A novel analytical ELISA-based methodology for pharmacologically active saikosaponins

A competitive enzyme-linked immunosorbent assay (ELISA) for saikosaponins was established using monoclonal antibody (MAb) 3G10. Hybridoma 3G10 prepared by fusing splenocytes immunized with saikosaponin a-BSA (SSa-BSA) conjugate and a hypoxanthine-aminopterin-thymidine (HAT)-sensitive mouse myeloma cell line, P3-X63-Ag8-U1, secreted monoclonal antibodies with wide cross-reactivity to saikosaponins including saikosaponin b(2) (SSb(2)), c (SSc) and d (SSd), which are stereo and/or regio isomers of SSa. The method, at an effective measuring range of 0.6 mug /ml to 2.3 mug/ml of SSa, successfully detected total saikosaponins in Bupleuri radix and Kampo medicines prescribed with Bupleuri radix. Good correlation between ELISA and HPLC analyses of total saikosaponin in a crude extract of Bupleuri radix was obtained after hydrolysis of acyl saikosaponins by treatment with a mild alkaline solution.     

Tithonia and senna green manures and inorganic fertilizers as phosphorus sources for maize in Western Kenya

Efforts to overcome declining soil fertility on small holder farms in western Kenya must be consistent with the reality of low utilization of inorganic fertilizers. Likewise organic inputs alone cannot supply adequate nutrients. The use of two organic resources, Tithonia diversifolia (tithonia) and Senna spectabilis (senna) leaves, and their combination with inorganic P for improving soil fertility and maize yields was investigated on a P limiting soil in Western Kenya. Treatments included: 1) control, no inputs; 2) 5 t ha⁻¹ (dry matter) tithonia leaves; 3) 5 t ha⁻¹ senna leaves; 4) 5 t ha⁻¹ tithonia leaves + 25 kg P ha⁻¹ as triple superphosphate (TSP); 5) 5 t ha⁻¹ senna leaves + 25 kg P ha⁻¹ (as TSP); and 6) 25 kg P ha⁻¹ of TSP. Maize was used as a test crop. Decomposition and P and N release of tithonia and senna leaves were determined in a litterbag study. Tithonia + TSP applications tripled maize yields compared to the control, senna + TSP and tithonia sole application doubled yields, while senna sole applications did not increase yields substantially. A large residual yield was produced in the tithonia treatments in a subsequent crop. These yield results were consistent with the higher quality and faster release of N and P from the tithonia leaves compared to senna. The tithonia biomass transfer system can improve yields in the short term but has limitations because of the large amount of biomass and the associated labor requirements. This revised version was published online in June 2006 with corrections to the Cover Date.     

Productivity of hedgerow shrubs and maize under alleycropping and block planting systems in semiarid Kenya

A field experiment was conducted for eight cropping seasons from 1988 to 1991 in semiarid Machakos, Kenya, to compare the productivity of shrubs and crops in intercropping (alleycropping) versus block (sole) planting systems. The study, conducted in a split-plot experiment with three replications, consisted of two tree species (Leucaena leucocephala andSenna siamea syn.Cassia siamea) in the main-plots and combinations of two planting systems (alleycropping and block planting of the shrubs and maize) with five different ratios of land allocation for the shrub and crop (1000, 1585, 2080, 2575, and 0100) in sub-plots. Thus, a sole maize and a sole tree were included in the subplot treatments. The spacing between hedgerows of shrubs in intercropping was 6.7, 5, and 4 m, respectively, corresponding to 15, 20, and 25% land allocation to shrubs. The trees were pruned to 0.5 m height four times a year. Intercropped hedgerows of senna and leucaena produced 10% and 24% more biomass than their respective block planting systems. On average, leucaena produced more biomass than senna. Maize alleycropped with leucaena yielded 16% less grain than sole-crop maize, whereas senna intercropping caused hardly any maize-yield reduction. Compared with the respective sole-crop systems, leucaena intercropping did not affect land equivalent ratio (LER), whereas LER increased by 28% with senna intercropping. The different tree:crop land occupancy ratios did not affect the production of either the component species or of the total system, except that LER declined with incrreased spacing between hedgerows. It is concluded that in semiarid highlands of Kenya, leucaena and crops should better be grown in sole blocks, not in alleycropping. In the case of slow-growing species such as senna, intercropping is worthwhile to consider only if the additional labour needed does not pose a serious problem for management, and the species has fodder value.     

Evaluation of anti-acne activity of selected Sudanese medicinal plants

Acne vulgaris is the most common skin disease in the world, and the number of antibiotics resistant to acne-inducing bacterial strains has been increasing in the past years. Natural substances from plants are promising candidates to treat this disease. The methanol and 50 % (v/v) ethanol extracts of 29 plant species traditionally used in Sudan for treatment of a variety of diseases were tested in vitro for their potential anti-acne activity. The activities of these extracts were determined using an antibacterial assay against Propionibacterium acnes, a lipase inhibitory assay, and l,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity assay. The results showed that methanol and 50 % ethanol extracts of Terminalia laxiflora Engl & Diels wood exhibited good antibacterial activity (minimum inhibitory concentration 0.13 mg/ml). The 50 % ethanol extracts of Abrus precatorius L. seed, T. laxiflora Engl & Diels and methanol extract of Acacia nilotica (L.) pods showed lipase inhibitory activity more than 70 % at 500 μg/ml. The methanol extracts of A. nilotica (L.) pods showed the best DPPH radical scavenging activity (IC₅₀ 1.32 μg/ml). Total phenolic, flavonoid and total tannin contents of selected plant extracts shown anti-acne activities were investigated. Almost all selected extracts contained phenolic compound. The highest level of flavonoids (38.87 μg/mg) was detected in T. brownii bark, whereas the highest amount of tannin was detected in A. nilotica (L.) bark (88.01 %).     

Purgative components in rhubarbs: Adrenergic receptor inhibitors linked with glucose carriers

Rhubarbs and their extractives have been used as cathartic for many years. There have been numerous breakthroughs in the pharmacological research of the drug. However, as the key point of the mechanism, the targets of the effective components still remain unclear. In this paper, with an in vitro system of isolated intestine, we found that both the rhubarb extractives and the anthraquinone derivatives can antagonize the adrenaline effectively. Furthermore, computer based docking provided the binding model of the anthraquinone derivatives and adrenergic receptor. Then, based on the results of the small intestinal promotion and purgative effect experiments in vivo, we built an “inhibitor–carrier” hypothesis to elucidate the mechanism of rhubarb. This work provided key massages for the pharmacological research of rhubarb, such a common and active medicinal plant, and might be of help for the development of new purgative drugs.     

Free radical scavenging activity of three Equisetum species from Fruska gora mountain

The scavenger activities of Equisetum arvense, Equisetum ramosissimum and Equisetum telmateia aboveground parts phosphate buffer (pH 7) extracts were evaluated using three different methods: DPPH assay, ESR and NO radical inhibition assay. Total reducing power was determined by FRAP assay. The E. telmateia extract demonstrated the most relevant scavenger and antioxidant properties. ESR signal of DMPO-OH radical adducts in the presence of E. telmateia phosphate buffer (pH 7) extract was reduced to 98.9% which indicated that E. telmateia could be a useful source of antioxidants with huge scavenger ability.     

Early detection of brown-rot decay in southern yellow pine using immunodiagnostic procedures

Summary Immunodiagnostic procedures have been used to detect incipient decay of southern yellow pine by six common brown-rot fungi. Rabbit polyclonal antibodies were raised to liquid culture fractions of the six fungi. The antibodies, after preadsorption to sawdust, were tested in particle agglutination assays, immuno-dot blot, and enzyme-linked immunosorbent assay (ELISA) for their ability to detect the decay organisms when they were grown in southern yellow pine. Results were correlated to wood-block weight loss.Each method specifically detected five of the six test organisms at very low wood-block weight loss. Agglutination assays were the least effective and lacked the sensitivity of the other assays, even though the latex particles were more sensitive and reproducible than charcoal particles. Cross-reactivity was noted only for one of the control organisms and only in the charcoal agglutination assay. Both dot blot and ELISA were sensitive test methods, but ELISA had the advantage of quantification. We feel that two of these assays, latex for presumptive results and ELISA for definitive results, could effectively detect incipient decay of common brown-rot fungi.     

养分胁迫对桉树幼苗叶片营养吸收的影响

通过设置Ca、Mg、Cu、Zn和B等中微量元素不同的浓度梯度(缺乏、低量和过量),调查养分胁迫对桉树幼苗矿质营养吸收的影响.试验结果表明:(1)各元素养分胁迫对桉树幼苗叶片营养元素含量的影响不同;(2)CK(未浇灌营养液)处理桉树营养元素总量最低,浇灌营养液各处理比CK(未浇灌营养液)处理提高36.03-43.55倍;...     

Antiproliferative activity of Saponaria vaccaria constituents and related compounds

Total methanolic extracts of Saponaria vaccaria seed derived from several varieties, as well as various purified components obtained through successive chromatographic separations of total extracts were evaluated for their growth inhibitory activity in WiDr (colon), MDA-MB-231 (breast), NCI-417 (lung) and PC-3 (prostate) human cancer cells as well as the non-tumorigenic fibroblast BJ (CRL-2522) cell line using MTT colorimetric assay. Purified bisdesmosidic saponins segetoside H and I were further examined using microscopy and apoptosis assays. Bisdesmosidic saponins exhibited dose-dependent growth inhibitory and selective apoptosis-inducing activity. Growth inhibitory effects were particularly strong in a breast (MDA-MB-231) and a prostate (PC-3) cancer cell line. Total extracts exhibited a different preference being most active against a colon cancer cell line (WiDr). In a comparison of varieties, all of the total seed extracts exhibited similar dose-dependent activities, but with some variation in potency. Monodesmosidic saponins vaccarosides A and B, phenolic vaccarin, and cyclopeptide segetalin A, co-occurring seed substituents, did not exhibit activity. The non-tumorigenic fibroblast cell line BJ (CRL 2522) was growth inhibited but did not undergo apoptosis when treated with bisdesmosidic saponins at low micromolar concentrations. Saponin-rich extracts from Kochia scoparia seed and Chenopodium quinoa were also evaluated alongside Saponaria saponins but did not exhibit activity. Closely related Quillaja saponins exhibited activity but were less potent.     

Identification of predator–prey relationships between coccinellids and <Emphasis Type="Italic">Saissetia oleae</Emphasis> (Hemiptera: Coccidae), in olive groves,using an enzyme-linked immunosorbent assay

A polyclonal antiserum (AS) was developed and characterized for the detection of immature stages of the black-scale, Saissetia oleae, in whole body homogenized field-collected coccinellid species, using an indirect enzyme-linked immunosorbent assay (ELISA). The indirect ELISA showed to be sensitive to the S. oleae AS, detecting a protein content between 0.118 and 0.0374 μg mL⁻¹. The specificity of the ELISA was tested by assaying a range of sympatric predators and alternative preys with the S. oleae AS. Coccinellid larvae obtained the highest cross-reaction and a positive–negative threshold was established at 0.674 μg mL⁻¹ protein. A total of 1,322 coccinellids were field-collected in three olive groves located in Trás-os-Montes (northeast of Portugal) by the beating technique and were analyzed to detect S. oleae proteins in their guts. Field-collected coccinellids which attained a S. oleae protein concentration equivalent higher than the threshold were considered as a positive reaction. In the overall collected coccinellids, 21.2% reacted positively with the S. oleae AS. Chilocorus bipustulatus and coccinellid larvae obtained the highest percentages of positives with 43.4 and 40.8%, respectively. The greatest frequency of positive responses occurred at the beginning of July, mid-August, and mid-October coinciding with the occurrence of the first, second and third instar nymphs of S. oleae, respectively. Thus, in this study, the role of coccinellids as natural control agents of S. oleae was highlighted by the number of individuals and species that tested positive for S. oleae AS.     

Irritant potential of triterpenoids from Ficus carica leaves

The irritant potential of total methanolic extract and five triterpenoids newly isolated from the leaves of Ficus carica investigated by open mouse ear assay. Total methanolic extract, calotropenyl acetate, methyl maslinate and lupeol acetate showed potent and persistent irritant effects.     

Response of <Emphasis Type="Italic">Sesbania sesban</Emphasis> (L.) Merr. to rhizobial inoculation in an N-deficient soil containing low numbers of effective indigenous rhizobia

Sesbania sesban (L.) Merr. (sesbania) is a fast growing N₂-fixing widely used as an improved fallow species by smallholder farmers in eastern and southern Africa to restore fertility of their N-deficient soils. In order to establish the need for inoculation, the population of sesbania rhizobia in soil collected from a site where the species is intended for introduction was assessed using the most probable number (MPN) plant infection assay. The MPN of sesbania rhizobia was low (21, 6–81 fiducial limits at P=0.05 g⁻¹soil) but with N₂-fixation potential comparable to sesbania inoculant strain KFR 651. Evaluation of an indigenous sesbania rhizobial isolate GSS 1 from the MPN assay in potted field soil showed that it was more effective than strain KFR 651 in terms of plant growth and shoot dry matter (DM) accumulation at 9 and 12 weeks after planting, respectively. Total shoot N content was also higher for plants inoculated with isolate GSS 1 than inoculant strain KFR 651 and uninoculated control treatments 12 weeks after planting. These results demonstrate that it is better to inoculate with effective indigenous than exogenous rhizobia where the need for inoculation has been established.     

Echinacoside promotes bone regeneration by increasing OPG/RANKL ratio in MC3T3-E1 cells

Echinacoside (ECH), isolated from Cistanche tubulosa (Schrenk) R. Wight stems, was subjected to in vitro experiments to investigate its bioactivities on proliferation, differentiation and mineralization of the osteoblastic cell line MC3T3-E1. MTT assay, the alkaline phosphatase (ALP) activity and calcium deposition were determined, and the secretion of collagen I (COL I), osteocalcin (OCN), osteoprotegerin (OPG) and receptor activator of nuclear factor-κB ligand (RANKL) were also assayed by enzyme-linked immunosorbent assay (ELISA). The results showed that ECH caused a significant increase in cell proliferation, ALP activity, COL I contents, OCN levels and an enhancement of mineralization in osteoblasts at the concentration range from 0.01 to 10 nmol·L^− 1 (p < 0.05), suggesting that ECH has a stimulatory effect on osteoblastic bone formation or has potential activity against osteoporosis. In addition, the ratio of OPG/RANKL also could be enhanced by ECH. These findings provide the potent evidence that ECH can promote bone regeneration in cultured osteoblastic MC3T3-E1 cells, which might be done by elevating the OPG/RANKL ratio, and potential evidence for echinacoside to be a promising drug or a lead compound in the development of disease-modifying drug to prevent osteoporosis.     

Determinants of biomass production in hybrid willows and prediction of field performance from pot studies

Pot experiments are often performed to assess plant physiological traits and relationships among growth traits under controlled environments. However, the reliability of pot studies for predicting the growth and performance of trees in the field has rarely been rigorously assessed. We evaluated the suitability of pot experiments for predicting field performance, measured as shoot biomass production, by investigating determinants of growth in hybrid willows (Salix spp.) grown under various environmental conditions in the field, and by comparing the data with the results from a corresponding pot study. Biomass production in six hybrid willow clones, bred for use as bio-fuels, was assessed in three field trials located in central and southeastern Sweden throughout the first 3-year cutting cycle. The determinants of biomass productivity, measured as biomass allocation and nitrogen (N) economy, were identified in one of the field trials. Key traits for shoot biomass production in the field were total leaf area and total amount of N; plant N losses by shed leaves were only partly controlled by leaf-litter N concentration. These key traits were also obtained from the pot study and related to shoot biomass production and abscission-leaf N loss in the field. Total leaf area and total N pool of plants grown in pot experiments were good predictors of long-term biomass production in the field, whereas shoot biomass production, specific leaf area and tissue N concentration of pot-grown plants were less suitable as predictors of field performance. Relationships between the key traits and shoot biomass production were clone-specific, indicating the need for analysis of growth traits at the clone level if field performance of trees is to be evaluated based on data from pot studies. Nutrient loss components are important for tree performance in the long term and evaluations of nutrient loss characteristics at the individual-tree level should focus on nutrient pools lost rather than on nutrient concentrations in abscised plant parts.     

陕西省药用大黄资源调查研究

本文报道陕西省药用大黄的种类、分布、生态环境及其资源概况,为中药大黄这一传统中药材的保护和利用提供基本资料。     

Applicability of semi-destructive method to derive allometric model for estimating aboveground biomass and carbon stock in the Hill zone of Bangladesh

Biomass estimation using allometric models is a nondestructive and popular method.Selection of an allometric model can influence the accuracy of biomass estimation.Bangladesh Forest Department initiated a nationwide forest inventory to assess biomass and carbon stocks in trees and forests.The relationship between carbon storage and sequestration in a forest has implications for climate change mitigation in terms of the carbon sink in Bangladesh.As part of the national forest inventory,we aimed to derive multi-species biomass models for the hill zone of Bangladesh and to determine the carbon concentration in tree components(leaves,branches,bark and stem).In total,175 trees of 14 species were sampled and a semi-destructive method was used to develop a biomass model,which included development of smaller branch(base dia 7 cm) biomass allometry and volume estimation of bigger branches and stems.The best model of leaf,branches,and bark showed lower values for adjusted R2(0.3152-0.8043) and model efficiency(0.436-0.643),hence these models were not recommended to estimate biomass.The best fit model of stem and total aboveground biomass(TAGB) showed higher model efficiency 0.948 and 0.837,respectively,and this model was recommended for estimation of tree biomass for the hill zone of Bangladesh.The best fit allometric biomass model for stem was Ln(Stem)=-10.7248+1.6094*Ln(D)+1.323*Ln(H)+1.1469*Ln(W);the best fit model for TAGB was Ln(TAGB)=-6.6937+0.809*Ln(D^2*H*W),where DBH=Diameter at Breast Height,H=Total Height,W=Wood density.The two most frequently used pan-tropical biomass models showed lower model efficiency(0.667 to 0.697) compared to our derived TAGB model.The best fit TAGB model proved applicable for accurate estimation of TAGB for the hill zone of Bangladesh.Carbon concentration varied significantly(p 0.05) by species and tree components.Higher concentration(48-49%) of carbon was recorded in the tree stem.     

ELISA Quantitation and GC-MS identification of abscisic acid in stigma, ovary and pedicel of pollinated poplar flowers (Populus nigra L.)

A method of fractionation, quantitation and identification of abscisic acid (ABA) in poplar flowers is presented in the context of intra- and interspecific pollination studies in Populus nigra x P. nigra and P. nigra x P. deltoides, respectively. Poplar flowers were dissected into stigma, ovary and pedicel. Abscisic acid concentrations were measured at receptivity and 5 days after pollination in each part of the flower. Abscisic acid was fractionated by reverse-phase HPLC followed by an enzyme-linked immunosorbent assay (ELISA) with anti-ABA polyclonal antibody and GC-MS identification of the immunoreactive ABA fractions. Samples were fractionated and ABA was identified by the presence of a cross-reactive metabolite of ABA in the extracts. The interspecific cross (P. nigra x P. deltoides) had a high concentration of ABA in the pedicel, where abscission occurred, whereas in the intraspecific cross, ABA concentration in the pedicel was low. The results are discussed with reference to the control ABA exerts on the abscission of poplar flowers after interspecific incompatible pollinations.     

不同采收期及不同部位的广东紫珠总黄酮含量分析

为确定广东紫珠适宜采收期和采收模式提供依据。定期采集同一基地广东紫珠药材样品,采用紫外分光光度法测定按芦丁计的总黄酮含量。广东紫珠总黄酮含量的积累与植物生长相一致,表现出随着植株的生长,总黄酮含量也随之增长的规律;不同部位器官总黄酮含量有显著差别,次序为：叶〉小茎枝〉果〉根〉主干茎,其中叶的总黄酮含量大大高于其他器官。     

Impact of mangrove vegetation on seasonal carbon burial and other sediment characteristics in the Vellar-Coleroon estuary,India

This work quantified the total carbon and 12 other sediment characteristics at 10 soil depths, in planted and or natural mangrove forests in comparison with non-vegetated soil for four seasons of the year 2009-2010 in the Vellar-Coleroon estuarine complex, India. The sedi- ment characteristics varied significantly between mangrove-vegetated and non-vegetated habitats or seasons of analysis, but not between soil depths. The mangrove sediments were rich in total carbon and total or- ganic carbon as compared to non-mangrove sediments （p 〈0.01）. Total carbon was 98.2% higher in mature mangroves and 41.8% in planted mangroves than that in non-mangrove soil. Total organic carbon was as much as 2.5 times greater in mature mangroves and 2 times greater in planted mangroves than that in unvegetated soil. Carbon contents also varied many fold by season. Total carbon content was 8.6 times greater during pre-monsoon, 4.1 times greater during post-monsoon and 2.5 times greater during monsoon than during summer （P〈0.01 in all cases）. Similarly, total organic carbon was 5.9 times greater during pre-monsoon, 3.1 times greater during post-monsoon and 69% greater during monsoon than during summer. In general, higher levels of sediment carbon were recorded during pre and post-monsoon seasons than during other seasons. Total carbon concentration was correlated negatively to temperature, sand and phosphorus （P 〈0.01）; positively correlated with redox potential, silt, clay, C/N ratio, potassium （P 〈0.01） and nitrogen （P〈0.05）; but not correlated with soil depth, pH or salinity. This work revealed that the carbon burial was rapid at the annual rate of 2.8% for total carbon, and 6.7% for total organic carbon in mangrove-planted sediment. Cleating of mangroves can result in significantly and rapidly reduced carbon stores.Our study highlights the importance of natural and plantation mangrove stands for conserving sediment carbon in the tropical coastal domain.     

利用混合抗血清检测苹果潜隐病毒

为寻求判断苗木是否带毒的简便方法，利用ＰＡＳ—ＥＬＩＳＡ检测方法，将ＡＣＬＳＶ和ＡＳＧＶ的抗血清混合，并同时加入同一检测孔中，定性检测苹果组培苗ＡＣＬＳＶ和ＡＳＧＶ的带毒情况。发现该方法在检测这两种病毒的混合毒源时，比采用单一抗血清检测灵敏度高，且费用较低。