In vitro metabolism,permeation, and brain availability of six major boswellic acids from Boswellia serrata gum resins

Boswellia serrata gum resin extracts (BSE) revealed potent anti-inflammatory actions in preclinical and clinical studies. In 2002 BSE was assigned an orphan drug status by the European Medicines Agency (EMA) for the treatment of peritumoral edema. In the past pharmacological effects of BSE were mainly attributed to 11-keto-β-boswellic acid (KBA) and 3-acetyl-11-keto-β-boswellic acid (AKBA). Therefore pharmacokinetic and pharmacodynamic studies focused mainly on these two boswellic acids (BAs). However, other BAs, like β-boswellic acid (βBA), might also contribute to the anti-inflammatory actions of BSE. Here, we determined the metabolic stability, permeability and brain availability of six major BAs, that is, KBA, AKBA, βBA, 3-acetyl-β-boswellic acid (AβBA), α-boswellic acid (αBA), and 3-acetyl-α-boswellic acid (AαBA). For permeability studies, the Caco-2 model was adapted to physiological conditions by the addition of bovine serum albumin (BSA) to the basolateral side and the use of modified fasted state simulated intestinal fluid (FaSSIF) on the apical side. Under these conditions the four BAs lacking the 11-keto moiety revealed moderate permeability. Furthermore the permeability of AKBA and KBA was improved compared to earlier studies. In contrast to Aα- and AβBA, βBA and αBA were intensively metabolized after incubation with human and rat liver microsomes. Finally, the availability of all six major BAs could be confirmed in rat brain 8 h after oral administration of 240 mg/kg BSE to rats showing mean concentrations of 11.6 ng/g for KBA, 37.5 ng/g for AKBA, 485.1 ng/g for αBA, 1066.6 ng/g for βBA, 43.0 ng/g for AαBA and 163.7 ng/g for AβBA.     

Permeability determination and pharmacokinetic study of nobiletin in rat plasma and brain by validated high-performance liquid chromatography method

In the present study, we are reporting permeability and pharmacokinetics of nobiletin in rat plasma and brain, using a validated reverse phase high performance liquid chromatographic method. Protein precipitation method was used for the extraction of nobiletin and coumarin (IS) from rat plasma and brain tissue. The system was run in isocratic mode with mobile phase consisting of potassium dihydrogen ortho-phosphate (pH 4.5; 0.04 mM) and acetonitrile in ratio of 50:50, v/v. The total chromatographic run time was 9.0 min. The method was proved to be accurate and precise at linearity range of 0.05–10 μg/mL with a correlation coefficient (r) of ≥ 0.994 in rat plasma and ≥ 0.995 in rat brain. The intra- and inter-day precision and accuracy values are found to be within the assay variability limits as per the FDA guidelines. Nobiletin was found stable in the battery of stability studies viz., bench-top, auto-sampler, freeze/thaw cycles and long term storage in a freezer at − 70 ± 10 °C. Maximum concentrations of nobiletin in both plasma and brain were observed at 1 h after single oral dosing (50 mg/kg). The maximum concentration in plasma and brain were 1.78 and 4.20 μg/mL, respectively. The AUC_0–t in plasma and brain were 7.49 and 20.66 μg·h/mL, respectively. The mean elimination half life (t_1/2) in plasma and brain were 1.80 and 11.42 h, respectively. The Parallel Artificial Membrane Permeability Assay (PAMPA) permeability of nobiletin was found to be high at both pH 4.0 and 7.0.     

Comparative pharmacokinetic study of paeoniflorin and albiflorin after oral administration of Radix Paeoniae Rubra in normal rats and the acute cholestasis hepatitis rats

A comparative study was designed and conducted to compare the pharmacokinetic difference of paeoniflorin and albiflorin after oral administration of Radix Paeoniae Rubra to normal rats and the acute cholestasis hepatitis rats induced by alpha-naphthylisothiocyanate (ANIT). UPLC-ESI-MS/MS method was employed to determine the level of paeoniflorin and albiflorin in rat plasma using geniposide as the internal standard (IS). Unpaired Student's t-test was used for the statistical comparison. The investigation showed that there were significant differences between the normal rats and the acute cholestasis hepatitis rat groups in calculated parameters, such as AUC(0-t), AUC(0-∞), T(max) and CLz/F. The results indicated that acute liver injury in rats could alter the pharmacokinetics of drug. Since patients are the final users of the drug, it is essential to investigate the pharmacokinetics of the drug in disease status. Therefore, we used normal rats and the acute cholestasis hepatitis rats to study pharmacokinetics of Radix Paeoniae Rubra with the purpose of examining the influence of disease on the metabolic course.     

Pharmacokinetics, tissue distribution, and the lactone/carboxylate equilibrium of hydroxycamptothecin delivered via aerosol in mice

Aerosol delivery is a route which is advantageous to the therapy of pulmonary diseases, such as lung cancer. The pharmacokinetics and tissue distribution after aerosol delivery of carboxylate form of hydroxycamptothecin (C-HCPT) were investigated. The concentrations of the three different types (lactone, carboxylate and the total of both forms) of HCPT were measured by HPLC analysis. The initial experiment showed no evident difference between lactone and carboxylate in the lungs during the aerosol treatment, compared with the HCPT content in plasma. The AUC(inf) value of lactone in the lungs was higher than that of carboxylate, which was 138,176.00minngg(-1) and 128,460.00minngg(-1), respectively. Meanwhile, AUC(inf) in the plasma during the entire treatment indicated that the lactone content was always at a lower level, and the carboxylate form tended to predominate, as shown by the lactone/carboxylate (L/C) equilibrium. The tissue distribution results showed that the lactone proportion in the liver increased up to the maximum value of 69.69% after aerosol administration, whereas the mean L/C equilibrium index for the liver was 2.07±1.06, and the C(max) and AUC(0-∞) values of the total HCPT were highest in the tissues. Based on these results we speculated that the initial wholly carboxylate form of the HCPT atomized liquid did not influence the transformation to lactone form. Moreover, the deposition of the total HCPT and lactone was higher in the lungs and other tissues than in the plasma after the aerosol treatment. This study will be beneficial to the therapy of pulmonary carcinoma.     

Ultra-performance liquid-chromatography with tandem mass spectrometry performing pharmacokinetic and biodistribution studies of croomine,neotuberostemonine and tuberostemonine alkaloids absorbed in the rat plasma after oral administration of Stemonae Radix

Stemonae Radix (Stemona tuberosa Lour, Bai Bu) is an important traditional Chinese medicinal (TCM) plant known for its antitussive activity. Croomine, neotuberostemonine and tuberostemonine alkaloids of Stemonae Radix are major components responsible for antitussive action. In this work, plasma pharmacokinetic and biodistribution characteristics of the three alkaloids after oral administration of Stemonae Radix are investigated using a rapid and sensitive UPLC-Q-TOF–HDMS method. Mass spectrometry (MS) was performed on a Waters Micromass high-definition technology with an electrospray ionization source in positive ion mode, with excellent MS mass accuracy and enhanced MS data acquisition. Separation of main alkaloids was achieved on a Waters BEH C₁₈ column by linear gradient elution. Data were analyzed and estimated by compartmental methods and pharmacokinetic parameters calculated using WinNonlin Professional version 5.1. It was found that croomine, neotuberostemonine and tuberostemonine had faster absorbed into the bloodstream, maintain the high plasma concentration, and pose a large AUC value. The biodistribution of neotuberostemonine and tuberostemonine showed that the higher levels were in liver, and lung. Croomine was discovered in brain and showed that it could cross the blood–brain barrier, indicating that croomine plays an antitussive effect as acting on the central nervous system. Neotuberostemonine and tuberostemonine were not discovered in brain, demonstrating that they play an antitussive effect as peripheral antitussive. This work suggests that the pharmacokinetics and biodistribution based-UPLC-Q-TOF–HDMS can provide a reliable tools for screening bioactive components contributing to pharmacological effects of medicinal herbs.     

In vitro regeneration of Pacific silver fir (Abies amabilis) plantlets and histological analysis of shoot formation

A protocol is described for the in vitro propagation of Abies amabilis (Dougl.) Forbes. Over 60% of the cotyledonary explants from 5-day-old cotyledons formed shoots when cultured for 7 days on Schenk and Hildebrandt's (SH) medium containing 10 micro M N(6)-benzyladenine (BA) followed by another 7 days on SH medium containing 10 micro M each of BA and zeatin. Shoot multiplication was unsuccessful. Seventeen percent rooting was obtained after pulsing for 4 h in 1 mM indole-3-butyric acid and planting pulsed shoots in 1/1 (v/v) perlite/vermiculite. Cell clusters (promeristemoids) of five to seven cells were observed on the cotyledonary explants after 7 days of culture in the presence of cytokinin. These cells developed further into meristematic domes and apical meristems. In the absence of cytokinin, stomata and resin canals reached maturity, whereas cells within the cortex became vacuolated and developed into palisade and spongy mesophyll.     

Floristic analysis and distribution pattern of alien plants in Shandong Province, eastern China

Alien plants, along with their ecological invasion and negative impacts on indigenous species diversity and ecosystems, are one of the major topics of current ecological research. The background investigation and floristic analysis of alien plants are very important and form an essential database for invasive species research, control and management. In our study these alien plants, mainly collected from the Flora of Shandong Province, combined with a field survey, were studied and analyzed. We also established a floristic database. Our findings are as follows: 1) there are a total of 827 alien species, belonging to 122 families and 416 genera of which 348 species were imported from other countries; 2) a high proportion, 39.0% of the flora in Shandong Province, is accounted for by alien species, of which 21 dominant families largely belong to the Rosaceae, Leguminosae, Asteraceae and Gramineae; 3) the diverse geographical distribution of the genera is characterized by species dominant in the temperate zone which accounts for 52.5% of the alien plants and 44.1% of the plants from the tropics; 4) the origins of alien species and their centralized distribution in Shandong together show the anthropogenic effect and unnatural impacts on the environment and 5) in Shandong Province, alien plants originate more from temperate zones than from any other areas of the world. __________ Translated from Chinese Journal of Ecology, 2007, 26(4): 489–494 [译自: 生态学杂志]     

福建省南方红豆杉的分布、生境和栽培技术研究

对南方红豆杉在福建省的自然分布区域、资源数量、生长环境条件、生物学特性，以及种子采集、贮藏催芽、繁育扦插嫁接栽培技术进行研究，并在试验示范基础上，全面系统地提出了南方红豆杉木材、药用、观赏方面的开发利用建议。     

In vitro antifungal activities of Allium cepa, Allium sativum and ketoconazole against some pathogenic yeasts and dermatophytes

By using an agar dilution assay, the antifungal activity of aqueous extracts prepared from Allium cepa (onion; AOE) and Allium sativum (garlic; AGE) were evaluated against Malassezia furfur (25 strains), Candida albicans (18 strains), other Candida sp. (12 strains) as well as 35 strains of various dermatophyte species and compared with the activity of a known antifungal drug, ketoconazole (KTZ). All the AOE, AGE and KTZ were found to be able to inhibit growth of all fungi tested in a dose-dependent manner with maximum of 100% at defined concentrations. The results indicate that onion and garlic might be promising in treatment of fungal-associated diseases from important pathogenic genera Candida, Malassezia and the dermatophytes.     

乳源木莲杉木马尾松混交林生物量分布格局分析

对乳源木莲、杉木和马尾松纯林及其三种不同混交林林分生物量和它们的空间分布状况进行分析,结果表明,乔木层的生物量占92%以上,混交林有利于干、枝、叶生物量的积累,不利于皮、草本、灌本及地下部分和凋落物层生物量的积累,但混交林中的半分解物含量比纯林有成倍增加,混交林有利于枯枝落叶的分解。     

In vitro ectomycorrhizal formation of Pinus patula, P. pseudostrobus, P. oocarpa and P. elliotii grown in southern India

In vitro syntheses of ectomycorrhizas of Pinus patula, P. pseudostrobus, P. oocarpa and P. elliotii with Amanita muscaria, Laccaria fraterna, L. laccata, Pisolithus tinctorius, Rhizopogon luteolus, Scleroderma citrinum and Thelephora terrestris were attempted. Most of the fungi initiated mycorrhizal formation within 2 weeks after inoculation. Laccaria fraterna and S. citrinum formed mycorrhizas after 6 weeks of inoculation. Amanita muscaria failed to form mycorrhizas during the 12 week period of observation. The degree of colonization of different mycorrhizal fungi varied with different species of Pinus.     

In vitro evaluation of fungicides against Cylindrocladium spp. causing diseases of Eucalyptus in Kerala,India1

A total of 22 fungicides were evaluated in vitro for their efficacy against Cylindrocladium quinqueseptatum (CQ), C. ilicicola (CI), C. floridanum (CF), C. parvum (CP), and C. camelliae (CC) causing various diseases of Eucalyptus in Kerala, India. Though there were a number of fungicides effective (ED₁₀₀) in the comdial germination and poisoned food techniques, only carbendazim provided complete inhibition of CQ, CI and CC in the soil-fungicide screening technique; carbendazim was also highly effective against CF and CP. C. ilicicola appeared to be more tolerant than other species of Cylindrocladium as only a few fungicides were found to be effective against it even in the conidial germination technique. On comparison of the three fungicidal evaluation techniques, it is concluded that, for a pathogen producing microsclerotia, like Cylindrocladium, the soil-fungicide screening technique is the most appropriate one for obtaining reliable results.     

In vitro cytotoxic activity of Brazilian plant extracts against human lung, colon and CNS solid cancers and leukemia

The cytotoxicity of extracts obtained from plants occurring in the Amazon and Atlantic rain forests against NCI-H460, KM-12, SF-268 and RPMI-8226 cancer cell lines was investigated. Expressive activity was observed in the extracts of Toulicia cf. pulvinata, Ampirrhox sp., Macoubea sprucei, Calophyllum brasiliense, Vismia guianensis, Caryocar microcarpum, Xylopia aromatica and Distictella magnoliifolia.     

农桐间作中光辐射分量实测分析

在田间实测基础上,分析了农桐间作中总辐射、反射辐射、净辐射、直接辐射、散射辐射及生理辐射的时空变化特征、分布规律及影响因素,为认识间作田中小气候形成原因和农田遮荫情况提供依据,对提高作物生产潜力,扩大农桐间作面积有参考意义。     

秦巴林区啮齿动物群落构成及区划分析

以秦巴林区森林的垂直分布带为依据,应用聚类分析法对秦巴林区不同地区啮齿动物分布情况进行分析。结果表明:中山桦木林带、亚高山针叶林带和高山灌丛草甸带聚为一类,说明在以上三个地理单元,啮齿动物种类和数量相似;而浅山农作区、低-中山栎林带聚为一类,说明它们的啮齿动物分布相似。将不同垂直带的平均海拔高度与其啮齿动物种数进行相关分析,表明海拔高度和啮齿动物种数之间呈负相关。     

Composition, distribution and supposed origin of mineral inclusions in sessile oak wood — consequences for microdensitometrical analysis

SEM and light-microscopical observations, supported by chemical microanalysis with an EDXA system, revealed that light-saturated pixels observed in X-ray negatives of sessile oak (Quercus petraea Liebl.) wood were caused by inorganic deposits present inside multiseriate ray and axial parenchyma cells. Calcium oxalate crystals, silica grains and amorphous granules with varied mineral compositions have been identified. The wood strips of three out of six sampled trees contained measurable amounts of mineral inclusions which were quantified using image analysis. Based on the variations of mineral content observed between trees and within and between annual rings of the same tree, some hypotheses were formulated concerning the factors involved in the formation of inorganic deposits in oak wood. Their occurrence varies depending on the mineral concerned and seems to be controlled largely by a tree effect. The time of formation appears to coincide with a shifting of the oak wood’s functions as a result of heartwood formation processes (inter-annual scale) or changes in leaf phenology and climate (intra-annual scale). In addition, the technical consequences of their presence as well as their effects on wood density measurements through microdensitometry are discussed.     

In vitro effects of four ectomycorrhizal fungi, Boletus edulis, Rhizopogon roseolus, Laccaria laccata and Lactarius deliciosus on Fusarium damping off in Pinus nigra seedlings

Species of Fusarium cause damping off of forest seedlings resulting in considerable losses and oftentimes fungicides are used to control the disease. Such chemicals may be detrimental to the environment. Protection of forest seedlings against this disease must be focused on integrated management for in which biological control is one of the most important tools. Mycorrhizal fungi may provide protection against pathogen invasion of seedlings. In this study, the interactions between the mycorrhizal fungi Boletus edulis, Rhizopogon roseolus, Laccaria laccata and Lactarius deliciosus and damping off pathogens (Fusarium oxysporum and F. moniliforme) were investigated in vitro and mechanisms involved in the protection of damping off in P. nigra seedlings are also discussed. The effect of filtrates from mycorrhizal species on spore germination of F. oxysporum varied throughout the tests. At the end of the assay, the inhibitory effect only could be observed in the L.␣deliciosus treatment. None of the filtrates reduced spore germination in F. moniliforme. Finally, three of the four mycorrhizal species grown for 2 months in the substrate, L. laccata, L.␣deliciosus and B. edulis, increased the survival of Fusarium inoculated P.␣nigra seedlings.     

激素、通气和pH值对四倍体刺槐和二乔刺槐离体生长的影响

以四倍体刺槐和二乔刺槐的不定芽为试验材料,通过比较培养前后的芽伸长量、增殖率和生物量等指标,研究了激素、通气状况(封口材料)、培养基pH值对刺槐离体生长的影响.结果表明,在BA0.5mg/L、NAA0.1mg/L时,四倍体刺槐不定芽的生物量最大;在BA0.5mg/L、NAA0.5mg/L时,二乔刺槐不定芽的生物量最大.在湿度为35%±5%环境条件下,四倍体刺槐封口材料以耐高温塑料最好,二乔刺槐以棉塞最好;在湿度为55%±5%条件下,四倍体刺槐和二乔刺槐均以棉塞最好.在pH值为5.6、5.8和6.0时四倍体刺槐的增殖系数大,二乔刺槐在pH为6.4时的增殖系数大,而两品种在不同pH下的质量增量百分率差异不显著.     

沙地云杉(Piceamongolica)在中国的形成、分布和特性(英文)

沙地云杉是中国的珍稀濒危树种。沙地云杉林是陆地上非常特殊的森林生态系统类型。它存在于我国森林草原过渡带,也是我国的农牧交错区。在本文中,我们讨论了沙地云杉的形成和分布,研究了沙地云杉的性质和特性。沙地云杉属于白扦云杉系列,是白扦云杉长期适应当地气候而演化的地方宗,然后形成异域的半种,最后形成分类种。沙地云杉林是森林草原过渡带上的超地带性顶极群落。     

秤锤树生态地理分布,生物学特性与繁殖的初步研究

研究了秤锤树的生态地理分布、生物学特性与繁殖技术。３种繁殖试验的结果表明：种子出苗率以随采随播为最高,出苗率２５％～３１％,比历年记录高近１０倍;硬枝与嫩枝扦插的成活率均在６０％以上;组培幼苗成活率达７５％。