus versicolor polysaccharopeptide has been reported to exert immunomodulatory and antitumor actions. The present study showed that it exhibits analgesic activity in the hot-plate test upon intraperitoneal administration to ICR mice. 2. It did not affect ovarian steroidogenesis, ovulation and midterm gestation in mice. It did not exert an adverse effect on mouse embryonic development either, as evidenced by the lack of an effect on somite number, axial length and the incidence of abnormalities in heartbeat, yolk sac circulation, optic vesicle, otic vesicle, shape of body axis, forelimb buds, branchial apparatus, cranial neural tube and head size. 3. Its analgesic activity would add to its attribute as an immunomodulatory and antitumor drug.

Tilapia (Oreochromis niloticus) kept in 15 ppt sea water (roughly iso-osmotic salinity) had higher growth rates than fish kept in 0 ppt (freshwater) or 30 ppt seawater, but circulating level of growth hormone was highest in fish exhibiting the poorest growth rate (30 ppt seawater). Serum thyroxine concentration was highest in 15 ppt seawater. Intestinal trypsin may play a role in promoting growth in iso-osmotic salinity since its activity was highest in fish cultured in 15 ppt seawater. The results indicate that changes in the digestive power, coupled with changes in thyroxine secretion, may account for the variations in growth rate in tilapia reared under different salinities.

A multiple screening approach to detect compounds inhibitory to various aspects of the human immunodeficiency virus type 1 (HIV-1) life-cycle has been applied to aqueous extracts of 19 herbs traditionally used in Chinese medicine as anti-viral agents. The extracts were tested for their ability to inhibit HIV-1 in a series of in vitro assays. The extracts were tested for inhibition of the interaction between HIV-1 gp120 and immobilized CD4 receptor, inhibition of recombinant HIV-1 reverse transcriptase and for inhibition of three glycohydrolase enzymes that contribute to viral protein glycosylation. Six of the herb extracts (30%) were potent inhibitors of the interaction between HIV-1 gp120 and the CD4 receptor (ID50 5.6 - 79.4 microg/ml), two extracts (10%) contained potent reverse transcriptase inhibitors (ID50 16.9 - 26.0 microg/ml) and 14 extracts (75%) were able to inhibit at least one of the glycohydrolase enzymes.

Bakuchiol was an active antifungal compound isolated from Psoraleae Fructus by means of bioassay-guided fractionation in our previous study. The present work aimed to investigate the underlying mechanisms and the therapeutic effect of bakuchiol in Trichophyton mentagrophytes-induced tinea pedis. After exposure to bakuchiol at 0.25-fold, 0.5-fold and 1-fold of minimum inhibitory concentration (MIC) (3.91 μg/ml) for 24h, the fungal conidia of T. mentagrophytes demonstrated a significant dose-dependent increase in membrane permeability. Moreover, bakuchiol at 1-fold MIC elicited a 187% elevation in reactive oxygen species (ROS) level in fungal cells after a 3-h incubation. However, bakuchiol did not induce DNA fragmentation. In a guinea pig model of tinea pedis, bakuchiol at 1%, 5% or 10% (w/w) concentration in aqueous cream could significantly reduce the fungal burden of infected feet (p

A β-mannanase gene (Man5) from Bacillus subtilis BS5 was cloned by PCR and integrated into the genome of Pichia pastoris GS115 via pPIC9 vector. The recombinant Man5 with a molecular mass of 43 kDa was successfully expressed and secreted into the culture medium. After methanol induction in a shake flask for 96 H, the recombinant Man5 protein reached 375 µg/mL in concentration, with an enzyme activity of 892 U/mL. The recombinant Man5 was purified 3.35-fold with 60% yield by using HiTrap DEAE FF and HiTrap Phenyl FF columns. The specific activity of the purified enzyme was 7,978 U/mg. The optimum temperature and pH of the recombinant Man5 were 50 °C and 6.0, respectively. Studies of substrate specificity showed that the optimum substrate for the Man5 was konjac flour, suggesting that it has great potential as an effective additive in the food industry.

Stress has been shown to suppress immune function and increase susceptibility to inflammatory and psychiatric diseases. This study sought to investigate the changes in erythrocyte immune functions and T-lymphocyte subsets and to explore the mechanism implicated in the process of stress-induced depression by employing a rat depression model induced by chronic unpredictable mild stress (CUMS). The body weights and behavioral changes of the rats were recorded, and plasma corticosterone levels were determined by radioimmunoassay. Erythrocyte immune function and T-lymphocytes subsets were respectively measured by the method of yeast rosette and flow cytometry at different time intervals, and their relationship was analyzed. Results indicated that a reduction was observed in the following: the rats' crossing and rearing movement times, the volume of sucrose intake and the preference for sucrose in the depression model group. Plasma corticosterone levels were elevated; the rate of E-C3bR decreased, and E-IC was increased. Some alterations in the percentage of T-lymphocytes and IL-2 appeared in the depression model group and some relationships existed between these parameters. Collectively, these findings disclose that long-time stress could induce changes in rat behavior and activities through an effect on erythrocyte immune functions and T-lymphocyte subsets.

In this study, a compound with antioxidant and anti-HIV activities designated as HEB was first isolated from the edible mushroom Pholiota adiposa by extraction with ethanol and ethyl acetate. HEB was then purified by high performance liquid chromatography (HPLC) and identified to be methyl gallate (C8H8O5, 184.1 Da) based on data from its mass spectrum (MS) and nuclear magnetic resonance (NMR) spectrum. HEB displayed strong antioxidant potency in inhibiting, at 1.36 mM concentration, erythrocyte hemolysis and scavenging DPPH radicals and superoxide anion (O2(-)) by 82.4%, 85.6% and 71.4%, respectively. Besides exhibiting a low cytotoxicity, compound HEB demonstrated significant anti-HIV activity in that it inhibited HIV-1 replication in TZM-BL cells infected by pseudovirus with an IC50 value of 11.9 μM. Further study disclosed that HEB inhibited the viral entry process and activities of key enzymes essential for the HIV-1 life cycle. HEB inhibited HIV-1 reverse transcriptase and integrase activities with an IC50 value of 80.1 μM and 228.5 μM, respectively, and at 10 mM concentration inhibited HIV-1 protease activity by 17.1% which was higher than that achieved by the positive control pepstatin A. Interestingly, this study first revealed that H2O2 stimulation not only activated cell oxidative stress responses, but also accelerated HIV-1 long terminal repeat (LTR) promotion in TZM-BL cells, which was significantly reduced by HEB from 18.2% to about 2%. It implied a direct relationship between the antioxidant and anti-HIV activities of the natural active constituent HEB. Nuclear transcription factor kappa B (NF-κB) signal pathways plays an important role in oxidative stress responses. Meanwhile, there is κB target sequence in HIV promoter LTR which is significant for virus replication and gene expression. In this study, Western Blot assay showed that HEB could inhibit the activation of NF-κB signal pathway stimulated by H2O2 in mouse spleen cells through suppressing NF-κB (p65) translocation into nucleus and NF-kappa-B inhibitor (IκB) degradation in cytoplasm. In summary, the antioxidant HEB from P. adiposa could inhibit HIV-1 replication through multiple target sites. The data suggest that natural antioxidant compounds might have a potential for treatment of AIDS.

A monomeric feruloyl esterase (FAE) with a molecular mass of 62 kDa was acquired from fresh fruiting bodies of the edible mushroom Russula virescens. The isolation procedure involved ion exchange chromatography on CM-cellulose, Q-Sepharose, and SP-Sepharose and finally fast protein liquid chromatography-gel filtration on Superdex 75. Two amino acid sequences were obtained after tryptic digestion, and they both showed some homology with the esterase of some fungi. Maximal activity was observed at pH 5.0 and at 50 °C. The enzyme displayed relatively high thermostability as evidenced by over 70 % residual activity at 70 °C and about 34 % residual activity at 80 °C. The K m and V max for this enzyme on methyl ferulate were 0.19 mM and 1.65 U/mg proteins, respectively. The purified FAE prefers methyl ferulate over methyl caffeate and is least active on methyl p-coumarate. The FAE activity was not significantly affected by the presence of cations such as Mn(2+), Ca(2+), Cd(2+), Zn(2+), Mg(2+), Cu(2+), and K(+) ions but inhibited by Al(3+), Hg(2+), Fe(2+), and Pb(2+) ions at a tested concentration of 2. 5 mM.

The defect of the melatonin signaling pathway has been proposed to be one of the key etiopathogenic factors in adolescent idiopathic scoliosis (AIS). A previous report showed that melatonin receptor, MT2, was undetectable in some AIS girls. The present study aimed to investigate whether the abnormal MT2 expression in AIS is quantitative or qualitative. Cultured osteoblasts were obtained from 41 AIS girls and nine normal controls. Semi-quantification of protein expression by Western blot and mRNA expression by TaqMan real-time PCR for both MT1 and MT2 were performed. Anthropometric parameters were also compared and correlated with the protein expression and mRNA expression of the receptors. The results showed significantly lower protein and mRNA expression of MT2 in AIS girls compared with that in normal controls (p = 0.02 and p = 0.019, respectively). No differences were found in the expression of MT1. When dichotomizing the AIS girls according to their MT2 expression, the group with low expression was found to have a significantly longer arm span (p = 0.036). The results of this study showed for the first time a quantitative change of MT2 in AIS that was also correlated with abnormal arm span as part of abnormal systemic skeletal growth.

In this study, a 60.8-kDa dimeric lectin was isolated from the Phaseolus vulgaris cv. jade bean and characterized. The lectin was bound on Blue Sepharose 6 and Q Sepharose and was finally purified by size exclusion chromatography on Superdex 200. Its hemagglutinating activity toward rabbit erythrocytes was dependent on divalent cations, especially calcium ions. Various carbohydrates tested were devoid of any effect on the hemagglutinating activity. The lectin was stable at pH between 4.5 and 9.4 and temperatures between 30 and 70 °C. It did not exert antifungal activity toward Valsa mali, Setosphaeria turcica, Mycosphaerella arachidicola, Fusarium oxysporum and Bipolaris maydis. The IC50 of the antiproliferative activity of the lectin toward MCF-7 human breast cancer cells was 174 μM. It did not inhibit proliferation of WRL-68 human normal embryonic hepatocytes. The lectin was dependent on calcium ions for hemagglutinating activity and possessed a blocked N-terminus. These two characteristics make the lectin unique among Phaseolus lectins.

Although Pleurotus abalonus is a well-known edible mushroom in Asia, there is a dearth of information on its antioxidant activity. The present report is the first one focused on the purification and characterization of 9-beta- d -ribofuranosidoadenine (ADO), 5′-deoxy-5′-(methylthio) adenosine (MTA) and a triterpenoid complex from P. abalonus . Different antioxidant activities including inhibitory effects on hemolysis and lipid peroxidation in brain and kidney homogenates as well as significant synergistic effect on scavenging of hydroxyl radicals were demonstrated, which lays a foundation for the development of P. abalonus as a natural antioxidant applied in medicine.

A 5.4-kDa antifungal peptide was purified from Phaseolus vulgaris L. cv. “northeast red bean” using a protocol that entailed affinity chromatography, ion exchange chromatography, and gel filtration. The molecular mass was determined by matrix-assisted laser desorption ionization time-of-flight. The N-terminal amino acid sequence of the peptide was highly homologous to defensins and defensin-like peptides from several plant species. The peptide impeded the growth of a number of pathogenic fungi, including Mycosphaerella arachidicola Khokhr. (IC50 = 1.7 μM), Setosphaeria turcica Luttr., Fusarium oxysporum Schltdl., and Valsa mali Miyabe & G. Yamada. Antifungal activity of the peptide was fully preserved at temperatures up to 100 °C and pH values from 0 to 12. Congo red deposition at the hyphal tip of M. arachidicola was detected after exposure to the peptide, signifying that the peptide had suppressed hyphal growth. The antifungal peptide did not manifest antiproliferative activity toward human breast cancer MCF7 cells and hepatoma HepG2 cells, in contradiction to the bulk of previously reported plant defensins. The data suggest distinct structural requirements for antifungal and antiproliferative activities.

A diversity of bioactive biomolecules present in soybean is covered in this chapter such as e-Phe-Leu and Trp-Leu, and Val-Leu-Ile-Val-Pro derived from glycinin; Bowman-Birk inhibitor, Kunitz trypsin inhibitor, hemagglutinin, isoflavone-deprived soy peptide, neu‐ tral PR-5 protein, SbPRP protein, ferritin, peroxisomal proteins, defense proteins such as calmodulin, disease resistance protein, beta-glucan-binding protein, l unasin, and glysoja‐ nin; enzymes such as phenylalanine ammonia-lyase and tyrosine ammonia-lyase, de‐ fense-related enzymes, cysteine proteinase, isoflavone synthase, isocitrate lyase, vestitone reductase and chalcone reductase, UDP-glucose: flavonoid 3-O-glucosyltransferase, betaglucosidase, isoflavone conjugate-hydrolyzing beta-glucosidase, genes related to 2-oxo‐ glutarate-dependent dioxygenases, 5'-adenylylsulfate reductase, and ATP sulfurylase, anticarcinogenic daidzein-rich fraction, polysaccharides, glyceollins, and isoflavones. The aforementioned soybean constituents manifest many uses and consumption of soybean promotes health.

Though eliciting a somewhat ‘bitter’ taste to our palate, Momordica charantia (MC) is ‘sweet’ for our health. As a popular vegetable indigenous to tropical countries, MC is frequently encountered in the fields of both culinary usage and folklore medicine, especially in Asia. The advance of scientific techniques helps build a platform for investigating the molecular basis of the pharmaceutical functions of medicinal herbs including MC. Over the past few years, a number of pure bioactive components with potential medicinal applications have been procured as a consequence of in vitro and in vivo investigations. In this review, some representative and promising medicinal constituents of MC have been categorized and their emerging medicinal effects such as antitumor, anti-HIV, and anti-diabetic activities are accompanied by the pertinent molecular mechanisms. In addition, the wide range of antimicrobial and pesticidal activities will further validate the medicinal value of MC. Further large-scale clinical trials for those components are needed to fully ascertain their medicinal effects and potential drawbacks, and a new horizon in the treatment of tumor, AIDS, and diabetes may then be opened.

Nature nourishes an inexhaustible drug bank which can be exploited to treat human diseases. On the way to find different natural drugs, significant strides have been made, including the discovery of antitumor drugs (such as paclitaxel/taxol), drugs against chronic diseases (such as ginsenosides, genipin), and drugs against infectious diseases (artemisinin) or even drugs that promote longevity (resveratrol, rapamycin, and spermidine). In this chapter we will give a brief description of the previous accomplishments and current problems on drug screening from natural sources. Further perspectives in this field will also be discussed.

1. Foreword 1. Joseph J.Y. Sung (The Chinese University of Hong Kong, Hong Kong). 2. Foreword: The most important human heritage: our knowledge of the use of plants as a resource for drug development. Rob Verpoorte (Leiden University, The Netherlands). PREFACE. Evandro Fei Fang, Tzi Bun Ng. PART I. Lectins: From biological recognition molecules to variable medicinal applications. 1. Lectins: Personal Comments of Nathan Sharon Taken from His Memoirs (Translation from Hebrew) . 2. The History of Lectinology. 3. Animal and Human Lectins. 4. Lectins of Marine Origin and their Clinical Applications. 5. Medicinal applications of plant lectins. PART II Structural and functional insights into medicinal ribonucleases. 6. Plant nucleases from nuclease I family: their properties and antitumor potential. 7. Bovine Seminal Ribonuclease and its special features: when two is better than one. PART III The perspectives for ribosome inactivating proteins: unveiling the working mechanisms and potential medicinal benefits. 8. The Discovery of MAP30 and Elucidation of its Medicinal Activities. 9. Ribosome Inactivating Proteins from Plants: Biological Properties and their Use in Experimental Therapy. 10. Current status & Biomedical applications of Ribosome-inactivating proteins. PART IV The research history of protease inhibitors and their health implications. 11. The Health Benefits of the Bowman-Birk Inhibitor. 12. Peptidic inhibitors of serine proteinases of plant origin. 13. Anti-tumor effect of synthetic serine protease inhibitor. 14. A landscape of the health benefits of different natural protease inhibitors. PART V Plethora of health benefits from small molecules: From bench-top to clinical settings. 15. Autophagy upregulation as a therapeutic strategy for neurodegenerative diseases. 16. Rapamycin - current and future uses. 17. Eating, drinking, smoking and cancer prevention: a focus on acetaldehyde. 18. The use of edible mushroom water soluble polysaccharides in the treatment and prevention of chronic diseases: a mechanistic approach.19. Essential Oil Constituents: Biodiversity and their Applicability for Cancer Therapy. PART VI Medicinal compounds from the garden: a bird's eye view of one mushroom, one fruit, one insect, and others.20. Recent research on pharmacological activities of the medicinal fungus Cordyceps sinensis. 21. The bitter fruit with sweet health benefits:A comprehensive synopsis of recent research progress on medicinal properties of Momordica charantia. 22. Bioactive constituents of the silk worm Bombyx mori. 23. Proteins with anticancer and antimicobial activities from mammals, submammalian vertebrates and invertebrates. 24. Defense proteins with antiproliferative and antimicrobial activities from fungi and bacteria. PART VII Modernizing investigations of traditional Chinese medicine at the molecular level. 25. The role of the GP-TCM Research Association to modernization and globalization of Traditional Chinese Medicine. 26. High-throughput screening in traditional Chinese medicine-based drug discovery. PART VIII Questions and future perspectives. 27. Achievements, questions arising and future outlook on the path to discover new medicinal compounds. Index.

Defensins are a group of proteins characterized by a conservedcysteine-stabilized alpha-helix and beta-sheet structural motif. They are widely distributed among vertebrates and invertebrates. In addition to the presence of defensins in mammals and submammalian vertebrates enumerated in Chap. 23, fungi and bacteria also produce a variety of defensins with antiproliferative activity toward tumor cells and anticancer activity in tumor bearing mice. The fungal proteins include ribonucleases, antifungal proteins, antibacterial proteins, ubiquitin-like peptides, ribosome inactivating proteins, hemolysins, hemagglutinins/lectins, laccases, and proteases. The bacterial proteins comprise ribonucleases and antifungal proteins. Some of the aforementioned proteins also display antimicrobial activities toward pathogenic microbes.

Porcine reproductive and respiratory syndrome (PRRS), caused by PRRS virus (PRRSV), is a serious contagious disease in the swine industry. At present, there are no effective control strategies against PRRSV. Thus, there is an urgent need for new treatment regimens that have efficacious antiviral activity to compensate for vaccines. The anti-infective effect of Cryptoporus volvatus has previously been demonstrated in Tradational Chinese Medicine. In this report, we expected to identify a new anti-PRRSV agent in the aqueous extract of C. volvatus, by employing a combination of modern chromatographic purification techniques and indirect immunofluorescence assay (IFA). Our results showed that C. volvatus extracts from every separation step differed in their inhibitory potency on PRRSV. One anti-PRRSV component designated as CM-H-L-5 was isolated from water-soluble fraction of C. volvatus. The inhibition induced by CM-H-L-5 occurred in a dose-dependent manner. CM-H-L-5 appeared to be a low-molecular-weight polyol fragment with amide groups and carboxylic acid groups. Collectively, our findings imply that CM-H-L-5 from the aqueous extract of C. volvatus has the potential to be used for anti-PRRSV therapy.

Plant lectins are a unique group of proteins and glycoproteins with potent biological activity and have received widespread attention for many years. They can be found in wheat, corn, tomatoes, peanuts, kidney beans, bananas, peas, lentils, soybeans, mushrooms, tubers, seeds, mistletoe and potatoes among many others. Due to their ability to bind reversibly with specific carbohydrate structures and their abundant availability, plant lectins have commonly been used as a molecular tool in various disciplines of biology and medicine. Whilst once thought of being a dietary toxin, the focus on plant lectins has since shifted to understanding the useful properties of these lectins and utilizing them in medicinal applications to advance human health. This chapter reviews the current and potential applications of plant lectins in various areas of medically related research.

The intent of this chapter is to review proteins with anticancer and antimicrobial activities from various animal species. The proteins comprise venom proteins from snakes, scorpions, and bees; peptides from frogs and toads; bovine pancreas and frogs; and mammalian proteins and peptides including lactoferrin, cathelicidins, tumor necrosis factor, interferons, interleukins, and hormonal peptides. To avoid duplication of efforts, the medicinal applications of mammalian lectins and ribonucleases fall in the realm of other chapters in this book. The current progress and future work on animal constituents with medicinal activity depicted in this chapter would help to shed light on their clinical potentials or applications.

The intent of this article is to review plant antifungal proteins that have been purified and characterized in the last 5–6 years. The antifungal proteins reported encompass 2S albumins, amidases and ureases, chitinases, defensins and defensin-like peptides, beta-1, 3-glucanases, beta-lactoglobulin-like protein, lectins, lipid transfer proteins, peroxidases, proteases, protease inhibitors, ribonucleases, thaumatin-like proteins, and other antifungal proteins. These proteins demonstrate a diversity of structures. Some of them possess additional activities such as antiproliferative activity toward cancer cells, inhibitory activity toward viral enzymes, and specific aforementioned enzyme activities. The antifungal mechanism may involve permeabilization of fungal cell membrane, induction of reactive oxygen species, and triggering of apoptosis.

Silkworm is economically important and also has a large number of activities encompassing anticancer, antibacterial, antibacterial, antiviral, antihypertensive, angiotensin-converting enzyme activity, hepatoprotective, superoxide dismutase enzyme activity, free radical scavenging and tyrosinase inhibitory, antidiabetic, antityrosinase, cognition enhancing, alternative complement pathway-inhibiting, carotenoid binding, chymotrypsin inhibiting, and chitinase activities. These activities may be beneficial to human health.

The Cordyceps sinensis (CS) fungus is an herbal medicinal product in China with a history of over two millenniums. It exhibits a spectacular array of pharmacological activities encompassing anticancer, antidiabetic, anti-adipogenic, analgesic, anti-inflammatory, neuroprotective, anti-aging, eryptosis inducing activities; ameliorating effect on pulmonary fibrosis, hypoxic pulmonary hypertension, and hypertension; protective action against viral myocarditis, stimulatory effect on corticosterone and androgen production, and enhancing effect on exercise performance in healthy older subjects. In this chapter we try to sketch out the recent progress on the pharmacological studies of CS, identify its current research hurdles, and recommend directions for future research.

A lectin, which exerted a hypotensive action in rats after intravenous injection via the jugular vein, was isolated from the mycelia of the edible mushroom Tricholoma mongolicum. The lectin possessed a molecular weight of 37 K and its hypotensive activity was dose-dependent. Administration of the lectin at a dose of 10 mg/kg body weight caused a mean arterial blood pressure reduction of 95.3 +/- 7.4 mmHg. The lectin's hypotensive action was not mediated via autonomic ganglion transmission, alpha-adrenoceptors, beta-adrenoceptors, cholinergic receptors, histaminergic receptors, nor the renin-angiotensin system, but it was probably mediated through vasorelaxation via adenosine A2 receptors and/or nitric oxide production.

Alpha- and beta-momorcharins, ribosome-inactivating proteins from Momordica charantia seeds, were utilized in this investigation. Ribonucleolytic cleavage was observed after naked rRNA was incubated with either momorcharin. Beta-momorcharin, and to a lesser extent alpha-momorcharin, also acted on tRNA to release acid-soluble UV-absorbing products. Such activity was optimal at pH around 5.5. Using polyhomoribonucleotides as substrate, it was found that the momorcharins preferentially acted on polyU, but exerted negligible effects on polyA, polyC and polyG. Chromatographic analysis of the reaction product indicated that mono and/or oligo-ribonucleotides, but not free base, were generated from polyU, suggesting that the enzymatic action involved ribonucleolytic cleavage. Similar to the results obtained with tRNA as substrate, beta-momorcharin was about 15-fold more active than alpha-momorcharin on polyU.

A new purification scheme, involving two successive ion exchange Chromatographic steps on DEAE-cellulose and Mono-S FPLC, was developed for the isolation of the ribosome-inactivating proteins, α- and β-momorcharins, from the Chinese herb Kuquazi (seeds of Momordica charantia). This simple and rapid procedure yielded 3.1 and 1.7 mg of α- and β-momorcharins, respectively, from 2.5 g of decorticated seeds in only two days. The N-terminal amino acid sequence of β-momorcharin was found to be DVNFDLSTATAKTYTKFIED. It differed from that of α-momorcharin (DVSFRLSGADPRSYGMFIKD) in 10 out of the 20 positions investigated. Like other ribosome-inactivating proteins, the purified momorcharins showed specific N-glycosidase activity at nanomolar concentrations, when rRNA from rabbit reticulocyte lysate was used as substrate. The N-glycosidase activity of both momorcharins was optimal at pH7, not inhibited by K+ and not appreciably affected by NH4+. The activity of α-momorcharin was not drastically altered by Mn2+ but (1–10mM) Mn2+ inhibited the activity of β-momorcharin by about 40%.