bluuebunny/crossref_metadata_2025 huggingface dataset:
Dicentrine Production from a Hairy Roots Culture of Stephania suberosa
Authors: Putalun Waraporn , Yusakul Gorawit , Patanasethanont Denpong
Published: January 2015

A hairy roots culture of Stephania suberosa was established using Agrobacterium rhizogenes ATCC15834. The production of dicentrine was found to be (8.92 ± 0.07) mg/g dry wt on day 35 of culture. Effects of sucrose content, tyrosine, and medium strength on growth and dicentrine production of S. suberosa were investigated. 6% (w/v) sucrose was an optimum content for the growth and dicentrine accumulation in S. suberosa hairy roots. The utilization of a precursor from tyrosine feeding enhanced the dicentrine production. The medium with 1.0 mM of tyrosine had the highest effect on dicentrine accumulation in hairy roots at day 40 of culture [(14.73 ± 0.47) mg/g dry wt]. In addition, ¼ Murashige and Skoog medium was suitable for biomass and dicentrine production in hairy roots. This culture system has a potential to produce dicentrine from hairy roots of S. suberosa
Enhanced Expression of Alternative Oxidase Genes Is Involved in the Tolerance of Rice (Oryza sativa L.) Seedlings to Drought Stress
Authors: Feng Han-Qing , Li Hong-Yu , Sun Kun
Published: January 2015

Drought stress significantly enhanced the capacity of the alternative respiratory pathway and induced AOX1a and AOX1b transcripts in rice seedling leaves. The drought-stressed seedlings pretreated with the inhibitor of the alternative respiratory pathway, 1 mM salicylhydroxamic acid, had a lower level of relative water content than the seedlings either subjected to drought or salicylhydroxamic acid treatment alone. This observation suggests that the alternative respiratory pathway could play a role in the tolerance of rice seedlings to drought stress. Pretreatment with exogenous hydrogen peroxide, salicylic acid, and abscisic acid alone mitigated the water loss of rice leaves exposed to drought stress. Exogenous application of hydrogen peroxide and salicylic acid increased the capacity of the alternative respiratory pathway and induced AOX1a and AOX1b transcripts, while exogenous abscisic acid failed to induce any expression of AOX1 genes. These observations suggest that rice AOX1a and AOX1b genes may be responsive especially to drought stress but not be induced by all of the stress signals related to drought
Heavy Metal Accumulation in Pseudevernia furfuracea (L.) Zopf from the Karabük Iron-Steel Factory in Karabük, Turkey
Authors: Cansaran-Duman Demet , Atakol Orhan , Atasoy Íknur , Kahya Didem , Aras Sümer , Beyaztaş Taylan
Published: January 2015

Pseudevernia furfuracea (L.) Zopf lichen specimens were collected every 5 km starting from around an iron-steel factory located in the central area of Karabük province, up to Yenice Forest. Zn, Cu, Mn, Fe, Pb, Ni, Cd, Cr contents were analyzed in the samples collected from polluted and unpolluted areas. A Pseudevernia furfuracea (L.) Zopf sample from Yenice Forest was used as a control. The reason for this choise was the abundance of species diversity, and therefore sample collection might cause a very low impact on natural population density. The forest is among the 100 forested areas that must be urgently taken under protection according to WWF (World Wildlife Fund) researches. Results of the current study manifested significant variations among the contents of these elements between stations. As expected, the pollution sources, such as iron-steel factory, roads and railroads, industry, heavy traffic, and waste treatment plants, have major impact on the heavy metal accumulation in P. furfuracea (L.) Zopf, and, in accordance to their location, samples 8 and 10 displayed high element accumulation. Surprisingly, although Yenice Forest is under protection, results of our study showed that the region is becoming polluted by the infl uence of many pollution sources in the area. The present study also confirms the efficient metal accumulation capacity of lichens
The Sex Pheromones of Two Pine Sawfly Species, Gilpinia frutetorum and Gilpinia socia: Chemical Identification, Synthesis and Biological Activity
Authors: Hedenström Erik , Edlund Helene , Wassgren Ann-Britt , Bergström Gunnar , Anderbrant Olle , Östrand Fredrik , Sierpinski Andrzej , Auger-Rozenberg Marie-Anne , Herz Annette , Heitland Werner , Varama Martti
Published: January 2015

3,7-Dimethylpentadecan-2-ol and 3-methylpentadecan-2-ol were identified in female whole body extracts from the two pine sawfly species Gilpinia frutetorum and Gilpinia socia. This is the first observation of 3-methylpentadecan-2-ol in extracts of a female pine sawfly species. Synthetic and highly pure stereoisomers of 3,7-dimethylpentadecan-2-ol and 3-methylpentadecan-2-ol were used to verify that the (2S,3R,7R)-isomer of 3,7-dimethylpentadecan- 2-ol and (2S,3R)-3-methylpentadecan-2-ol were present in the extracts. The four stereoisomers of 3-methylpentadecan-2-ol and their biologically active esters were produced via chemoenzymatic methods and the synthesis is described in detail. Male G. socia antennae responded strongly in EAG recordings to the (2S,3R)-isomer of the acetate and propionate of 3-methylpentadecan-2-ol. Male antennae of both G. frutetorum and G. socia also responded to the (2S,3R,7R)- and (2S,3R,7S)-acetates of 3,7-dimethylpentadecan-2-ol
Antibacterial Activities of Ramalin, Usnic Acid and its Three Derivatives Isolated from the Antarctic Lichen Ramalina terebrata
Authors: Paudel Babita , Bhattarai Hari Datta , Lee Hong Kum , Oh Hyuncheol , Shin Hyun Woung , Yim Joung Han
Published: January 2015

The development of new antibacterial compounds is an urgent issue to meet the evolution of resistivity of pathogenic bacteria against the available drugs. The objective of this study was to investigate the antibacterial compounds from the Antarctic lichen species Ramalina terebrata. A total of fi ve compounds, usnic acid, usimine A, usimine B, usimine C, and ramalin, were isolated by bioactivity guided-fractionation of the methanol extract of R. terebrata after several chromatographic procedures. The qualitative antibacterial activities of the crude extract and isolated compounds were determined by the disk diffusion method while the minimum inhibitory concentration (MIC) determination assay gave the quantitative strength of the test samples. All the test samples showed antibacterial activity against Bacillus subtilis. The crude extract and usnic acid showed antibacterial activity against Staphylococcus aureus. The MIC values of the isolated compounds against B. subtilis were in the range of 1 to 26 μg/mL. These observed experimental data showed the strong antibacterial potential of these compounds against B. subtilis.
Degradation of Native Feathers by a Novel Keratinase-Producing, Thermophilic Isolate, Brevibacillus thermoruber T1E
Authors: Bihari Zoltán , Vidéki Diána , Mihalik Erzsébet , Szvetnik Attila , Szabó Zsolt , Balázs Margit , Kesserű Péter , Kiss István
Published: January 2015

Strain T1E, isolated and identifi ed as Brevibacillus thermoruber, and evolutionally distant from the known keratinolytic isolates, proved to have feather-degrading ability. During the 7-day fermentation period, T1E consumed 10 g/l native goose feathers as the sole source of carbon and energy at 50 oC under aerobic conditions. The isolate secreted a thermostable, keratinolytic protease, which exhibited activity optimally at pH 6.5, whilst it was inhibited at alkaline pH. The keratin cleavage and catabolism resulted in the accumulation of free aspartic acid and soluble peptides with maximum values of 31.6 and 720 mg/l, respectively. The majority of the fermentation end-products were found to be small oligopeptides with an average molecular mass of 2275 Da.
Chemical Composition and Antimicrobial Activity of the Essential Oils from Chloranthus japonicus Sieb. and Chloranthus multistachys Pei
Authors: Kang Jie-fang , Zhang Yuan , Du Yong-liang , Wang Zhe-zhi
Published: January 2015

We examined the composition and antimicrobial activity of two essential oils from Chloranthus japonicus Sieb. and Chloranthus multistachys Pei. GC-FID and GC-MS analyses identifi ed 48 and 39 compounds, which represented 95.56% and 94.58%, respectively, of all components in these oils. Of these, 28 compounds were common to both, with a relatively high amount of oxygenated monoterpenes (50.95% and 39.97%). Antimicrobial properties were evaluated in vitro via disc diffusion and microbroth dilution assays. Activities were strong against most tested microorganisms, with inhibition zones ranging from 8.1 to 22.2 mm. For both species, minimum values for inhibitory and bactericidal concentrations were 0.39 to 12.50 mg/mL and 0.78 to 50.00 mg/mL, respectively. These results suggest that these essential oils are potent natural sources of antimicrobial agents for the medicinal and pharmaceutical industries
A Green Paramecium Strain with Abnormal Growth of Symbiotic Algae
Authors: Irie Kanami , Furukawa Shunsuke , Kadono Takashi , Kawano Tomonori
Published: January 2015

Some hundred cells of Chlorella-like green algae are naturally enclosed within the cytoplasm of a single cell of green paramecia (Paramecium bursaria). Therefore, P. bursaria serves as an experimental model for studying the nature of endo-symbiosis made up through chemical communication between the symbiotic partners. For studying the mechanism of symbiotic regulations, the materials showing successful symbiosis are widely used. Apart from such successful model materials, some models for symbiotic distortion would be of great interest in order to understand the nature of successful symbiosis. Here, we describe a case of unsuccessful symbiosis causing unregulated growth of algae inside the hosting ciliates. Recently, we have screened some cell lines, from the mass of P. bursaria cells survived after paraquat treatment. The resultant cell lines (designated as KMZ series) show novel and unusual morphological features with heavily darker green colour distinguishable from the original pale green-coloured paramecia. In this type of isolates, endo-symbiotic algae are restricted within one or two dense spherical structures located at the center of the host cells’ cytoplasm. Interestingly, this isolate maintains the host cells’ circadian mating response which is known as an alga-dependent behaviour in the host cells. In contrast, we discuss that KMZ lacks the host-dependent regulation of algal growth, thus the algal complex often over-grows obviously exceeding the original size of the normal hosting ciliates. Additionally, possible use of this isolate as a novel model for symbiotic cell-to-cell communication is discussed
Characterization and Induction of Two Cytochrome P450 Genes, CYP6AE28 and CYP6AE30, in Cnaphalocrocis medinalis: Possible Involvement in Metabolism of Rice Allelochemicals
Authors: Liu Xiaoli , Chen Jun , Yang Zhifan
Published: January 2015

Two cDNAs specific for P450 genes, CYP6AE28 and CYP6AE30, have been isolated from the rice leaf folder Cnaphalocrocis medinalis Guenée (Lepidoptera: Pyralidae). Both cDNApredicted proteins have 504 amino acid residues in length, but with molecular masses of 60177 Dalton for CYP6AE28 and 60020 Dalton for CYP6AE30, and theoretical pI values of 8.49 for CYP6AE28 and 8.56 for CYP6AE30, respectively. Both putative proteins contain the conserved structural and functional domains characteristic of all CYP6 members. CYP6AE28 and CYP6AE30 show 52% amino acid identity to each other; both of them have 49 - 56% identities with CYP6AE1, Cyp6ae12, and CYP6AE14. Phylogenetic analysis showed that the two P450s are grouped in the lineage containing some of the CYP6AE members, CYP6B P450s and CYP321A1. The transcripts of CYP6AE28 and CYP6AE30 were found to be induced in response to TKM-6, a rice variety with high resistance to C. medinalis. The results suggest that the two P450s may play important roles in adaptation to the host plant rice. This is the first report of P450 genes cloned in C. medinalis
Steroidal Alkaloids of Veratrum lobelianum Bernh. and Veratrum nigrum L.
Authors: Christov Veselin , Mikhova Bozhanka , Ivanova Antoaneta , Serly Julianna , Molnar Joseph , Selenge Dangaa , Solongo Amgalan , Kostova Nadezhda , Gerelt-Od Yadamsuren , Dimitrov Dimitar
Published: January 2015

Twelve steroidal alkaloids were isolated from four populations of Veratrum lobelianum Bernh. and Veratrum nigrum L. Full NMR data for veralosinine (1), and extensive 1H NMR data for veralosine (3) and teinemine (5) are presented here for the first time. (±)-15-O-(2- Methylbutyroyl)germine (10) is undescribed up to now. The antiproliferative activities of veranigrine, veralosinine, and neogermitrine have shown that they are a perspective for further studies.
Nematicidal Activity of Beauvericin Produced by the Fungus Fusarium bulbicola
Authors: Shimada Atsumi , Fujioka Shozo , Koshino Hiroyuki , Kimura Yasuo
Published: January 2015

A nematicide, beauvericin (1), was isolated from cultures of the fungus Fusarium bulbicola, and its structure was identified by spectroscopic analysis. Compound 1 showed nematicidal activities against the pine wood nematode Bursaphelenchus xylophilus and the freeliving nematode Caenorhabditis elegans.
Isolation and Identification of Poplar Isoplastocyanins
Authors: Dimitrov Mitko I. , Donchev Anthony A. , Shosheva Alexandra C. , Getov Vladimir I. , Terezova Nedyalka P. , Stoichev Svetozar D.
Published: January 2015

An improved four-stage isolation and purification procedure for preparing poplar isoplastocyanins is described in detail. Absorbance (UV-VIS) spectroscopy and isoelectric focusing (IEF) are used to determine the protein purity and identity. The present procedure increases twice the total plastocyanin (PC) yield. Four PC isoform fractions are consecutively isolated at the third chromatographic step: oxidized PCa(II) and PCb(II) and reduced PCb(I) and PCa(I). PCa(II) and PCb(II) obtained at the fourth chromatographic step are highly purified PC isoforms which show the purity index (p.i.) A278/A597 ≤ 0.85. Isoelectric points (pI values) of the PC isoforms are found to be at pH 3.92 ± 0.04 for PCa and at pH 3.85 ± 0.02 for PCb. The results of appropriate biological experiments that include the highly purified poplar PC isoforms could give answers to the questions about the physiological significance of PC dimorphism for photosynthesis.
Physicochemical Characteristics of a Thermostable Gellan Lyase from Geobacillus stearothermophilus 98
Authors: Derekova Anna , Atanassova Miroslava , Christova Petya , Tchorbanov Bojidar , Shosheva Alexandra , Mandeva Rossitsa , Rodríguez-Alonso Patricia , Garabal Jose I. , Kambourova Margarita
Published: January 2015

A purified thermostable gellan lyase, produced by a thermophilic bacterium, Geobacillus stearothermophilus 98, was characterized in relation to its physicochemical properties. The gellan lyase was established to have a molecular weight of 216 kDa, defined by capillary gel electrophoresis. Amino acid analysis revealed high quantities of Lys, His, Ala, Val, Ile, Glx, and Pro residues. The circular dichroism revealed 45% β-structure and practically lack of α-spiral domains. Kinetic studies showed high affinity of the enzyme to gellan as a substrate (Km = 0.21 μM). The thermal denaturation investigated by cicular dichroism showed a highly cooperative transition with a midpoint (Tm) at about 75 °C. A single product was identified after enzyme action on gellan. Large exothermic aggregation near Tm was observed by differential scanning calorimetry. Two types of gellan lyase crystals were reproducibly isolated.
Progesterone and β-Estradiol Stimulate Seed Germination in Chickpea by Causing Important Changes in Biochemical Parameters
Authors: Erdal Serkan , Dumlupinar Rahmi
Published: January 2015

Effects of progesterone and β-estradiol on morphologic (germination velocity, root and shoot length) and biochemical (activities of α-amylase, superoxide dismutase, peroxidase and catalase, H2O2 content, lipid peroxidation) parameters during germination and post-germination stages of chickpea seeds were studied. The seeds germinated at various hormone concentrations (10 - 4, 10 - 6, 10 - 9, 10 - 12, 10 - 15 M) were harvested at the end of the 1st, 3rd, and 5th day. With comparison to the control, these hormones caused an increment in the number of germinating seeds at the end of days 1 and 3 by accelerating the seed germination. Root and shoot lengths were augmented by both hormones at all hormone concentrations tested. The highest elongation was recorded in 10 - 6 M progesterone and 10 - 9 - 10 - 12 M β-estradiol. Similarly, activities of α-amylase and superoxide dismutase were increased by all concentrations of both hormones, and maximum increases were obtained with 10 - 6 M progesterone and 10- 9-10- 12 M β-estradiol. In the case of superoxide dismutase activity, not only the H2O2 content but also the peroxidase and catalase activities increased. Lipid peroxidation decreased depending on an increase in the antioxidant enzyme activities. In the present study, it was demonstrated that progesterone and β-estradiol even at low concentrations increase the germination velocity and resistance to stress conditions by changing the activities of some biochemical pathways.
Essential Region for 3-N Methylation in N-Methyltransferases Involved in Caffeine Biosynthesis
Authors: Mizuno Kouichi , Kurosawa Shin-ichi , Yoshizawa Yuko , Kato Misako
Published: January 2015

The caffeine biosynthetic pathway is composed of three methylation steps, and N-methyltransferase catalyzing each step has high substrate specificity. Since the amino acid sequences among coffee 7-methylxanthosine synthase (CmXRS1), theobromine synthase, and caffeine synthase are highly homologous to each other, these substrate specificities seem to be determined in a very restricted region. The analysis of site-directed mutants for CmXRS1 that naturally acts at the initial step, i.e. 7-N methylation of xanthosine, revealed that the activity of 3-N methylation needs a histidine residue at corresponding position 161 in the CmXRS1 sequence. We succeeded in producing the mutant enzyme which can catalyze the first and second methylation steps in caffeine biosynthesis.
Differential Expression of Thaumatin-Like Proteins in Sorghum Infested with Greenbugs
Authors: Chou Juan , Huang Yinghua
Published: January 2015

This study was designed to quantitatively analyze the expression of thaumatin-like protein (TLP) at the transcriptional level in different sorghum lines when they were infested with greenbugs. Three sorghum lines, Tx7000, PI550607, and PI550610, were used. RNAs were isolated from the different sorghum lines that were infested with greenbugs at different infestation times. The resultant mRNA was reverse transcribed into cDNA, and the RT-PCR products were separated by agarose gel. Then, real-time PCR data of the TLP gene expression were analyzed in comparison with the β-actin gene as a reference. The expression levels of the TLP gene were also compared between samples. The results showed that the transcripts of the TLP were induced by greenbug feeding and the increased levels were time-dependent. In the susceptible line, the TLP’s transcripts increased several thousand-fold at 120 hours post infestation, while for the two resistant sorghum lines the TLP expression level increased less than one hundred-fold compared to the controls. This is the first demonstration that thaumatin-like proteins are involved in plant defense response against insects.
Molecular Cloning, Expression and Characterization of BmIDGF Gene from Bombyx mori
Authors: Pan Ye , Chen Keping , Xia Hengchuan , Yao Qin , Gao Lu , Lü Peng , Huojuan  , He Yuanqing , Wang Lin
Published: January 2015

Imaginal disc growth factors (IDGF) play a key role in insect development, but their mechanism remains unclear. In this study, we cloned a novel IDGF gene in Bombyx mori and designated it as BmIDGF. We found that the BmIDGF gene contains eight exons and seven introns, encoding a peptide of 434 amino-acid residues. The protein was predicted to contain one conserved motif of the glycosyl hydrolases family 18 and fall into group V chitinases. Sequence alignment showed that BmIDGF shares extensive homology with other invertebrate IDGF. RT-PCR analysis showed that BmIDGF is expressed in all developmental stages of silkworm larvae and various larvae tissues, which was further confirmed by Western blot analysis. Subcellular localization analysis indicated that BmIDGF is located in the extracellular space. We also successfully expressed it in E. coli and further characterized it by SDS-PAGE and mass spectrometry. Taken together, our data suggests that BmIDGF is a chitinase-like extracellular protein, and provides an excellent platform for subsequent studies on its enzyme activity and role in B. mori development
Polymorphism of Insulin-Like Growth Factor I Gene among Chicken Breeds in Egypt
Authors: Gouda Eman M. , Essawy Gamal S.
Published: January 2015

The insulin-like growth factor I (IGF-I) regulates growth, protein synthesis, and cell proliferation and differentiation in vertebrates. Polymorphisms of IGF-I gene transcripts of three breeds of chicken were assessed. The associations of these polymorphisms with the growth rate of the studied breeds were also evaluated. Total RNA was isolated from chicken livers, and the IGF-I gene was amplified from each breed RNA by RT-PCR using specific primers flanking a certain region of the gene. The amplified RT-PCR products were formed to identify the transcripts and to correlate them to the phenotype of growth, by performing single stranded conformation polymorphism (SSCP) analysis for genotype identification. In this report, we describe how SSCP analysis of RT-PCR products can be used to evaluate the transcript expression pattern of avian IGF-I polymorphism, and their effect on the growth traits of chickens.
In vitro Effects of Chlorpyrifos on the Acetylcholinesterase Activity of Euryhaline Fish, Oreochromis mossambicus
Authors: Venkateswara Rao Janapala , Kavitha Pathakoti
Published: January 2015

The in vitro effect of a widely used organophosphorus insecticide, chlorpyrifos (CPP), on the acetylcholinesterase (AChE) activity was studied in vitro. The kinetic constants Km and Vmax and the bimolecular constant ki were determined in vitro. The in vitro AChE study indicated that CPP is neurotoxic and that it alters the apparent Km values widely in a concentration-dependent manner, resulting in a competitive type of inhibition. Based on the ki values, the sensitivity of AChE in brain is greater than that in gill tissue, at 7.3 · 10 - 5 M and 11.92 · 10 - 5 M, respectively. The study points to the importance of kinetic studies and the results suggest that in biomonitoring programmes brain AChE activity can be a good diagnostic tool for CPP toxicity.
Toxic Effects of Crotocaudin Extracted from the Medicinal Plant Croton tiglium
Authors: Yadav Ram P. , Singh Ajay
Published: January 2015

The compound crotocaudin extracted from the stem bark of the medicinal plant Croton tiglium Linn. was administered for 24 h or 96 h to the freshwater vector snail Lymnaea (Radix) acuminata Lamarck in order to test its toxicity. L. acuminata is the intermediate host of Fasciola hepatica and Fasciola gigantica which cause immense harm to man and his domestic animals. It was observed that the molluscicidal activity of crotocaudin against L. acuminata is time- as well as dose-dependent. There was a significant negative correlation among LC50 values and exposure periods, i.e. increasing the exposure time, the LC50 value of crotocaudin decreased from 5.37 μM (24 h) > 2.08 μM (48 h) > 1.36 μM (72 h) to 1.01 μM (96 h), respectively, against L. acuminata. The toxicological experiments to proof for environmental toxicity, if any, have also been carried out on the non-target freshwater fish Channa punctatus (Bloch) [Channidae (Ophicephalidae)], which shares the habitat with L. acuminata. The sublethal doses of crotocaudin (40% and 80% of LC50) administered over 24 h caused significant changes in the carbohydrate and nitrogenous metabolisms in nervous, hepatopancreas, and ovotestis tissues of Lymnaea acuminata. Channa punctatus was also exposed to sublethal doses of crotocaudin (40% and 80% of 24-h LC50 of L. acuminata) for 96 h which showed significant alterations in the metabolism in muscle, liver, and gonad tissues. After withdrawal of crotocaudin the snail tissues recovered in part after 7 days and the fish tissues completely.
Evaluation of Silymarin as a Promising Radioprotector
Authors: Adhikari Manish , Arora Rajesh , Chawla Raman , Sharma Jyoti , Dhaker Atlar Singh , Gupta Damodar , Dubey Neha , Kumar Raj , Ivanov Veselin , Gevrenova Renata , Sharma Rakesh Kumar
Published: January 2015

Silymarin, a purified extract of seeds of Silybum marianum L. and well known for its hepatoprotective abilities, has been evaluated for inherent utility as a radioprotective agent. A fraction (INM-7035) was authenticated by characterizing the percentage composition of silybin A and B (39.9% and 57.4%). Free radical scavenging activities of INM-7035 against superoxide radicals (>68%), hydroxyl radicals (>33.75%), DPPH (67.2%), and ABTS (32.4%) were also evaluated. The fraction chelated (>30%) ferrous ions, thereby able to restrict amplification. INM-7035 exhibited >50% peroxyl radical scavenging activity in the lipid phase along with dose-dependent (R2 = 0.990) reducing power in the aqueous phase. Radiation-induced free radical flux can lead to disruption of biomolecules like membrane lipids. INM-7035 completely inhibited lipid peroxidative stress in case of membranes against supralethal radiation stress in the liposomal system. The ability of INM-7035 to modulate the levels of NF-κB, indicated its inherent potential as a radioprotective bioactive constituent.
Different Functions and Expression Profiles of Curcin and Curcin-L in Jatropha curcas L.
Authors: Qin Xiaobo , Shao Caixia , Hou Pei , Gao Jihai , Lei Ningfei , Jiang Luding , Ye Shengliang , Gou Chunbao , Luo Shaoyin , Zheng Xiaojiang , Gu Xiaoping , Zhu Xihong , Xu Ying , Chen Fang
Published: January 2015

To date, two types of ribosome-inactivating proteins (RIPs) have been found in Jatropha curcas. One is curcin, which has been isolated from the endosperm, and the other is curcin-L, which is expressed in leaves upon stress treatment. Phylogenetic analysis of the predicted amino acid sequences of the RIPs in plants revealed that these belong to a major subfamily and are close to trichosanthin (TCS). Studies on the mRNA and protein levels showed that both curcin and curcin-L have an organ-specific expression pattern. Curcin is only expressed and accumulated in the endosperm; its expression begins in the globular embryo period and peaks during the mature embryo period. In contrast, curcin-L is only expressed in the leaves, but its expression is induced by certain conditions such as treatment with phytohormones or polyethylene glycol, exposure to high and low temperatures, and fungal infection. Analysis of the 5’ flanking regions of curcin and curcin-L revealed that the 5’ flanking region of curcin-L has three major inserted fragments, which are not present in the corresponding region of curcin. Comparison of characteristic cis-elements suggests the presence of several motifs that are involved in the endosperm-specific expression in the 5’ flanking region of curcin, while in curcin-L some stress- and defense-responsive motifs are found to be mainly located in the three inserted fragments. Comparison of the antifungal activity of the two RIPs showed that the one of curcin-L is higher than that of curcin. Differences in the expression and activity of curcin and curcin-L suggest that these two RIPs have different functions.
Deleterious Activity of Natural Products on Postures of Spodoptera frugiperda (Lepidoptera: Noctuidae) and Diatraea saccharalis (Lepidoptera: Pyralidae)
Authors: Tavares Wagner S. , Cruz Ivan , Fonseca G. , Gouveia Natalia L. , Serrão José E. , Zanuncio José C.
Published: January 2015

The control of Lepidoptera pests should be carried out before hatching of their caterpillars to avoid damage to the crops. The aim of this work was to assess the activity of neem (trade name: Natuneem®, producer: Base Fértil, Chapadão do Sul, Brazil) and pyroligneous extracts (trade name: Biopirol 7M®, producer: Biocarbo, Itabirito, Brazil) at 10 mL/L (1%) and 20 mL/L (2%) contents on egg masses of different ages of Spodoptera frugiperda Smith (Lepidoptera: Noctuidae) and of Diatraea saccharalis F. (Lepidoptera: Pyralidae) at Embrapa Corn and Sorghum in Sete Lagoas, Minas Gerais State, Brazil. The tests took place in an unbiased casualized design with 12 treatments and four replications. The insecticides were diluted in water, and 0.04 mL of the solution was applied to recently laid and one- and two-day-old eggs of S. frugiperda and D. saccharalis. Caterpillars hatching from recently laid egg masses of S. frugiperda was lower with 2% pyroligneous extract [(0.02 ± 0.00)%]. Recently laid eggs and one- or two-day-old eggs of D. saccharalis presented lower caterpillar hatching with 1% neem extract [(0.00 ± 0.00)%, (0.00 ± 0.00)%, and (1.00 ± 0.01)%] and 2% neem extract [(0.00 ± 0.00)%], compared to 1% pyroligneous extract [(27.30 ± 3.22)%, (28.40 ± 3.32)%, and (37.80 ± 4.14)%] and 2% pyroligneous extract [(42.20 ± 4.49)%, (48.70 ± 4.97)%, and (56.60 ± 5.52)%], respectively. Neem and pyroligneous extracts had impact on hatching of S. frugiperda and D. saccharalis caterpillars.
Investigation of Aminomethyl Indole Derivatives as Hyaluronidase Inhibitors
Authors: Ölgen Süreyya , Kaessler Andre , Zühal Kılıç-Kurt Zühal , Jose Joachim
Published: January 2015

Hyaluronidase inhibitors are of potential therapeutic value for the treatment of a variety of diseases, such as cancer, arthrosis, or bacterial infections. Potent and selective hyaluronidase inhibitors are not known so far, and current approaches to the development of hyaluronidase inhibitors are limited. Elevated levels of hyaluronan (HA) are connected with most malignant tumours. Therefore, the search for drugs modulating the hyaluronidase activity became very important. In the present study, a new series of aminomethyl indole derivatives (AMIDs) were tested for inhibition of bovine testes hyaluronidase (BTH). In vitro assays were performed using stains-all at pH 7 and Morgan-Elson reaction at pH 3.5. Among the AMIDs, 3-[(4-methylpiperazin-1-yl)methyl]-5-phenyl-1H-indole (9) was found to be active with 23% inhibition at 50 μM and pH 7. All the other inhibitors showed less activity at pH 3.5 and pH 7. These activity results demonstrated that compounds with phenyl substitution at position 5 have higher activity. The results confirmed that more lipophilic compounds have better inhibition against the hyaluronidase enzyme.
Devil’s Claw Hairy Root Culture in Flasks and in a 3-L Bioreactor: Bioactive Metabolite Accumulation and Flow Cytometry
Authors: Homova Vediha , Weber Jost , Schulze Josef , Alipieva Kalina , Bley Thomas , Georgiev Milen
Published: January 2015

Phenylethanoids are a group of natural water-soluble compounds with high biological value, which could potentially be commercially produced by hairy root cultures. Thus, we have examined the capacity of transformed root cultures of Devil’s claw (Harpagophytum procumbens) to accumulate four phenylethanoid glycosides - β-OH-verbascoside, verbascoside, leucosceptoside A, and martynoside - in shake-flasks and a 3-L stirred tank reactor. Verbascoside was found to be the major phenylethanoid, and its maximal contents were the same (1.12 mg/g dry weight) in both kinds of culture. However, peak leucosceptoside A contents were 1.6-times higher in bioreactor cultures than in shake-flask cultures. Flow cytometry analysis revealed that G0 + G1-phase cells predominated throughout the growth of the cultures, which was in accordance with the very high proportion of quiescent cells in the transformed roots. The results provide the first demonstration of the potential utility of Devil’s claw hairy roots as biofactories for producing high-value phenylethanoid glycosides.
A New Laboratory Cultivation of Paramecium bursaria Using Non-Pathogenic Bacteria Strains
Authors: Bator Tomasz
Published: January 2015

In most studies dealing with the laboratory cultivation of paramecia (Paramecium bursaria), Klebsiella pneumoniae bacteria are used to inoculate the medium. However, Klebsiella pneumoniae is a typical pathogen, and its use is always associated with a risk of infection. The aim of the present research was to examine non-pathogenic bacteria strains as components of the medium for Paramecium bursaria. The paramecia were incubated on lettuce infusions bacterized with different bacteria strains: Bacillus subtilis DSM 10, Bacillus megaterium DSM 32, Escherichia coli DSM 498, Micrococcus luteus DSM 348. A strain derived from the natural habitat of Paramecium bursaria was used as the control one. Experiments were conducted under constant light and in the dark. Paramecia cells were counted under a stereomicroscope on consecutive days of incubation. The obtained results show that the most intensive growth of Paramecium bursaria occurs in the presence of Escherichia coli DSM 498. The use of this strain as a component of the medium allows one to obtain a high number of ciliates regardless of the light conditions. It can be concluded that the Paramecium bursaria cultivation procedure can be modified by using the non-pathogenic bacteria strain Escherichia coli DSM 498 instead of Klebsiella pneumoniae.
Analysis and Identification of Astaxanthin and its Carotenoid Precursors from Xanthophyllomyces dendrorhous by High-Performance Liquid Chromatography
Authors: Lu Mingbo , Zhang Yang’e , Zhao Chunfang , Zhou Pengpeng , Yu Longjiang
Published: January 2015

This study presents an HPLC method for simultaneous analysis of astaxanthin and its carotenoid precursors from Xanthophyllomyces dendrorhous. The HPLC method is accomplished by employing a C18 column and the mobile phase methanol/water/acetonitrile/ dichloromethane (70:4:13:13, v/v/v/v). Astaxanthin is quantified by detection at 480 nm. The carotenoid precursors are identified by LC-APCI-MS and UV-vis absorption spectra. Peaks showed in the HPLC chromatogram are identified as carotenoids in the monocyclic biosynthetic pathway or their derivatives. In the monocyclic carotenoid pathway, 3,3’-dihydroxy- β,ψ-carotene-4,4’-dione (DCD) is produced through γ-carotene and torulene.
Changes in Liver Gene Expression of Azin1 Knock-out Mice
Authors: Wan Tao , Hu Yuan , Huang Ailong , Yamamura Ken-ichi , Tang Hua
Published: January 2015

The ornithine decarboxylase antizyme inhibitor (AZI) was discovered as a protein that binds to the regulatory protein antizyme and inhibits the ability of antizyme to interact with the enzyme ornithine decarboxylase (ODC). Several studies showed that the AZI protein is important for cell growth in vitro. However, the function of this gene in vivo remained unclear. In our study, we analyzed the transcriptional profiles of livers on the 19th day of pregnancy of Azin1 knock-out mice and wild-type mice using the Agilent oligonucleotide array. Compared to the wild-type mice, in the liver of Azin1 knock-out mice 1812 upregulated genes (fold change ≥ 2) and 1466 downregulated genes (fold change ≤ 0.5) were showed in the microarray data. Altered genes were then assigned to functional categories and mapped to signaling pathways. These genes have functions such as regulation of the metabolism, transcription and translation, polyamine biosynthesis, embryonic morphogenesis, regulation of cell cycle and proliferation signal transduction cascades, immune response and apoptosis. Real-time PCR was used to confirm the differential expression of some selected genes. Overall, our study provides novel understanding of the biological functions of AZI in vivo.
Optimization and Purifi cation of L-Asparaginase Produced by Streptomyces tendae TK-VL_333
Authors: Kavitha Alapati , Vijayalakshmi Muvva
Published: January 2015

Cultural factors affecting the production of L-asparaginase by Streptomyces tendae isolated from laterite soil samples of Guntur region were investigated on glycerolasparagine- salts (modified ISP-5) broth. Optimal yields of L-asparaginase were recorded in the culture medium with the initial pH 7.0 incubated at 30 °C for 72 h. The strain utilized sucrose (2%) and yeast (2%) extract as carbon and nitrogen sources for L-asparaginase production. The productivity of L-asparaginase was slightly enhanced when the strain was treated with cell-disrupting agents like EDTA. The crude enzyme was purifi ed to homogeneity by ammonium sulfate precipitation, Sephadex G-100 and CM-Sephadex G-50 gel filtration. By employing sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the molecular weight of the enzyme was recorded as 97.4 kDa. This is the first report on production and purification of L-asparaginase from S. tendae.
Antinociceptive Activity of Atranorin in Mice Orofacial Nociception Tests
Authors: Siqueira Rosana S. , Bonjardim Leonardo R. , Araújo Adriano A. S. , Araújo Bruno E. S. , Melo Marcélia G. D. , Oliveira Marília G. B. , Gelain Daniel P. , Silva Francilene A. , Albuquerque-Júnior Ricardo L. C. , Rocha Ricardo F. , Moreira José C. F. , Antoniolli Angelo R. , Quintans-Júniora Lucindo J. , Santana Josimari M. De
Published: January 2015

Physicochemical characterization and antinociceptive and anti-inflammatory activities of atranorin (AT) extracted from Cladina kalbii Ahti in formalin- and capsaicin-induced orofacial pain and anti-inflammatory tests in rodents were studied. Physicochemical characterization showed that AT has the general formula C19H18O8. Male Swiss mice were pretreated with AT (100, 200, and 400 mg/kg, i.p.), morphine (3 mg/kg, i.p.), or vehicle (0.9% saline with two drops of 0.2% Tween 80) before formalin (20 μl, 2%) or capsaicin (20 μl, 2.5 μg) were injected into the right vibrissa. Our results showed that i.p. treatment with AT displayed marked inhibitory effects in different orofacial pain tests in mice. AT (400 mg/kg, i.p.) was effective in reducing the nociceptive face-rubbing behavioural response in both phases of the formalin test, which was also naloxone-sensitive. Additionally, AT produced a significant antinociceptive effect at all doses in the capsaicin test. Such results were unlikely to be provoked by motor abnormality, since AT-treated mice exhibited no performance alteration on the rota rod apparatus. AT exhibited significant anti-inflammatory activity in the acute model of inflammation (leukocyte migration to the peritoneal cavity), carrageenan- and arachidonic acid-induced hind paw edema in rats. Additionally, AT exhibited a dose-dependent antioxidant activity in vitro, as assessed by total radical-trapping antioxidant parameter and total antioxidant reactivity assays. All these findings suggest that AT might represent an important tool for the management of orofacial pain and/or inflammatory disorders
Bioassay-Guided Separation of an α-Amylase Inhibitor Anthocyanin from Vaccinium arctostaphylos Berries
Authors: Nickavar Bahman , Amin Gholamreza
Published: January 2015

Vaccinium arctostaphylos is a traditional medicinal plant in Iran used for the treatment of diabetes mellitus. In our search for antidiabetic compounds from natural sources, we found that the extract obtained from V. arctostaphylos berries showed an inhibitory effect on pancreatic α-amylase in vitro [IC50 = 1.91 (1.89 -1.94) mg/mL]. The activity-guided purification of the extract led to the isolation of malvidin-3-O-β-glucoside as an α-amylase inhibitor. The compound demonstrated a dose-dependent enzyme inihibitory activity [IC50 = 0.329 (0.316 - 0.342) mM].
A New Class of Antimetabolites: Pyridine Thioglycosides as Potential Anticancer Agents
Authors: Elgemeie Galal H. , Mahdy Elsayed M. , Elgawish Mona A. , Ahmed Mohammad M. , Shousha Wafaa G. , Eldin Mohammad E.
Published: January 2015

The present study was designed for highlighting and focusing on the cytotoxic activity of a new class of antimetabolites both on human cell lines, namely liver carcinoma cell line (Hepg2), lung carcinoma cell line (H460), breast carcinoma cell line (MCF7), brain carcinoma cell line (U251), and animal cell line EAC (Ehrlich ascites carcinoma cells). The results revealed that some of these modified deazapyrimidine thioglycosides have significant cytotoxic activity against EAC cells with growth inhibition percentage ranged between 80% to 90%. The possible inhibitory mechanism of the pyridine thioglycosides was explored by studying the cell cycle perturbation of thioglycosides against human cell lines (in vitro) as well as the most suitable time for maximum compound cytotoxic activity after 6, 18, and 24 h of incubation. To confirm the cytotoxic activity of these compounds, they have been tested for their apoptotic and antiproliferative activity in vivo against solid Ehrlich tumours using five groups of Swiss albino mice for 37 days from inoculation and three treatments, 250, 500 and 1,000 μg/kg body weight. There was signifi cant reduction in Ehrlich tumour size in case of the 500 and 1,000 μg/kg body weight group but mild significant tumour reduction in the 250 μg/kg body weight group. Histograms of DNA per cell for each treatment group indicated that there was a dose-dependent increase in the preG1 phase with a corresponding complete arrest of cells from entering the G2/M phase compared to the untreated EAC group. In conclusion, pyridine thioglycosides have proven good cytotoxic effects against EAC cells and also significant cytotoxic activity against the four tested human cell lines. Flow cytometric DNA ploidy analysis of pyridine thioglcyosides against the Hepg2 and U251 cell lines revealed that the postulated mechanism of action of pyridine thioglcyosides is cell cycle arrest in the S phase. This is similar to antimetabolites and cell cycle arrest in the G2/M phase (M phase) in the same way as microtubule inhibitors like pyridine thioglycosides are cell-cycle-specific in the S phase and the M phase (in case of human cell lines) and have apoptotic effects (in case of animal cell line).
Antibacterial Activity of the Essential Oil from Rosmarinus offi cinalis and its Major Components against Oral Pathogens
Authors: Bernardes Wagner A. , Lucarini Rodrigo , Tozatti Marcos G. , Bocalon Flauzino Lúzio G. , Souza Maria G. M. , Turatti Isabel C. C. , Andrade e Silva Marcio L. , Martins Carlos H. G. , Silva Filho Ademar A. da , Cunha Wilson R.
Published: January 2015

The essential oil of Rosmarinus officinalis L. (rosemary) was obtained by hydro-distillation and analysed by gas chromatography-mass spectrometry. Sixty-two constituents were identified, representing 98.06% of the total oil content. Oxygenated monoterpenes were the predominant components. The rosemary oil was characterized as having prominent (> 5%) contents of camphor (18.9%), verbenone (11.3%), α-pinene (9.6%), β-myrcene (8.6%), 1,8-cineole (8.0%), and β-caryophyllene (5.1%). The antimicrobial activity of the oil as well as of its major constituents was tested against the following microorganisms: Streptococcus mutans, Streptococcus mitis, Streptococcus sanguinis, Streptococcus salivarius, Streptococcus sobrinus, and Enterococcus faecalis, which are potentially responsible for the formation of dental caries in humans. The microdilution method was used for determination of the minimum inhibitory concentration (MIC) during evaluation of the antibacterial activity. The essential oil displayed low activity against the selected microorganisms. In the present study, the pure major compounds were more active than the essential oil. Among all the microorganisms tested, the pathogen S. mitis was the most susceptible and E. faecalis was the most resistant to the evaluated samples. This is the first report on antimicrobial activity of the major components of rosemary oil against oral pathogens.
Biotransformation of Progesterone by Cultured Cells of Marchantia polymorpha
Authors: Haridy Mamdouh S. A. , Hegazy Mohamed-Elamir F. , Mohamed Abou El-Hamd H. , Paré Paul W. , Hirata Toshifumi
Published: January 2015

Cell suspensions of Marchantia polymorpha hydrogenate progesterone to 5α-pregnane-3,20- dione. Structure elucidation of the product was achieved by comprehensive NMR analyses.
Biotransformation of the Phytoestrogen 8-Prenylnaringenin
Authors: Bartmańska Agnieszka , Tronina Tomasz , Huszcza Ewa
Published: January 2015

In order to select microorganisms capable of transforming the potent phytoestrogen 8-prenylnaringenin, preliminary screening tests on 30 fungal cultures were performed. No reports concerning successful biotransformation of 8-prenylnaringenin by this class of organisms have been known so far. Fusarium equiseti converted 8-prenylnaringenin into 2”-(2”’-hydroxyisopropyl)-dihydrofuran-[2”,3”:7,8]-5,4’-dihydroxyfl avanon in high yield.
Plastid Signals Confer Arabidopsis Tolerance to Water Stress
Authors: Cheng Jian , He Chun-Xia , Zhang Zhong-Wei , Xu Fei , Zhang Da-Wei , Wang Xiao , Yuan Shu , Lin Hong-Hui
Published: January 2015

Plastid-to-nucleus retrograde signalling coordinates nuclear gene expression with chloroplast function and is essential for the photoautotrophic life-style of plants. The relationship between plastid signalling and water stress response was investigated with genome uncoupled (gun) mutants, gun1, gun3, and gun5, and an abscisic acid (ABA)-responsible transcription factor mutant, abi4. The results showed that gun1, gun3, gun5, and abi4 mutants suffered from more oxidative damages than the wild-type plants under the water stress and the water stress + herbicide (norflurazon, NF) co-treatment. Superoxide dismutase (SOD), peroxidase (POD), and ascorbate peroxidase (APX) activities could not be prompted in the plastidsignalling defective mutants under the stress conditions. At the same time, Lhcb expression was not repressed in the plastid-signalling defective mutants by the NF treatment or water stress. Therefore, the photosynthetic apparatus in the mutant cells could not be closed during the stresses and the excessive light caused more photodamages on the mutant leaves. The roles of GUN1, GUN3, GUN5 and ABI4 proteins in environmental stress adaptation have been discussed.
WWOX Oxidoreductase – Substrate and Enzymatic Characterization
Authors: Sałuda-Gorgul Anna , Seta Karolina , Nowakowska Magdalena , Bednarek Andrzej K.
Published: January 2015

WWOX is a tumour suppressor gene that spans the common fragile site FRA16D. Analysis of the WWOX expression pattern in normal human tissues showed the highest expression in testis, prostate, and ovary. Its altered expression has been demonstrated in different tissues and tumour types. The WWOX gene encodes a 414-amino acids protein, which is the first discovered protein with a short-chain dehydrogenase/reductase (SDR) central domain and two WW domains at the NH2 terminus. Due to its potential role in sex-steroid metabolism, using two bacterial expression systems, we have cloned WWOX fusion proteins showing oxidoreductase activity in a crude extract, defined a course of enzymatic reactions for selected steroid substrates, and determined related Km values. Our results show that the SDR domain of the WWOX protein has dehydrogenase activity and is reactive both in the presence of NAD+ and NADP+ for all examined steroid substrates. On the other hand, with the same substrates and reduced cofactors (NADH and NADPH) reduction activity was not observed.