Harmaline and hispidin from Peganum harmala and Inonotus hispidus with binding affinity to Candida rugosa lipase: In silico and in vitro studies

The inhibitory effect of phenolic compounds and alkaloids of Inonotus hispidus and Peganum harmala on Candida rugosa lipase was investigated, also, their antioxidant activities using DPPH, ABTS and phosphomolybdenum were studied in this paper. The phenolic extracts have shown a stronger antiradical activity than the alkaloids extracts. The enzymatic inhibition produced by these extracts is described here for the first time. The results have shown that the phenolic and the alkaloid extracts are good inhibitors of C. rugosa lipase. Thus, the inhibitor molecules (harmaline and hispidin) have been isolated from P. harmala and I. hispidus. Their structures were elucidated by ¹H NMR analysis. Molecular docking has been achieved using AutoDock Vina program to discuss the nature of interactions and the mechanism of inhibition. Therefore, these isolated molecules could be used in the treatment of candidiasis.     

Partial characterization of a phenolic pigment from sporocarps of Phellinus igniarius

A dark brown phenolic polymer, isolated from mature sporocarps of Phellinus igniarius (DC. ex Fr.) Quél., is responsible for the darkening of sporocarp tissue when a drop of base is applied. The pigment is rich in aliphatic as well as phenolic hydroxyl groups, and also contains significant numbers of carboxyls and carbonyls. The presence of 3,4-dihydroxyphenyl moieties was documented by the identification of veratric and metahemipinic acids from oxidative degradation of methylated samples. Despite a 2% methoxyl content, the polymer contains no guaiacyl or syringyl moieties and therefore contains no lignin. It appears to be formed by the oxidative polymerization of a molecule containing a 3,4-dihydroxystyryl moiety. Hispidin was identified as its trimethyl ether in methylated extracts of pigmented mycelial mats of Phellinus igniarius grown in liquid culture.     

Hispidin biosynthesis in cultures of Polyporus hispidus

Cultures of Polyporus hispidus grown on a liquid medium with glucose as the principal carbon source produced the yellow pigment hispidin (4-hydroxy-6-(3,4-dihydroxystyryl)-2-pyrone) (II). Tracer studies showed that DL-phenylalanine, DL-tyrosine, cinnamate, p-coumarate and caffeate were incorporated into the hispidin molecule without scrambling of the label. Good incorporation of acetate and roalonate into the pyrone portion of the molecule was observed. The related styrylpyrone, bis-noryangonin (4-hydroxy-6-(4-hydroxystyryl)-2-pyrone) (1) was also detected in extracts of cultured mycelium and a cell-free enzyme preparation was obtained which catalyzed the hydroxylation of bis-noryangonin to hispidin.     

Synthetic hispidin, a PKC inhibitor, is more cytotoxic toward cancer cells than normal cells in vitro

The trypanocidal activity of naturally occurring 6-(3,4-dihydroxystyryl)-4-hydroxy-2-pyrone (hispidin) prompted us to examine its cytotoxic activity toward normal and cancerous cells in culture. Hispidin synthesized in our laboratory to a high degree of purity (checked by 1H and 13C NMR spectroscopy) was shown to be cytotoxic (between 10-3 mol/L and 10-7 mol/L) toward normal human MRC-5 fibroblasts, human cancerous keratinocytes (SCL-1 cell line), and human cancerous pancreatic duct cells (Capan-1 cell line). Interestingly, addition of hispidin in three successive doses (between 10-5 mol/L and 10-7 mol/L) led to a 100-fold increase in activity with an enhanced activity on cancer cells compared to normal cells (50%). Synthetic hispidin was found to inhibit isoform of protein kinase C (IC50 of 2 × 10-6 mol/L), but not E. coli and placental type XV alkaline phosphatases. The enhanced activity of hispidin toward the cancerous cell lines is discussed.     

茉莉酸和双氧水对斜生褐孔菌一株黑色素缺失突变体产生酚类化合物的影响

黑色素合成是桦褐孔菌Phaeoporus obliquus次生代谢过程中的一个重要事件。黑色素的过多合成势必影响其他酚类的积累。本文以茉莉酸和双氧水为调控因子,开展了该突变体深层发酵过程酚类化合物积累研究。正常培养条件下,每升发酵液积累酚类化合物最高可达317.45mg/L,其中包括苯甲酸衍生物(45.3mg/L),黄酮苷元(74.4mg/L),黄酮苷(160mg/L)以及硬毛素类化合物(18.5mg/L)。加入茉莉酸后,总酚最大值增加到620.45mg/L,其中苯甲酸衍生物增加到230.26mg/L,黄酮苷元增加到166.86mg/L,而黄酮苷和硬毛素类化合物却分别降低到98.5mg/L和7.1mg,双氧水的加入没有提高总酚的最大积累(315.69mg/L)。因此,与野生型相比,黑色素缺失使总酚的积累有所增加,随茉莉L酸的加入总酚积累最大值增加近一倍。虽然双氧水的加入提高了苯丙氨酸裂解酶活性,但总酚的积累却没有增加,这可能是由于一部分酚类化合物在抗双氧水氧化胁迫过程中被消耗。     

深层培养的斜生褐孔菌酚类化合物对铅处理小鼠耐力的增强作用

本文研究了不同条件下深层培养的斜生褐孔菌Phaeoporus obliquus与野生桦褐孔菌在酚类化合物组成及其含量上的差异以及对铅处理小鼠常压缺氧和游泳耐力影响。野生桦褐孔菌主要含有水溶性黑色素、硬毛素衍生物,两者各占总酚类化合物的12.1%和42.2%。而深层培养的桦褐孔菌则主要积累黄酮类化合物,其中正常生理条件下积累量达到总酚的35.5%,加入H2O2后积累量达到总酚的92.5%,同时加入H2O2和熊果苷后则为总酚的76.9%。正常培养基中水溶性黑色素的积累量达到总酚的20.2%,而加入H2O2或同时加入H2O2和熊果苷后水溶性黑色素则很少合成。口服野生斜生褐孔菌或不同条件下深层培养的斜生褐孔菌酚类化合物都能使铅处理小鼠忍耐缺氧和游泳的时间延长12–15min。同时,口服不同来源的斜生褐孔菌酚类化合物可提升铅处理小鼠外周淋巴细胞的活力和总超氧化物歧化酶活性,并降低丙二醛的积累。因此,斜生褐孔菌酚类化合物对铅处理小鼠耐缺氧和游泳的提升作用在于抑制了脂质过氧化反应,增强了过氧化物歧化酶的表达,从而保护了铅处理小鼠的细胞和器官免于氧化胁迫的损伤。     

Antioxidant polyphenols from the mycelial culture of the medicinal fungi Inonotus xeranticus and Phellinus linteus

Aims:  The medicinal fungi Inonotus xeranticus and Phellinus linteus in the family Hymenochaetaceae have been used as traditional medicines for the treatment of various diseases. However, the compound responsible for the antioxidant activity is still unknown. Therefore, this study was conducted to characterize the antioxidant substances present in cultured broths made from these fungi.
Methods and Results:  Antioxidant fractions of the cultured broths obtained from I. xeranticus and P. linteus were analysed using reversed-phase HPLC, which revealed several peaks that exhibited a potent free radical scavenging activity. To identify these antioxidant peaks, an I. xeranticus strain was mass-cultured, and the cultured broth was separated using antioxidant activity-guided fractionation. Four major active substances were purified and identified as hispidin and its dimers, 3,14'-bihispidinyl, hypholomine B, and 1,1-distyrylpyrylethan based on spectroscopic analyses. All compounds exhibited a significant scavenging activity against these radical species in a concentration-dependent manner.
Conclusions:  Antioxidant substances found in the cultured broths of the medicinal fungi I. xeranticus and P. linteus were identified as hispidin and its dimers, 3,14'-bihispidinyl, hypholomine B, and 1,1-distyrylpyrylethan.
Significance and Impact of the Study:  Polyphenol antioxidants were isolated from the cultured broth of the medicinal fungi I. xeranticus and P. linteus and identified based on extensive spectroscopic analyses. These compounds exhibited a strong antioxidant activity.     

桦褐孔菌酚类化合物及其免疫促进作用

Phenolic compounds from field-grown Inonotus obliquus sclerotia (Chaga) consist mainly of hispidin analogs and melanins, and are thought to be the active constituents to treat several human diseases. In submerged cultures of the fungus, however, no information is currently available on the production of phenolic compounds and their corresponding pharmacological functions. In this study, phenolic compounds from Chaga and submerged cultures of the fungus were assayed for their composition and immune-stimulating effects. Phenolic compounds produced by I. obliquus in submerged cultures mostly consist of flavonoids, together with small amounts of hispidin analogs and melanins. This is quite contrary to the situation in Chaga, where flavonoids are determined as trace elements. Furthermore, phenolic compounds from Chaga show capacity about two-fold higher than those produced in submerged cultures in inhibiting cyclophosphamide-induced reduction of bodyweight, spleen index and viability of peripheral lymphocytes in test mice. Thus less production of hispidin analogs and melanins is likely to be responsible for less immune-stimulating effects in phenolic compounds from submerged cultures, and additional factors should be imposed during submerged cultures of I. obliquus to regulate biosynthesis of phenolic compounds directed to the composition similar to Chaga.     

鲍氏层孔菌子实体的化学成分研究

采用硅胶和Sephadex LH20柱层析方法,从鲍氏层孔菌子实体提取物中分离得到8个化合物。运用NMR和MS等波谱法分析和鉴定为7(8),22(23)-二烯-3-酮-麦角甾烷、4,6,8(14),22(23)-四烯-3-酮-麦角甾烷、麦角甾醇、过氧化麦角甾醇、三十烷酸对羟基苯乙酯、4-(3,4-二羟苯基)-3-丁烯-2-酮、hispolon、hispidin。     

Recent advances in the study of Kaposi's sarcoma-associated herpesvirus replication and pathogenesis

It has now been over twenty years since a novel herpesviral genome was identified in Kaposi's sarcoma biopsies. Since then, the cumulative research effort by molecular biologists, virologists, clinicians, and epidemiologists alike has led to the extensive characterization of this tumor virus, Kaposi's sarcoma-associated herpesvirus(KSHV; also known as human herpesvirus 8(HHV-8)), and its associated diseases. Here we review the current knowledge of KSHV biology and pathogenesis, with a particular emphasis on new and exciting advances in the field of epigenetics. We also discuss the development and practicality of various cell culture and animal model systems to study KSHV replication and pathogenesis.     

The structure, reproduction and taxonomy of Vidalia and Osmundaria (Rhodophyta, Rhodomelaceae)

Comprises species occurring mostly in subtidal habitats in tropical, subtropical and warm-temperate areas of the world. An analysis of the type species, V. spiralis (Sonder) Lamouroux ex J. Agardh, a species from Australia, establishes basic characters for distinguishing species in the genus. These characters are (1) branching patterns of thalli, (2) flat blades that may be spiralled on their axis, (3) width of the blade, (4) primary or secondary derivation of sterile and fertile branchlets and (5) position of sterile and fertile branchlets on the thalli. Application of the latter two characters provides an important basic method for separation of species into three major groups. Osmundaria , a genus known only in southern Australia, was studied in relation to Vidalia , and its separation from the Vidalia assemblage is not accepted. Species of Vidalia therefore are transferred to the older genus name, Osmundaria. Two new species, Osmundaria papenfussii and Osmundaria oliveae are described from Natal. Confusion in the usage of the epithet, Vidalia fimbriala Brown ex Turner has been clarified, and Vidalia gregaria Falkenberg, described as an epiphyte on Osmundaria pro/ifera Lamouroux, is revealed to be young branches of the host, Osmundaria prolifera.     

New or rare chromosome counts in Artemisia L. (Asteraceae, Anthemideae) and related genera from Kazakhstan

Fifteen chromosome counts of six Artemisia taxa and one species of each of the genera Brachanthemum, Hippolytia, Kaschgaria, Lepidolopsis and Turaniphytum are reported from Kazakhstan. Three of them are new reports, two are not consistent with previous counts and the remainder are confirmations of very scarce (one to four) earlier records. All the populations studied have the same basic chromosome number, x = 9, with ploidy levels ranging from 2x to 6x. Some correlations between ploidy level, morphological characters and distribution are noted.     

肝癌中HBV和HCV基因和抗原的分布及意义

采用原位分子杂交方法检测HCV RNA及HBV X基因;采用免疫组织化学方法研究HCV核心抗原,非结构区C33c抗原及HBxAg在肝细胞肝癌中的定位及分布.结果表明(1)HCV RNA、HBV X基因在肝细胞肝癌组织检出率分别为40%(55/136)和82%(112/136).HCV RNA定位于癌细胞的胞浆内,阳性细胞呈散在、灶状及弥漫分布三种形式;HBV X基因在肝癌细胞中的分布呈胞浆型、核型及核浆型,阳性细胞也呈上述三种分布形式;(2)HCV C33c抗原、核心抗原在肝细胞肝癌中的阳性率为81%(133/164)及86%(141/164).C33c抗原定位于癌细胞及肝细胞的胞浆内;核心抗原既定位于癌细胞核中,又可定位于胞浆中.C33c抗原阳性细胞以灶状分布为主;而核心抗原阳性细     

Selection with two alleles and complete dominance

For a plant selection model with frequency-independent viabilities, fertilities and selfing rates, it is shown that apart from global fixation, for certain parameter combinations a protected polymorphism and facultative fixation (either allele may become fixed according to initial frequencies) may both occur. Facultative fixation requires different selling rates for the dominant and recessive type. Protection of the polymorphism requires resource allocation for male and female function. In this connection the problem of purely genetically caused population extinction is discussed.
For general frequency dependence and regular segregation, the chances for establishment of a completely recessive gene are compared to those of a completely dominant gene. It is proven that the process of establishment of the recessive gene, despite a fitness advantage, may be considerably endangered by drift effects if random mating prevails. The recessive gene may reach the same effectivity in establishment as a dominant gene, only if the recessive homozygote mates exclusively with its own type during the period of establishment.