Search results for: conserved forage
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 257

Search results for: conserved forage

77 Effect of Mannitol on in Vitro Conservation of Local and Exotic Taro-Genotypes (Colocasia Esculenta Var Esculenta)

Authors: Benjamin Bonsu Bruce, Marian Dorcas Quain David Appiah-Kubi, Gertrude Osei-Diko, Harrison Kwame Dapaah

Abstract:

Taro [Colocasia esculenta (L.) Schott] is a major staple food and remains a significant crop to many cultural and agricultural customs worldwide. In Ghana, taro is mostly propagated using vegetative material, which is conserved in field collection and recycled from their farms to establish new fields. However, this practice promotes the accumulation of systemic pathogens. Prior exposure to pests and subsequent expression of disease symptoms can also be a huge constraint to sustainable conservation and utilization of taro genetic resources. In vitro, slow growth is one of the most promising techniques to be utilized for conservation. The objective of this study was to find a medium-term in vitro conservation protocol for local and exotic taro genotypes. The medium-term conservation study was conducted using actively growing shoots obtained from in vitro cultures. Explants were cultured to full strength in complete Murashige and Skoog medium supplemented with Mannitol at different concentrations (0g/l, 20g/l, 25g/l, and 30g/l). Another medium that was tested as an additional treatment is the White’s medium. The highest number of shoots (6.33) and leaves (22.67) occurred on medium containing 20 and 25g/l mannitol in genotype SAO 006 as compared to other genotypes, whereas 30g/l mannitol was the best to restrict growth for the entire 6 months period in terms of shoot height (22.50cm). The study reveals that mannitol supplemented culture media could reduce the growth of Colocasia plantlets, especially in stem height. Culture growth following 6 months of conservation, showed that healthy shoot cultures of Taro were obtained after 6 months of storage in a medium containing 20gl⁻¹ and 25gl⁻¹ mannitol.

Keywords: complete murashige, skoog medium, culture conditions, mannitol, slow growth conservation

Procedia PDF Downloads 141
76 Archaeological Study of Statues of King Thutmosis III from Luxor

Authors: Mahmoud Abualsoud

Abstract:

The era of Thutmosis III represents a transitional period between the art of the Thutmoside art and the Amarna period, so we intend to declare that it serves as the cradle of Amarna art. The study will examine the Statues of king Thutmose III that was discovered in Luxor by an Egyptian mission. These Statues have been transferred to the Conservation Center of the Grand Egyptian Museum (GEM) to be conserved and made ready to be displayed at the new museum (the project of the century). We focus on three Statues chosen because they relate to different years of the king's reign. These Statues were all made of granite. The first one is a Kneeling statue representing the god Amun showing king Thutmose III offering to the goddess Hathor. The second is decorated with king Thutmose III with the red crown, between the goddess Hathor and the royal wife, Nefertari. The third shows the king offering NW vessels and bread to the god Seker. Each statue is divided into registers containing a description and decorated with scenes of the king presenting offerings to gods. The proposed study will focus on the development which happened sequentially according to differences that occur in each statue. We will use comparative research to determine the workshops of these statues, whether one or several, and what are the distinguishing features of each one. We will examine what innovations the artisans added to royal art. The description and the texts will be translated with linguistic comments. This research focuses on text analyses and technology. Paleographic information found on these objects includes the names and titles of the king. This research focuses on text analyses and technology. The study aims to create a manual that may help in dating the artwork of Thutmosis III. This research will be beneficial and useful for heritage and ancient civilizations, particularly when we talk about opening museums like the Grand Egyptian Museum, which will exhibit a collection of statues. Indeed, this kind of study will open a new destination in order to know how to identify these collections and how to exhibit them commensurate with the nature of ancient Egyptian history and heritage.

Keywords: archaeological study, Giza, new kingdom, statues, royal art

Procedia PDF Downloads 44
75 Structural and Functional Characterization of the Transcriptional Regulator Rv1176 of Mycobacterium tuberculosis H37Rv

Authors: Vikash Yadav, Ashish Arora

Abstract:

Microorganisms have self-defense mechanisms to protect themselves from toxic environments. Phenolic acid decarboxylase(pad) is responsible for the defense against toxicity caused by phenolic acids, converting them into less toxic vinyl derivatives. The transcription of the pad gene is regulated by a negative transcription factor, phenolic acid decarboxylase regulators (PadR), in a substrate-inducible manner. The PadR family members share the conserved DNA-binding features and interact with the operator DNA using a winged helix-turn-helix (wHTH) motif, which contains a three-helix motif and a β-stranded wing. The members of this family function as transcriptional regulators that are involved in various cellular survival processes, such as toxin production, detoxification, multidrug resistance, antibiotic biosynthesis, and carbon catabolism. Rv1176 of Mycobacterium tuberculosis H37Rv has been assigned to the PadR family protein that remains to be structurally and functionally uncharacterized. To reveal the structural mechanism by which Rv1176 could regulates effector-responsive transcription, several experiments were performed, including Electrophoretic Mobility Shift Assay (EMSA) for DNA protein interaction, differential scanning calorimetry (DSC) and Differential Scanning Fluorimetry (DSF) for temperature and ligand-dependent protein stability, Circular Dichroism (CD) spectroscopy for secondary structure analysis. Further, to evaluate the functional role of Rv1176, the intracellular survival of recombinant M. smegmatis was examined in murine macrophage cell line J774A.1 and different stressed conditions like oxidative, pH, and nutritive stress. All these studies demonstrated that Rv1176 could behave as a transcription regulator and its expression in recombinant M. smegmatis increases intracellular survival.

Keywords: EMSA, Mycobacterium tuberculosis, PadR family protein, transcriptional regulator

Procedia PDF Downloads 48
74 The Subcellular Localisation of EhRRP6 and Its Involvement in Pre-Ribosomal RNA Processing in Growth-Stressed Entamoeba histolytica

Authors: S. S. Singh, A. Bhattacharya, S. Bhattacharya

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 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.

Keywords: entamoeba histolytica, exosome complex, rRNA processing, Rrp6

Procedia PDF Downloads 173
73 The Interplay between Autophagy and Macrophages' Polarization in Wound Healing: A Genetic Regulatory Network Analysis

Authors: Mayada Mazher, Ahmed Moustafa, Ahmed Abdellatif

Abstract:

Background: Autophagy is a eukaryotic, highly conserved catabolic process implicated in many pathophysiologies such as wound healing. Autophagy-associated genes serve as a scaffolding platform for signal transduction of macrophage polarization during the inflammatory phase of wound healing and tissue repair process. In the current study, we report a model for the interplay between autophagy-associated genes and macrophages polarization associated genes. Methods: In silico analysis was performed on 249 autophagy-related genes retrieved from the public autophagy database and gene expression data retrieved from Gene Expression Omnibus (GEO); GSE81922 and GSE69607 microarray data macrophages polarization 199 DEGS. An integrated protein-protein interaction network was constructed for autophagy and macrophage gene sets. The gene sets were then used for GO terms pathway enrichment analysis. Common transcription factors for autophagy and macrophages' polarization were identified. Finally, microRNAs enriched in both autophagy and macrophages were predicated. Results: In silico prediction of common transcription factors in DEGs macrophages and autophagy gene sets revealed a new role for the transcription factors, HOMEZ, GABPA, ELK1 and REL, that commonly regulate macrophages associated genes: IL6,IL1M, IL1B, NOS1, SOC3 and autophagy-related genes: Atg12, Rictor, Rb1cc1, Gaparab1, Atg16l1. Conclusions: Autophagy and macrophages' polarization are interdependent cellular processes, and both autophagy-related proteins and macrophages' polarization related proteins coordinate in tissue remodelling via transcription factors and microRNAs regulatory network. The current work highlights a potential new role for transcription factors HOMEZ, GABPA, ELK1 and REL in wound healing.

Keywords: autophagy related proteins, integrated network analysis, macrophages polarization M1 and M2, tissue remodelling

Procedia PDF Downloads 121
72 Design of New Baby Food Product Using Whey

Authors: Henri El Zakhem, Anthony Dahdah, Lara Frangieh, Jessica Koura

Abstract:

Nowadays, the removal of whey produced in the dairy processes has been the most important problem in the dairy industry. Every year, about 47% of the 115 million tons of whey produced world-wide are disposed in the environment. Whey is a nutritious liquid, containing whey proteins (β-lactoglobulin, α-lactalbumin, immunoglobulin-G, proteose pepton), lactose, vitamins (B5, B2, C, and B6), minerals (Calcium, Magnesium, Phosphorous, Potassium, Chloride, and Sodium), and trace elements (Zinc, Iron, Iodine, and Copper). The first objective was to increase the economical and commercial value of whey which is considered as by-product. The second objective of this study was to formulate a new baby food with good nutritional, sensory and storage properties and acceptable to consumers using the cheese whey. The creation of the new product must pass through the following stages: idea stage, development stage which includes the business planning and the product development prototype, packaging stage, production stage, test marketing stage, quality control/sanitation. Three types of whey-based food were selected and prepared by mixing whey and apple, whey and banana as well as whey, apple, and banana.To compile with the recommended dietary allowances (RDA) and adequate intakes (AI) for vitamins and minerals, each sample is formed from 114g of sliced and smashed fruits mixed with 8 mL of whey. Mixtures are heated to 72oC for 15 seconds, and filled in pasteurized jars. Jars were conserved at 4oC. Following the experimental part, sensory evaluation made by an experienced panel took place. Hedonic tests results show that the mixture of whey, apple, and banana has the most delicious and sweetness taste followed by the mixture of whey and banana, and finally the mixture of whey and apple. This study was concluded with a managerial and engineering study that reveals that the project is economically profitable to be executed in Lebanon.

Keywords: baby food, by-product, cheese whey, formulation

Procedia PDF Downloads 248
71 Molecular Characterization of Grain Storage Proteins in Some Hordeum Species

Authors: Manar Makhoul, Buthainah Alsalamah, Salam Lawand, Hassan Azzam

Abstract:

The major storage proteins in endosperm of 33 cultivated and wild barley genotypes (H.vulgare, H. spontaneum, H. bulbosum, H. murinum, H. marinum) were analyzed to demonstrate the variation in the hordein polypeptides encoded by multigene families in grains. The SDS-PAGE revealed 13 and 17 alleles at the Hor1 and the Hor2 loci respectively, with frequencies from 0.83 to 14 and 0.56 to 13.41% respectively, while seven alleles at the Hor3 locus with frequencies from 3.63 to 30.91% were recognized. The phylogenetic analysis indicated to relevance of the polymorphism in hordein patterns as successful tool in identifying the individual genotypes and discriminating the species according to genome type. We also reported in this research complete nucleotide sequence B-hordein genes of seven wild and cultivated barley genotypes. A 152bp upstream sequence of B-hordein promoter contained a TATA box, CATC box, AAAG motif, N-motif and E-motif. In silico analysis of B-Hordein sequences demonstrated that the coding regions were not interrupted by any intron, and included the complete ORF which varied between 882 and 906 bp, and encoded mature proteins with 293-301 residues characterized by high contents of glutamine (29%), and proline (18%). Comparison of the predicted polypeptide sequences with the published ones suggested that all S-rich prolamins genes are descended from common ancestor. The sequence started at N-terminal with a signal peptide, and then followed directly by two domains; a repetitive one based on the repetition of the repeat unit PQQPFPQQ and C-terminal domain. Also, it was found that positions of the eight cysteine residues were highly conserved in all the B-hordein sequences, but Hordeum bulbosum had additional unpaired one. The phylogenetic tree of B-hordein polypeptide separated the genotypes in distinct seven subgroups. In general, the high homology between B-hordeins and LMW glutenin subunits suggests similar bread-making influences for these B-hordeins.

Keywords: hordeum, phylogenetic tree, sequencing, storage protein

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70 Allelopathic Action of Diferents Sorghum bicolor [L.] Moench Fractions on Ipomoea grandifolia [Dammer] O'Donell

Authors: Mateus L. O. Freitas, Flávia H. de M. Libório, Letycia L. Ricardo, Patrícia da C. Zonetti, Graciene de S. Bido

Abstract:

Weeds compete with agricultural crops for resources such as light, water, and nutrients. This competition can cause significant damage to agricultural producers, and, currently, the use of agrochemicals is the most effective method for controlling these undesirable plants. Morning glory (Ipomoea grandifolia [Dammer] O'Donell) is an aggressive weed and significantly reduces agricultural productivity making harvesting difficult, especially mechanical harvesting. The biggest challenge in modern agriculture is to preserve high productivity reducing environmental damage and maintaining soil characteristics. No-till is a sustainable practice that can reduce the use of agrochemicals and environmental impacts due to the presence of plant residues in the soil, which release allelopathic compounds and reduce the incidence or alter the growth and development of crops and weeds. Sorghum (Sorghum bicolor [L.] Moench) is a forage with proven allelopathic activity, mainly for producing sorgholeone. In this context, this research aimed to evaluate the allelopathic action of sorghum fractions using hexane, dichloromethane, butanol, and ethyl acetate on the germination and initial growth of morning glory. The parameters analyzed were the percentage of germination, speed of germination, seedling length, and biomass weight (fresh and dry). The bioassays were performed in Petri dishes, kept in an incubation chamber for 7 days, at 25 °C, with a 12h photoperiod. The experimental design was completely randomized, with five replicates of each treatment. The data were evaluated by analysis of variance, and the averages between each treatment were compared using the Scott Knott test at a 5% significance level. The results indicated that the dichloromethane and ethyl acetate fractions showed bioherbicidal effects, promoting effective reductions on germination and initial growth of the morning glory. It was concluded that allelochemicals were probably extracted in these fractions. These secondary metabolites can reduce the use of agrochemicals and environmental impact, making agricultural production systems more sustainable.

Keywords: allelochemicals, secondary metabolism, sorgoleone, weeds

Procedia PDF Downloads 124
69 Archaeological Study of Statues of King Thutmosis III from Luxor

Authors: Ahmed Mamdouh

Abstract:

Introduction: The era of Thutmosis III represents a transitional period between the art of the Thutmoside art and the Amarna period, so we intend to declare that it serves as the cradle of Amarna art. The study will examine the Statues of king Thutmose III that was discovered in Luxor by an Egyptian mission. These Statues have been transferred to the Conservation Center of the Grand Egyptian Museum (GEM) to be conserved and made ready to bedisplayed at the new museum (the project of the century). We focus upon three Statues (GEM numbers 45863, 45864, 45865), chosen because they relate to different years of the king's reign. These Statues were all made of granite. The first one is a Kneeling statue representing the god Amun showing king Thutmose III offering to the goddess Hathor. The second is decorated with king Thutmose III with the red crown, between the goddess Hathor and the royal wife, Nefertari. The third shows the king offering NW vessels and bread to the god Seker. Each Statue is divided into registers containing a description and decorated with scenes of the king presenting offerings to gods. Methodology: The proposed study will focus on the development which happened sequentially according to differences that occur in each Statue. We will use comparative research to determine the workshops of these statues, whether one or several, and what are the distinguishing features of each one. We will examine what innovations the artisans added to royal art. The description and the texts will be translated with linguistic comments. This research focuses on text analyses and technology. Paleographic information found on these objects includes the names and titles of the king. Conclusion: This research focuses on text analyses and technology. The study aims to create a manual that may help in dating the artwork of Thutmosis III. This research will be beneficial and useful for heritage and ancient civilizations, particularly when we talk about opening museums like the Grand Egyptian museum, which will exhibit a collection of statues. Indeed this kind of study will open a new destination in order to know how to identify these collections and how to exhibit them commensurate with the nature of ancient Egyptian history and heritage.

Keywords: archaeological study, Giza, new kingdom, statues, royal art

Procedia PDF Downloads 43
68 Impact of Land Ownership on Rangeland Condition in the Gauteng Province, South Africa

Authors: N. L. Letsoalo, H. T. Pule, J. T. Tjelele, N. R. Mkhize, K. R. Mbatha

Abstract:

Rangelands are major feed resource for livestock farming in South Africa, despite being subjected to different forms of degradation. These forms of degradation are as a result of inappropriate veld and livestock management practices such as excessive stocking rates. While information on judicious veld management is available, adoption of appropriate practices is still unsatisfactory and seems to depend partly on the type of land ownership of farmers. The objectives of this study were to; (I) compare rangeland condition (species richness, basal cover, veld condition score, and herbaceous biomass) among three land ownership types (leased land, communal land and private land), and (II) determine the relationships between veld condition score (%) and herbaceous biomass (kg DM/ha) production. Vegetation was assessed at fifty farms under different land use types using nearest plant technique. Grass species composition and forage value were estimated using PROC FREQ procedure of SAS 9.3. A one-way ANOVA was used to determine significant differences (P < 0.05) in species richness, basal cover, veld condition (%) large stock units, grazing capacity and herbaceous biomass production among the three grazing systems. A total of 28 grass species were identified, of which 95% and 5% were perennials and annuals, respectively. The most commonly distributed and highly palatable grass species, Digitaria eriantha had significantly higher frequency under private owned lands (32.3 %) compared to communal owned lands (12.3%). There were no significant difference on grass species richness and basal cover among land ownership types (P > 0.05). There were significant differences on veld condition score and biomass production (P < 0.05). Private lands had significantly higher (69.63%) veld condition score than leased (56.07%) and communal lands (52.55%). Biomass production was significantly higher (± S.E.) 2990.30 ± 214 kg DM/ha on private owned lands, compared to leased lands 2069.85 ± 196 kg DM/ha and communal lands 1331.04 ± 102 kg DM/ha. Biomass production was positively correlated with rangeland condition (r = 0.895; P < 0.005). These results suggest that rangeland conditions on communal and leased lands are in poor condition than those on private lands. More research efforts are needed to improve management of rangelands in communal and leased land in Gauteng province.

Keywords: grazing, herbaceous biomass, management practices, species richness

Procedia PDF Downloads 138
67 Large-scale Foraging Behaviour of Free-ranging Goats: Influence of Herd Size, Landscape Quality and Season

Authors: Manqhai Kraai, Adrian M. Shrader, Peter F. Scogings

Abstract:

For animals living in herds, competition between group members increases as herd size increases. The intensity of this competition is likely greater across poor quality landscapes and during the dry season. In contrast to wild herbivores, herd size in domestic livestock is determined by their owners. This then raises the question, how do domestic livestock, like goats, reduce competition for food within these defined herds? To explore this question, large-scale foraging behaviour of both small (12 to 28 individuals) and large (42 to 83 individuals) herds of free-ranging goats were recorded in Tugela Ferry, KwaZulu-Natal, South Africa. The study was conducted on three different landscapes that varied in both food quality and availability, during the wet and dry seasons of 2013-2014. The goats were housed in kraals overnight and let out in the mornings to forage unattended. Thus, foraging decisions were made by the goats and not by herders. The large-scale foraging behaviours focussed on included, (i) total distance travelled by goats while foraging, (ii) distance travelled before starting to feed, (iii) travel speed, and (iv) feeding duration. This was done using Garmin Foretrex 401 GPS devices harnessed to two goats per herd. Irrespective of season, there was no difference in the total distance travelled by the different sized herds across the different quality landscapes. However, both small and large herds started feeding farther from the kraal in the dry compared to the wet season. Despite this, there was no significant seasonal difference in total amount of time the herds spent feeding across the different landscapes. Finally, both small and large herds increased their travel speed across all the landscapes in the dry season, but large herds travelled faster than small herds. This increase was likely to maximise the time that large herds could spend feeding in good areas. Ultimately, these results indicate that both small and large herds were affected by declines in food quality and quantity during the dry season. However, as large herds made greater behavioural adjustments compared to smaller herds (i.e., feeding farther away from the kraal and travelling faster), it appeared that they were more affected by the seasonal increases in intra-herd competition.

Keywords: distance, feeding duration, food availability, food quality, travel speed

Procedia PDF Downloads 94
66 Modulation of Fish Allergenicity towards the Production of a Low Allergen Farmed Fish

Authors: Denise Schrama, Claudia Raposo, Pedro Rodrigues

Abstract:

Background: Food allergies are conducted by a hypersensitive response of the immune system. These allergies are a global concern for the public health. Consumption of fish is increasing worldwide as it is a healthy meat with high nutritional value. Unfortunately, fish can cause adverse immune-mediate reactions, affecting part of the population with higher incidence in children. β-parvalbumin, a small, highly conserved stable, calcium or magnesium binding muscle protein is the main fish allergen. In fish-allergic patients, cross-reactivity between different fish species exist due to recognition of highly identical protein regions. Enolases, aldolases, or fish gelatin are other identified fish allergens in some fish species. With no available cure for fish allergies, clinical management is only based on an avoidance diet aiming at the total exclusion of offending food. Methods: Mediterranean fish (S. aurata and D. labrax) were fed specifically designed diets, enriched in components that target the expression or inactivation of parvalbumin (creatine and EDTA, respectively). After 90 days fish were sampled and biological tissues were excised. Proteomics was used to access fish allergens characterization and expression in muscle while IgE assays to confirm the lower allergenic potential are conducted in patients with history of fish allergies. Fish welfare and quality of flesh were established with biochemical, texture and sensorial analysis. Results: Fish welfare shows no major impact between diets. In case of creatine supplementation in D. labrax proteomic analysis show a slight decrease in parvalbumin expression. No accumulation of this compound was found in muscle. For EDTA supplementation in S. aurata IgE assay show a slight decrease in allergenicity when using sera of fish allergic patients. Conclusion: Supplementation with these two compounds seems to change slightly the allergenicity of the two mean Mediterranean species.

Keywords: fish allergies, fish nutrition, proteomics, aquaculture

Procedia PDF Downloads 125
65 The Mapping of Pastoral Area as a Basis of Ecological for Beef Cattle in Pinrang Regency, South Sulawesi, Indonesia

Authors: Jasmal A. Syamsu, Muhammad Yusuf, Hikmah M. Ali, Mawardi A. Asja, Zulkharnaim

Abstract:

This study was conducted and aimed in identifying and mapping the pasture as an ecological base of beef cattle. A survey was carried out during a period of April to June 2016, in Suppa, Mattirobulu, the district of Pinrang, South Sulawesi province. The mapping process of grazing area was conducted in several stages; inputting and tracking of data points into Google Earth Pro (version 7.1.4.1529), affirmation and confirmation of tracking line visualized by satellite with a variety of records at the point, a certain point and tracking input data into ArcMap Application (ArcGIS version 10.1), data processing DEM/SRTM (S04E119) with respect to the location of the grazing areas, creation of a contour map (a distance of 5 m) and mapping tilt (slope) of land and land cover map-making. Analysis of land cover, particularly the state of the vegetation was done through the identification procedure NDVI (Normalized Differences Vegetation Index). This procedure was performed by making use of the Landsat-8. The results showed that the topography of the grazing areas of hills and some sloping surfaces and flat with elevation vary from 74 to 145 above sea level (asl), while the requirements for growing superior grass and legume is an altitude of up to 143-159 asl. Slope varied between 0 - > 40% and was dominated by a slope of 0-15%, according to the slope/topography pasture maximum of 15%. The range of NDVI values for pasture image analysis results was between 0.1 and 0.27. Characteristics of vegetation cover of pasture land in the category of vegetation density were low, 70% of the land was the land for cattle grazing, while the remaining approximately 30% was a grove and forest included plant water where the place for shelter of the cattle during the heat and drinking water supply. There are seven types of graminae and 5 types of legume that was dominant in the region. Proportionally, graminae class dominated up 75.6% and legume crops up to 22.1% and the remaining 2.3% was another plant trees that grow in the region. The dominant weed species in the region were Cromolaenaodorata and Lantana camara, besides that there were 6 types of floor plant that did not include as forage fodder.

Keywords: pastoral, ecology, mapping, beef cattle

Procedia PDF Downloads 316
64 Human Par14 and Par17 Isomerases Bind Hepatitis B Virus Components Inside and Out

Authors: Umar Saeed

Abstract:

Peptidyl-prolyl cis/trans isomerases Par14 and Par17 in humans play crucial roles in diverse cellular processes, including protein folding, chromatin remodeling, DNA binding, ribosome biogenesis, and cell cycle progression. However, the effects of Par14 and Par17 on viral replication have been explored to a limited extent. We first time discovered their influential roles in promoting Hepatitis B Virus replication. In this study, we observed that in the presence of HBx, either Par14 or Par17 could upregulate HBV replication. However, in the absence of HBx, neither Par14 nor Par17 had any effect on replication. Their mechanism of action involves binding to specific motifs within HBc and HBx proteins. Notably, they target the conserved 133Arg-Pro134 (RP) motif of HBc and the 19RP20-28RP29 motifs of HBx. This interaction is fundamental for the stability of HBx, core particles, and HBc. Par14 and Par17 exhibit versatility by binding both outside and inside core particles, thereby facilitating core particle assembly through their participation in HBc dimer-dimer interactions. NAGE and immunoblotting analyses unveiled the binding of Par14/Par17 to core particles. Co-immunoprecipitation experiments further demonstrated the interaction of Par14/Par17 with core particle assembly-defective and dimer-positive HBc-Y132A. It's essential to emphasize that R133 is the key residue in the HBc RP motif that governs their interaction with Par14/Par17. Chromatin immunoprecipitation conducted on HBV-infected cells elucidated the participation of residues S19 and E46/D74 in Par14 and S44 and E71/D99 in Par17 in the recruitment of 133RP134 motif-containing HBc into cccDNA. Depleting PIN4 in liver cell lines results in a significant reduction in cccDNA levels, pgRNA, sgRNAs, HBc, core particle assembly, and HBV DNA synthesis. Notably, parvulin inhibitors like juglone and PiB have proven to be effective in substantially reducing HBV replication. These inhibitors weaken the interaction between HBV core particles and Par14/Par17, underscoring the dynamic nature of this interaction. It's also worth noting that specific Par14/Par17 inhibitors hold promise as potential therapeutic options for chronic hepatitis B.

Keywords: Par14Par17, HBx, HBc, cccDNA, HBV

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63 Arbuscular Mycorrhizal Symbiosis in Trema orientalis: Effect of a Naturally-Occurring Symbiosis Receptor Kinase Mutant Allele

Authors: Yuda Purwana Roswanjaya, Wouter Kohlen, Rene Geurts

Abstract:

The Trema genus represents a group of fast-growing tropical tree species within the Cannabaceae. Interestingly, five species nested in this lineage -known as Parasponia- can establish rhizobium nitrogen-fixing root nodules, similar to those found in legumes. Parasponia and legumes use a conserved genetic network to control root nodule formation, among which are genes also essential for mycorrhizal symbiosis (the so-called common symbiotic pathway). However, Trema species lost several genes that function exclusively in nodulation, suggesting a loss-of the nodulation trait in Trema. Strikingly, in a Trema orientalis population found in Malaysian Borneo we identified a truncated SYMBIOSIS RECEPTOR KINASE (SYMRK) mutant allele lacking a large portion of the c-terminal kinase domain. In legumes this gene is essential for nodulation and mycorrhization. This raises the question whether Trema orientalis can still be mycorrhized. To answer this question, we established quantitative mycorrhization assay for Parasponia andersonii and Trema orientalis. Plants were grown in closed pots on half strength Hoagland medium containing 20 µM potassium phosphate in sterilized sand and inoculated with 125 spores of Rhizopagus irregularis (Agronutrion-DAOM197198). Mycorrhization efficiency was determined by analyzing the frequency of mycorrhiza (%F), the intensity of the mycorrhizal colonization (%M) and the arbuscule abundance (%A) in the root system. Trema orientalis RG33 can be mycorrhized, though with lower efficiency compared to Parasponia andersonii. From this we conclude that a functional SYMRK kinase domain is not essential for Trema orientalis mycorrhization. In ongoing experiments, we aim to investigate the role of SYMRK in Parasponia andersonii mycorrhization and nodulation. For this two Parasponia andersonii symrk CRISPR-Cas9 mutant alleles were created. One mimicking the TorSYMRKRG33 allele by deletion of exon 13-15, and a full Parasponia andersonii SYMRK knockout.

Keywords: endomycorrhization, Parasponia andersonii, symbiosis receptor kinase (SYMRK), Trema orientalis

Procedia PDF Downloads 135
62 Cryopreservation of Ring-Necked Pheasant (Phasianus colchicus) Semen for Establishing Cryobank

Authors: Rida Pervaiz, Bushra Allah Rakha, Muhammad Sajjad Ansari, Shamim Akhter, Kainat Waseem, Sumiyyah Zuha, Tooba Javed

Abstract:

Ring-necked pheasant (Phasianus colchicus) belongs to order Galliformes and family Phasianidae. It has been recognized as the most hunted bird due to its attractive colorful appearance and meat. Loss of habitat and hunting pressure has caused population fluctuations in the native range. Under these circumstances, this species can be conserved by employing ex-situ in vitro conservation techniques. Captive breeding, in combination with semen cryobanking is the most appropriate option to conserve/propagate this species without deteriorating the genetic diversity. Cryopreservation protocols of adequate efficiency are necessary to establish semen cryobanking for a species. Therefore, present study was designed to devise an efficient extender for cryopreservation of ring-necked pheasant semen. For this purpose, a range of extenders (Beltsville Poultry, red fowl, Lake, EK, Tselutin Poultry and Chicken semen extenders) were evaluated for cryopreservation of ring-necked pheasant semen. Semen collected from 10 cocks, diluted in the Beltsville Poultry (BPSE), Red Fowl (RFE), Lake (LE), EK (EKE), Tselutin Poultry (TPE) and Chicken Semen (CSE) extenders and cryopreserved. Glycerol (10%) was added to semen at 4°C, equilibrated for 10 min, filled in 0.5 mL French straws, kept over liquid nitrogen vapors for 10 min, cryopreserved in LN2 and stored. Sperm motility (%), viability (%), live/dead ratio (%), plasma membrane (%) and DNA Integrity (%) were evaluated at post-dilution, post-cooling, post-equilibration and post-thawing stage of cryopreservation. Sperm motility (83.8 ± 3.1; 81.3 ± 3.8; 73.8 ± 2.4; 62.5 ± 1.4), viability (79.0 ± 1.7; 75.5 ± 1.6; 69.5 ± 2.3; 65.5 ± 2.4), live/dead ratio (80.5 ± 5.7; 77.3 ± 4.9; 76.0 ± 2.7; 68.3 ± 2.3), plasma membrane (74.5 ± 2.9; 73.8 ± 3.4; 71.3 ± 2.3; 75.0 ± 3.4) and DNA integrity (78.3 ± 1.7; 73.0 ± 1.2; 68.0 ± 2.0; 63.0 ± 2.5) at all four stages of cryopreservation were recorded higher (P < 0.05) in red fowl extender compared to all experimental extenders. It is concluded that red fowl extender is the best extender for cryopreservation of ring-necked pheasant semen and can be used in establishing cryobank for ex situ conservation.

Keywords: ring-necked pheasant; extenders; cryopreservation; semen quality; DNA integrity

Procedia PDF Downloads 114
61 Role of Autophagic Lysosome Reformation for Cell Viability in an in vitro Infection Model

Authors: Muhammad Awais Afzal, Lorena Tuchscherr De Hauschopp, Christian Hübner

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Introduction: Autophagy is an evolutionarily conserved lysosome-dependent degradation pathway, which can be induced by extrinsic and intrinsic stressors in living systems to adapt to fluctuating environmental conditions. In the context of inflammatory stress, autophagy contributes to the elimination of invading pathogens, the regulation of innate and adaptive immune mechanisms, and regulation of inflammasome activity as well as tissue damage repair. Lysosomes can be recycled from autolysosomes by the process of autophagic lysosome reformation (ALR), which depends on the presence of several proteins including Spatacsin. Thus ALR contributes to the replenishment of lysosomes that are available for fusion with autophagosomes in situations of increased autophagic turnover, e.g., during bacterial infections, inflammatory stress or sepsis. Objectives: We aimed to assess whether ALR plays a role for cell survival in an in-vitro bacterial infection model. Methods: Mouse embryonic fibroblasts (MEFs) were isolated from wild-type mice and Spatacsin (Spg11-/-) knockout mice. Wild-type MEFs and Spg11-/- MEFs were infected with Staphylococcus aureus (multiplication of infection (MOI) used was 10). After 8 and 16 hours of infection, cell viability was assessed on BD flow cytometer through propidium iodide intake. Bacterial intake by cells was also calculated by plating cell lysates on blood agar plates. Results: in-vitro infection of MEFs with Staphylococcus aureus showed a marked decrease of cell viability in ALR deficient Spatacsin knockout (Spg11-/-) MEFs after 16 hours of infection as compared to wild-type MEFs (n=3 independent experiments; p < 0.0001) although no difference was observed for bacterial intake by both genotypes. Conclusion: Suggesting that ALR is important for the defense of invading pathogens e.g. S. aureus, we observed a marked increase of cell death in an in-vitro infection model in cells with compromised ALR.

Keywords: autophagy, autophagic lysosome reformation, bacterial infections, Staphylococcus aureus

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60 Structure and Mechanics Patterns in the Assembly of Type V Intermediate-Filament Protein-Based Fibers

Authors: Mark Bezner, Shani Deri, Tom Trigano, Kfir Ben-Harush

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Intermediate filament (IF) proteins-based fibers are among the toughest fibers in nature, as was shown by native hagfish slime threads and by synthetic fibers that are based on type V IF-proteins, the nuclear lamins. It is assumed that their mechanical performance stems from two major factors: (1) the transition from elastic -helices to stiff-sheets during tensile load; and (2) the specific organization of the coiled-coil proteins into a hierarchical network of nano-filaments. Here, we investigated the interrelationship between these two factors by using wet-spun fibers based on C. elegans (Ce) lamin. We found that Ce-lamin fibers, whether assembled in aqueous or alcoholic solutions, had the same nonlinear mechanical behavior, with the elastic region ending at ~5%. The pattern of the transition was, however, different: the ratio between -helices and -sheets/random coils was relatively constant until a 20% strain for fibers assembled in an aqueous solution, whereas for fibers assembled in 70% ethanol, the transition ended at a 6% strain. This structural phenomenon in alcoholic solution probably occurred through the transition between compacted and extended conformation of the random coil, and not between -helix and -sheets, as cycle analyses had suggested. The different transition pattern can also be explained by the different higher order organization of Ce-lamins in aqueous or alcoholic solutions, as demonstrated by introducing a point mutation in conserved residue in Ce-lamin gene that alter the structure of the Ce-lamins’ nano-fibrils. In addition, biomimicking the layered structure of silk and hair fibers by coating the Ce-lamin fiber with a hydrophobic layer enhanced fiber toughness and lead to a reversible transition between -helix and the extended conformation. This work suggests that different hierarchical structures, which are formed by specific assembly conditions, lead to diverse secondary structure transitions patterns, which in turn affect the fibers’ mechanical properties.

Keywords: protein-based fibers, intermediate filaments (IF) assembly, toughness, structure-property relationships

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59 Inviscid Steady Flow Simulation Around a Wing Configuration Using MB_CNS

Authors: Muhammad Umar Kiani, Muhammad Shahbaz, Hassan Akbar

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Simulation of a high speed inviscid steady ideal air flow around a 2D/axial-symmetry body was carried out by the use of mb_cns code. mb_cns is a program for the time-integration of the Navier-Stokes equations for two-dimensional compressible flows on a multiple-block structured mesh. The flow geometry may be either planar or axisymmetric and multiply-connected domains can be modeled by patching together several blocks. The main simulation code is accompanied by a set of pre and post-processing programs. The pre-processing programs scriptit and mb_prep start with a short script describing the geometry, initial flow state and boundary conditions and produce a discretized version of the initial flow state. The main flow simulation program (or solver as it is sometimes called) is mb_cns. It takes the files prepared by scriptit and mb_prep, integrates the discrete form of the gas flow equations in time and writes the evolved flow data to a set of output files. This output data may consist of the flow state (over the whole domain) at a number of instants in time. After integration in time, the post-processing programs mb_post and mb_cont can be used to reformat the flow state data and produce GIF or postscript plots of flow quantities such as pressure, temperature and Mach number. The current problem is an example of supersonic inviscid flow. The flow domain for the current problem (strake configuration wing) is discretized by a structured grid and a finite-volume approach is used to discretize the conservation equations. The flow field is recorded as cell-average values at cell centers and explicit time stepping is used to update conserved quantities. MUSCL-type interpolation and one of three flux calculation methods (Riemann solver, AUSMDV flux splitting and the Equilibrium Flux Method, EFM) are used to calculate inviscid fluxes across cell faces.

Keywords: steady flow simulation, processing programs, simulation code, inviscid flux

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58 Rangeland Monitoring by Computerized Technologies

Authors: H. Arzani, Z. Arzani

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Every piece of rangeland has a different set of physical and biological characteristics. This requires the manager to synthesis various information for regular monitoring to define changes trend to get wright decision for sustainable management. So range managers need to use computerized technologies to monitor rangeland, and select. The best management practices. There are four examples of computerized technologies that can benefit sustainable management: (1) Photographic method for cover measurement: The method was tested in different vegetation communities in semi humid and arid regions. Interpretation of pictures of quadrats was done using Arc View software. Data analysis was done by SPSS software using paired t test. Based on the results, generally, photographic method can be used to measure ground cover in most vegetation communities. (2) GPS application for corresponding ground samples and satellite pixels: In two provinces of Tehran and Markazi, six reference points were selected and in each point, eight GPS models were tested. Significant relation among GPS model, time and location with accuracy of estimated coordinates was found. After selection of suitable method, in Markazi province coordinates of plots along four transects in each 6 sites of rangelands was recorded. The best time of GPS application was in the morning hours, Etrex Vista had less error than other models, and a significant relation among GPS model, time and location with accuracy of estimated coordinates was found. (3) Application of satellite data for rangeland monitoring: Focusing on the long term variation of vegetation parameters such as vegetation cover and production is essential. Our study in grass and shrub lands showed that there were significant correlations between quantitative vegetation characteristics and satellite data. So it is possible to monitor rangeland vegetation using digital data for sustainable utilization. (4) Rangeland suitability classification with GIS: Range suitability assessment can facilitate sustainable management planning. Three sub-models of sensitivity to erosion, water suitability and forage production out puts were entered to final range suitability classification model. GIS was facilitate classification of range suitability and produced suitability maps for sheep grazing. Generally digital computers assist range managers to interpret, modify, calibrate or integrating information for correct management.

Keywords: computer, GPS, GIS, remote sensing, photographic method, monitoring, rangeland ecosystem, management, suitability, sheep grazing

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57 Growth Stimulating Effects of Aspilia africana Fed to Female Pseudo-Ruminant Herbivores (Rabbits) at Different Physiological States

Authors: Nseabasi Nsikakabasi Etim

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In recent times, there has been a significant shortfall in between the production and supply of animal protein to meet the ever increasing population. To meet the increasing demand for animal protein, there is a need to focus attention on the production of livestock whose nutritional requirement does not put much strain on the limited sources of feed ingredients to which men subscribe. An example of such livestock is the rabbit. Rabbit is a pseudo-ruminant herbivore which utilizes much undigested and unabsorbed feed materials as sources of nutrient for maintenance and production. Thus, this study was conducted to investigate the effects of feeding Aspilia africana as forage on the growth rates of female pseudo-ruminant herbivores (rabbits) at different physiological states. Thirty (30) Dutch breed rabbit does of 5–6 months of age were used for the experiment which was conducted in a completely randomized design for four months. The rabbits were divided into three treatment groups, ten does per treatment group; which consisted of mixed forages (Centrosema pubescent (200g), Panicum maximum (200g) and Ipomea batatas leaves (100g) without Aspilia africana (T1; control), fresh Aspilia africana (500g/dose/day) (T2) and wilted Aspilia africana (500g/dose/day) (T3). Rabbits in all treatment groups received the same concentrate (300g/animal/day) throughout the period of the study and mixed forages from the commencement of the experiment till the does kindled. After parturition, fresh and wilted Aspilia africana were introduced in treatments 2 and three respectively, whereas the control group continued on mixed forages throughout the study. The result of the study revealed that the initial average body weight of the rabbit does was 1.74kg. At mating and gestation periods, the body weights of the does in T2 was significantly higher (P<0.05) than the rest. There were no significant differences (P<0.05) in the body weights of does at kindling between the various treatment groups. During the physiological states of lactation, weaning and re-mating, the control group (T1) had significantly lower body weight than those of the treated groups (T2 and T3). Furthermore, T2 had significantly higher body weight than T3. The study revealed that Aspilia africana; mainly the fresh leaves have greater growth stimulating effects when fed to pseudo-ruminants (rabbits), thereby enhancing body weights of does during lactation and weaning.

Keywords: Aspilia africana, herbivores, pseudo-ruminants, physiological states

Procedia PDF Downloads 654
56 Modeling Spatio-Temporal Variation in Rainfall Using a Hierarchical Bayesian Regression Model

Authors: Sabyasachi Mukhopadhyay, Joseph Ogutu, Gundula Bartzke, Hans-Peter Piepho

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Rainfall is a critical component of climate governing vegetation growth and production, forage availability and quality for herbivores. However, reliable rainfall measurements are not always available, making it necessary to predict rainfall values for particular locations through time. Predicting rainfall in space and time can be a complex and challenging task, especially where the rain gauge network is sparse and measurements are not recorded consistently for all rain gauges, leading to many missing values. Here, we develop a flexible Bayesian model for predicting rainfall in space and time and apply it to Narok County, situated in southwestern Kenya, using data collected at 23 rain gauges from 1965 to 2015. Narok County encompasses the Maasai Mara ecosystem, the northern-most section of the Mara-Serengeti ecosystem, famous for its diverse and abundant large mammal populations and spectacular migration of enormous herds of wildebeest, zebra and Thomson's gazelle. The model incorporates geographical and meteorological predictor variables, including elevation, distance to Lake Victoria and minimum temperature. We assess the efficiency of the model by comparing it empirically with the established Gaussian process, Kriging, simple linear and Bayesian linear models. We use the model to predict total monthly rainfall and its standard error for all 5 * 5 km grid cells in Narok County. Using the Monte Carlo integration method, we estimate seasonal and annual rainfall and their standard errors for 29 sub-regions in Narok. Finally, we use the predicted rainfall to predict large herbivore biomass in the Maasai Mara ecosystem on a 5 * 5 km grid for both the wet and dry seasons. We show that herbivore biomass increases with rainfall in both seasons. The model can handle data from a sparse network of observations with many missing values and performs at least as well as or better than four established and widely used models, on the Narok data set. The model produces rainfall predictions consistent with expectation and in good agreement with the blended station and satellite rainfall values. The predictions are precise enough for most practical purposes. The model is very general and applicable to other variables besides rainfall.

Keywords: non-stationary covariance function, gaussian process, ungulate biomass, MCMC, maasai mara ecosystem

Procedia PDF Downloads 260
55 Characterization of a Putative Type 1 Toxin-Antitoxin System in Shigella Flexneri

Authors: David Sarpong, Waleed Khursheed, Ernest Danquah, Erin Murphy

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Shigella is a pathogenic bacterium responsible for shigellosis, a severe diarrheal disease that claims the lives of immunocompromised individuals worldwide. To develop therapeutics against this disease, an understanding of the molecular mechanisms underlying the pathogen’s physiology is crucial. Small non-coding RNAs (sRNAs) have emerged as important regulators of bacterial physiology, including as components of toxin-antitoxin systems. In this study, we investigated the role of RyfA in S. flexneri physiology and virulence. RyfA, originally identified as an sRNA in Escherichia coli, is conserved within the Enterobacteriaceae family, including Shigella. Whereas two copies of ryfA are present in S. dysenteriae, all other Shigella species contain only one copy of the gene. Additionally, we identified a putative open reading frame within the RyfA transcript, suggesting that it may be a dual-functioning gene encoding a small protein in addition to its sRNA function. To study ryfA in vitro, we cloned the gene into an inducible plasmid and observed the effect on bacterial growth. Here, we report that RyfA production inhibits the growth of S. flexneri, and this inhibition is dependent on the contained open reading frame. In-silico analyses have revealed the presence of two divergently transcribed sRNAs, RyfB1 and RyfB2, which share nucleotide complementarity with RyfA and thus are predicted to function as anti-toxins. Our data demonstrate that RyfB2 has a stronger antitoxin effect than RyfB1. This regulatory pattern suggests a novel form of a toxin-antitoxin system in which the activity of a single toxin is inhibited to varying degrees by two sRNA antitoxins. Studies are ongoing to investigate the regulatory mechanism(s) of the antitoxin genes, as well as the downstream targets and mechanism of growth inhibition by the RyfA toxin. This study offers distinct insights into the regulatory mechanisms underlying Shigella physiology and may inform the development of new anti-Shigella therapeutics.

Keywords: sRNA, shigella, toxin-antitoxin, Type 1 toxin antitoxin

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54 Unravelling of the TOR Signaling Pathway in Human Fungal Pathogen Cryptococcus neoformans

Authors: Yee-Seul So, Guiseppe Ianiri, Alex Idnurm, Yong-Sun Bahn

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Tor1 is a serine/threonine protein kinase that is widely conserved across eukaryotic species. Tor1 was first identified in Saccharomyces cerevisiae as a target of rapamycin (TOR). The TOR pathway has been implicated in regulating cellular responses to nutrients, proliferation, translation, transcription, autophagy, and ribosome biogenesis. Here we identified two homologues of S. cerevisiae Tor proteins, CNAG_06642 (Tor1) and CNAG_05220 (Tlk1, TOR-like kinase 1), in Cryptococcus neoformans causing a life-threatening fungal meningoencephalitis. Both Tor1 and Tlk1 have rapamycin-binding (RB) domains but Tlk1 has truncated RB form. To study the TOR-signaling pathway in the fungal pathogen, we attempt to construct the tor1Δ and tlk1Δ mutants and phenotypically analyze them. Although we failed to construct the tor1Δ mutant, we successfully construct the tlk1Δ mutant. The tlk1Δ mutant does not exhibit any discernable phenotypes, suggesting that Tlk1 is dispensable in C. neoformans. The essentiality of TOR1 is independently confirmed by constructing the TOR1 promoter replacement strain by using a copper transporter 4 (CTR4) promoter and the TOR1/tor1 heterozygous mutant in diploid C. neoformans strain background followed by sporulation analysis. To further analyze the function of Tor1, we construct TOR1 overexpression mutant using a constitutively active histone H3 in C. neoformans. We find that the Tor1 overexpression mutant is resistant to rapamycin but the tlk1Δ mutant does not exhibit any altered resistance to rapamycin, further confirming that Tor1, but not Tlk1, is critical for TOR signaling. Furthermore, we found that Tor1 is involved in response to diverse stresses, including genotoxic stress, oxidative stress, thermo-stress, antifungal drug treatment, and production of melanin. To identify any TOR-related transcription factors, we screened C. neoformans transcription factor library that we constructed in our previous study and identified several potential downstream factors of Tor1, including Atf1, Crg1 and Bzp3. In conclusion, the current study provides insight into the role of the TOR signaling pathway in human fungal pathogens as well as C. neoformans.

Keywords: fungal pathogen, serine/threonine kinase, target of rapamycin, transcription factor

Procedia PDF Downloads 195
53 A Rapid Colorimetric Assay for Direct Detection of Unamplified Hepatitis C Virus RNA Using Gold Nanoparticles

Authors: M. Shemis, O. Maher, G. Casterou, F. Gauffre

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Hepatitis C virus (HCV) is a major cause of chronic liver disease with a global 170 million chronic carriers at risk of developing liver cirrhosis and/or liver cancer. Egypt reports the highest prevalence of HCV worldwide. Currently, two classes of assays are used in the diagnosis and management of HCV infection. Despite the high sensitivity and specificity of the available diagnostic assays, they are time-consuming, labor-intensive, expensive, and require specialized equipment and highly qualified personal. It is therefore important for clinical and economic terms to develop a low-tech assay for the direct detection of HCV RNA with acceptable sensitivity and specificity, short turnaround time, and cost-effectiveness. Such an assay would be critical to control HCV in developing countries with limited resources and high infection rates, such as Egypt. The unique optical and physical properties of gold nanoparticles (AuNPs) have allowed the use of these nanoparticles in developing simple and rapid colorimetric assays for clinical diagnosis offering higher sensitivity and specificity than current detection techniques. The current research aims to develop a detection assay for HCV RNA using gold nanoparticles (AuNPs). Methods: 200 anti-HCV positive samples and 50 anti-HCV negative plasma samples were collected from Egyptian patients. HCV viral load was quantified using m2000rt (Abbott Molecular Inc., Des Plaines, IL). HCV genotypes were determined using multiplex nested RT- PCR. The assay is based on the aggregation of AuNPs in presence of the target RNA. Aggregation of AuNPs causes a color shift from red to blue. AuNPs were synthesized using citrate reduction method. Different sets of probes within the 5’ UTR conserved region of the HCV genome were designed, grafted on AuNPs and optimized for the efficient detection of HCV RNA. Results: The nano-gold assay could colorimetrically detect HCV RNA down to 125 IU/ml with sensitivity and specificity of 91.1% and 93.8% respectively. The turnaround time of the assay is < 30 min. Conclusions: The assay allows sensitive and rapid detection of HCV RNA and represents an inexpensive and simple point-of-care assay for resource-limited settings.

Keywords: HCV, gold nanoparticles, point of care, viral load

Procedia PDF Downloads 180
52 Active Packaging Films Based on Chitosan Incorporated with Thyme Essential Oil and Cross Linkers and Its Effect on the Quality Shelf Life of Food

Authors: Aiman Zehra, Sajad Mohd Wani

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Packaging has a vital role as it contains and protects the food that moves from the supply chain to the consumer. Chitosan (CH) has been extensively used in food packaging applications among the plentiful natural macromolecules, including all the polysaccharide class, owing to its easy film-forming capacity, biodegradability, better oxygen and water vapour barrier ability and good mechanical strength. Compared to synthetic films, the films produced from chitosan present poor barrier and mechanical properties. To overcome its deficient qualities, a number of modification procedures are required to enhance the mechanical and physical properties. Various additives such as plasticizers (e.g., glycerol and sorbitol), crosslinkers (e.g.,CaCl₂, ZnO), fillers (nanoclay), and antimicrobial agents (e.g. thyme essential oil) have been used to improve the mechanical, thermal, morphological, antimicrobial properties and emulsifying agents for the stability and elasticity of chitosan-based biodegradable films. Different novel biocomposite films based on chitosan incorporated with thyme essential oil and different additives (ZnO, CaCl₂, NC, and PEG) were successfully prepared and used as packaging material for carrot candy. The chitosan film incorporated with crosslinkers was capable of forming a protective barrier on the surface of the candy to maintain moisture content, water activity, TSS, total sugars, and titratable acidity. ZnO +PEG +NC +CaCl₂ remarkably promotes a synergistic effect on the barrier properties of the film. The combined use of ZnO +PEG +NC +CaCl₂ in CH-TO films was more effective in preventing the moisture gain in candies. The lowest a𝓌 (0.624) was also observed for the candies stored in treatment. The color values L*, a*, b* of the candies were also retained in the film containing all the additives during the 6th month of storage. The value for L*, a*, and b* observed for T was 42.72, 9.89, and 10.84, respectively. The candies packaged in film retained TSS and acidity. The packaging film significantly p≤0.05 conserved sensory qualities and inhibited microbial activity during storage. Carrot candy was found microbiologically safe for human consumption even after six months of storage in all the packaging materials.

Keywords: chitosan, biodegradable films, antimicrobial activity, thyme essential oil, crosslinkers

Procedia PDF Downloads 72
51 Organic Farming for Sustainable Production of Some Promising Halophytic Species in Saline Environment

Authors: Medhat Tawfik, Ezzat Abd El Lateef, Bahr Amany, Mohamed Magda

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Applying organic farming systems in biosaline agriculture is unconventional approach for sustainable use of marginal soil and desert land for planting non-traditional halophytic crops such as Leptochloa fusca, Kochia indica, Sporobolus virginicus and Spartina patens. These plants are highly salt tolerant C4 halophytic forage plants grown well in coastal salt marsh. These halophytic plant will take important place in the farming system, especially in the coastal areas and salt-affected land. We can call it environmentally smart crops because they ensure food security, contribute to energy security, guarantee environmental sustainability, and mitigate the negative impacts of climate change. Organic Agriculture is the most important and widely practiced agro-ecological farming system. It is claimed to be the most sustainable approach and long term adaptation strategy. It promotes soil fertility and diversity at all levels and makes soils less susceptible to erosion. It is also reported to be climate change resilience farming systems as it promotes the proper management of soil, water, biodiversity and local knowledge and provides producers with ecologically sound management decisions. A field experiment was carried out at the Model Farm of National Research Centre, El Tour, South Sinai to study the impact of (Mycorrhiza 1kg/fed., charcoal 4 tons/fed., chicken manure 5 tons/fed., in addition to control treatment) on some growth characters, photosynthetic pigments content, and some physiological aspects i.e. prolind and soluble carbohydrates content, succulence and osmotic pressure values, as well as nutritive values i.e. Crude fat (CF), Acid detergent fiber (ADF), Neutral detergent fiber (NDF), Ether extract (EE) and Nitrogen-free extract (NFE) of five halophytic plant species (Leptochloa fusca, Kochia indica, Sporobolus virginicus and Spartina patens). Our results showed that organic fertilizer treatment enhanced all the previous character as compared with control with superiority to chicken manure over the other treatments.

Keywords: organic agriculture, halophytic plants, saline environment, water security

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50 Significance of Molecular Autophagic Pathway in Gaucher Disease Pathology

Authors: Ozlem Oral, Emre Taskin, Aysel Yuce, Serap Dokmeci, Devrim Gozuacik

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Autophagy is an evolutionary conserved lysosome-dependent catabolic pathway, responsible for the degradation of long-lived proteins, abnormal aggregates and damaged organelles which cannot be degraded by the ubiquitin-proteasome system. Lysosomes degrade the substrates through the activity of lysosomal hydrolases and lysosomal membrane-bound proteins. Mutations in the coding region of these proteins cause malfunctional lysosomes, which contributes to the pathogenesis of lysosomal storage diseases. Gaucher disease is a lysosomal storage disease resulting from the mutation of a lysosomal membrane-associated glycoprotein called glucocerebrosidase and its cofactor saposin C. The disease leads to intracellular accumulation of glucosylceramide and other glycolipids. Because of the essential role of lysosomes in autophagic degradation, Gaucher disease may directly be linked to this pathway. In this study, we investigated the expression of autophagy and/or lysosome-related genes and proteins in fibroblast cells isolated from patients with different mutations. We carried out confocal microscopy analysis and examined autophagic flux by utilizing the differential pH sensitivities of RFP and GFP in mRFP-GFP-LC3 probe. We also evaluated lysosomal pH by active lysosome staining and lysosomal enzyme activity. Beside lysosomes, we also performed proteasomal activity and cell death analysis in patient samples. Our data showed significant attenuation in the expression of key autophagy-related genes and accumulation of their proteins in mutant cells. We found decreased the ability of autophagosomes to fuse with lysosomes, associated with elevated lysosomal pH and reduced lysosomal enzyme activity. Proteasomal degradation and cell death analysis showed reduced proteolytic activity of the proteasome, which consequently leads to increased susceptibility to cell death. Our data indicate that the major degradation pathways are affected by multifunctional lysosomes in mutant patient cells and may underlie in the mechanism of clinical severity of Gaucher patients. (This project is supported by TUBITAK-3501-National Young Researchers Career Development Program, Project No: 112T130).

Keywords: autophagy, Gaucher's disease, glucocerebrosidase, mutant fibroblasts

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49 Sequence Analysis and Molecular Cloning of PROTEOLYSIS 6 in Tomato

Authors: Nurulhikma Md Isa, Intan Elya Suka, Nur Farhana Roslan, Chew Bee Lynn

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The evolutionarily conserved N-end rule pathway marks proteins for degradation by the Ubiquitin Proteosome System (UPS) based on the nature of their N-terminal residue. Proteins with a destabilizing N-terminal residue undergo a series of condition-dependent N-terminal modifications, resulting in their ubiquitination and degradation. Intensive research has been carried out in Arabidopsis previously. The group VII Ethylene Response Factor (ERFs) transcription factors are the first N-end rule pathway substrates found in Arabidopsis and their role in regulating oxygen sensing. ERFs also function as central hubs for the perception of gaseous signals in plants and control different plant developmental including germination, stomatal aperture, hypocotyl elongation and stress responses. However, nothing is known about the role of this pathway during fruit development and ripening aspect. The plant model system Arabidopsis cannot represent fleshy fruit model system therefore tomato is the best model plant to study. PROTEOLYSIS6 (PRT6) is an E3 ubiquitin ligase of the N-end rule pathway. Two homologs of PRT6 sequences have been identified in tomato genome database using the PRT6 protein sequence from model plant Arabidopsis thaliana. Homology search against Ensemble Plant database (tomato) showed Solyc09g010830.2 is the best hit with highest score of 1143, e-value of 0.0 and 61.3% identity compare to the second hit Solyc10g084760.1. Further homology search was done using NCBI Blast database to validate the data. The result showed best gene hit was XP_010325853.1 of uncharacterized protein LOC101255129 (Solanum lycopersicum) with highest score of 1601, e-value 0.0 and 48% identity. Both Solyc09g010830.2 and uncharacterized protein LOC101255129 were genes located at chromosome 9. Further validation was carried out using BLASTP program between these two sequences (Solyc09g010830.2 and uncharacterized protein LOC101255129) to investigate whether they were the same proteins represent PRT6 in tomato. Results showed that both proteins have 100 % identity, indicates that they were the same gene represents PRT6 in tomato. In addition, we used two different RNAi constructs that were driven under 35S and Polygalacturonase (PG) promoters to study the function of PRT6 during tomato developmental stages and ripening processes.

Keywords: ERFs, PRT6, tomato, ubiquitin

Procedia PDF Downloads 217
48 The Effect of Alternative Organic Fertilizer and Chemical Fertilizer on Nitrogen and Yield of Peppermint (Mentha peperita)

Authors: Seyed Ali Mohammad, Modarres Sanavy, Hamed Keshavarz, Ali Mokhtassi-Bidgoli

Abstract:

One of the biggest challenges for the current and future generations is to produce sufficient food for the world population with the existing limited available water resources. Peppermint is a specialty crop used for food and medicinal purposes. Its main component is menthol. It is used predominantly for oral hygiene, pharmaceuticals, and foods. Although drought stress is considered as a negative factor in agriculture, being responsible for severe yield losses; medicinal plants grown under semi-arid conditions usually produce higher concentrations of active substances than same species grown under moderate climates. Nitrogen (N) fertilizer management is central to the profitability and sustainability of forage crop production. Sub-optimal N supply will result in poor yields, and excess N application can lead to nitrate leaching and environmental pollution. In order to determine the response of peppermint to drought stress and different fertilizer treatments, a field experiment with peppermint was conducted in a sandy loam soil at a site of the Tarbiat Modares University, Agriculture Faculty, Tehran, Iran. The experiment used a complete randomized block design, with six rates of fertilizer strategies (F1: control, F2: Urea, F3: 75% urea + 25% vermicompost, F4: 50% urea + 50% vermicompost, F5: 25% urea + 75% vermicompost and F6: vermicompost) and three irrigation regime (S1: 45%, S2: 60% and S3: 75% FC) with three replication. The traits such as nitrogen, chlorophyll, carotenoids, anthocyanin, flavonoid and fresh biomass were studied. The results showed that the treatments had a significant effect on the studied traits as drought stress reduced photosynthetic pigment concentration. Also, drought stress reduced fresh yield of peppermint. Non stress condition had the greater amount of chlorophyll and fresh yield more than other irrigation treatments. The highest concentration of chlorophyll and the fresh biomass was obtained in F2 fertilizing treatments. Sever water stress (S1) produced decreased photosynthetic pigment content fresh yield of peppermint. Supply of N could improve photosynthetic capacity by enhancing photosynthetic pigment content. Perhaps application of vermicompost significantly improved the organic carbon, available N, P and K content in soil over urea fertilization alone. To get sustainable production of peppermint, application of vermicompost along with N through synthetic fertilizer is recommended for light textured sandy loam soils.

Keywords: fresh yield, peppermint, synthetic nitrogen, vermicompost, water stress

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