Screening of Botanical Extracts for Repellence against Western Corn Rootworm Larvae

The ability of western corn rootworm (WCR) (Diabrotica virgifera virgifera LeConte) to develop resistance to various management practices requires the development of new management options. Repellent extracts can act as efficacy enhancing agents in WCR control. The present study investigated the potential repellence of garlic (Allium sativum L.), pepper (Capsicum sp. L), cape aloe (Aloe ferox Mill.), neem (Azadirachta indica A. Juss) and turmeric (Curcuma longa L.) derived extracts against WCR larvae in a no-choice bioassays. In initial screenings garlic and turmeric were evaluated as the most promising extracts as WCR larvae avoided garlic / turmeric treated soil. Turmeric was further evaluated in a rhizotron set up to quantify larval distribution and behavioral changes over time. WCR larvae avoided turmeric treated soil up to 0.05 m from its application in the root system, resulting in an increased dispersal and the formation of multiple clusters in the rhizotron. The proportion of larvae actively moving in the soil subsequently increased, whereas larval feeding significantly decreased. The spatial and behavioral alterations in the soil lead to the conclusion that turmeric is a repellent extract for WCR larvae. Turmeric repellence might further be exploited for WCR management through a synergy with insecticidal substances or microbial antagonists.     

Altered somatic mutation level and DNA repair gene expression in <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis> exposed to ultraviolet C,salt, and cadmium stresses

It is known that somatic mutations arising during animal growth and ageing contribute to the development of neurodegenerative and other animal diseases. For plants, several studies showed that small-scale somatic DNA mutations accumulated during Arabidopsis life cycle. However, there is a lack of data on the influence of environmental stresses on somatic DNA mutagenesis in plants. In this study, we analyzed the effects of ultraviolet C (UV-C) irradiation, high soil salinity, and cadmium (CdI₃) stresses on the level of small-scale somatic DNA mutations in Arabidopsis thaliana. The number of DNA mutations was examined in the Actin2 3′UTR (Actin-U1), ITS1-5.8rRNA-ITS2 (ITS), and ribulose-1,5-biphosphate carboxylase/oxygenase (rbcL) DNA regions. We found that somatic mutation levels considerably increased in CdI₃-treated Arabidopsis plants, while the mutation levels declined in the UV-C- and NaCl-treated A. thaliana. Cadmium is a mutagen that is known to inhibit DNA repair processes. The detected stress-induced alterations in somatic DNA mutation levels were accompanied by markedly increased expression of base excision repair genes (AtARP, AtDME, AtDML2, AtDML3, AtMBD4, AtROS, AtUNG, and AtZDP), nucleotide excision repair genes (AtDDB1a, AtRad4, and AtRad23a), mismatch repair genes (AtMSH2, AtMSH3, and AtMSH7), and photoreactivation genes (AtUVR2, AtUVR3). Thus, the results demonstrated that UV-C, high soil salinity, and cadmium stresses influence both the level of DNA mutations and expression of DNA repair genes. Salt- and UV-induced activation of DNA repair genes could contribute to the stress-induced decrease in somatic mutation level.     

Reliability of the search for 19 common mutations in the <Emphasis Type="Italic">CFTR</Emphasis> gene in Russian cystic fibrosis patients and the calculated frequency of the disease in Russian Federation

A study of Russian cystic fibrosis (CF) patient DNA was conducted to assess the incidence frequency of 19 mutations, namely CFTRdele2,3(21kb), F508del, I507del, 1677delTA, 2143delT, 2184insA, 394delTT, 3821delT, L138ins, 604insA, 3944delGT, G542X, W1282X, N1303K, R334W, and 3849 + 10kbC > T, S1196X, 621 + 1g > t, and E92K of the CFTR gene. We also sought to determine the estimated CF frequency in Russian Federation. In addition, we determined the total information content of the approach for 19 common mutations registration in the CFTR gene, 84.6%, and the allelic frequencies of the examined mutations: three mutations were observed with a frequency exceeding 5% (F508del, 53.98%, E92K, 6.47%, CFTRdele2,3(21kb), 5.35%); other mutations were observed with frequencies ranging from 0.13 to 3.0%. The CF population carrier frequency was 1 in 38 subjects, while the predicted CF frequency was 1 in 5776 newborns.     

Allelopathic Potential of <Emphasis Type="Italic">Koelreuteria bipinnata</Emphasis> var. <Emphasis Type="Italic">integrifoliola</Emphasis> on Germination of Three Turf Grasses

Allelopathy is very important for the scientific disposition of garden plants. To understand the allelopathic potential of Koelreuteria bipinnata Franch. var. integrifoliola, the germination of Agrostis tenuis Sibth., Festuca arundinacea Schreb. and Lolium perenne L. were determined under laboratory conditions. The results showed that root, stem and leaf aqueous extracts of K. bipinnata var. integrifoliola had allelopathic effects on all three turf grasses, and the allelopathic activity varied according to extract concentrations, test species, and extract sources. Lower extract concentrations did not affect or promoted the germination and initial seedling growth of turf grasses, but the highest concentrations almost had inhibitory effect. The order of allelopathic potentials of the three organs on germination of these receptors was root < stem < leaf. And at the highest concentration of leaf extract, the most strongly inhibition was found in A. tenuis, followed by F. arundinaces and then L. perenne. In addition, according to gas chromatography–mass spectrometry (GC–MS) analysis, the allelopathic potential compounds and their abundance in root, stem and leaf were obviously different. Therefore, the allelopathic compounds may responsible for allelopathy of K. bipinnata var. integrifoliola. These findings suggested that more attention should be paid to the leaf of K. bipinnata var. integrifoliola for the relative higher allelopathic effects.     

Genotoxicity of Thermopsis turcica on Allium cepa L. roots revealed by alkaline comet and random amplified polymorphic DNA assays

This study was undertaken to evaluate genotoxic potential of Thermopsis turcica aqueous extracts on the roots of onion bulb (Allium cepa L.) by comet assay and random amplified polymorphic DNA technique. The Allium root growth inhibition test indicated that the EC₅₀ and 2×EC₅₀ values were 8 and 16 mg/ml concentrations of T. turcica aqueous extracts, respectively. The negative control (distilled water), positive control (methyl methane sulfonate, 10 mg/l) and 8 and 16 mg/ml concentrations of T. turcica extracts were introduced to the roots of onion bulbs for 24 and 96 h. The root growth, DNA damage in root cells and randomly amplified polymorphic DNA (RAPD) profiles of root tissue were used as endpoints of the genotoxicity. The comet assay clearly indicated that dose-dependent single strand DNA breaks in the root nuclei of onions were determined for the treatment concentrations of T. turcica extracts. In comparison to RAPD profile of negative control group, RAPD polymorphisms became evident as disappearance and/or appearance of RAPD bands in treated roots. The diagnostic and phenetic numerical analyses of RAPD profiles obviously indicated dose-dependent genotoxicity induced by Thermopsis extracts. In conclusion, the results clearly indicated that water extract of T. turcica has genotoxic potential on the roots of onion bulbs as shown by comet assay and RAPD technique.     

Extraction of sterols in brown macroalgae from Antarctica and their identification by liquid chromatography coupled with tandem mass spectrometry

This study reports for the first time the extraction and quantification of sterols in six species of brown macroalgae from Antarctica: Adenocystis utricularis, Ascoseira mirabilis, Cystosphaera jacquinotii, Desmarestia anceps, Desmarestia antarctica, and Himantothallus grandifolius. Ultrasound irradiation was used as a promotor to extract sterols from algal biomass. The extracts were analyzed by high-performance liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) for sterol quantization (ergosterol, brassicasterol, fucosterol, β-sitosterol, campesterol, cholesterol, and stigmasterol). In general, fucosterol was the most abundant (6.60 to 48.13 mg kg^?1), followed by β-sitosterol (5.29 to 16.49 mg kg^?1), stigmasterol (2.69 to 14.84 mg kg^?1) in five of the six examined algae. The sterol campesterol was found in lower concentrations (0.07 to 0.15 mg kg^?1) in all studied samples.     

Bone marrow mesenchymal stem cell transplantation improves radiation-induced heart injury through DNA damage repair in rat model

Radiotherapy is an effective form of therapy for most thoracic malignant tumors. However, myocardial injury resulting from the high doses of radiation is a severe complication. Here we aimed to study the possibility of reducing radiation-induced myocardial injury with mesenchymal stem cell (MSC) transplantation. We used MSCs extracted from bone marrow (BMSCs) to transplant via the tail vein into a radiation-induced heart injury (RIHI) rat model. The rats were divided into six groups: a Sham group, an IRR (irradiation) group, and four IRR + BMSCs transplantation groups obtained at different time points. After irradiation, BMSC transplantation significantly enhanced the cardiac function in rats. By analyzing the expression of PPAR-α, PPAR-γ, TGF-β, IL-6, and IL-8, we found that BMSC transplantation alleviated radiation-induced myocardial fibrosis and decreased the inflammatory reaction. Furthermore, we found that expression of γ-H2AX, XRCC4, DNA ligase4, and TP53BP1, which are associated with DNA repair, was up-regulated, along with increased secretion of growth factors SDF-1, CXCR4, VEGF, and IGF in rat myocardium in the IRR + BMSCs transplantation groups compared with the IRR group. Thus, BMSC transplantation has the potential to improve RIHI via DNA repair and be a new therapeutic approach for patients with myocardial injury.     

Development of Microemulsions and Microemulgels for Enhancing Transdermal Delivery of <Emphasis Type="Italic">Kaempferia parviflora</Emphasis> Extract

The purpose of this research was to develop microemulsions (ME) and microemulgels (MG) for enhancing transdermal delivery of Kaempferia parviflora (KP) extract. The methoxyflavones were used as markers. Various formulations of ME and MG containing 10% w/v KP extract were prepared, and the in vitro skin permeation and deposition were investigated. The potential ME system containing oleic acid (5% w/v), Tween 20 (20% w/v), PG (40% w/v), and water (35% w/v) was successfully formulated. ME with 10% w/v limonene (ME-L10%) showed higher methoxyflavones flux than ME-L5%, ME-L1%, ME without limonene, and KP extract in water, respectively. ME-L10% was selected for adding a gelling agent to form microemulgels (MG-L10%). However, the high viscosity of the gel formulation might control the diffusion of the compound from gel layer into the skin. Therefore, the liquid formulation provided potential ME droplets to deliver KP extract through the skin. Limonene also plays an effective role on the skin permeation, in which the histological image of the skin treated with ME-L10% exhibited larger space of each flattened keratinocyte layer in the stratum corneum compared to the skin treated with KP extract in water. Moreover, ME-L10% showed good stability. Therefore, ME-L10% was a potential formulation for improving transdermal delivery of KP extract.     

Molecular mechanisms of <Emphasis Type="Italic">Bombyx batryticatus</Emphasis> ethanol extract inducing gastric cancer SGC-7901 cells apoptosis

Bombyx batryticatus is a traditional Chinese medicine. To understand apoptotic effect of B. batryticatus ethanol extract (BBE), we investigated the role of BBE in inducing apoptosis of human gastric cancer cells SGC-7901. Cells treated with BBE and apoptosis was assessed by methyl thiazolyl tetrazolium (MTT) assay, morphological changes, DNA fragmentation and flow cytometry assays. The expression of Bcl-2, Bax and P21 were evaluated by western blot analysis and real time polymerase chain reaction. MTT assay showed that the cytotoxicity of BBE extract on SGC-7901 cells was correlated with treatment time and concentration. After treatment with 6 mg/mL of BBE the microscopy showed that, the majority of SGC-7901 cells were obviously reduced, distorted and grew slowly. Annexin-V/propidium iodide double-staining assay emerge the early apoptosis and the late apoptosis after treatment with different times by laser confocal fluorescence microscopy and flow cytometer. Cell cycle analysis of SGC 79 cells showed that BBE induced cell cycle arrest in the G1 and G2 phases. DNA fragmentation indicated the trend of BBE inducing apoptosis on SGC-7901 cells. The qRT-PCR and western blot analysis indicated that the mRNA and protein expressions of Bax and P21 were significantly up-regulated whereas that of Bc1-2 was down-regulated after treatment with BBE for 24 h. Our results revealed a correlation between gene regulation and BBE-induced apoptosis, which might indicate the potential of BBE in cancer therapy.     

Role of Gene-Gene Interactions in the Chromosomal Instability in Workers at Coal Thermal Power Plants

The 23 polymorphic variants in genes encoding the enzymes of xenobiotics biotransformation (CYP1A1 (rs4646903), CYP1A2 (rs762551), GSTP1 (rs1138272, rs1695), GSTM1 (del), and GSTT1 (del)), DNA repair (XRCC1 (rs25489, rs25487), APEX1 (rs1130409), hOGG1 (rs1052133), ADPRT (rs1136410), XPD (rs13181), XPG (rs17655), XPC (rs2228001), ATM (rs1801516), NBS (rs1805794), XRCC2 (rs3218536), and XRCC3 (rs861539)), antioxidant system (MnSOD (rs4880) and GPx1 (rs1050450)), cell cycle control and apoptosis (TP53 (rs1042522)), DNA methylation (MTHFR (rs1801133) and MTR (rs1805087)), and chromosomal aberrations in lymphocytes in the workers at thermal power plants were analyzed. We found that allelic variants in the CYP1A1 (rs4646903), hOGG1 (rs1052133), XRCC1 (rs25487), and APEX1 (rs1130409) genes were associated with an increased level of chromosomal aberrations in workers. Informative models of gene-gene interactions including CYP1A1 (rs4646903, T>C), CYP1A2 (rs762551, C>A), GSTT1 (del); XRCC1 (rs25487, G>A), MTHFR (rs1801133, C>T), GSTT1 (del); XRCC1 (rs25487, G>A), APEX1 (rs1130409, T>G), TP53 (rs1042522, G>C) determining the formation of the increased frequency of chromosomal aberrations in the workers at coal thermal power plants were discovered.     

Culture Optimization Strategy for 1-Deoxynojirimycin-producing <Emphasis Type="Italic">Bacillus methylotrophicus</Emphasis> K26 Isolated from Korean Fermented Soybean Paste, <Emphasis Type="Italic">Doenjang</Emphasis>

1-Deoxynojirimycin (1-DNJ) is an α-glucosidase inhibitor that is used for the treatment of type 2 diabetes. In this study, we isolated Bacillus methylotrophicus K26 with α-glucosidase inhibition (AGI) activity from Korean fermented soybean paste (Doenjang) and confirmed that the genome harbored the DNJ biosynthesis genes including gabT1, yktc1, and gutB1 by PCR screening, while 1-DNJ production was confirmed by ultra-performance liquid chromatography–quadrupole time-of-flight–mass spectrometry. To increase 1-DNJ production by B. methylotrophicus K26, culture conditions were optimized with one-factor-ata- time (OFAT) and response surface methodology (RSM) approaches. Screen of 11 carbon and 9 nitrogen sources by the OFAT method identified sucrose and yeast extract as optimal culture components. Sucrose concentration (X₁), yeast extract concentration (X₂), and culture temperature (X₃) were selected as independent variables for central composite design. The coefficient of determination (R²) for the model was 0.927, and the probability value of the regression model was highly significant. RSM predicted the optimal conditions for 1-DNJ production by B. methylotrophicus K26 as sucrose and yeast extract concentrations of 4.61% and 7.03%, respectively, at a temperature of 34°C. Under these conditions, AGI activity was experimentally measured as 89.3%, which was close to the predicted value of 91.9%.     

DNA barcoding of selected UAE medicinal plant species: a comparative assessment of herbarium and fresh samples

It is commonly difficult to extract and amplify DNA from herbarium samples as they are old and preserved using different compounds. In addition, such samples are subjected to the accumulation of intrinsically produced plant substances over long periods (up to hundreds of years). DNA extraction from desert flora may pause added difficulties as many contain high levels of secondary metabolites. Herbarium samples from the Biology Department (UAE University) plant collection and fresh plant samples, collected from around Al-Ain (UAE), were used in this study. The three barcode loci for the coding genes matK, rbcL and rpoC1-were amplified. Our results showed that T. terresteris, H. robustum,T. pentandrus and Z. qatarense were amplified using all three primers for both fresh and herbaium samples. Both fresh and herbarium samples of C. comosum, however, were not amplified at all, using the three primers. Herbarium samples from A. javanica, C. imbricatum, T. aucherana and Z. simplex were not amplified with any of the three primers. For fresh samples 90, 90 and 80% of the samples were amplified using matK, rbcL and rpoC1, respectively. In short, fresh samples were significantly better amplified than those from herbarium sources, using the three primers. Both fresh and herbarium samples from one species (C. comosum), however, were not successfully amplified. It is also concluded that the rbcL regions showed real potentials to distinguish the UAE species under investigation into the appropriate family and genus.     

Antioxidative properties of phenolic compounds isolated from the fungal endophytes of <Emphasis Type="Italic">Zingiber nimmonii</Emphasis> (J. Graham) Dalzell.

Background

The microbes living in planta termed ‘endophytes’ is bestowed with the potential to produce bioactive substances. The aim of this investigation was focused on the isolation and molecular identification of the fungal endophytes from Zingiber nimmonii (J. Graham) Dalzell., an endemic medicinal plant species of the ‘Western ghats’, a hotspot location in southern India and characterization of the secondary metabolites responsible for the antioxidant and DNA protective capacity using chromatography and mass spectrometry techniques.

Methods

Endophytic fungi were isolated and identified by sequencing the Internal Transcribed Spacer (ITS). The secondary metabolites were extracted with ethyl acetate and evaluated for the total phenolic, flavonoid and antioxidant capacities. The isolates with potential antioxidative property were further analyzed for the DNA protection ability and the presence of bioactive phenolic compounds by High Performance Liquid Chromatography (HPLC) and Electrospray Ionization-Mass Spectroscopy/Mass Spectroscopy (ESI-MS/MS) techniques.

Results

Endophytic fungi belonging to 11 different taxa were identified. The total phenolic content of the extracts ranged from 10.8±0.7 to 81.6±6.0 mg gallic acid equivalent/g dry extract. Flavonoid was present in eight extracts in the range of 5.2± 0.5 to 24.3±0.9 mg catechin equivalents/g dry extract. Bipolaris specifera, Alternaria tenuissima, Aspergillus terreus, Nectria haematococca and Fusarium chlamydosporum extracts exhibited a potentially high antioxidant capacity. Characterization of the extracts revealed an array of phenolic acids and flavonoids. N. haematococca and F. chlamydosporum extracts contained quercetin and showed DNA protection ability.

Conclusion

This study is the first comprehensive report on the fungal endophytes from Z. nimmonii, as potential sources of antioxidative and DNA protective compounds. The study indicates that Z. nimmonii endophytes are potential sources of antioxidants over the plant itself.

     

Interferon-mediated antiviral activities of Angelica tenuissima Nakai and its active components

Angelica tenuissima Nakai is a widely used commodity in traditional medicine. Nevertheless, no study has been conducted on the antiviral and immune-modulatory properties of an aqueous extract of Angelica tenuissima Nakai. In the present study, we evaluated the antiviral activities and the mechanism of action of an aqueous extract of Angelica tenuissima Nakai both in vitro and in vivo. In vitro, an effective dose of Angelica tenuissima Nakai markedly inhibited the replication of Influenza A virus (PR8), Vesicular stomatitis virus (VSV), Herpes simplex virus (HSV), Coxsackie virus, and Enterovirus (EV-71) on epithelial (HEK293T/HeLa) and immune (RAW264.7) cells. Such inhibition can be described by the induction of the antiviral state in cells by antiviral, IFNrelated gene induction and secretion of IFNs and pro-inflammatory cytokines. In vivo, Angelica tenuissima Nakai treated BALB/c mice displayed higher survivability and lower lung viral titers when challenged with lethal doses of highly pathogenic influenza A subtypes (H1N1, H5N2, H7N3, and H9N2). We also found that Angelica tenuissima Nakai can induce the secretion of IL-6, IFN-λ, and local IgA in bronchoalveolar lavage fluid (BALF) of Angelica tenuissima Nakai treated mice, which correlating with the observed prophylactic effects. In HPLC analysis, we found the presence of several compounds in the aqueous fraction and among them; we evaluated antiviral properties of ferulic acid. Therefore, an extract of Angelica tenuissima Nakai and its components, including ferulic acid, play roles as immunomodulators and may be potential candidates for novel anti-viral/anti-influenza agents.     

Participation of <Emphasis Type="Italic">SRM5/CDC28</Emphasis>, <Emphasis Type="Italic">SRM8/NET1</Emphasis>, and <Emphasis Type="Italic">SRM12/HFI1</Emphasis> genes in checkpoint control in yeast <Emphasis Type="Italic">Saccharomyces cerevisiae</Emphasis>

About twenty genes participating in checkpoint control are known in yeast Saccharomyces cerevisiae. The involvement of SRM genes in the cell cycle arrest under the action of DNA damaging agents was studied in this work. These genes were earlier defined as genes affecting genetic stability and radiosensitivity. It was shown that mutations srm5/cdc28-srm, srm8/net1-srm, and srm12/hfi1-srm fail the cell cycle arrest in the presence of DNA damage and influence the checkpoint arrest in G0/S (srm5, srm8), G1/S (srm5, srm8, srm12), S (srm5, srm12), and G₂/M (srm5). It seems likely that genes SRM5/CDC28, SRM12/HFI1/ADA1, and SRM8/NET1 are involved in a cell response to DNA damage, and in checkpoint regulation in particular.     

The mitochondrial DNA of <Emphasis Type="Italic">Dictyostelium discoideum</Emphasis>: complete sequence,gene content and genome organization

We present an overview of the gene content and organization of the mitochondrial genome of Dictyostelium discoideum. The mitochondria genome consists of 55,564?bp with an A + T content of 72.6%. The identified genes include those for two ribosomal RNAs (rnl and rns), 18 tRNAs, ten subunits of the NADH dehydrogenase complex (nad1, 2, 3, 4, 4L, 5, 6, 7, 9 and 11), apocytochrome b (cytb), three subunits of the cytochrome oxidase (cox1/2 and 3), four subunits of the ATP synthase complex (atp1, 6, 8 and 9), 15 ribosomal proteins, and five other ORFs, excluding intronic ORFs. Notable features of D. discoideum mtDNA include the following. (1) All genes are encoded on the same strand of the DNA and a universal genetic code is used. (2) The cox1 gene has no termination codon and is fused to the downstream cox2 gene. The 13 genes for ribosomal proteins and four ORF genes form a cluster 15.4?kb long with several gene overlaps. (3) The number of tRNAs encoded in the genome is not sufficient to support the synthesis of mitochondrial protein. (4) In total, five group I introns reside in rnl and cox1/2, and three of those in cox1/2 contain four free-standing ORFs. We compare the genome to other sequenced mitochondrial genomes, particularly that of Acanthamoeba castellanii.