Search results for: protein complementation assay
Commenced in January 2007
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Paper Count: 3222

Search results for: protein complementation assay

642 Interaction of GCN5L1 with WHAMM and KIF5B Regulates Autolysosome Tubulation

Authors: Allen Seylani

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Lysosome-dependent autophagy is a nutrient-deprivation-induced evolutionarily conserved intracellular recycling program that sequestrates intracellular cargo into autophagosomes (AP), which then fuse with lysosomes to form autolysosomes (ALs) for cargo digestion. To restore free lysosomes, autophagic lysosome reformation (ALR) is initiated by extrusion of tubular structures from autolysosomes at the final stage of autophagy, in a process called lysosomal tubulation (LT). This project aimed to investigate the molecular mechanism of GCN5L1 in LT and the following lysosomal signaling. GCN5L1 belongs to the BORC multiprotein complexes and is involved in controlling lysosomal trafficking; however, the effect of GCN5L1 on lysosome tubulation remains largely unknown. Genetic ablation of GCN5L1 in the mouse primary hepatocytes showed dramatically increased autolysosomes (ALs), decreased lysosome regeneration and absence of lysosomal tubulation. This phenotype suggests the possibility of disruption in lysosome tubulation, which results in the disturbance of the overall lysosome homeostasis. The formation of tubulars from ALs requires kinesin motor protein KIF5B. Immunoprecipitation was employed and confirmed the interaction of GCN5L1 with the ARL8B-KIF5B complex, which recruited KIF5B to ALs. At the same time, GCN5L1 interacted with WHAMM, which promotes the actin nucleation factor, which brings actin cytoskeleton to ALs and initiates LT. Furthermore, impaired LT in GCN5L1 deficient hepatocytes was restored by overexpression of GCN5L1, and this rescue effect was attenuated by knockdown of KIF5B. Additionally, lysosomal mTORC1 activity was upregulated in GCN5L1-/- hepatocytes, while inhibition of mTORC1 abrogated the GCN5L1 mediated rescue of LT in knockout hepatocytes. Altogether these findings revealed a novel mechanism of ALR, in which a simultaneous interaction of GCN5L1 with KIF5B and WHAMM is required for LT and downstream mTORC1 signaling.

Keywords: autophagy, autolysosome, GCN5L1, lysosome

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641 Influence of Dietary Herbal Blend on Crop Filling, Growth Performance and Nutrient Digestibility in Broiler Chickens

Authors: S. Ahmad, M. Rizwan, B. Ayub, S. Mehmood, P. Akhtar

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This experiment was conducted to investigate the effect of supplementation of pure herbal blend on growth performance of boilers. One hundred and twenty birds were randomly distributed into 4 experimental units of 3 replicates (10 birds/replicate) as: negative control (basal diet), positive control (Lincomycin at the rate of 5g/bag), pure herbal blend at the rate of 150g/bag and pure herbal blend at the rate of 300g/bag. The data regarding weekly feed intake, body weight gain and feed conversion ratio were recorded, and fecal samples were collected at the end of starter and finisher phase for nutrient digestibility trial. The results of feed intake showed significant (P < 0.05) results in 1st (305g), 2nd (696.88g), 3rd (1046.9g) and 4th (1173.2g) week and feed conversion ratio indicated significant (P < 0.05) variations in 1st (2.54) and 4th (2.28) week of age. Also, both starter and finisher phase indicated significant (P < 0.05) differences among all treatment groups in feed intake (2023.4g) and (2302.6g) respectively. The statistical analysis indicated significant (P < 0.05) results in crop filling percentage (86.6%) after 2 hours of first feed supplementation. In case of nutrient digestibility trial, results showed significant (P < 0.05) values of crude protein and crude fat in starter phase as 69.65% and 56.62% respectively, and 69.57% and 48.55% respectively, in finisher phase. Based on overall results, it was concluded that the dietary inclusion of pure herbal blend containing neem tree leaves powder, garlic powder, ginger powder and turmeric powder increase the production performance of broilers.

Keywords: neem tree leave, garlic, ginger, herbal blend, broiler

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640 Management of Pressure Ulcer with a Locally Constructed Negative Pressure Device (NPD) in Traumatic Paraplegia Patients: A Randomized Controlled Clinical Trial

Authors: Mukesh K. Dwivedi, Rajeshwar N. Srivastava, Amit K. Bhagat, Saloni Raj

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Introduction: Management of Pressure Ulcer (PU) is an ongoing clinical challenge particularly in traumatic paraplegia patients in developing countries where socio economic conditions often dictate treatment modalities. When negative pressure wound therapy (NPWT) was introduced, there were a series of devices (V.A.C., KCI, San Antonio, TX) manufactured. These devices for NPWT are costly and hard to afford by patients in developing countries like India. Considering this limitation, this study was planned to design an RCT to compare NPWT by an indigenized locally constructed NPD and conventional gauze dressing for the treatment of PU. Material and Methods: This RCT (CTRI/2014/09/0050) was conducted in the Department of Orthopaedic Surgery at King George’s Medical University (KGMU), India. Thirty-four (34) subjects of traumatic paraplegia having PU of stage 3 or 4, were enrolled and randomized in two treatment groups (NPWT Group & Conventional dressing group). The outcome measures of this study were surface area and depth of PU, exudates, microorganisms and matrix metalloproteinase-8 (MMP-8) during 0 to 9 weeks follow-ups. Levels of MMP-8 were analyzed in the tissues of PU at week 0, 3, 6 and week 9 by Enzyme Linked Immuno Sorbent Assay (ELISA). Results: Significantly reduced length of PU in NPWT group was observed at week 6 (p=0.04) which further reduced at week 9 (p=0.001) as compared to conventionally treated group. Similarly significant reduction of width and depth of PU was observed in NPWT at week 9 (p<0.05). The exudate became significantly (p=0.001) lower in NPWT group as compared with conventionally treated group from 6th to 9th week. Clearance and conversion of slough into red granulation tissue was significantly higher in NPWT group (p=0.001). At week 9, the wound culture was negative in all the subjects of NPWT group, while it was positive in 10 (41⋅6%) subjects of conventional group. Significantly lower level of MMP-8 was observed in subjects of NPWT group at week 6 (0.006**), and continually more reduction was observed at week 9 (<0.0001**) as compared to the conventional group. Conclusion: NPWT by locally constructed NPD is better wound care procedure for management of PU. Our device gave similar results as commercially available devices. Reduction of level of MMP-8 and increased rate of healing was achieved by negative pressure wound therapy (NPWT) as compared to conventional dressing.

Keywords: NPWT, NPD, MMP8, ELISA

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639 Cerebrum Maturity Damage Induced by Fluoride in Suckling Mice

Authors: Hanen Bouaziz, Françoise Croute, Najiba Zeghal

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In order to investigate the toxic effects of fluoride on cerebrum maturity of suckling mice, we treated adult female mice of Swiss Albinos strain by 500 ppm NaF in their drinking water from the 15th day of pregnancy until the day 14 after delivery. All mice were sacrificed on day 14 after parturition. During treatment, levels of thiobarbituric acid reactive substances, the marker of lipid peroxidation extend, increased, while the activities of the antioxidant enzymes such as glutathione peroxidase, superoxide dismutase and catalase and the level of glutathione decreased significantly in cerebellum compared with those of the control group. These results suggested that fluoride enhanced oxidative stress, thereby disturbing the antioxidant defense of nursing pups. In addition, acetylcholinesterase activity in cerebellum was inhibited after treatment with fluoride. In cerebellum of mice, migration of neurons from the external granular layer to the internal granular layer occurred postnatally. Key guidance signals to these migrating neurons were provided by laminin, an extracellular matrix protein fixed to the surface of astrocytes. In the present study, we examined the expression and distribution of laminin in cerebellum of 14-day-old mice. Immunoreactive laminin was disappeared by postnatal day 14 in cerebellum parenchyma of control pups and was restricted to vasculature despite the continued presence of granular cells in the external granular layer. In contrast, in cerebellum of NaF treated pups, laminin was deposited in organised punctuate clusters in the molecular layer. These data indicated that the disruption of laminin distribution might play a major role in the profound derangement of neuronal migration observed in cerebellum of NaF treated pups.

Keywords: acetylcholinesterase activity, cerebellum, laminin, oxidative stress, suckling mice

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638 Impact of Length of Straw by the Use of a Straw Mill on the Selective Feeding of Young Cattle and Their Effects for the Cattle

Authors: Heiko Scholz

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When feeding high qualitysilagetoheifersfromthe age of two, there is a riskofenergyoversupply. Depending on the feeding valueorscarceavailability ofsilageorcorn silage diets withhighproportionsof straw is often incorporated. Foran energetically standardized young cattle supply of strawproportion can be more than 20% of dry matter. It was investigated whether the grinding of straw with the strawmillselective feeding significantly limits. The investigation has been carried out with young cattle in the second year. 78 animals were kept and fed under similar conditions in two groups. The experimental group (EG) consisted of cattle 12 to 15 months, and in the control group (CG), the cattle were 15 to 20 months old. The experimental feeding took place in five days of feed distribution, and residual feed were weighed. The ration of EG contained ground with the straw mill straw, and CG was further fed rotor-cut pressed straw. To determine the selective seizure samples of feed distributionandtheremainingfood with the particle separator boxandthecrude protein-and energy-content have been determined. The grinding of the straw increased the daily feed intake.IntheEGan increase infeed intakewas observedby grinding of the straw. Feed intakedirectlyon the day for changing the dietoflongonground straw increased by more than 2.0 kgofDMper animal. In the following days, the feed intakewasincreasedby 0.9kg DMper animal and day on average (7.4 vs. 8.3 kg DM per day). The results of the screen distribution of residual feed point to a differentiated feeding behavior between the groups. In the EG, the particle length of the residual feed to a large extent with the template matches. The acid-base-balance (NSBA)valuesofEGarewithin normal limits. Ifstrawsharesof25% and more are federations to young cattle (heifers), the theparticlelengthof straw has significant impact ontheselectivefeeding behavior. Aparticlelength of 1.5cmcompared to7.5 cmlongpreventedstrawcertainly discarding of the straw on the feeding barn. The feed intake increases whenshortstrawis mixed into theTMR.

Keywords: straw mill, heifer, feed selection, dry matter intake

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637 Antibacterial and Anti-Biofilm Activity of Vaccinium meridionale S. Pomace Extract Against Staphylococcus aureus, Escherichia coli and Salmonella Enterica

Authors: Carlos Y. Soto, Camila A. Lota, G. Astrid Garzón

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Bacterial biofilms cause an ongoing problem for food safety. They are formed when microorganisms aggregate to form a community that attaches to solid surfaces. Biofilms increase the resistance of pathogens to cleaning, disinfection and antibacterial products. This resistance gives rise to problems for human health, industry, and agriculture. At present, plant extracts rich in polyphenolics are being investigated as natural alternatives to degrade bacterial biofilms. The pomace of the tropical Berry Vaccinium meridionale S. contains high amounts of phenolic compounds. Therefore, in the current study, the antimicrobial and antibiofilm effects of extracts from the pomace of Vaccinium meridionale S. were tested on three foodborne pathogens: Enterohaemorrhagic Escherichia coli O157:H7 (ATCC®700728TM), Staphylococcus aureus subsp. aureus (ATCC® 6538TM), and Salmonella enterica serovar Enteritidis (ATCC® 13076TM). Microwave-assisted extraction was used to extract polyphenols with aqueous methanol (80% v/v) at a solid to solvent ratio of 1:10 (w/v) for 20 min. The magnetic stirring was set at 400 rpm, and the microwave power was adjusted to 400 W. The antimicrobial effect of the extract was assessed by determining the half maximal inhibitory concentration (IC50) against the three food poisoning pathogens at concentrations ranging from 50 to 2,850 μg gallic acid equivalents (GAE)/mL of the extract. Biofilm inhibition was assessed using a crystal violet assay applying the same range of concentration. Three replications of the experiments were carried out, and all analyses were run in triplicate. IC50 values were determined using the GraphPad Prism8® program. Significant differences (P<0.05) among means were identified using one-factor analysis of variance (ANOVA) and the post-hoc least significant difference (LSD) test using the Statgraphics plus program, version 2.1.There was significant difference among the mean IC50 values for the tested bacteria. The IC50 for S. aureus was 48 ± 9 μg GAE/mL, followed by 123 ± 49 μg GAE/mL for Salmonella and 376 ± 32 μg GAE/mL for E. coli. The percent inhibition of the extract on biofilm formation was significantly higher for S. aureus (85.8  0.3), followed by E. coli (74.5  1.0) and Salmonella (53.6  9.7). These findings suggest that polyphenolic extracts obtained from the pomace of V. meridionale S. might be used as natural antimicrobial and anti-biofilm natural agents, effective against S. aureus, E. coli and Salmonella enterica.

Keywords: antibiofilm, antimicrobial, E. coli, S. aureus, salmonella, IC50, pomace, V. meridionale

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636 Immuno-Modulatory Role of Weeds in Feeds of Cyprinus Carpio

Authors: Vipin Kumar Verma, Neeta Sehgal, Om Prakash

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Cyprinus carpio has a wide spread occurrence in the lakes and rivers of Europe and Asia. Heavy losses in natural environment due to anthropogenic activities, including pollution as well as pathogenic diseases have landed this fish in IUCN red list of vulnerable species. The significance of a suitable diet in preserving the health status of fish is widely recognized. In present study, artificial feed supplemented with leaves of two weed plants, Eichhornia crassipes and Ricinus communis were evaluated for their role on the fish immune system. To achieve this objective fish were acclimatized to laboratory conditions (25 ± 1 °C; 12 L: 12D) for 10 days prior to start of experiment and divided into 4 groups: non-challenged (negative control= A), challenged [positive control (B) and experimental (C & D)]. Group A, B were fed with non-supplemented feed while group C & D were fed with feed supplemented with 5% Eichhornia crassipes and 5% Ricinus communis respectively. Supplemented feeds were evaluated for their effect on growth, health, immune system and disease resistance in fish when challenged with Vibrio harveyi. Fingerlings of C. carpio (weight, 2.0±0.5 g) were exposed with fresh overnight culture of V. harveyi through bath immunization (concentration 2 Χ 105) for 2 hours on 10 days interval for 40 days. The growth was monitored through increase in their relative weight. The rate of mortality due to bacterial infection as well as due to effect of feed was recorded accordingly. Immune response of fish was analyzed through differential leucocyte count, percentage phagocytosis and phagocytic index. The effect of V. harveyi on fish organs were examined through histo-pathological examination of internal organs like spleen, liver and kidney. The change in the immune response was also observed through gene expression analysis. The antioxidant potential of plant extracts was measured through DPPH and FRAP assay and amount of total phenols and flavonoids were calculates through biochemical analysis. The chemical composition of plant’s methanol extracts was determined by GC-MS analysis, which showed presence of various secondary metabolites and other compounds. Investigation revealed immuno-modulatory effect of plants, when supplemented with the artificial feed of fish.

Keywords: immuno-modulation, gc-ms, Cyprinus carpio, Eichhornia crassipes, Ricinus communis

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635 Design and Optimisation of 2-Oxoglutarate Dioxygenase Expression in Escherichia coli Strains for Production of Bioethylene from Crude Glycerol

Authors: Idan Chiyanzu, Maruping Mangena

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Crude glycerol, a major by-product from the transesterification of triacylglycerides with alcohol to biodiesel, is known to have a broad range of applications. For example, its bioconversion can afford a wide range of chemicals including alcohols, organic acids, hydrogen, solvents and intermediate compounds. In bacteria, the 2-oxoglutarate dioxygenase (2-OGD) enzymes are widely found among the Pseudomonas syringae species and have been recognized with an emerging importance in ethylene formation. However, the use of optimized enzyme function in recombinant systems for crude glycerol conversion to ethylene is still not been reported. The present study investigated the production of ethylene from crude glycerol using engineered E. coli MG1655 and JM109 strains. Ethylene production with an optimized expression system for 2-OGD in E. coli using a codon optimized construct of the ethylene-forming gene was studied. The codon-optimization resulted in a 20-fold increase of protein production and thus an enhanced production of the ethylene gas. For a reliable bioreactor performance, the effect of temperature, fermentation time, pH, substrate concentration, the concentration of methanol, concentration of potassium hydroxide and media supplements on ethylene yield was investigated. The results demonstrate that the recombinant enzyme can be used for future studies to exploit the conversion of low-priced crude glycerol into advanced value products like light olefins, and tools including recombineering techniques for DNA, molecular biology, and bioengineering can be used to allowing unlimited the production of ethylene directly from the fermentation of crude glycerol. It can be concluded that recombinant E.coli production systems represent significantly secure, renewable and environmentally safe alternative to thermochemical approach to ethylene production.

Keywords: crude glycerol, bioethylene, recombinant E. coli, optimization

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634 Molecular Characterization of Two Thermoplastic Biopolymer-Degrading Fungi Utilizing rRNA-Based Technology

Authors: Nuha Mansour Alhazmi, Magda Mohamed Aly, Fardus M. Bokhari, Ahmed Bahieldin, Sherif Edris

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Out of 30 fungal isolates, 2 new isolates were proven to degrade poly-β-hydroxybutyrate (PHB). Enzyme assay for these isolates indicated the optimal environmental conditions required for depolymerase enzyme to induce the highest level of biopolymer degradation. The two isolates were basically characterized at the morphological level as Trichoderma asperellum (isolate S1), and Aspergillus fumigates (isolate S2) using standard approaches. The aim of the present study was to characterize these two isolates at the molecular level based on the highly diverged rRNA gene(s). Within this gene, two domains of the ribosome large subunit (LSU) namely internal transcribed spacer (ITS) and 26S were utilized in the analysis. The first domain comprises the ITS1/5.8S/ITS2 regions ( > 500 bp), while the second domain comprises the D1/D2/D3 regions ( > 1200 bp). Sanger sequencing was conducted at Macrogen (Inc.) for the two isolates using primers ITS1/ITS4 for the first domain, while primers LROR/LR7 for the second domain. Sizes of the first domain ranged between 594-602 bp for S1 isolate and 581-594 bp for S2 isolate, while those of the second domain ranged between 1228-1238 bp for S1 isolate and 1156-1291 for S2 isolate. BLAST analysis indicated 99% identities of the first domain of S1 isolate with T. asperellum isolates XP22 (ID: KX664456.1), CTCCSJ-G-HB40564 (ID: KY750349.1), CTCCSJ-F-ZY40590 (ID: KY750362.1) and TV (ID: KU341015.1). BLAST of the first domain of S2 isolate indicated 100% identities with A. fumigatus isolate YNCA0338 (ID: KP068684.1) and strain MEF-Cr-6 (ID: KU597198.1), while 99% identities with A. fumigatus isolate CCA101 (ID: KT877346.1) and strain CD1621 (ID: JX092088.1). Large numbers of other T. asperellum and A. fumigatus isolates and strains showed high level of identities with S1 and S2 isolates, respectively, based on the diversity of the first domain. BLAST of the second domain of S1 isolate indicated 99 and 100% identities with only two strains of T. asperellum namely TR 3 (ID: HM466685.1) and G (ID: KF723005.1), respectively. However, other T. species (ex., atroviride, hamatum, deliquescens, harzianum, etc.) also showed high level of identities. BLAST of the second domain of S2 isolate indicated 100% identities with A. fumigatus isolate YNCA0338 (ID: KP068684.1) and strain MEF-Cr-6 (ID: KU597198.1), while 99% identities with A. fumigatus isolate CCA101 (ID: KT877346.1) and strain CD1621 (ID: JX092088.1). Large numbers of other A. fumigatus isolates and strains showed high level of identities with S2 isolate. Overall, the results of molecular characterization based on rRNA diversity for the two isolates of T. asperellum and A. fumigatus matched those obtained by morphological characterization. In addition, ITS domain proved to be more sensitive than 26S domain in diversity profiling of fungi at the species level.

Keywords: Aspergillus fumigates, Trichoderma asperellum, PHB, degradation, BLAST, ITS, 26S, rRNA

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633 Effect of Diet Inulin Prebiotic on Growth, Reproductive Performance, Carcass Composition and Resistance to Environmental Stresses in Zebra Danio (Danio rerio)

Authors: Ehsan Ahmadifar

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In this research, the effects of different levels (control group (T0), (T1)1, (T2)2 and (T3)3 gr Inulin per Kg diet) of prebiotic Inulin as nutritional supplement on Danio rerio were investigated for 4 month. Since the beginning of feeding larvae until adult (average weight: 67.1 g, length: 4.5 cm) were fed with experimental diets. The survival rate of fish had no significant effect on rate survival (P > 0.05). The highest food conversion ratio (FCR) was in control group and the lowest was observed in T3. Treatment of T3 significantly caused the best feed conversion ratio in Zebra fish (P < 0.05). By increasing the inulin diet during the experiment, specific growth rate increased. The highest and the lowest body weight gain and condition factor were observed in T3 and control, respectively (P < 0.05). Adding 3 gr inulin in Zebra fish diet can improve the performance of the growth indices and final biomass, also this prebiotic can be considered as a suitable supplement for Cyprinidae diet. In the first sampling stage for feeding fish, fat and muscle protein was significantly higher than the second sampling stage (P < 0.05). Given that the second stage fish were full sexual maturity, the amount of fat in muscle decreased (P < 0.05). Moisture and ash levels were significantly (P < 0.05) higher in the second stage sampling than the first stage. Overall, different stage of living affected on muscle chemical composition muscle. Reproductive performance in treatment T2 and T3 were significantly higher than other treatments (P < 0.05). According to the results, the prebiotic inulin does not have a significant impact on the sex ratio in zebrafish (P > 0.05). Based on histology of the gonads, the use of dietary inulin accelerates the process of gonad development in zebrafish.

Keywords: inulin, zebrafish, reproduction, histology

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632 Inadequate Intake of Energy and Nutrients: A Comparative Cross-Sectional Study Between Sport and Non-sport Science University Students of Southern Ethiopia

Authors: Beruk Berhanu Desalegn, Kebede Awgechew, Addisalem Mesfin

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Introduction: This study aimed to investigate and compare the energy and selected nutrient intakes of sport science and non-sport science University students of Southern Ethiopia. Method: Multiple-day dietary data were collected from 166 university students (76 sport science and 90 non-sport sciences). Average daily energy and nutrient intake, and inadequate intakes were calculated using NutriSurvey (NS). Results: There were significant differences (p < 0.05) in the median intakes of energy, total carbohydrate, and vitamin B1 between female students from the sport science and non-sport science groups, but only the median intake of iron was significantly different (p < 0.05) between the male sport and non-sport science students’ group. The prevalence of inadequate intake of vitamin B1 were significantly (p<0.05) higher in the male and female from the non-sport science groups compared to the male and female students’ groups in the sport science, respectively. Whereas, the prevalence of inadequate iron intake by the male sport science students’ group was significantly (p<0.05) higher compared to their counterparts. Similarly, the prevalence of inadequate energy among the females from the sport science group was significantly (p<0.05) higher compared to the female students from the non-sport science department group. The prevalence of inadequate intakes of dietary energy, and the majority of the nutrients (protein, fat, vitamin A, B1, B2, and magnesium) were high (>50%) in selected University students. Conclusion: The energy and majority of nutrient intakes by the students in the selected universities of southern Ethiopia were sub-optimal. Therefore, activities that will improve the dietary intake of University students should include weekly meal plan revision considering their average recommended nutrient intake (RNI).

Keywords: dietary intake, sport science, University students, Ethiopia

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631 Acupoint Injection of High Concentration of Glucose Attenuates Mice Chronic Pain and Depression Comorbidity

Authors: Chanya Inprasit, Yi-Wen Lin

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Inflammation causes changes of peripheral and central nervous system properties, affecting both neuronal and non-neuronal cells, resulting in inflammatory pain. Acupoint injection (AI) was developed in the 1950s and has been widely used for relieving pain. It is an acupoint-stimulating technique that utilizes anatomically based meridians derived from Chinese medicine theory. AI has been accepted as an effective treatment and is thought to display superior results when compared to traditional acupuncture methods. However, the mechanism of AI needs to be ratified by more scientific evidence in order to support the theory and its therapeutic development. In this study, we explored the effect of AI on the comorbidity of chronic pain and depression. Mice hindpaw was injected by complete Freund’s adjuvant (CFA) to induce the condition of chronic pain. Measurements of mechanical and thermal hyperalgesia and depression-like behavior were analyzed. The results indicated a positive tendency to AI treatment. The comorbidity of chronic pain and depression was investigated with relation to transient receptor potential V1 (TRPV1) mechanism through the use of TRPV1 gene deletion. The expression of nociceptors such as voltage-gated sodium channels (Navs) or TRPV1, was significantly down-regulated by AI. The expression of inflammation-activated molecules: astrocytic marker glial fibrillary acidic protein (GFAP), the microglial marker Iba-1, S100B, and related kinases, were reversed by AI in both the peripheral and central nervous system. Taken together, these data provided a detailed molecular mechanism of AI-induced analgesia and anti-inflammatory properties. This finding may be utilized for clinical practice to treat chronic pain and depression comorbidity.

Keywords: inflammatory pain, acupoint injection, TRPV1, GFAP, S100B

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630 Hyparrhenia hirta: A Potential Protective Agent against DNA Damage and Liver Toxicity of Sodium Nitrate in Adult Rats

Authors: Hanen Bouaziz-Ketata, Ghada Ben Salah, Hichem Ben Salah, Kamel Jamoussi, Najiba Zeghal

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The present study investigated the protective role of Hyparrhenia hirta on nitrate-induced liver damage. Experiments were carried out on adult rats divided into 3 groups, a control group and two treated groups. NaNO3 was administered daily by oral gavage at a dose of 400 mg/kg bw in treated groups either alone or coadministered with Hyparrhenia hirta methanolic extract via drinking water at a dose of 200 mg/kg bw for 50 days. Liver toxicity induced by NaNO3 was characterized by higher serum levels of glucose, total cholesterol and triglyceride and lower serum total protein than those of controls. Transaminases and lactate deshydrogenase activities in serum were elevated indicating hepatic cells’ damage after treatment with NaNO3. The hyperbilirubinemia and the increased serum gamma glutamyl transferase activities suggested the presence of cholestasis in NaNO3 exposed rats. In parallel, NaNO3 caused oxidant/antioxidant imbalance in the liver as reflected by the increased lipid peroxidation, the decreased total glutathione content and superoxide dismutase, catalase and glutathione peroxidase activities. Nitrate caused also a significant induction of DNA fragmentation as evidenced by the presence of a smear without ladder formation on agarose gel. Hyparrhenia hirta supplementation showed an improvement of all parameters cited above. We conclude that the present work provides ethnopharmacological relevance of Hyparrhenia hirta against the toxic effect of nitrate, suggesting its role as a potential antioxidant.

Keywords: Hyparrhenia hirta, liver, nitrate toxicity, oxidative stress, rat

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629 Evaluation of the Influence of Graphene Oxide on Spheroid and Monolayer Culture under Flow Conditions

Authors: A. Zuchowska, A. Buta, M. Mazurkiewicz-Pawlicka, A. Malolepszy, L. Stobinski, Z. Brzozka

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In recent years, graphene-based materials are finding more and more applications in biological science. As a thin, tough, transparent and chemically resistant materials, they appear to be a very good material for the production of implants and biosensors. Interest in graphene derivatives also resulted at the beginning of research about the possibility of their application in cancer therapy. Currently, the analysis of their potential use in photothermal therapy and as a drug carrier is mostly performed. Moreover, the direct anticancer properties of graphene-based materials are also tested. Nowadays, cytotoxic studies are conducted on in vitro cell culture in standard culture vessels (macroscale). However, in this type of cell culture, the cells grow on the synthetic surface in static conditions. For this reason, cell culture in macroscale does not reflect in vivo environment. The microfluidic systems, called Lab-on-a-chip, are proposed as a solution for improvement of cytotoxicity analysis of new compounds. Here, we present the evaluation of cytotoxic properties of graphene oxide (GO) on breast, liver and colon cancer cell line in a microfluidic system in two spatial models (2D and 3D). Before cell introduction, the microchambers surface was modified by the fibronectin (2D, monolayer) and poly(vinyl alcohol) (3D, spheroids) covering. After spheroid creation (3D) and cell attachment (2D, monolayer) the selected concentration of GO was introduced into microsystems. Then monolayer and spheroids viability/proliferation using alamarBlue® assay and standard microplate reader was checked for three days. Moreover, in every day of the culture, the morphological changes of cells were determined using microscopic analysis. Additionally, on the last day of the culture differential staining using Calcein AM and Propidium iodide were performed. We were able to note that the GO has an influence on all tested cell line viability in both monolayer and spheroid arrangement. We showed that GO caused higher viability/proliferation decrease for spheroids than a monolayer (this was observed for all tested cell lines). Higher cytotoxicity of GO on spheroid culture can be caused by different geometry of the microchambers for 2D and 3D cell cultures. Probably, GO was removed from the flat microchambers for 2D culture. Those results were also confirmed by differential staining. Comparing our results with the studies conducted in the macroscale, we also proved that the cytotoxic properties of GO are changed depending on the cell culture conditions (static/ flow).

Keywords: cytotoxicity, graphene oxide, monolayer, spheroid

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628 Drying Effect on the Proximate Composition and Functional Properties of Cocoyam Flour

Authors: K. Maliki, A. Ajayi, O. M. Makanjuola, O. J. Adebowale

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Cocoyam is herbaceous perennial plant which belongs to the family Araceae and genus xanthosoma or cococasia is mostly cultivated as food crop. It is very rich in Vitamin B6, Magnesium and also in dietary fiber. Matured cocoyam is eaten boiled, Fried or roasted in Nigeria. It can also be dried and used to make flour. Food drying is a method of food preservation in which food is dried, thus inhibit the growth of bacteria yeast and mold through the removal of water. Drying effect on the proximate composition and functional properties of cocoyam flour were investigated. Freshly harvested cocoyam cultivars at matured level were washed with portable water, peeled, sliced into 0.3mm thickness blanch in boiling water at 100°C for 15 minutes and dried using sun drying oven and cabinet dryers. The blanched slices were divided into three lots and were subjected to different drying methods. The dried cocoyam slices were milled into flour using Apex mill and packed into Low Density Polyethylene Film (LDPE) 75 Micron 4 thickness and kept for four months under ambient temperature before analysis. The results showed that the moisture content, ash, crude fiber, fat, protein and carbohydrate ranged from 7.35% to 13.89%, 1.45% to 3.3%, 1.2% to 3.41%, 2.1% to 3.1%, 6.30% to 9.1% and 66% to 82% respectively. The functional properties of the cocoyam flour ranged from 1. 65ml/g to 4.24ml/g water absorption capacity, 0.85ml/g to 2.11ml/g oil absorption capacity 0.56ml/g and 0.78ml/g bulk density and 4.91% to 6.80% swelling capacity. The result showed that there was not significant difference (P ≥ 0.5) across the various drying methods used. Cabinet drying method was found to have the best quality characteristic values than the other drying methods. In conclusion, drying of cocoyam could be used for value addition and provide extension to shelf-life.

Keywords: cocoyam flour, drying, cabinet dryer, oven dryer

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627 Biogenic Amines Production during RAS Cheese Ripening

Authors: Amr Amer

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Cheeses are among those high-protein-containing foodstuffs in which enzymatic and microbial activities cause the formation of biogenic amines from amino acids decarboxylation. The amount of biogenic amines in cheese may act as a useful indicator of the hygienic quality of the product. In other words, their presence in cheese is related to its spoilage and safety. Formation of biogenic amines during Ras cheese (Egyptian hard cheese) ripening was investigated for 4 months. Three batches of Ras cheese were manufactured using Egyptian traditional method. From each batch, Samples were collected at 1, 7, 15, 30, 60, 90 and 120 days after cheese manufacture. The concentrations of biogenic amines (Tyramine, Histamine, Cadaverine and Tryptamine) were analyzed by high performance liquid chromatography (HPLC). There was a significant increased (P<0.05) in Tyramine levels from 4.34± 0.07 mg|100g in the first day of storage till reached 88.77± 0.14 mg|100g at a 120-day of storage. Also, Histamine and Cadaverine levels had the same increased pattern of Tyramine reaching 64.94± 0.10 and 28.28± 0.08 mg|100g in a 120- day of storage, respectively. While, there was a fluctuation in the concentration of Tryptamine level during ripening period as it decreased from 3.24± 0.06 to 2.66± 0.11 mg|100g at 60-day of storage then reached 5.38±0.08 mg|100g in a 120- day of storage. Biogenic amines can be formed in cheese during production and storage: many variables, as pH, salt concentration, bacterial activity as well as moisture, storage temperature and ripening time, play a relevant role in their formation. Comparing the obtained results with the recommended standard by Food and Drug Administration "FDA" (2001), High levels of biogenic amines in various Ras cheeses consumed in Egypt exceeded the permissible value (10 mg%) which seemed to pose a threat to public health. In this study, presence of high concentrations of biogenic amines (Tyramine, Histamine, cadaverine and Tryptamine) in Egyptian Ras cheeses reflects the bad hygienic conditions under which they produced and stored. Accordingly, the levels of biogenic amines in different cheeses should be come in accordance with the safe permissible limit recommended by FDA to ensure human safety.

Keywords: Ras cheese, biogenic amines, tyramine, histamine, cadaverine

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626 Studies on the Proximate Composition and Functional Properties of Extracted Cocoyam Starch Flour

Authors: Adebola Ajayi, Francis B. Aiyeleye, Olakunke M. Makanjuola, Olalekan J. Adebowale

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Cocoyam, a generic term for both xanthoma and colocasia, is a traditional staple root crop in many developing countries in Africa, Asia and the Pacific. It is mostly cultivated as food crop which is very rich in vitamin B6, magnesium and also in dietary fiber. The cocoyam starch is easily digested and often used for baby food. Drying food is a method of food preservation that removes enough moisture from the food so bacteria, yeast and molds cannot grow. It is a one of the oldest methods of preserving food. The effect of drying methods on the proximate composition and functional properties of extracted cocoyam starch flour were studied. Freshly harvested cocoyam cultivars at matured level were washed with portable water, peeled, washed and grated. The starch in the grated cocoyam was extracted, dried using sun drying, oven and cabinet dryers. The extracted starch flour was milled into flour using Apex mill and packed and sealed in low-density polyethylene film (LDPE) 75 micron thickness with Nylon sealing machine QN5-3200HI and kept for three months under ambient temperature before analysis. The result showed that the moisture content, ash, crude fiber, fat, protein and carbohydrate ranged from 6.28% to 12.8% 2.32% to 3.2%, 0.89% to 2.24%%, 1.89% to 2.91%, 7.30% to 10.2% and 69% to 83% respectively. The functional properties of the cocoyam starch flour ranged from 2.65ml/g to 4.84ml/g water absorption capacity, 1.95ml/g to 3.12ml/g oil absorption capacity, 0.66ml/g to 7.82ml/g bulk density and 3.82% to 5.30ml/g swelling capacity. Significant difference (P≥0.5) was not obtained across the various drying methods used. The drying methods provide extension to the shelf-life of the extracted cocoyam starch flour.

Keywords: cocoyam, extraction, oven dryer, cabinet dryer

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625 Protective Impact of Some Natural Extracts Against Acute Hepatotoxicity in Wistar Rats: DNA Protecting, Antioxidant and Anti-Inflammatory Effects

Authors: Yara Mohamed Taha, Mohamed Ali El Desouky, Heba Kamal Abdel Hakim, Maha Hanafy Mahmoud

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Hepatotoxicity due to drugs and toxic chemicals constitutes a crucial health problem nowadays. Medicinal plants are widely used recently for protecting against many liver disorders and inflammatory conditions. This study aims to evaluate hepatoprotective impact of green tea extract (GTE), rosemary extract (RE) and rosmarinic acid (RA) against hepatotoxins; ferric nitrilotriacetate (Fe-NTA) and diethylnitrosamine (DEN) in rats. Five groups of male Wistar rats were included; one control negative, while the other groups were treated intraperitoneally with DEN as 160 mg.kg-1 b.w. on 15th day and Fe-NTA as 5 mg.kg-1 b.w. on 33rd day. One of them was control positive. The other three groups were pre-administered with daily protective oral doses of either 200 mg.kg-1 b.w. of RE or 1 g.kg- 1 b.w. of GTE or 50 mg.kg-1 b.w. of RA two weeks prior to DEN exposure and continued till the end of the experimental period. The obtained data revealed a highly significant increase of MDA, 8-OHdG, DNA damage percent, a significant depletion of GSH and elevated Gr-1 protein expression in hepatocytes with liver tissue histopathological changes of rats exposed to DEN+Fe-NTA. Pre-administration of protective doses of RE, GTE and RA to DEN+Fe-NTA treated rats could normalize the altered biochemical, histopathological and immunohistochemical parameters. In conclusion, RE, GTE and RA showed a hepatoprotective effect against liver toxicity induced by DEN+Fe-NTA, with the best antioxidant and anti-inflammatory impact were for RA and GTE. Therefore, the current study declared that rosemary, green tea and products enriched with rosmarinic acid should be involved daily in diet of people who are exposed to chemicals and environmental toxins to protect themselves from hepatotoxicity.

Keywords: hepatotoxicity, diethylnitrosamine and ferric nitrilotriacetate, rosemary extract (RE), green tea extract (GTE), rosmarinic acid (RA)

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624 Environmental Effect on Yield and Quality of French Bean Genotypes Grown in Poly-Net House of India

Authors: Ramandeep Kaur, Tarsem Singh Dhillon, Rajinder Kumar Dhall, Ruma Devi

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French bean (Phaseolous vulgaris L.) is an economically potential legume vegetable grown at high altitude (>1000 ft.). More recently, its cultivation in Northern Indian plans is gaining popularity but there is severe reduction in its yield and quality due to low temperature during extreme winter conditions of December-January in open field conditions. Therefore, present study was undertaken to evaluate 29 indeterminate French bean genotypes for various yield and quality traits in poly-net house with the objective to identify best performing genotypes during winter conditions. The significant variation was observed among all the genotypes for all the studied traits. The green pod yield was significantly higher in genotype Lakshmi (992.33 g/plant) followed by Star-I (955.50 g/plant) and FBK-4 (911.17 g/plant). However, the genotypes FBK-10 (105.50 days) and Lakshmi (106.83 days) took least number of days to first harvest and were significantly better than all other genotypes (109.00-136.83 days). The maximum numbers of 10 pickings were recorded in genotype Lakshmi whereas maximum harvesting span as also observed in Lakshmi (60.50 days) which was significantly higher than all other genotypes (31.17-56.50 days). Regarding quality traits, maximum dry matter was observed in FBK-13 (13.87%), protein content in FBK-1 (9.67%), sugar content in FBK-5 (9.60%) and minimum fiber content in FBK-12 (0.69%). It is hereby concluded that high productivity and better quality of French bean (genotypes: Lakshmi, Star-I, FBK-4) was produced in poly-net house conditions of Punjab, India and these pods fetches premium price in the market as there is no availability of green pods at that time in high altitudes. Hence, there is a great scope of cultivation of indeterminate French bean under poly-net house conditions in Punjab.

Keywords: earliness, pod, protected environment, quality, yield

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623 Assessment of Soil Contamination on the Content of Macro and Microelements and the Quality of Grass Pea Seeds (Lathyrus sativus L.)

Authors: Violina R. Angelova

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Comparative research has been conducted to allow us to determine the content of macro and microelements in the vegetative and reproductive organs of grass pea and the quality of grass pea seeds, as well as to identify the possibility of grass pea growth on soils contaminated by heavy metals. The experiment was conducted on an agricultural field subjected to contamination from the Non-Ferrous-Metal Works (MFMW) near Plovdiv, Bulgaria. The experimental plots were situated at different distances of 0.5 km and 8 km, respectively, from the source of pollution. On reaching commercial ripeness the grass pea plants were gathered. The composition of the macro and microelements in plant materials (roots, stems, leaves, seeds), and the dry matter content, sugars, proteins, fats and ash contained in the grass pea seeds were determined. Translocation factors (TF) and bioaccumulation factor (BCF) were also determined. The quantitative measurements were carried out through inductively-coupled plasma (ICP). The grass pea plant can successfully be grown on soils contaminated by heavy metals. Soil pollution with heavy metals does not affect the quality of the grass pea seeds. The seeds of the grass pea contain significant amounts of nutrients (K, P, Cu, Fe Mn, Zn) and protein (23.18-29.54%). The distribution of heavy metals in the organs of the grass pea has a selective character, which reduces in the following order: leaves > roots > stems > seeds. BCF and TF values were greater than one suggesting efficient accumulation in the above ground parts of grass pea plant. Grass pea is a plant that is tolerant to heavy metals and can be referred to the accumulator plants. The results provide valuable information about the chemical and nutritional composition of the seeds of the grass pea grown on contaminated soils in Bulgaria. The high content of macro and microelements and the low concentrations of toxic elements in the grass pea grown in contaminated soil make it possible to use the seeds of the grass pea as animal feed.

Keywords: Lathyrus sativus L, macroelements, microelements, quality

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622 Biophysical Characterization of the Inhibition of cGAS-DNA Sensing by KicGAS, Kaposi's Sarcoma-Associated Herpesvirus Inhibitor of cGAS

Authors: D. Bhowmik, Y. Tian, Q. Yin, F. Zhu

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Cyclic GMP-AMP synthase (cGAS), recognises cytoplasmic double-stranded DNA (dsDNA), indicative of bacterial and viral infections, as well as the leakage of self DNA by cellular dysfunction and stresses, to elicit the host's immune responses. Viruses also have developed numerous strategies to antagonize the cGAS-STING pathway. Kaposi's sarcoma-associated herpesvirus (KSHV) is a human DNA tumor virus that is the causative agent of Kaposi’s sarcoma and several other malignancies. To persist in the host, consequently causing diseases, KSHV must overcome the host innate immune responses, including the cGAS-STING DNA sensing pathway. We already found that ORF52 or KicGAS (KSHV inhibitor of cGAS), an abundant and basic gamma herpesvirus-conserved tegument protein, directly inhibits cGAS enzymatic activity. To better understand the mechanism, we have performed the biochemical and structural characterization of full-length KicGAS and various mutants in regarding binding to DNA. We observed that KicGAS is capable of self-association and identified the critical residues involved in the oligomerization process. We also characterized the DNA-binding of KicGAS and found that KicGAS cooperatively oligomerizes along the length of the double stranded DNA, the highly conserved basic residues at the c-terminal disordered region are crucial for DNA recognition. Deficiency in oligomerization also affects DNA binding. Thus DNA binding by KicGAS sequesters DNA and prevents it from being detected by cGAS, consequently inhibiting cGAS activation. KicGAS homologues also inhibit cGAS efficiently, suggesting inhibition of cGAS is evolutionarily conserved mechanism among gamma herpesvirus. These results highlight the important viral strategy to evade this innate immune sensor.

Keywords: Kaposi's sarcoma-associated herpesvirus, KSHV, cGAS, DNA binding, inhibition

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621 Chemical Composition and Nutritional Value of Leaves and Pods of Leucaena Leucocephala, Prosopis Laevigata and Acacia Farnesiana in a Xerophyllous Shrubland

Authors: Miguel Mellado, Cecilia Zapata

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Goats can be exploited in harsh environments due to their capacity to adjust to limited quantity and quality forage sources. In these environments, leguminous trees can be used as supplementary feeds as foliage and fruits of these trees can contribute to maintain or improve production efficiency in ruminants. The objective of this study was to determine the nutritional value of three leguminous trees heavily selected by goats in a xerophyllous shrubland. Chemical composition and in vitro dry matter disappearance (IVDMD) of leaves and pods from leucaena (Leucaena leucocephala), mesquite (Prosopis laevigata) and huisache (Acacia farnesiana) is presented. Crude protein (CP) ranged from 17.3% for leaves of huisache to 21.9% for leucaena. The neutral detergent fiber (NDF) content ranged from 39.0 to 40.3 with no difference among fodder threes. Across tree species, mean IVDMD was 61.6% for pods and 52.2% for leaves. IVDMD for leaves was highest (P < 0.01) for leucaena (54.9%) and lowest for huisache (47.3%). Condensed tannins in an acetonic extract were highest for leaves of huisache (45.3 mg CE/g DM) and lowest for mesquite (25.9 mg CE/g DM). Pods and leaves of huisache presented the highest number of secondary metabolites, mainly related to hydrobenzoic acid and flavonols; leucaena and mesquite presented mainly flavonols and anthocyanins. It was concluded that leaves and pods of leucaena, mesquite and huisache constitute valuable forages for ruminant livestock due to its low fiber, high CP levels, moderate in vitro fermentation characteristics and high mineral content. Keywords: Fodder tree; ruminants; secondary metabolites; minerals; tannins

Keywords: fodder tree, ruminants, secondary metabolites, minerals, tannins

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620 Evaluation of Biological Seed Coating Technology On-Field Performance of Wheat in Regenerative Agriculture and Conventional Systems

Authors: S. Brain, P. J. Storer, H. Strydom, Z. M. Solaiman

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Increasing farmer awareness of soil health, the impact of agricultural management practices, and the requirement for high-quality agricultural produce are major factors driving the rapid adoption of biological seed treatments - currently valued globally at USD 1.5 billion. Biological seed coatings with multistrain plant beneficial microbial technology have the capability to affect plant establishment, growth, and development positively. These beneficial plant microbes can potentially increase soil health, plant yield, and nutrition – acting as bio fertilisers, rhizoremediators, phytostimulators, and stress modulators, and can ultimately reduce the overall use of agrichemicals. A field trial was conducted on MACE wheat in the central wheat belt of Western Australia to evaluate a proprietary seed coating technology (Langleys Bio-EnergeticTM Microbe blend (BMB)) on a conventional program (+/- BMB microbes) and a Regenerative Biomineral fertiliser program (+/- BMB microbes). The Conventional (+BMB) and Biomineral (+BMB) treated plants had no fungicide treatments and had no disease issues. Control (No fertiliser, No microbes), Conventional (No Microbes), and Biomineral (No Microbes) were treated with fungicides (seed dressing and foliar). From the research findings, compared to control and no microbe treatments, both the Conventional (+ BMB) and Biomineral (+ BMB) showed significant increases in Soil Carbon (SOC), Seed germination, nutrient use efficiency (NUE) of nitrogen, phosphate and mineral nutrients, grain mineral nutrient uptake, protein %, hectolitre weight, and fewer screenings, yield, and gross margins.

Keywords: biological seed coating, biomineral fertiliser, plant nutrition, regenerative and conventional agriculture

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619 Anticancer Lantadene Derivatives: Synthesis, Cytotoxic and Docking Studies

Authors: A. Monika, Manu Sharma, Hong Boo Lee, Richa Dhingra, Neelima Dhingra

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Nuclear factor-κappa B serve as a molecular lynchpin that links persistent infections and chronic inflammation to increased cancer risk. Inflammation has been recognized as a hallmark and cause of cancer. Natural products present a privileged source of inspiration for chemical probe and drug design. Herbal remedies were the first medicines used by humans due to the many pharmacologically active secondary metabolites produced by plants. Some of the metabolites like Lantadene (pentacyclic triterpenoids) from the weed Lantana camara has been known to inhibit cell division and showed anti-antitumor potential. The C-3 aromatic esters of lantadenes were synthesized, characterized and evaluated for cytotoxicity and inhibitory potential against Tumor necrosis factor alpha-induced activation of Nuclear factor-κappa B in lung cancer cell line A549. The 3-methoxybenzoyloxy substituted lead analogue inhibited kinase activity of the inhibitor of nuclear factor-kappa B kinase in a single-digit micromolar concentration. At the same time, the lead compound showed promising cytotoxicity against A549 lung cancer cells with IC50 ( half maximal inhibitory concentration) of 0.98l µM. Further, molecular docking of 3-methoxybenzoyloxy substituted analogue against Inhibitor of nuclear factor-kappa B kinase (Protein data bank ID: 3QA8) showed hydrogen bonding interaction involving oxygen atom of 3-methoxybenzoyloxy with the Arginine-31 and Glutamine-110. Encouraging results indicate the Lantadene’s potential to be developed as anticancer agents.

Keywords: anticancer, lantadenes, pentacyclic triterpenoids, weed

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618 Protective Effect of Diosgenin against Silica-Induced Tuberculosis in Rat Model

Authors: Williams A. Adu, Cynthia A. Danquah, Paul P. S. Ossei, Selase Ativui, Michael Ofori, James Asenso, George Owusu

Abstract:

Background Silicosis is an occupational disease of the lung that is caused by chronic exposure to silica dust. There is a higher frequency of co-existence of silicosis with tuberculosis (TB), ultimately resulting in lung fibrosis and respiratory failure. Chronic intake of synthetic drugs has resulted in undesirable side effects. Diosgenin is a steroidal saponin that has been shown to exert a therapeutic effect on lung injury. Therefore, we investigated the ability of diosgenin to reduce the susceptibility of silica-induced TB in rats. Method Silicosis was induced by intratracheal instillation of 50 mg/kg crystalline silica in Sprague Dawley rats. Different doses of diosgenin (1, 10, and 100 mg/kg), Mycobacterium smegmatis and saline were administered for 30 days. Afterwards, 5 of the rats from each group were sacrificed, and the 5 remaining rats in each group, except the control, received Mycobacterium smegmatis. Treatment of diosgenin continued until the 50th day, and the rats were sacrificed at the end of the experiment. The result was analysed using a one-way analysis of variance (ANOVA) with a Graph-pad prism Result At a half-maximal inhibition concentration of 48.27 µM, diosgenin inhibited the growth of Mycobacterium smegmatis. There was a marked decline in the levels of immune cell infiltration and cytokines production. Lactate dehydrogenase and total protein levels were significantly reduced compared to control. There was an increase in the survival rate of the treatment group compared to the control. Conclusion Diosgenin ameliorated silica-induced pulmonary tuberculosis by declining the levels of inflammatory and pro-inflammatory cytokines and, in effect, significantly reduced the susceptibility of rats to pulmonary TB.

Keywords: silicosis, tuberculosis, diosgenin, fibrosis, crystalline silica

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617 Inf-γ and Il-2 Asses the Therapeutic Response in Anti-tuberculosis Patients at Jamot Hospital Yaounde, Cameroon

Authors: Alexandra Emmanuelle Membangbi, Jacky Njiki Bikoï, Esther Del-florence Moni Ndedi, Marie Joseph Nkodo Mindimi, Donatien Serge Mbaga, Elsa Nguiffo Makue, André Chris Mikangue Mbongue, Martha Mesembe, George Ikomey Mondinde, Eric Walter Perfura-yone, Sara Honorine Riwom Essama

Abstract:

Background: Tuberculosis (TB) is one of the top lethal infectious diseases worldwide. In recent years, interferon-γ (INF-γ) release assays (IGRAs) have been established as routine tests for diagnosing TB infection. However, produced INF-γ assessment failed to distinguish active TB (ATB) from latent TB infection (LTBI), especially in TB epidemic areas. In addition to IFN-γ, interleukin-2 (IL-2), another cytokine secreted by activated T cells, is also involved in immune response against Mycobacterium tuberculosis. The aim of the study was to assess the capacity of IFN-γ and IL2 to evaluate the therapeutic response of patients on anti-tuberculosis treatment. Material and Methods: We conducted a cross-sectional study in the Pneumonology Departments of the Jamot Hospital in Yaoundé between May and August 2021. After signed the informed consent, the sociodemographic data, as well as 5 mL of blood, were collected in the crook of the elbow of each participant. Sixty-one subjects were selected (n= 61) and divided into 4 groups as followed: group 1: resistant tuberculosis (n=13), group 2: active tuberculosis (n=19), group 3 cured tuberculosis (n=16), and group 4: presumed healthy persons (n=13). The cytokines of interest were determined using an indirect Enzyme-linked Immuno-Sorbent Assay (ELISA) according to the manufacturer's recommendations. P-values < 0.05 were interpreted as statistically significant. All statistical calculations were performed using SPSS version 22.0 Results: The results showed that men were more 14/61 infected (31,8%) with a high presence in active and resistant TB groups. The mean age was 41.3±13.1 years with a 95% CI = [38.2-44.7], the age group with the highest infection rate was ranged between 31 and 40 years. The IL-2 and INF-γ means were respectively 327.6±160.6 pg/mL and 26.6±13.0 pg/mL in active tuberculosis patients, 251.1±30.9 pg/mL and 21.4±9.2 pg/mL in patients with resistant tuberculosis, while it was 149.3±93.3 pg/mL and 17.9±9.4 pg/mL in cured patients, 15.1±8.4 pg/mL and 5.3±2.6 pg/mL in participants presumed healthy (p <0.0001). Significant differences in IFN-γ and IL-2 rates were observed between the different groups. Conclusion: Monitoring the serum levels of INF-γ and IL-2 would be useful to evaluate the therapeutic response of anti-tuberculosis patients, particularly in the both cytokines association case, that could improve the accuracy of routine examinations.

Keywords: antibiotic therapy, interferon gamma, interleukin 2, tuberculosis

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616 Design of a Fuzzy Expert System for the Impact of Diabetes Mellitus on Cardiac and Renal Impediments

Authors: E. Rama Devi Jothilingam

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Diabetes mellitus is now one of the most common non communicable diseases globally. India leads the world with largest number of diabetic subjects earning the title "diabetes capital of the world". In order to reduce the mortality rate, a fuzzy expert system is designed to predict the severity of cardiac and renal problems of diabetic patients using fuzzy logic. Since uncertainty is inherent in medicine, fuzzy logic is used in this research work to remove the inherent fuzziness of linguistic concepts and uncertain status in diabetes mellitus which is the prime cause for the cardiac arrest and renal failure. In this work, the controllable risk factors "blood sugar, insulin, ketones, lipids, obesity, blood pressure and protein/creatinine ratio" are considered as input parameters and the "the stages of cardiac" (SOC)" and the stages of renal" (SORD) are considered as the output parameters. The triangular membership functions are used to model the input and output parameters. The rule base is constructed for the proposed expert system based on the knowledge from the medical experts. Mamdani inference engine is used to infer the information based on the rule base to take major decision in diagnosis. Mean of maximum is used to get a non fuzzy control action that best represent possibility distribution of an inferred fuzzy control action. The proposed system also classifies the patients with high risk and low risk using fuzzy c means clustering techniques so that the patients with high risk are treated immediately. The system is validated with Matlab and is used as a tracking system with accuracy and robustness.

Keywords: Diabetes mellitus, fuzzy expert system, Mamdani, MATLAB

Procedia PDF Downloads 264
615 Site-Specific Delivery of Hybrid Upconversion Nanoparticles for Photo-Activated Multimodal Therapies of Glioblastoma

Authors: Yuan-Chung Tsai, Masao Kamimura, Kohei Soga, Hsin-Cheng Chiu

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In order to enhance the photodynamic/photothermal therapeutic efficacy on glioblastoma, the functionized upconversion nanoparticles with the capability of converting the deep tissue penetrating near-infrared light into visible wavelength for activating photochemical reaction were developed. The drug-loaded nanoparticles (NPs) were obtained from the self-assembly of oleic acid-coated upconversion nanoparticles along with maleimide-conjugated poly(ethylene glycol)-cholesterol (Mal-PEG-Chol), as the NP stabilizer, and hydrophobic photosensitizers, IR-780 (for photothermal therapy, PTT) and mTHPC (for photodynamic therapy, PDT), in aqueous phase. Both the IR-780 and mTHPC were loaded into the hydrophobic domains within NPs via hydrophobic association. The peptide targeting ligand, angiopep-2, was further conjugated with the maleimide groups at the end of PEG adducts on the NP surfaces, enabling the affinity coupling with the low-density lipoprotein receptor-related protein-1 of tumor endothelial cells and malignant astrocytes. The drug-loaded NPs with the size of ca 80 nm in diameter exhibit a good colloidal stability in physiological conditions. The in vitro data demonstrate the successful targeting delivery of drug-loaded NPs toward the ALTS1C1 cells (murine astrocytoma cells) and the pronounced cytotoxicity elicited by combinational effect of PDT and PTT. The in vivo results show the promising brain orthotopic tumor targeting of drug-loaded NPs and sound efficacy for brain tumor dual-modality treatment. This work shows great potential for improving photodynamic/photothermal therapeutic efficacy of brain cancer.

Keywords: drug delivery, orthotopic brain tumor, photodynamic/photothermal therapies, upconversion nanoparticles

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614 Genetic Variability in Advanced Derivatives of Interspecific Hybrids in Brassica

Authors: Yasir Ali, Farhatullah

Abstract:

The present study was conducted to estimate the genetic variability, heritability and genetic advance in six parental lines and their 56 genotypes derived from five introgressed brassica populations on the basis of morphological and biochemical traits. The experiment was laid out in a randomized complete block design with two replications at The University of Agriculture Peshawar-Pakistan during growing season of 2015-2016. The ANOVA of all traits of F5:6 populations showed highly significant differences (P ≤ 0.01) for all morphological and biochemical traits. Among F5:6 populations, the genotype 2(526) was earlier in flowering (108.65 days), and genotype 14(485) was earlier in maturity (170 days). Tallest plants (182.5 cm), largest main raceme (91.5 cm) and maximum number of pods (80.5) on main raceme were recorded for genotype 17(34). Maximum primary branches plant-1(6.2) and longest pods (10.26 cm) were recorded for genotype 15, while genotype 16(171) had more seeds pod⁻¹ (22) and gave maximum yield plant-1 (30.22 g). The maximum 100-seed weight (0.60 g) was observed for genotype 10(506) while high protein content (22.61%) was recorded for genotype 4(99). Maximum oil content (54.08 %) and low linoleic acid (7.07 %) were produced by genotype (12(138) and low glucosinolate (59.01 µMg⁻¹) was recorded for genotype 21(113). The genotype 27(303) having high oleic acid content (51.73 %) and genotype 1(209) gave low erucic acid (35.97 %). Among the F5:6 populations moderate to high heritability observed for all morphological and biochemical traits coupled with high genetic advance. Cluster analysis grouped the 56 F5:6 populations along their parental lines into seven different groups. Each group was different from the other group on the basis of morphological and biochemical traits. Moreover all the F5:6 populations showed sufficient variability. Genotypes 10(506) and 16(171) were superior for high seed yield⁻¹, 100-seeds weight, and seed pod⁻¹ and are recommended for future breeding program.

Keywords: Brassicaceae, biochemical characterization, introgression, morphological characterization

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613 Fluorescence Resonance Energy Transfer in a Supramolecular Assembly of Luminescent Silver Nanoclusters and Cucurbit[8]uril Based Host-Guest System

Authors: Srikrishna Pramanik, Sree Chithra, Saurabh Rai, Sameeksha Agrawal, Debanggana Shil, Saptarshi Mukherjee

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The understanding of interactions between organic chromophores and biologically useful luminescent noble metal nanoclusters (NCs) leading to an energy transfer process that has applications in light-harvesting materials is still in its nascent stage. This work describes a photoluminescent supramolecular assembly, made in two stages, employing an energy transfer process between silver (Ag) NCs as the donor and a host-guest system as the acceptor that can find potential applications in diverse fields. Initially, we explored the host-guest chemistry between a cationic guest, Ethidium Bromide and the anionic host Cucurbit[8]uril using spectroscopic and calorimetric techniques to decipher their interaction mechanism in modulating photophysical properties of the chromophore. Next, we synthesized a series of blue-emitting AgNCs using different templates such as protein, peptides, and cyclodextrin. The as-prepared AgNCs were characterized by various spectroscopic techniques. We have established that these AgNCs can be employed as donors in the FRET process with the above acceptor for FRET-based emission color tuning. Our in-depth studies revealed that surface ligands play a key role in modulating FRET efficiency. Overall, by employing a non-covalent strategy, we have tried to develop FRET pairs using blue-emitting NCs and a host-guest complex, which could find potential applications in constructing advanced white light-emitting, anti-counterfeiting materials, and developing biosensors.

Keywords: absorption spectroscopy, cavities, energy transfer, fluorescence, fluorescence resonance energy transfer

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