International Science Index

International Journal of Bioengineering and Life Sciences

1148
58191
Eco-Friendly Control of Bacterial Speck on Solanum lycopersicum by Azadirachta indica Extract
Abstract:
Tomato (Solanum lycopersicum) is attacked by Pseudomonas syringae pv. tomato causing speck lesions on the leaves leading to severe economic casualty. In the present study, aqueous fruit extracts of Azadirachta indica (neem) were sprayed on a single node of tomato plants grown under controlled contamination-free conditions. The treatment of plants was performed with neem fruit extract either alone or along with the pathogen. The parameters of observation were activities of polyphenol oxidase (PPO) and lysozyme, and isoform analysis of PPO; both at the treated leaves as well as untreated leaves away from the site of extract application. Polyphenol oxidase initiates phenylpropanoid pathway resulting in the synthesis of quinines from cytoplasmic phenols and production of reactive oxygen species toxic to broad spectrum microbes. Lysozyme is responsible for the breakdown of bacterial cell wall. The results indicate the upregulation of PPO and lysozyme activities in both the treated and untreated leaves along with de novo expression of newer PPO isoenzymes (which were absent in control samples). The appearance of additional PPO isoenzymes in bioelicitor-treated plants indicates that either the isoenzymes were expressed after bioelicitor application or the already expressed but inactive isoenzymes were activated by it. Lysozyme activity was significantly increased in the plants when treated with the bioelicitor or the pathogen alone. However, no new isoenzymes of lysozyme were expressed upon application of the extract. Induction of resistance by neem fruit extract could be a potent weapon in eco-friendly plant protection strategies.
1147
60055
Structural and Histochemical Alterations in the Development of the Stigma in Vibirnum Tinus
Abstract:
This study presents the structural and cytochemical alterations of stigma at the stages of pre-anthesis, anthesis and post-anthesis in Vibirnum tinus. Capitate stigma continues with a closed style. The receptive surface of stigma is composed of unicellular papillae which are short and flattened at pre-anthesis stage. The papillae in this stage have dense cytoplasm with small vacuoles and a centrally located nucleus. With the start of anthesis, the stigma widens, papillae lengthen and become cylindrical. At anthesis stage, vacuoles enlarge, and nucleus moves to the base of the cell. At post-anthesis stage, the boundaries of the papillae become less noticeable. As proved by Periodic Acid Schiff procedure, the cytoplasm of papillae is rich in insoluble polysaccharides at all stages of development but it becomes remarkable at post-anthesis, particularly at the sub-papillar area. Although there is no significant difference in the content of protein in all stages of the development, it is more abundant at post-anthesis stage, as in Coomassie Brillant Blue stained sections. The surface of papillae is covered by a cuticle which becomes thicker at post-anthesis, and it gives positive reaction with Sudan Black B and Auramine O. The cuticle is covered by a pellicle stained by Coomassie Brillant Blue, indicating dry type of stigma.
1146
59414
Screening Microalgae Strains Which Were Isolated from Agriculture and Municipal Wastewater Drain, Reno, Nevada and Reuse of Effluent Water from Municipal Wastewater Treatment Plant in Microalgae Cultivation for Biofuel Feedstock
Abstract:
The aim of this study is to select microalgae strains, which were isolated from agriculture and municipal wastewater drain, Reno, Nevada that has highest growth rate and lipid contents. The experiments in this study were carried out in two consecutive stages. The first stage is aimed at testing the survival capability of all isolated microalgae strains and determining the best candidates to grow in centrate cultivation system. The second stage was targeted at determination the highest growth rate and highest lipid content of the selected top performing algae strain when cultivated on centrate wastewater. 26 microalgae strains, which were isolated from municipal and agriculture waste water, were analyzed using Flow cytometer for FACS of lipid with BODIPY and Nile Red as a lipid dyes and they grew on 96 wells plate for 31 days to determine growth rate as a based line data for growth rate. The result showed that microalgae strains which showed a high mean of fluorescence for BODIPY and Nile Red were F3.BP.1, F3.LV.1, T1.3.1, and T1.3.3. Five microalgae strains which have high growth rate were T1.3.3, T2.4.1. F3.LV.1, T2.12.1 and T3.3.1. In conclusion, microalgae strain which showed the highest starch content was F3.LV.1. T1.3.1 had the highest mean of fluorescence for Nile Red and BODIPY. Microalgae strains were potential for biofuel feedstock such as F3.LV.1 and T1.3.1, those microalgae strains showed a positive correlation between growth rate at stationary phase, biomass and meant of fluorescence for Nile Red and BODIPY.
1145
60303
A Genetic Identification of Candida Species Causing Intravenous Catheter-Associated Candidemia in Heart Failure Patients
Abstract:
Introduction: Intravenous catheter-associated fungal infection as nosocomial infection continue to be a deep problem among hospitalized patients, decreasing quality of life and adding healthcare costs. The capacity of catheters in the spread of candidemia in heart failure patients is obvious. The aim of this study was to evaluate the prevalence and genetic identification of Candida species in heart disorder patients. Material and Methods: This study was conducted in Tehran Hospital of Cardiology Center (Tehran, Iran, 2014) during 1.5 years on the patients hospitalized for at least 7 days and who had central or peripheral vein catheter. Culture of catheters, blood and skin of the location of catheter insertion were applied for detecting Candida colonies in 223 patients. Identification of Candida species was made on the basis of a combination of various phenotypic methods and confirmed by sequencing the ITS1-5.8S-ITS2 region amplified from the genomic DNA using PCR and the NCBI BLAST. Results: Of the 223 patients samples tested, we identified totally 15 Candida isolates obtained from 9 (4.04%) catheter cultures, 3 (1.35%) blood cultures and 2 (0.90%) skin cultures of the catheter insertion areas. On the base of ITS region sequencing, out of nine Candida isolates from catheter, 5(55.6%) C. albicans, 2(22.2%) C. glabrata, 1(11.1%) C. membranifiaciens and 1 (11.1%) C. tropicalis were identified. Among three Candida isolates from blood culture, C. tropicalis, C. carpophila and C. membranifiaciens were identified. Non-candida yeast isolated from one blood culture was Cryptococcus albidus. One case of C. glabrata and one case of Candida albicans were isolated from skin culture of the catheter insertion areas in patients with positive catheter culture. In these patients, ITS region of rDNA sequence showed a similarity between Candida isolated from the skin and catheter. However, the blood samples of these patients were negative for fungal growth. We report two cases of catheter-related candidemia caused by C. membranifiaciens and C. tropicalis on the base of genetic similarity of species isolated from blood and catheter which were treated successfully with intravenous fluconazole and catheter removal. In phenotypic identification methods, we could only identify C. albicans and C. tropicalis and other yeast isolates were diagnosed as Candida sp. Discussion: Although more than 200 species of Candida have been identified, only a few cause diseases in humans. There is some evidence that non-albicans infections are increasing. Many risk factors, including prior antibiotic therapy, use of a central venous catheter, surgery, and parenteral nutrition are considered to be associated with candidemia in hospitalized heart failure patients. Identifying the route of infection in candidemia is difficult. Non-albicans candida as the cause of candidemia is increasing dramatically. By using conventional method, many non-albicans isolates remain unidentified. So, using more sensitive and specific molecular genetic sequencing to clarify the aspects of epidemiology of the unknown candida species infections is essential. The positive blood and catheter cultures for candida isolates and high percentage of similarity of their ITS region of rDNA sequence in these two patients confirmed the diagnosis of intravenous catheter-associated candidemia.
1144
60350
Multilocus Phylogenetic Approach Reveals Informative DNA Barcodes for Studying Evolution and Taxonomy of Fusarium Fungi
Abstract:
Fusarium fungi are among the most devastating plant pathogens distributed all over the world. Significant reduction of grain yield and quality caused by Fusarium leads to multi-billion dollar annual losses to the world agricultural production. These organisms can also cause infections in immunocompromised persons and produce the wide range of mycotoxins, such as trichothecenes, fumonisins, and zearalenone, which are hazardous to human and animal health. Identification of Fusarium fungi based on the morphology of spores and spore-forming structures, colony color and appearance on specific culture media is often very complicated due to the high similarity of these features for closely related species. Modern Fusarium taxonomy increasingly uses data of crossing experiments (biological species concept) and genetic polymorphism analysis (phylogenetic species concept). A number of novel Fusarium sibling species has been established using DNA barcoding techniques. Species recognition is best made with the combined phylogeny of intron-rich protein coding genes and ribosomal DNA sequences. However, the internal transcribed spacer of (ITS), which is considered to be universal DNA barcode for Fungi, is not suitable for genus Fusarium, because of its insufficient variability between closely related species and the presence of non-orthologous copies in the genome. Nowadays, the translation elongation factor 1 alpha (TEF1α) gene is the “gold standard” of Fusarium taxonomy, but the search for novel informative markers is still needed. In this study, we used two novel DNA markers, frataxin (FXN) and heat shock protein 90 (HSP90) to discover phylogenetic relationships between Fusarium species. Multilocus phylogenetic analysis based on partial sequences of TEF1α, FXN, HSP90, as well as intergenic spacer of ribosomal DNA (IGS), beta-tubulin (β-TUB) and phosphate permease (PHO) genes has been conducted for 120 isolates of 19 Fusarium species from different climatic zones of Russia and neighboring countries using maximum likelihood (ML) and maximum parsimony (MP) algorithms. Our analyses revealed that FXN and HSP90 genes could be considered as informative phylogenetic markers, suitable for evolutionary and taxonomic studies of Fusarium genus. It has been shown that PHO gene possesses more variable (22 %) and parsimony informative (19 %) characters than other markers, including TEF1α (12 % and 9 %, correspondingly) when used for elucidating phylogenetic relationships between F. avenaceum and its closest relatives – F. tricinctum, F. acuminatum, F. torulosum. Application of novel DNA barcodes confirmed the fact that F. arthrosporioides do not represent a separate species but only a subspecies of F. avenaceum. Phylogeny based on partial PHO and FXN sequences revealed the presence of separate cluster of four F. avenaceum strains which were closer to F. torulosum than to major F. avenaceum clade. The strain F-846 from Moldova, morphologically identified as F. poae, formed a separate lineage in all the constructed dendrograms, and could potentially be considered as a separate species, but more information is needed to confirm this conclusion. Variable sites in PHO sequences were used for the first-time development of specific qPCR-based diagnostic assays for F. acuminatum and F. torulosum. This work was supported by Russian Foundation for Basic Research (grant № 15-29-02527).
1143
60533
Studies of Single Nucleotide Polymorphism of Proteosomal Gene Complex and Their Association with HBV Infection Risk in India
Abstract:
Single Nucleotide polymorphism (SNP) of proteosomal gene complex is involved in the pathogenesis of hepatitis B Virus (HBV) infection. Some of such proteosomal gene complex are large multifunctional proteins (LMP) and antigen associated transporters that help in antigen presentation. Both are involved in intracellular processing and presentation of viral antigens in association with Major Histocompatability Complex (MHC) Class I molecules. A total of hundred each of hepatitis B virus infected and control samples from northern India were studied. Genomic DNA was extracted from all studied samples and PCR-RFLP method was used for genotyping at different positions of LMP genes. Genotypes at a given position were inferred from pattern of bands and genotype frequencies and haplotype frequencies were also calculated. Homozygous SNP {A>C} was observed at codon 145 of LMP7 gene and having a protective role against HBV as there was statistically significant high distribution of this SNP among controls than cases. Heterozygous SNP {A>C} was observed at codon 145 of LMP7 gene and made individuals more susceptible to HBV infection as there was statistically significant high distribution of this SNP among cases than control. SNP {T>C} was observed at codon 60 of LMP2 gene but statistically significant differences were not observed among controls and cases. For codon 145 of LMP7 and codon 60 of LMP2 genes, four haplotypes were constructed. Haplotype I (LMP2 ‘C’ and LMP7 ‘A’) made individuals carrying it more susceptible to HBV infection as there was statistically significant high distribution of this haplotype among cases than control. Haplotype II (LMP2 ‘C’ and LMP7 ‘C’) made individuals carrying it more immune to HBV infection as there was statistically significant high distribution of this haplotype among control than cases. Thus it can be concluded that homozygous SNP {A>C} at codon 145 of LMP7 and Haplotype II (LMP2 ‘C’ and LMP7 ‘C’) has a protective role against HBV infection whereas heterozygous SNP {A>C} at codon 145 of LMP7 and Haplotype I (LMP2 ‘C’ and LMP7 ‘A’) made individuals more susceptible to HBV infection.
1142
59044
TARF: Web Toolkit for Annotating RNA-Related Genomic Features
Abstract:
Genomic features, the genome-based coordinates, are commonly used for the representation of biological features such as genes, RNA transcripts and transcription factor binding sites. For the analysis of RNA-related genomic features, such as RNA modification sites, a common task is to correlate these features with transcript components (5'UTR, CDS, 3'UTR) to explore their distribution characteristics in terms of transcriptomic coordinates, e.g., to examine whether a specific type of biological feature is enriched near transcription start sites. Existing approaches for performing these tasks involve the manipulation of a gene database, conversion from genome-based coordinate to transcript-based coordinate, and visualization methods that are capable of showing RNA transcript components and distribution of the features. These steps are complicated and time consuming, and this is especially true for researchers who are not familiar with relevant tools. To overcome this obstacle, we develop a dedicated web app TARF, which represents web toolkit for annotating RNA-related genomic features. TARF web tool intends to provide a web-based way to easily annotate and visualize RNA-related genomic features. Once a user has uploaded the features with BED format and specified a built-in transcript database or uploaded a customized gene database with GTF format, the tool could fulfill its three main functions. First, it adds annotation on gene and RNA transcript components. For every features provided by the user, the overlapping with RNA transcript components are identified, and the information is combined in one table which is available for copy and download. Summary statistics about ambiguous belongings are also carried out. Second, the tool provides a convenient visualization method of the features on single gene/transcript level. For the selected gene, the tool shows the features with gene model on genome-based view, and also maps the features to transcript-based coordinate and show the distribution against one single spliced RNA transcript. Third, a global transcriptomic view of the genomic features is generated utilizing the Guitar R/Bioconductor package. The distribution of features on RNA transcripts are normalized with respect to RNA transcript landmarks and the enrichment of the features on different RNA transcript components is demonstrated. We tested the newly developed TARF toolkit with 3 different types of genomics features related to chromatin H3K4me3, RNA N6-methyladenosine (m6A) and RNA 5-methylcytosine (m5C), which are obtained from ChIP-Seq, MeRIP-Seq and RNA BS-Seq data, respectively. TARF successfully revealed their respective distribution characteristics, i.e. H3K4me3, m6A and m5C are enriched near transcription starting sites, stop codons and 5’UTRs, respectively. Overall, TARF is a useful web toolkit for annotation and visualization of RNA-related genomic features, and should help simplify the analysis of various RNA-related genomic features, especially those related RNA modifications.
1141
60425
Do Life Histories Differ between Members of a Species Complex (Retropinna Spp.)?
Abstract:
Population connectivity plays an important role in the conservation and recovery of declining species. It affects genetic diversity, adaptive potential and resilience of species in nature. Loss of genetic variation can affect populations by limiting their ability to persist in stressful environmental conditions. Generally, freshwater fishes show higher levels of genetic structuring and subdivision among populations than those inhabiting estuarine or marine environments due to the presence of artificial (e.g. dams) and natural (e.g. mountain ranges) barriers to dispersal in freshwater ecosystems. The Australian smelt (Retropinnidae: Retropinna spp.) is a common freshwater fish species which is widely distributed throughout coastal and inland drainages in South - eastern Australia. These fish are found in a number of habitats from headwaters to lowland sites. They form large shoals in the mid to upper water column and inhabit deep slow – flowing pools as well as shallow fast flowing riffle-runs. Previously, Australian smelt consisted of two described taxa (Retropinna semoni and Retropinna tasmanica), but recently a complex of five or more species has been recognized based on an analysis of allozyme variation. In many area, they spend their entire life cycle within freshwater. However, it has been reported that they can tolerate very high salinities. Although most populations of the species are thought to be non-diadromous, a recent otolith chemistry study showed that the majority of individuals sampled from a coastal river in southern Victoria had spent their early life history in saline habitats, suggesting these fish were most likely catadromous or amphidromous. It is currently unclear whether individuals within coastal populations of Australian Retropinna exhibit diadromous migrations or whether fish collected from marine/estuarine environments are vagrants that have strayed out of the freshwater reaches. In this current study, the population structure and genetic differentiation of Australian smelt fish were investigated among eight rivers of South-East Queensland (SEQ), Australia. 11 microsatellite loci were used to examine genetic variation within and among populations. Genetic diversity was very high. Number of alleles ranged from three to twenty. Expected heterozygosity averaged across loci ranged from 0.572 to 0.852. There was a high degree of genetic differentiation among rivers (FST = 0.23), although low levels of genetic differentiation among populations within rivers. These extremely high levels of genetic differentiation suggest that the all smelt in SEQ complete their life history within freshwater, or, if they go to the estuary, they do not migrate to sea. This hypothesis is being tested further with a micro-chemical analysis of their otoliths.
1140
59255
Bifunctional Activity and Stability of Fused Plasmodium Falciparum Orotate Phosphoribosyltransferase and Orotidine 5′-Monophosphate Decarboxylase
Abstract:
Fusion of the last two enzymes in the pyrimidine biosynthetic pathway in the inversed order by having COOH-terminal orotate phosphoribosyltransferase (OPRT) and NH2-terminal orotidine 5'-monophosphate decarboxylase (OMPDC), as OMPDC-OPRT, are described in many organisms. Here, we produced gene fusions of Plasmodium falciparum OMPDC-OPRT and expressed the bifunctional protein in Escherichia coli. The enzyme was purified to homogeneity using affinity and anion-exchange chromatography, exhibited enzymatic activities and functioned as a dimer. The activities, although unstable, can be stabilized by its substrate and product during purification and long-term storage. Furthermore, the enzyme expressed a perfect catalytic efficiency (kcat/Km). The kcat was selectively enhanced up to 3 orders of magnitude, while the Km was not much affected and remained at low µM levels when compared to the monofunctional enzymes. The fusion of the two enzymes, creating a “super-enzyme” with perfect catalytic power and more flexibility, reflects cryptic relationship of enzymatic reactivaties and metabolic functions on molecular evolution.
1139
59256
Expression of Fused Plasmodium falciparum Orotate Phosphoribosyltransferase and Orotidine 5'-Monophosphate Decarboxylase in Escherichia coli
Abstract:
Fusion of the last two enzymes in the pyrimidine biosynthetic pathway in the inversed order by having COOH-terminal orotate phosphoribosyltransferase (OPRT) and NH2-terminal orotidine 5'-monophosphate decarboxylase (OMPDC), as OMPDC-OPRT, are described in many organisms. In this study, we constructed gene fusions of Plasmodium falciparum OMPDC-OPRT (1,836 bp) in pTrcHisA vector and expressed as an 6xHis-tag bifunctional protein in three Escherichia coli strains (BL21, Rosetta, TOP10) at 18 °C, 25 °C and 37 °C. The recombinant bifunctional protein was partially purified by Ni-Nitrilotriacetic acid-affinity chromatography. Specific activities of OPRT and OMPDC domains in the bifunctional enzyme expressed in E. coli TOP10 cells were approximately 3-4-fold higher than those in BL21 cells. There were no enzymatic activities when the construct vector expressed in Rosetta cells. Maximal expression of the fused gene was observed at 18 °C and the bifunctional enzyme had specific activities of OPRT and OMPDC domains in a ratio of 1:2. These results provide greater yields and better catalytic activities of the bifunctional OMPDC-OPRT enzyme for further purification and kinetic study.
1138
59982
In vitro Culture of Node Stem Segments of Maerua crassifolia
Abstract:
The stem node segments were cultured on Murashige and Skoog (MS) medium. In the case of using MS+ Zeatin (1 mg/l), small shoot buds were formed directly in 70% of explants after 15 days, their length range between 0.1 to 0.3 cm after two weeks and reached 0.3 cm in length and three shoots in numbers after 4 weeks. When those small shoots were sub cultured on the same medium, they increased in length, number and reached 0.4 cm with 4 shoots, 0.4 cm with 5 shoots after six, eight and ten weeks respectively. In the case of using MS free hormones, MS+IAA (0.2mg/l) +BA (0.5mg/l), MS + kin(0.5mg/l), MS + kin (3mg/l) and MS +NAA (3mg/l) +BA (1mg/l), no sign of responses were noticed and only change in color in some cases. Different types of parenchyma cells and many layers of thick wall sclerenchyma cells were observed on MS+BA (1mg/l).
1137
58691
Caged Compounds as Light-Dependent Initiators for Enzyme Catalysis Reactions
Abstract:
By using light as trigger, it is possible to study many biological processes, such as the activity of genes, proteins, and other molecules, with precise spatiotemporal control. Caged compounds, where biologically active molecules are generated from an inert precursor upon laser photolysis, offer the potential to initiate such biological reactions with high temporal resolution. As light acts as the trigger for cleaving the protecting group, the ‘caging’ technique provides a number of advantages as it can be intracellular, rapid and controlled in a quantitative manner. We are developing caging strategies to study the catalytic cycle of a number of enzyme systems, such as nitric oxide synthase and ethanolamine ammonia lyase. These include the use of caged substrates, caged electrons and the possibility of caging the enzyme itself. In addition, we are developing a novel freeze-quench instrument to study these reactions, which combines rapid mixing and flashing capabilities. Reaction intermediates will be trapped at low temperatures and will be analysed by using electron paramagnetic resonance (EPR) spectroscopy to identify the involvement of any radical species during catalysis. EPR techniques typically require relatively long measurement times and very often, low temperatures to fully characterise these short-lived species. Therefore, common rapid mixing techniques, such as stopped-flow or quench-flow are not directly suitable. However, the combination of rapid freeze-quench (RFQ) followed by EPR analysis provides the ideal approach to kinetically trap and spectroscopically characterise these transient radical species. In a typical RFQ experiment, two reagent solutions are delivered to the mixer via two syringes driven by a pneumatic actuator or stepper motor. The new mixed solution is then sprayed into a cryogenic liquid or surface, and the frozen sample is then collected and packed into an EPR tube for analysis. The earliest RFQ instrument consisted of a hydraulic ram unit as a drive unit with direct spraying of the sample into a cryogenic liquid (nitrogen, isopentane or petroleum). Improvements to the RFQ technique have arisen from the design of new mixers in order to reduce both the volume and the mixing time. In addition, the cryogenic isopentane bath has been coupled to a filtering system or replaced by spraying the solution onto a surface that is frozen via thermal conductivity with a cryogenic liquid. In our work, we are developing a novel RFQ instrument which combines the freeze-quench technology with flashing capabilities to enable the studies of both thermally-activated and light-activated biological reactions. This instrument also uses a new rotating plate design based on magnetic couplings and removes the need for mechanical motorised rotation, which can otherwise be problematic at cryogenic temperatures.
1136
58028
Growth and Anatomical Responses of Lycopersicon esculentum (Tomatoes) under Microgravity and Normal Gravity Conditions
Abstract:
Microgravity is known to be a major abiotic stress in space which affects plants depending on the duration of exposure. In this work, tomatoes seeds were exposed to long hours of simulated microgravity condition using a one-axis clinostat. The seeds were sown on a 1.5% combination of plant nutrient and agar-agar solidified medium in three Petri dishes. One of the Petri dishes was mounted on the clinostat and allowed to rotate at the speed of 20 rpm for 72 hours while the others were subjected to the normal gravity vector. The anatomical sections of both clinorotated and normal gravity plants were made after 72 hours and observed using a Phase contrast digital microscope. The percentage germination as well as the growth rate of the normal gravity seeds were higher than the clinorotated ones. The orientations of the clinorotated roots during germination were in different directions unlike the normal gravity which all germinated towards the direction of gravity vector. The clinostat was able to switch off gravistimulation. Distinct cellular arrangement was observed for tomatoes under normal gravity condition unlike those of clinorotated ones. The thickness of the epidermis and cortex of the roots of normal gravity are higher than those of clinorotated. This implied that under long-term microgravity influence, plants do alter their anatomical features as a way of adapting to the stress condition.
1135
57130
Illegal Anthropogenic Activity Drives Large Mammal Population Declines in An African Protected Area
Abstract:
High levels of anthropogenic activity such as habitat destruction, poaching and encroachment into natural habitat have resulted in significant global wildlife declines. In order to protect wildlife, many protected areas such as national parks have been created. However, it is argued that many protected areas are only protected in name and are often exposed to continued, and often illegal, anthropogenic pressure. In West African protected areas, declines of large mammals have been documented between 1962 and 2008. This study aimed to produce occupancy estimates of the remaining large mammal fauna in the third largest National Park in Nigeria, Old Oyo, and to compare the estimates with historic estimates while also attempting to quantify levels of illegal anthropogenic activity using a multi-disciplinary approach. Large mammal populations and levels of illegal anthropogenic activity were assessed using empirical field data (camera trapping and transect surveys) in combination with data from questionnaires completed by local villagers and park rangers. Four of the historically recorded species in the park, lion (Panthera leo), hunting dog (Lycaon pictus), elephant (Loxodonta africana) and buffalo (Syncerus caffer) were not detected during field studies nor were they reported by respondents. In addition, occupancy estimates of hunters and illegal grazers were higher than the majority of large mammal species inside the park. This finding was reinforced by responses from the villagers and rangers who’s perception was that large mammal densities in the park were declining, and that a large proportion of the local people were entering the park to hunt wild animals and graze their domestic livestock. Our findings also suggest that widespread poverty and a lack of alternative livelihood opportunities, culture of consuming bushmeat, lack of education and awareness of the value of protected areas, and weak law enforcement are some of the reasons for the illegal activity. Law enforcement authorities were often constrained by insufficient on-site personnel and a lack of modern equipment and infrastructure to deter illegal activities. We conclude that there is a need to address the issue of illegal hunting and livestock grazing, via provision of alternative livelihoods, in combination with community outreach programmes that aim to improve conservation education and awareness and develop the capacity of the conservation authorities in order to achieve conservation goals. Our findings have implications for the conservation management of all protected areas that are available for exploitation by local communities.
1134
60315
High Expression Levels and Amplification of rRNA Genes in a Mentally Retarded Child with 13p+: A Familial Case Study
Abstract:
A cytogenetic and molecular genetic study of the family with a male child who had mental retardation and autistic features revealed an abnormal chromosome 13 bearing an enlarged p-arm with amplified ribosomal DNA (rDNA) in a boy and his father. Cytogenetic analysis using standard G-banding and FISH with labeled rDNA probes revealed an abnormal chromosome 13 with an enlarged p-arms due to rDNA amplification in a male child, who had clinically confirmed mental retardation and an autistic behavior. This chromosome is evidently inherited from the father, who has morphologically the same chromosome, but is healthy. The karyotype of the mother was normal. Ag-NOR staining showed brightly stained large whole-p-arm nucleolus organizer regions (NORs) in a child and normal-sized NORs in his father with 13p+-NOR-amount mosaicism. qRT-PCR with specific primers showed highly increased levels of 18S, 28S and 5,8 S ribosomal RNA (rRNA) in the patient’s blood samples compared to a normal healthy control donor. Both patient’s father and mother had no elevated levels of rRNAs expression. Thus, in this case, rRNA level seems to correlate with mental retardation in familial individuals with 13p+. Our findings of rRNA overexpression in a patient with mental retardation and his parents may show a possible link between the karyotype (p-arm enlargement due to rDNA amplification), rDNA functionality (rRNA overexpression), functional changes in the brain and mental retardation. The study is supported by Russian Science Foundation Grant 15-15-10001.
1133
58046
Determination and Quatification of Asiaticoside in an Endophytic Fungus from Centella asiatica (L.) Urban
Abstract:
Centella asiatica (L.) Urban is a highly regarded medicinal plant owing to its active constituent’s viz., asiaticoside, madecassoside, asiatic acid, madecassic acid etc. Asiaticoside, one of the most important constituents of the plant, is a triterpenoid saponin having memory enhancement property. In the present study, a total of 8 strains of endophytic fungi were isolated from leaves of C. asiatica out of which only one of the isolates produced asiaticoside. The asiaticoside producing isolate was identified as Colletotrichum gloeosporioides by ITS r DNA sequencing. The fungal extracts were chromatographed using a C18 column (100 X 2.1, 1.7 µm particle size, on Waters ACQUITY UPLC-TQD) and then analysed through electrospray-mass spectroscopy using the electrospray technique with an Agilent LC/ MSD trap with a spray flow of 2μL/min and spray voltage of 2.2 kV by the loop injection method. The production of asiaticoside measured in relation to incubation time and subculture generation revealed the presence of 62.29±3.36 µg/100mL of asiaticoside by C. gloeosporioides on 15th day in first subculture generation. To the best of our knowledge this is the very first report regarding the production of asiaticoside by an endophytic fungus. The present findings definitely provide an impetus to the production of asiaticoside by utilizing the endophytic source.
1132
56771
Influence of Bias Voltage on Amorphous Silicon Deposition as Seed Layer in the Tribological Properties and Corrosion Resistance of Amorphous Hydrogenated Carbon Thin Films
Abstract:
Failures in amorphous hydrogenated carbon (a-C:H) coatings in biomedical applications are mainly attributed to different mechanism at the interface substrate-coating. This paper focuses on the variation of bias voltage (5 kV, 8 kV, and 10 kV) during amorphous silicon deposition as seed layer for amorphous hydrogenated carbon (a-C:H) coatings on AISI 316L substrates. An Active Screen Plasma Enhanced Chemical Vapor Deposition process was used to carry out the experiment. Surface characterization was performed with contact profilometry, obtaining an average roughness from 19 nm up to 30 nm. Mechanical properties were measured using nanoindentation, obtaining the Young’s modulus (170–210 GPa) and hardness (25–30 GPa). Using Raman spectroscopy, the intensity ratio between D and G peaks was calculated in order to stablish the amorphous carbon classification. Friction coefficient was obtained through a Pin on Disk test performed in Hanks solution and dry atmosphere, obtaining a friction coefficient under 0.1 during 30000 cycles using 20 N applied load. Adhesion was evaluated calculating the critical load in a Scratch test. The Electrochemical study was evaluated using Tafel tests obtaining the corresponding polarization curve. The surface tension (30-35 nJ/cm2) was studied with the angle contact test. Adhesion forces between live cell and coating surface were measured by Atomic Force Microscopy.
1131
57993
Functional Variants Detection by RNAseq
Abstract:
RNAseq represents an attractive methodology for the detection of functional genomic variants. RNAseq results obtained from polyA+ RNA selection protocol (POLYA) and from exonic regions capturing protocol (ACCESS) indicate that ACCESS detects 10% more coding SNV/INDELs with respect to POLYA. ACCESS requires less reads for coding SNV detection with respect to POLYA. However, if the analysis aims at identifying SNV/INDELs also in the 5’ and 3’ UTRs, POLYA is definitively the preferred method. No particular advantage comes from ACCESS or POLYA in the detection of fusion transcripts.
1130
56468
3D Label-Free Bioimaging of Native Tissue with Selective Plane Illumination Optical Microscopy
Abstract:
Biomedical imaging of native tissue using light offers the potential to obtain excellent structural and functional information in a non-invasive manner with good temporal resolution. Image contrast can be derived from intrinsic absorption, fluorescence, or scatter, or through the use of extrinsic contrast. A major challenge in applying optical microscopy to in vivo tissue imaging is the effects of light attenuation which limits light penetration depth and achievable imaging resolution. Recently Selective Plane Illumination Microscopy (SPIM) has been used to map the 3D distribution of fluorophores dispersed in biological structures. In this approach, a focused sheet of light is used to illuminate the sample from the side to excite fluorophores within the sample of interest. Images are formed based on detection of fluorescence emission orthogonal to the illumination axis. By scanning the sample along the detection axis and acquiring a stack of images, 3D volumes can be obtained. The combination of rapid image acquisition speeds with the low photon dose to samples optical sectioning provides SPIM is an attractive approach for imaging biological samples in 3D. To date all implementations of SPIM rely on the use of fluorescence reporters be that endogenous or exogenous. This approach has the disadvantage that in the case of exogenous probes the specimens are altered from their native stage rendering them unsuitable for in vivo studies and in general fluorescence emission is weak and transient. Here we present for the first time to our knowledge a label-free implementation of SPIM that has downstream applications in the clinical setting. The experimental set up used in this work incorporates both label-free and fluorescent illumination arms in addition to a high specification camera that can be partitioned for simultaneous imaging of both fluorescent emission and scattered light from intrinsic sources of optical contrast in the sample being studied. This work first involved calibration of the imaging system and validation of the label-free method with well characterised fluorescent microbeads embedded in agarose gel. 3D constructs of mammalian cells cultured in agarose gel with varying cell concentrations were then imaged. A time course study to track cell proliferation in the 3D construct was also carried out and finally a native tissue sample was imaged. For each sample multiple images were obtained by scanning the sample along the axis of detection and 3D maps reconstructed. The results obtained validated label-free SPIM as a viable approach for imaging cells in a 3D gel construct and native tissue. This technique has the potential use in a near-patient environment that can provide results quickly and be implemented in an easy to use manner to provide more information with improved spatial resolution and depth penetration than current approaches.
1129
55263
Quantum Dots Incorporated in Biomembrane Models for Cancer Marker
Abstract:
Quantum dots (QD) are semiconductor nanocrystals that can be employed in biological research as a tool for fluorescence imagings, having the potential to expand in vivo and in vitro analysis as cancerous cell biomarkers. Particularly, cadmium selenide (CdSe) magic-sized quantum dots (MSQDs) exhibit stable luminescence that is feasible for biological applications, especially for imaging of tumor cells. For these facts, it is interesting to know the mechanisms of action of how such QDs mark biological cells. For that, simplified models are a suitable strategy. Among these models, Langmuir films of lipids formed at the air-water interface seem to be adequate since they can mimic half a membrane. They are monomolecular films formed at liquid-gas interfaces that can spontaneously form when organic solutions of amphiphilic compounds are spread on the liquid-gas interface. After solvent evaporation, the monomolecular film is formed, and a variety of techniques, including tensiometric, spectroscopic and optic can be applied. When the monolayer is formed by membrane lipids at the air-water interface, a model for half a membrane can be inferred where the aqueous subphase serve as a model for external or internal compartment of the cell. These films can be transferred to solid supports forming the so-called Langmuir-Blodgett (LB) films, and an ampler variety of techniques can be additionally used to characterize the film, allowing for the formation of devices and sensors. With these ideas in mind, the objective of this work was to investigate the specific interactions of CdSe MSQDs with tumorigenic and non-tumorigenic cells using Langmuir monolayers and LB films of lipids and specific cell extracts as membrane models for diagnosis of cancerous cells. Surface pressure-area isotherms and polarization modulation reflection-absorption spectroscopy (PM-IRRAS) showed an intrinsic interaction between the quantum dots, inserted in the aqueous subphase, and Langmuir monolayers, constructed either of selected lipids or of non-tumorigenic and tumorigenic cells extracts. The quantum dots expanded the monolayers and changed the PM-IRRAS spectra for the lipid monolayers. The mixed films were then compressed to high surface pressures and transferred from the floating monolayer to solid supports by using the LB technique. Images of the films were then obtained with atomic force microscopy (AFM) and confocal microscopy, which provided information about the morphology of the films. Similarities and differences between films with different composition representing cell membranes, with or without CdSe MSQDs, was analyzed. The results indicated that the interaction of quantum dots with the bioinspired films is modulated by the lipid composition. The properties of the normal cell monolayer were not significantly altered, whereas for the tumorigenic cell monolayer models, the films presented significant alteration. The images therefore exhibited a stronger effect of CdSe MSQDs on the models representing cancerous cells. As important implication of these findings, one may envisage for new bioinspired surfaces based on molecular recognition for biomedical applications.
1128
56658
Giant Cancer Cell Formation: A Link between Cell Survival and Morphological Changes in Cancer Cells
Abstract:
Introduction: Giant cancer cells (GCC) are common in all types of cancer, especially after poor therapy. Some specific features of such cells include ~10-fold enlargement, drug resistance, and the ability to propagate similar daughter cells. We used murine NK/Ly lymphoma, an aggressive and fast growing lymphoma model that has already shown drastic changes in GCC comparing to parental cells (chromatin condensation, nuclear fragmentation, tighter OXPHOS/cellular respiration coupling, multidrug resistance). Materials and methods: In this study, we compared morpho-functional changes of GCC that predominantly show either a cytostatic or a cytotoxic effect after treatment with drugs. We studied the effect of a combined cytostatic/cytotoxic drug treatment to determine the correlation of drug efficiency and GCC formation. Doses of G1/S-specific drug paclitaxel/PTX (G2/M-specific, 50 mg/mouse), vinblastine/VBL (50 mg/mouse), and DNA-targeting agents doxorubicin/DOX (125 ng/mouse) and cisplatin/CP (225 ng/mouse) on C57 black mice. Several tests were chosen to estimate morphological and physiological state (propidium iodide, Rhodamine-123, DAPI, JC-1, Janus Green, Giemsa staining and other), which included cell integrity, nuclear fragmentation and chromatin condensation, mitochondrial activity, and others. A single and double factor ANOVA analysis were performed to determine correlation between the criteria of applied drugs and cytomorphological changes. Results: In all cases of treatment, several morphological changes were observed (intracellular vacuolization, membrane blebbing, and interconnected mitochondrial network). A lower gain in ascites (49.97% comparing to control group) and longest lifespan (22+9 days) after tumor injection was obtained with single VBL and single DOX injections. Such ascites contained the highest number of GCC (83.7%+9.2%), lowest cell count number (72.7+31.0 mln/ml), and a strong correlation coefficient between increased mitochondrial activity and percentage of giant NK/Ly cells. A high number of viable GCC (82.1+9.2%) was observed compared to the parental forms (15.4+11.9%) indicating that GCC are more drug resistant than the parental cells. All this indicates that the giant cell formation and its ability to obtain drug resistance is an expanding field in cancer research.
1127
59264
Follicular Fluid Proteins and Cells Study on Large White Pig for Use as Biotechnology
Abstract:
Our study was aimed at morphology of oocytes, follicle cells and follicular fluid proteins of Large White pig. Porcine ovaries collected from local slaughter house in Nakhon Pathom Province. The porcine oocytes and follicular fluid of healthy small follicles (1-2 and 3-4 mm), medium follicles (5-6 mm in diameters) and large follicles (7-8 mm and 10 mm in diameter) were aspirated and collected from the ovaries by sterile technique. Then, the oocytes and the follicle cells were separated from the fluid. The oocytes were round shape and surrounded by zona pellucida with numerous layers of cumulus cells. Based on the number of cumulus cell layers surrounding oocytes, the oocytes were classified into 5 types, which were intact-, multi-, partial- cumulus layer oocyte, completely denuded oocyte and degenerative oocyte. The collected oocytes showed high percentages of intact- and multi- cumulus cell layers in the small follicles (54.05%) medium follicles (56.94%) and large follicles (56.52%) which have high potential to develop into mature oocytes in vitro. The proteins from follicular fluid of 3 size follicles were separated by SDS-PAGE and LC/MS/MS. The molecular weight of follicular fluid proteins from the small follicles were ranging from 27, 62, 70, 80, 115, 140, >220 kDa. Interestingly, protein bands of 70 and 80 kDa played an important role in decreasing metaphase I of oocyte maturation. The follicular fluid protein from medium and large follicle contained 50, 65, 75, 90, 95, 110, 120, 160, 180, 190 and >220 kDa. All proteins played important roles in promoting and regulating growth and development of oocytes and ovulation. This finding was an initial tool for in vitro testing and applied biotechnology research. Acknowledgements: The project was funded by a grant from Silpakorn University Research & Development Institute (SURDI) and Faculty of Science, Silpakorn University, Thailand.
1126
58923
Differential Proteomics Expression in Purple Rice Supplemented Type 2 Diabetic Rats’ Skeletal Muscle
Abstract:
Type 2 diabetes is one of the most common metabolic diseases all over the world. The pathogenesis of type 2 diabetes is not the only dysfunction of pancreatic beta cells but also insulin resistance in muscle, liver and adipose tissue. High levels of circulating free fatty acids, an increased lipid content of muscle cells, impaired insulin-mediated glucose uptake and diminished mitochondrial functioning are pathophysiological hallmarks of diabetic skeletal muscles. Purple rice (Oryza sativa L. indica) has been shown to have antidiabetic effects. However, the underlying mechanism(s) of antidiabetic activity of purple rice is still unraveled. In this research, to explore in-depth cellular mechanism(s), proteomic profile of purple rice supplemented type 2 diabetic rats’ skeletal muscle were analyzed contract with non-supplemented rats. Diabetic rats were induced high-fat diet combined with streptozotocin injection. By using one- dimensional gel electrophoresis (1-DE) and LC-MS/MS quantitative proteomic method, we analyzed proteomic profiles in skeletal muscle of normal rats, normal rats with purple rice supplementation, type 2 diabetic rats, and type 2 diabetic rats with purple rice supplementation. Total 2676 polypeptide expressions were identified. Among them, 24 peptides were only expressed in type 2 diabetic rats, and 24 peptides were unique peptides in type 2 diabetic rats with purple rice supplementation. Acetyl CoA carboxylase 1 (ACACA) found as unique protein in type 2 diabetic rats which is the major enzyme in lipid synthesis and metabolism. Interestingly, DNA damage response protein, heterogeneous nuclear ribonucleoprotein K [Mus musculus] (Hnrnpk), was upregulated in type 2 diabetic rats’ skeletal muscle. Meanwhile, unique proteins of type 2 diabetic rats with purple rice supplementation (bone morphogenetic 7 protein preproprotein, BMP7; and forkhead box protein NX4, Foxn4) involved with muscle cells growth through the regulation of TGF-β/Smad signaling network. Moreover, BMP7 may effect on insulin signaling through the downstream signaling of protein kinase B (Akt) which acts in protein synthesis, glucose uptake, and glycogen synthesis. In conclusion, our study supports that type 2 diabetes impairs muscular lipid metabolism. In addition, purple rice might recover the muscle cells growth and insulin signaling.
1125
57055
Ethnomedicinal Uses of Plants in Bridim Village Development Committee in Langtang National Park, Nepal
Authors:
Abstract:
Bridim Village Development Committee (VDC) is one of the medicinal plants hot spots of Nepal. It is located on a ridge above the lower Langtang Khola, steep and narrow spot in between 1944 m to 4833 m altitude. The study area is homogeneously inhabited by Tamang communities. An investigation on folk herbal medicine on the basis of traditional uses of medicinal plants was done in 2014. The local traditional healers, elder men and women, traders and teachers, were consulted as key informants for documentation of indigenous knowledge on the medicinal plants. It was found that altogether seventy-one medicinal plant species belonging to sixty genera and thirty-three families were used by local people for twenty-seven diseases. Roots of thirty-four species were the most frequently used plant parts and bigger numbers of species were found to be used in fever of ten species. Most medicines were prepared in the form of juice of forty species. The attempt of the study was to document ethno medicinal practices to treat different diseases in the study area for conservation of indigenous knowledge.
1124
58901
Micropropagation Protocols for Conservation, Reforestation and Commercial Propagation of Critically Endangered Economically Valuable Mangroves
Abstract:
Micropropagation protocols were developed for conservation, reforestation and commercial propagation of economically valuable critically endangered mangrove species (less than 50 individuals left) in Singapore. Mangroves are a group of specialized tropical trees and shrubs well adopted to grow in salty muddy intertidal regions between land and sea. Mangrove forests are rich in biodiversity and provide home to various animals and birds like fishes, prawns, monkeys, deer, tiger, herons, storks, eagles etc. Mangroves prevent soil erosion and keep shore-lines intact against tides and storms. As mangroves are wide spread in tropical regions and are highly productive, they are considered huge carbon sinks and help mitigate climate change. They support human communities by providing food, biomass, wood and other forest products. Mangrove forests are diminishing at alarming rate due to over exploitation for natural resources, deforestation in favor of alternate land use, pollution, storms and hurricanes. In wild, small saplings are often washed away in strong water currents or get eaten by wild animals. Efforts to artificially propagate and restore degraded mangrove forests by propagules are hindered by seasonal production, low availability, short viability and poor germination rates of propagules. Thus in vitro propagation technology holds great potential towards mass production of high quality, pathogen-free plantation materials. In our current project highly efficient tissue culture protocols were developed for large scale regeneration of three economically valuable species – Bruguiera hainesii (firewood, charcoal, construction timber), Dolichandrone spothacea (wood for saddles, float, medicinal value) and Sonneratia caseolaris (medicinal value). This asexual production method of micropropagation has advantages of producing same genetic material off-springs as the elite mother plant, the vegetative starting material is available throughout the year and plants can be regenerated in shorter time and space as compared to those obtained from sexually produced propagules. In our protocols, Apical and Axillary bud explants were used to initiate sterile cultures. The initial problem of lower survival rates due to high contamination and exodus of phenolic compounds was overcome by addition of anti-leaching compounds. The shoots multiplied and elongated on cytokinin medium and rooted on auxin supplemented medium. In vitro propagated plants acclimatized well under green-house conditions. The hardened plants in nursery are undergoing salt and high water tolerance assessments. This project will help to restore these endangered species; preserve bio-diversity and counter deforestation. Mangroves are of great scientific interest as they serve as pool of candidate genes for traits like salt tolerance, flood tolerance etc. These genes when transferred to other crops via protoplast fusion, Agrobacterium or nanoparticles will render them tolerant to abiotic stress and adaptable to climate change.
1123
55711
Biochemical Characterization of the Functionally Conserved, Exosome-Associated 3'-5' Exonuclease Rrp6 Homolog (EhRrp6) from the Early-Branching Human Parasite Entamoeba histolytica
Abstract:
The eukaryotic exosome complex plays a pivotal role in RNA biogenesis, maturation, surveillance and differential expression of various RNAs in response to varying environmental signals. The exosome is composed of evolutionary conserved nine core subunits and the associated exonucleases Rrp6 and Rrp44. Rrp6p is crucial for the processing of rRNAs, other non-coding RNAs, regulation of polyA tail length and termination of transcription. Rrp6p, a 3’-5’ exonuclease is required for degradation of 5’-external transcribed spacer (ETS) released from the rRNA precursors during the early steps of pre-rRNA processing. In the parasitic protist Entamoeba histolytica in response to growth stress, there occurs the accumulation of unprocessed pre-rRNA and 5’ ETS sub fragment. To understand the processes leading to this accumulation, we looked for Rrp6 and the exosome subunits in E. histolytica, by in silico approaches. Of the nine core exosomal subunits, seven had a high percentage of sequence similarity with the yeast and human. The EhRrp6 homolog contained exoribonuclease and HRDC domains like yeast but its N- terminus lacked the PMC2NT domain. EhRrp6 complemented the temperature sensitive phenotype of yeast rrp6Δ cells suggesting conservation of biological activity. We showed 3’-5’ exoribonuclease activity of EhRrp6p with in vitro-synthesized appropriate RNAs substrates. Like the yeast enzyme, EhRrp6p degraded unstructured RNA, but could degrade the stem-loops slowly. Furthermore, immunolocalization revealed that EhRrp6 was nuclear-localized in normal cells but was diminished from nucleus during serum starvation, which could explain the accumulation of 5’ETS during stress. Our study shows functional conservation of EhRrp6p in E. histolytica, an early-branching eukaryote, and will help to understand the evolution of exosomal components and their regulatory function.
1122
57291
Adjustment and Scale-Up Strategy of Pilot Liquid Fermentation Process of Azotobacter Sp.
Abstract:
Azotobacter sp. has been widely used as bio-fertilizer due to its significant effects on the stimulation and promotion of plant growth in various agricultural species of commercial interest. In order to obtain the biomass for the development of a biofertilizer product, a scale-up strategy for a liquid fermentation process (SmF) in co-culture with two strains of A. chroococcum (named Ac1 and Ac10) was validated and adjusted at laboratory and pilot level. A batch fermentation process was carried out on a bioreactor Infors, model Minifors of 3.5L, which served as a baseline for this research. In order to increase process efficiency, the effect of the reduction of stirring speed was evaluated, maintaining the air flow and the culture time under conditions of fed-batch type fermentation (pulse type). In this process, an increase of 2.73 times in cell concentration of Ac1 strain and 1.8 times for AC10 strain were achieved; with a 96.7% reduction in energy consumption. Additionally, the magnitudes of efficiency parameters, biomass concentration, and fermentation productivity in fed-batch fermentation were increased twice compared with baseline. To reproduce efficiency parameters at the pilot level, a scale-up strategy with two criteria, geometric and fluid dynamic behavior similarities (Reynolds number), was evaluated. According to the analysis of variance, this scale-up strategy had not significant effect on efficiency parameters and cell concentration, between laboratory and pilot fermentation (Tukey, p > 0.05). Regarding to air consumption, fermentation process at pilot level showed a reduction of 23% in the baseline. The percentage of energy consumption reduction under pilot level conditions was 96.9% compared with baseline.
1121
57293
Biomass Production Improvement of Beauveria bassiana at Laboratory Level for the Development of a Biopesticide
Abstract:
Beauveria sp. has been used as entomopathogenic microorganism for biological control of various plant plagues such as whitefly, thrips, aphids and chrysomelidaes such as Cerotoma tingomariana species, which affect soybean production in Colombia´s Altillanura region. Therefore, a biopesticide prototype based on B. bassiana strain Bv060 at Corpoica laboratories was developed. For the production B. bassiana conidia, a baseline fermentation was performed at laboratory in a solid medium using broken rice as a substrate, a temperature of 25±2 °C and a humidity of 60±10%. The experimental design was completely random, with three-time repetition. Under these culture conditions, an average conidial concentration of 1.48x1010 conidia/g, a yield of 13.07 g/kg dry substrate and a productivity of 8.83x107 conidia/g*h were achieved. Consequently, the objective of this study was to evaluate the influence of the particular size reduction of rice (
1120
58692
Anecic and Epigeic Earthworms as Potential Biocontrol Agents of Fusarium graminearum, Causal Agent of Fusarium Head Blight on Wheat
Abstract:
Fusarium Head Blight (FHB) is one of the most important Fusarium-caused diseases, which affects cereals with serious detrimental effects on yield and grain quality worldwide. Earthworms have been suggested as an alternative to control this disease, which requires a combination of preventive methods to reduce level of damage, although it has been proven that their effect is species dependent. Our objective was to evaluate the effect of the earthworms Aporrectodea longa and Lumbricus rubellus, on the inoculum of Fusarium graminearum on wheat straw. To test this we kept earthworms in vessels with soil, and F. graminearum-inoculated straw covering the surface, under controlled conditions for 6 weeks. Two factors were evaluated with a complete factorial design: earthworms (three levels: without earthworms, A. longa, and L. rubellus), and straw (two levels: inoculated with the pathogen, and sterile). The presence of L. rubellus significantly (P
1119
56654
Multiscale Model of Blast Explosion Human Injury Biomechanics
Abstract:
Bomb blasts from Improvised Explosive Devices (IEDs) account for vast majority of terrorist attacks worldwide. Injuries caused by IEDs result from a combination of the primary blast wave, penetrating fragments, and human body accelerations and impacts. This paper presents a multiscale computational model of coupled blast physics, whole human body biodynamics and injury biomechanics of sensitive organs. The disparity of the involved space- and time-scales is used to conduct sequential modeling of an IED explosion event, CFD simulation of blast loads on the human body and FEM modeling of body biodynamics and injury biomechanics. The paper presents simulation results for blast-induced brain injury coupling macro-scale brain biomechanics and micro-scale response of sensitive neuro-axonal structures. Validation results on animal models and physical surrogates are discussed. Results of our model can be used to 'replicate' filed blast loadings in laboratory controlled experiments using animal models and in vitro neuro-cultures.