Search results for: protein adsorption

Authors: Ekta, G. K. Darbha

Abstract:

Nanotechnology offers the potential of simultaneously increasing efficiency as compared to their bulk material as well as reducing harmful environmental impacts of pesticides in field of agriculture. The term nanopesticide covers different pesticides that are cumulative of several surfactants, polymers, metal ions, etc. of nanometer size ranges from 1-1000 nm and exhibit abnormal behavior (high efficacy and high specific surface area) of nanomaterials. Commercial formulations of pesticides used by farmers nowadays cannot be used effectively due to a number of problems associated with them. For example, more than 90% of applied formulations are either lost in the environment or unable to reach the target area required for effective pest control. Around 20−30% of pesticides are lost through emissions. A number of factors (application methods, physicochemical properties of the formulations, and environmental conditions) can influence the extent of loss during application. It is known that among various formulations, polymer-based formulations show the greatest potential due to their greater efficacy, slow release and protection against premature degradation of active ingredient as compared to other commercial formulations. However, the nanoformulations can have a significant effect on the fate of active ingredient as well as may release some new ingredients by reacting with existing soil contaminants. Environmental fate of these newly generated species is still not explored very well which is essential to field scale experiments and hence a lot to be explored in the field of environmental fate, nanotoxicology, transport properties and stability of such formulations. In our preliminary work, we have synthesized polymer based nanoformulation of commercially used weedicide atrazine. Atrazine belongs to triazine class of herbicide, which is used in the effective control of seed germinated dicot weeds and grasses. It functions by binding to the plastoquinone-binding protein in PS-II. Plant death results from starvation and oxidative damage caused by breakdown in electron transport system. The stability of the suspension of nanoformulation containing herbicide has been evaluated by considering different parameters like polydispersity index, particle diameter, zeta-potential under different environmental relevance condition such as pH range 4-10, temperature range from 25°C to 65°C and stability of encapsulation also have been studied for different amount of added polymer. Morphological characterization has been done by using SEM.

Keywords: atrazine, nanoformulation, nanopesticide, nanotoxicology

Procedia PDF Downloads 257

Authors: Fatimah Abd Elghani, Raymonde Szargel, Vered Shani, Hazem Safory, Haya Hamza, Mor Savyon, Ruth Rott, Rina Bandopadhyay, Simone Engelender

Abstract:

Background: PINK1 is a mitochondrial kinase mutated in some familial cases of Parkinson’s disease. Down regulation of PINK1 results in abnormal mitochondrial morphology and altered membrane potential. Although PINK1 has a predicted mitochondrial import sequence, it’s cellular, and submitochondrial localization remains unclear, in part because it is rapidly degraded. In this work, we investigated the mechanisms involved in PINK1 degradation and how this may affect PINK1 stability and function, with implications for mitochondrial function in PD. In addition, pharmacological inhibition of proteasome activity was shown to lead to PINK1 accumulation, indicating that PINK1 degradation depends on the ubiquitin-proteasome system (UPS). Methods: Using co-immunoprecipitation assays, we identified E3 ubiquitin ligase SIAH1 as a PINK1-interacting protein in HEK293 cells as well as on rat brain tissues. In addition, we determined the effect of SIAH 1, SIAH2 and Parkin on PINK1 steady-state levels by Western blot analysis, and checked their possibility to ubiquitinate and mediate PINK1 degradation through the proteasome carried out in vivo ubiquitination experiments. Results: We have obtained results showing that SIAH-1 interacts with and ubiquitinates PINK1. The ubiquitination promoted by SIAH-1 leads to the proteasomal degradation of PINK1. We confirmed these findings by knocking down SIAH-1 and observing important accumulation of PINK1 in cells. Besides, we found that SIAH-1 decreases PINK1 steady-state levels but not the E3 ligase Parkin. We also investigated the interaction of SIAH-1 with PINK1 disease mutants and its ability to promote their ubiquitination and degradation. Although, no clear difference in the ability of SIAH-1 to promote the degradation of PINK1 disease mutants was observed. It is possible that dysfunction of proteasomal activity in the disease may lead to the accumulation and aggregation of ubiquitinated PINK1 in patients with PINK1 mutations, with possible implications to the pathogenesis of PD. Conclusions: Here, we demonstrated that SIAH-1 ubiquitinates and promotes the degradation of PINK1. In addition, SIAH-1 represents now a target that may help the improvement of mitophagy in PD. Further investigations needed to understand how mitophagy is regulated by PINK1-SIAH-1 axis to provide targets for future therapeutics.

Keywords: PD, Parkinson's disease, PINK1, PTEN-induced kinase1, SIAH, seven in absentia homolog, SN, substantia nigra

Procedia PDF Downloads 146

Authors: Kevin Kellner, Nga Lao, Orla Coleman, Paula Meleady, Niall Barron

Abstract:

Chinese Hamster Ovary (CHO) cells are the prominent cell line used in biopharmaceutical production. To improve yields and find beneficial bioprocess phenotypes genetic engineering plays an essential role in recent research. The miR-23 cluster, specifically miR-24 and miR-27, was first identified as differentially expressed during hypothermic conditions suggesting a role in proliferation and productivity in CHO cells. In this study, we used sponge decoy technology to stably deplete the miRNA expression of the cluster. Furthermore, we implemented the CRISPR/Cas9 system to knockdown miRNA expression. Sponge constructs were designed for an imperfect binding of the miRNA target, protecting from RISC mediated cleavage. GuideRNAs for the CRISPR/Cas9 system were designed to target the seed region of the miRNA. The expression of mature miRNA and precursor were confirmed using RT-qPCR. For both approaches stable expressing mixed populations were generated and characterised in batch cultures. It was shown, that CRISPR/Cas9 can be implemented in CHO cells with achieving high knockdown efficacy of every single member of the cluster. Targeting of one miRNA member showed that its genomic paralog is successfully targeted as well. The stable depletion of miR-24 using CRISPR/Cas9 showed increased growth and specific productivity in a CHO-K1 mAb expressing cell line. This phenotype was further characterized using quantitative label-free LC-MS/MS showing 186 proteins differently expressed with 19 involved in proliferation and 26 involved in protein folding/translation. Targeting miR-27 in the same cell line showed increased viability in late stages of the culture compared to the control. To evaluate the phenotype in an industry relevant cell line; the miR-23 cluster, miR-24 and miR-27 were stably depleted in a Fc fusion CHO-S cell line which showed increased batch titers up to 1.5-fold. In this work, we highlighted that the stable depletion of the miR-23 cluster and its members can improve the bioprocess phenotype concerning growth and productivity in two different cell lines. Furthermore, we showed that using CRISPR/Cas9 is comparable to the traditional sponge decoy technology.

Keywords: Chinese Hamster ovary cells, CRISPR/Cas9, microRNAs, sponge decoy technology

Procedia PDF Downloads 201

Authors: Päärn Paiste, Margit Kõiv, Riho Mõtlep, Kalle Kirsimäe

Abstract:

For small-scale wastewater management, the treatment wetlands (TWs) as a low cost alternative to conventional treatment facilities, can be used. However, P removal capacity of TW systems is usually problematic. P removal in TWs is mainly dependent on the physico–chemical and hydrological properties of the filter material. Highest P removal efficiency has been shown trough Ca-phosphate precipitation (i.e. active filtration) in Ca-rich alkaline filter materials, e.g. industrial by-products like hydrated oil shale ash (HOSA), metallurgical slags. In this contribution we report preliminary results of a full-scale TW system using HOSA material for P removal for a municipal wastewater at Nõo site, Estonia. The main goals of this ongoing project are to evaluate: a) the long-term P removal efficiency of HOSA using real waste water; b) the effect of high organic loading rate; c) variable P-loading effects on the P removal mechanism (adsorption/direct precipitation); and d) the form and composition of phosphate precipitates. Onsite full-scale experiment with two concurrent filter systems for treatment of municipal wastewater was established in September 2013. System’s pretreatment steps include septic tank (2 m2) and vertical down-flow LECA filters (3 m2 each), followed by horizontal subsurface HOSA filters (effective volume 8 m3 each). Overall organic and hydraulic loading rates of both systems are the same. However, the first system is operated in a stable hydraulic loading regime and the second in variable loading regime that imitates the wastewater production in an average household. Piezometers for water and perforated sample containers for filter material sampling were incorporated inside the filter beds to allow for continuous in-situ monitoring. During the 18 months of operation the median removal efficiency (inflow to outflow) of both systems were over 99% for TP, 93% for COD and 57% for TN. However, we observed significant differences in the samples collected in different points inside the filter systems. In both systems, we observed development of preferred flow paths and zones with high and low loadings. The filters show formation and a gradual advance of a “dead” zone along the flow path (zone with saturated filter material characterized by ineffective removal rates), which develops more rapidly in the system working under variable loading regime. The formation of the “dead” zone is accompanied by the growth of organic substances on the filter material particles that evidently inhibit the P removal. Phase analysis of used filter materials using X-ray diffraction method reveals formation of minor amounts of amorphous Ca-phosphate precipitates. This finding is supported by ATR-FTIR and SEM-EDS measurements, which also reveal Ca-phosphate and authigenic carbonate precipitation. Our first experimental results demonstrate that organic pollution and loading regime significantly affect the performance of hydrated ash filters. The material analyses also show that P is incorporated into a carbonate substituted hydroxyapatite phase.

Keywords: active filtration, apatite, hydrated oil shale ash, organic pollution, phosphorus

Procedia PDF Downloads 276

Authors: Aswini M. S.

Abstract:

Tomato (Solanum lycopersicum L.) is one of the most significant vegetable crops in terms of its economic benefits. Both fresh and processed tomatoes are consumed. Tomatoes have a limited genetic base, which makes breeding extremely challenging. Plant breeding has become much simpler and more effective with genome editing tools of CRISPR and CRISPR-associated 9 protein (CRISPR/Cas9), which address the problems with traditional breeding, chemical/physical mutagenesis, and transgenics. With the use of CRISPR/Cas9, a number of tomato traits have been functionally distinguished and edited. These traits include plant architecture as well as flower characters (leaf, flower, male sterility, and parthenocarpy), fruit ripening, quality and nutrition (lycopene, carotenoid, GABA, TSS, and shelf-life), disease resistance (late blight, TYLCV, and powdery mildew), tolerance to abiotic stress (heat, drought, and salinity) and resistance to herbicides. This study explores the potential of CRISPR/Cas9 genome editing for enhancing yield in tomato plants. The study utilized the CRISPR/Cas9 genome editing technology to functionally edit various traits in tomatoes. The de novo domestication of elite features from wild cousins to cultivated tomatoes and vice versa has been demonstrated by the introgression of CRISPR/Cas9. The CycB (Lycopene beta someri) gene-mediated Cas9 editing increased the lycopene content in tomato. Also, Cas9-mediated editing of the AGL6 (Agamous-like 6) gene resulted in parthenocarpic fruit development under heat-stress conditions. The advent of CRISPR/Cas has rendered it possible to use digital resources for single guide RNA design and multiplexing, cloning (such as Golden Gate cloning, GoldenBraid, etc.), creating robust CRISPR/Cas constructs, and implementing effective transformation protocols like the Agrobacterium and DNA free protoplast method for Cas9-gRNAs ribonucleoproteins (RNPs) complex. Additionally, homologous recombination (HR)-based gene knock-in (HKI) via geminivirus replicon and base/prime editing (Target-AID technology) remains possible. Hence, CRISPR/Cas facilitates fast and efficient breeding in the improvement of tomatoes.

Keywords: CRISPR-Cas, biotic and abiotic stress, flower and fruit traits, genome editing, polygenic trait, tomato and trait introgression

Procedia PDF Downloads 72

Authors: Joanna Wozniak-Holecka, Sylwia Jaruga-Sekowska

Abstract:

There are 45 statutory spa and treatment areas in Poland, and the demand for spa and treatment services increases year by year. Within each type of spa treatment facilities, nutritional education services are provided. During spa treatment, the patient learns the principles of rational nutrition and applied diet therapy. It should help him develop proper eating habits, which will also follow at home. However, the nutrition education system of spa resort patients should be considered as very imperfect and requiring a definite systemic correction. It has, at the same time, a wide human and infrastructure base, which guarantees to obtain positive reinforcement in the scope of undertaken activities and management. Unfortunately, this advantage is not fully used. The aim of the project was to assess the quality of implemented nutritional education and to assess the diet of patients in spa treatment entities from a nationwide perspective. The material for the study was data obtained as part of an in-depth interview conducted among nutrition department managers (25 interviews) and a survey addressed to patients (600 questionnaires) of a selected group of spa resorts from across the country about the implementation of nutritional education in institutions. Also, decade menus for the basic diet, easily digestible diet and diet with limitation of easily digestible carbohydrates (a total of 1,120 menus) were obtained for the study. Almost 2/3 of respondents (73.2%) were overweight or obese, but only 32.8% decided on an easily digestible or low-energy diet during the treatment. Most of the surveyed patients rated the nutrition in spa resorts as satisfactory. Classes on nutrition education were carried out mainly by a dietitian (65% of meetings), the other educators were doctors and nurses. The meetings (95%) were of a group nature and lasted only 30 minutes on average. The subjects of the classes concerned the principles of proper nutrition and composition of meals, a nutrition pyramid and a diet adapted to a given disease. The assessed menus did not meet the nutrition standards and, therefore, did not provide patients with the correct quality of nutrition. The norm of protein, fat, vitamin A, B12, phosphorus, iron and sodium was exceeded, while vitamin D, folic acid, magnesium and zinc were not enough than recommended. The study allowed to conclude that there is a large discrepancy between the recommendations presented during the nutrition education classes and the quality of diet implemented in the examined institutions. The project may contribute to the development of effective educational tools in nutrition, especially about a specific group of chronically ill patients.

Keywords: diet, management, nutritional education, spa resort

Procedia PDF Downloads 146

Authors: Tadesse Kebede Dabsu

Abstract:

Insect pest is one the major limiting factor for sustainable food production. To overtake insect pest problem, since Second World War, producers have used excessive insecticide for insect pest management. However, in the era of 21st Century, the excessive use of insecticide caused insect resistant, insecticide bioaccumulation, insecticide hazard to environment, human health problem, and the like. Due to these problems, research efforts have been focused on the development of environmental free sustainable insect pest management method. To minimize all above mentioned risk utilizing of biological control such as entomopathogenicmicroorganism include bacteria, virus, fungus, and their productsare the best option for suppress insect population below certain density level. The objective of this review was to review the updated available studies and recent developments on the entomopathogenic bacteria (EPB) as biological control of insect pest and challenge of using them for control of insect pest. EPB’s mechanisms of insecticidal activities, type, taxonomy, and history are included in this paper body. EPB has been successfully used for the suppression of populations of insect pests. Controlling of harmful insect by entomopathogenic bacteria is an effective, low bioaccumulation in environment and food, very specific, reduce resistance risk in insect pest, economically and sustainable method of major insect pest management method. Identified and reported as potential major common type of entomopathogenic bacteria include Bacillus thuringiensis, Photorhabdus sp., Xenorhabdus spp.Walbachiaspp, Actinomycetesspp.etc. These bacteria being enter into insect body through natural opening or by vector release toxin protein inside of insect and disrupt the cell’s content cause natural mortality under natural condition. As per reported by different scientists, insect orders like Lepidoptera, Hemiptera, Hymenoptera, Coleoptera, and Dipterahave been successful controlled by entomopathogenic bacteria. As per coming across in different scientific research journals, much of the work was emphasised on Bacillus thuringiensisbsp. Therefore, for commercial production like Bacillus thuringiensi, detail research should be done on other bacteria species. The efficacy and practical application of EPB are restricted to some crops and greenhouse area, but their field application at farmers’ level very less. So still much work needs to be done to the practical application of the EPB at widely application. Their efficacy, pathogenicity, and host range test should be tested under environmental condition.

Keywords: insect pest, entomopathogenic bacteria, biological control, agent

Procedia PDF Downloads 140

Authors: Yumeho Obata, Mariko Era, Takayoshi Kawahara, Takahide Kanyama, Hiroshi Morita

Abstract:

Dermatophagoides farinae is major mite allergens in indoors. D. farinae is often swarm over powder products (e.g. wheat flour), because it feeds on starch or protein that are included in them. Eating powder products which are mixed D.farinae causes various allergic symptoms. Therefore, the creation of food additive agents with high safety and control of mite effect is required. Fatty acid salts and spices are known that have pesticidal activities. This study describes the effects of fatty acid salts and spices against Dermatophagoides farinae. Materials and Methods: Potassium salts of 9 fatty acids (C4:0, C6:0, C8:0, C10:0, C12:0, C14:0, C18:1, C18:2, C18:3) were prepared by mixing the fatty acid with the appropriate amount of KOH solution to a concentration of 175 mM and pH 10.5. C12Cu and C12Zn were selected as other fatty acid salts. Cayenne pepper, habanero, Japanese pepper, mustard, jalapeno pepper, curry aroma and cinnamon were selected as spices. D. farina, have been cultured in laboratory. To rear the mites, double-soled dishes containing of sterilized food were put on the big plastic container (30.0 × 20.0 × 20.0cm) which had 100% ammonium nitrate solution in the bottom. Plastic container was placed on incubator at 25 °C and 64 % relative humidity (RH) under dark condition. Sterilized food composed of dried bonito flakes and dried yeast (Ebios), 1:1 by weight. The antiproliferative method, sample and medium culture were mixed in double-soled dish and kept at 25 °C and 64 % RH. Decrease rates were determined 1 week and 4 week after treatment under microscope. D. farina was considered to be dead if appendages did not move when prodded with a pin. Results and Conclusions: The results show that the fatty acids potassium showed no antiproliferative effects against D. farinae. On the other hand, Japanese pepper, mustard, curry aroma and cinnamon were effective to decrease propagative rate (over 80 %) after treatment for 1 week against D. farina. Japanese pepper, curry aroma and cinnamon were effective to decrease propagative rate (approximately 100 %) after treatment for 4 weeks against D. farina. Especially, Japanese pepper and cinnamon showed the fasted and the most consecutive antiproliferative effects. These results indicate that Japanese pepper and cinnamon have high antiproliferative effects against D. farina and suggest spices will be used as a food additive agent.

Keywords: fatty acid salts, spices, antiproliferative effects, dermatophagoides farinae

Procedia PDF Downloads 236

Authors: R. L. Shirole, N. L. Shirole, R. B. Patil, M. N. Saraf

Abstract:

The present investigation aimed to elucidate the probable mechanism of antiasthmatic action of essential oil of Pistacia integerrima J.L. Stewart ex Brandis galls (EOPI). EOPI was investigated for its potential antiasthmatic action using in vitro antiallergic assays mast cell degranulation and soyabean lipoxidase enzyme activit, and spasmolytic action using isolated guinea pig ileum preparation. In vivo studies included lipopolysaccharide-induced bronchial inflammation in rats and airway hyperresponsiveness in ovalbumin in sensitized guinea pigs using spirometry. Data was analysed by GraphPad Prism 5.01 and results were expressed as means ± SEM. P < 0.05 was considered to be significant. EOPI inhibits 5-lipoxidase enzyme activity, DPPH scavenging activity and erythropoietin- induced angiogenesis. It showed dose dependent anti-allergic activity by inhibiting compound 48/80 induced mast cell degranulation. The finding that essential oil induced inhibition of transient contraction of acetylcholine in calcium free medium, and relaxation of S-(-)-Bay 8644-precontracted isolated guinea pig ileum jointly suggest that suggesting that the L-subtype Cav channel is involved in spasmolytic action of EOPI. Treatment with EOPI dose dependently (7.5, 15 and 30 mg/kg i.p.) inhibited lipopolysaccharide- induced increased in total cell count, neutrophil count, nitrate-nitrite, total protein, albumin levels in bronchoalveolar fluid and myeloperoxidase levels in lung homogenates. Mild diffused lesions involving focal interalveolar septal, intraluminal infiltration of neutrophils were observed in EOPI (7.5 &15 mg/kg) pretreated while no abnormality was detected in EOPI (30 mg/kg) and roflumilast (1mg/kg) pretreated rats. Roflumilast was used as standard. EOPI reduced the respiratory flow due to gasping in ovalbumin sensitized guinea pigs. The study demonstrates the effectiveness of EOPI in bronchial asthma possibly related to its ability to inhibit L-subtype Cav channel, mast cell stabilization, antioxidant, angiostatic and through inhibition of 5-lipoxygenase enzyme.

Keywords: asthma, lipopolysaccharide, spirometry, Pistacia integerrima J.L. Stewart ex Brandis, essential oil

Procedia PDF Downloads 285

Authors: Boris Nemzer, Tania Reyes-Izquierdo, Zbigniew Pietrzkowski

Abstract:

Glucosinolate is a family of glucosides that can be found in a family of brassica vegetables. Upon the damage of the plant, glucosinolate breakdown by an internal enzyme myrosinase (thioglucosidase; EC 3.2.3.1) into isothiocyanates, such as sulforaphane. Sulforaphane is formed by glucoraphanin cleaving the sugar off by myrosinase and rearranged. Sulforaphane nitrile is formed in the same reaction as sulforaphane with the active of epithiospecifier protein (ESP). Most common food processing procedure would break the plant and mix the glucoraphanin and myrosinase together, and the formed sulforaphane would be further degraded. The purpose of this study is to understand the glucoraphanin/sulforaphane and the myrosinase activity of broccoli seeds germinated at a different time and technological processing conditions that keep the activity of the enzyme to form sulforaphane. Broccoli seeds were germinated in the house. Myrosinase activities were tested as the glucose content using glucose assay kit and measured UV-Vis spectrophotometer. Glucosinolates were measured by HPLC/DAD. Sulforaphane was measured using HPLC-DAD and GC/MS. The 6 hr germinated sprouts have a myrosinase activity 32.2 mg glucose/g, which is comparable with 12 and 24 hour germinated seeds and higher than dry seeds. The glucoraphanin content in 6 hour germinated sprouts is 13935 µg/g which is comparable to 24 hour germinated seeds and lower than the dry seeds. GC/MS results show that the amount of sulforaphane is higher than the amount of sulforaphane nitrile in seeds, 6 hour and 24 hour germinated seeds. The ratio of sulforaphane and sulforaphane nitrile is high in 6 hour germinated seeds, which indicates the inactivated ESP in the reaction. After evaluating the results, the short time germinated seeds can be used as the source of glucoraphanin and myrosinase supply to form potential higher sulforaphane content. Broccoli contains glucosinolates, glucoraphanin (4-methylsulfinylbutyl glucosinolate), which is an important metabolite with health-promoting effects. In the pilot clinical study, we observed the effects of a glucosinolates/glucoraphanin-rich extract from short time germinated broccoli seeds on blood adenosine triphosphate (ATP), reactive oxygen species (ROS) and lactate levels. A single dose of 50 mg of broccoli sprouts extract increased blood levels of ATP up to 61% (p=0.0092) during the first 2 hours after the ingestion. Interestingly, this effect was not associated with an increase in blood ROS or lactate. When compared to the placebo group, levels of lactate were reduced by 10% (p=0.006). These results indicate that broccoli germinated seed extract may positively affect the generation of ATP in humans. Due to the preliminary nature of this work and promising results, larger clinical trials are justified.

Keywords: broccoli glucosinolates, glucoraphanin, germinated seeds, myrosinase, adenosine triphosphate

Procedia PDF Downloads 291

Authors: Jia-Ling Ho, Xiu-Ru Zhang, Zwe-Ling Kong

Abstract:

Diabetes mellitus (DM) is a major health problem that affects patients’ life quality throughout the world due to its many complications. It characterized by chronic hyperglycemia with oxidative stress, which impaired male reproductive function. Fibroblast growth factor 21 (FGF21) is a metabolic regulator that is required for normal spermatogenesis and protects against diabetes-induced germ cell apoptosis. Echinacea purpurea ethanol extract (EE), which contain phenolic acid and isobutylamide, had been proven to have antidiabetic property. Silica-chitosan nanoparticles (Nano-CS) has drug delivery and controlled release properties. This study aims to investigate whether silica-chitosan nanoparticles encapsulated EE (Nano-EE) had more ameliorating male infertility by analyzing the effect of testicular FGF21. The Nano-EE was characterized before used to treatment the diabetic rat model. Male Sprague-Dawley (SD) rats were obtained and divided into seven groups. A group was no induced Streptozotocin (STZ), marked as normal group. Diabetic rats were induced into diabetes by STZ (33 mg/kg). A diabetic group was no treatment with sample (diabetic control group), and other groups were treatment by Nano-CS (465 mg/kg), Nano-EE (93, 279, 465 mg/kg), and metformin (Met) (200 mg/kg) used as reference drug for 7 weeks. Our results indicated that the average nanoparticle size and zeta potential of Nano-EE were 2630 nm and -21.3 mV, respectively. The encapsulation ratio of Nano-EE was about 70%. It also confirmed the antioxidative activity was unchanged by comparing the DPPH and ABTS scavenging of Nano-EE and EE. In vivo test, Nano-EE can improve the STZ induced hyperglycemia, insulin resistance, and plasma FGF21 levels. Nano-EE has increased sperm motility, mitochondria membrane potential (MMP), plasma testosterone level, and reduction of abnormal sperm, nitric oxide (NO), superoxide production as well as reactive oxygen species (ROS). In addition, in plasma antioxidant enzymes glutathione peroxidase (GPx) and superoxide dismutase (SOD) was increased whereas pro-inflammatory cytokines TNF-α, and IL-1β were decreased. Further, in testis, protein content of FGF21, PGC-1α, and SIRT1 were improved. Nano-EE might improve diabetes-induced down-regulation of testicular FGF21 and SIRT1/PGC-1α signaling hence maintain spermatogenesis.

Keywords: diabetes mellitus, Echinacea purpurea, reproductive dysfunction, silica-chitosan nanoparticles

Procedia PDF Downloads 194

Authors: Thabiso M. Sebolai, Victor Mlambo, Solomon Tefera, Othusitse R. Madibela

Abstract:

In some tree species, sustained herbivory can induce changes in biosynthetic pathways resulting in overproduction of anti-nutritional secondary plant compounds. This inductive mechanism, which has not been demonstrated in semi-arid rangelands of South Africa, may result in browse leaves of lower nutritive value. In this study we investigate the interactive effect of browsing pressure and tree species on chemical composition, buffer nitrogen solubility index (NSI), in vitro ruminal dry matter degradability (IVDMD) and in vitro ruminal N degradability (IVND) of leaves. Leaves from Maytenus capitata, Olea africana, Coddia rudis, Carissa macrocarpa, Rhus refracta, Ziziphus mucronata, Boscia oliedes, Grewia robusta, Phyllanthus vessucosus and Ehretia rigida trees growing in a communal grazing area were harvested at two heights: browsable ( < 1.5 m) and non-browsable ( > 1.5 m), representing high and low browsing pressure, respectively. The type of animals utilizing the communal rangeland includes cattle at 1 livestock unit (450kg)/12 to 15 hectors and goats at 1 livestock unit/4 ha. Harvested leaves were dried, milled and analysed for proximate components, soluble phenolics, condensed tannins, minerals and in vitro ruminal fermentation. A significant plant species and harvesting height interaction effect (P < 0.05) was observed for total nitrogen (N) and soluble phenolics concentration. Tree species and harvesting height affected (P < 0.05) condensed tannin (CTs) content where samples harvested from the non-browsable height had higher (0.61 AU550 nm/200 mg) levels than those harvested at browsable height (0.55 AU550 nm/200 mg) while their interaction had no effects. Macro and micro-minerals were only influenced (P < 0.05) by browse species but not harvesting height. Species and harvesting height interacted (P < 0.05) to influence IVDMD and IVND of leaves at 12, 24 and 36 hours of incubation. The different browse leaves contained moderate to high protein, moderate level of phenolics and minerals, suggesting that they have the potential to provide supplementary nutrients for ruminants during the dry seasons.

Keywords: browse plants, chemical composition, harvesting heights, phenolics

Procedia PDF Downloads 147

Authors: Omar Pillaca-Pullo, Arturo Alejandro-Paredes, Carol Flores-Fernandez, Marijuly Sayuri Kina, Amparo Iris Zavaleta

Abstract:

Aqueous two-phase system (ATPS) is an interesting alternative for separating industrial enzymes due to it is easy to scale-up and low cost. Polyethylene glycol (PEG) mixed with potassium phosphate or magnesium sulfate is one of the most frequently polymer/salt ATPS used, but the consequences of its use is a high concentration of phosphates and sulfates in wastewater causing environmental issues. Citrate could replace these inorganic salts due to it is biodegradable and does not produce toxic compounds. On the other hand, statistical design of experiments is widely used for ATPS optimization and it allows to study the effects of the involved variables in the purification, and to estimate their significant effects on selected responses and interactions. The 24 factorial design with four central points (20 experiments) was employed to study the partition and purification of proteases produced by Pseudomonas sp. in PEG/citrate ATPS system. ATPS was prepared with different sodium citrate concentrations [14, 16 and 18% (w/w)], pH values (7, 8 and 9), PEG molecular weight (2,000; 4,000 and 6,000 g/mol) and PEG concentrations [18, 20 and 22 % (w/w)]. All system components were mixed with 15% (w/w) of the fermented broth and deionized water was added to a final weight of 12.5 g. Then, the systems were mixed and kept at room temperature until to reach two-phases separation. Volumes of the top and bottom phases were measured, and aliquots from both phases were collected for subsequent proteolytic activity and total protein determination. Influence of variables such as PEG molar mass (MPEG), PEG concentration (CPEG), citrate concentration (CSal) and pH were evaluated on the following responses: purification factor (PF), activity yield (Y), partition coefficient (K) and selectivity (S). STATISTICA program version 10 was used for the analysis. According to the obtained results, higher levels of CPEG and MPEG had a positive effect on extraction, while pH did not influence on the process. On the other hand, the CSal could be related with low values of Y because of the citrate ions have a negative effect on solubility and enzymatic structure. The optimum values of Y (66.4 %), PF (1.8), K (5.5) and S (4.3) were obtained at CSal (18%), MPEG (6,000 g/mol), CPEG (22%) and pH 9. These results indicated that the PEG/citrate system is accurate to purify these Pseudomonas sp. proteases from fermented broth as a first purification step.

Keywords: citrate, polyethylene glycol, protease, Pseudomonas sp

Procedia PDF Downloads 198

Authors: T. Rakhi, Thrivikram Shenoy

Abstract:

Esophageal atresia is a disorder of maldevelopment of esophagus with or without a connection to the trachea. Radiological reviews are needed in consultation with the pediatric surgeon and neonatologist and we report a rare case of esophageal atresia associated with atrial septal defect-patent ductus arteriosus complex. A 2-day old female baby born at term, weighing 3.010kg, admitted to the Neonatal Intensive Care Unit with respiratory distress and excessive oral secretions. On examination, continuous murmur and cyanosis were seen. Esophageal atresia was suspected, after a failed attempt to pass a nasogastric tube. Chest radiograph showed coiling of the nasogastric tube and absent gas shadow in the abdomen. Echocardiography confirmed Patent Ductus Arteriosus with Atrial Septal Defect not in failure and was diagnosed with esophageal atresia with suspected fistula posted for surgical repair. After preliminary management with oxygenation, suctioning in prone position and antibiotics, investigations revealed Hb 17gms serum biochemistry, coagulation profile and C-Reactive Protein Test normal. The baby was premedicated with 5mcg of fentanyl and 100 mcg of midazolam and a rapid awake laryngoscopy was done to rule out difficult airway followed by induction with o2 air, sevo and atracurium 2 mg. Placement of a 3.5 tube was uneventful at first attempt and after confirming bilateral air entry positioned in the lateral position for Right thoracotomy. A pulse oximeter, Echocardiogram, Non-invasive Blood Pressure, temperature and a precordial stethoscope in left axilla were essential monitors. During thoracotomy, both the ends of the esophagus and the fistula could not be located after thorough search suggesting an on table finding of type A esophageal atresia. The baby was repositioned for gastrostomy, and cervical esophagostomy ventilated overnight and extubated uneventful. Absent gas shadow was overlooked and the purpose of this presentation is to create an awareness between the neonatologist, pediatric surgeons and anesthesiologist regarding variation of typing of Tracheoesophageal fistula pre and intraoperatively. A need for imaging modalities warranted for a definitive diagnosis in the presence of a gasless stomach.

Keywords: anesthetic, atrial septal defects, esophageal atresia, patent ductus arteriosus, perioperative, chest x-ray

Procedia PDF Downloads 179

Authors: M. Saleem, M. Shahnawaz, A. W. Gandahi, S. M. Bhatti

Abstract:

The micronutrients boron and zinc deficiencies are extensive in the areas of rice-wheat cropping system. Optimum levels of these nutrients in soil are necessary for healthy crop growth. Since rice and wheat are major staple food of worlds’ populace, the higher yields and nutrition status of these crops has direct effect on the health of human being and economy of the country. A field study was conducted to observe the direct and residual effect of two selected micronutrients boron (B) and zinc (Zn)) on rice and wheat crop growth and its grain nutrient status. Each plot received either B or Zn at the rates of 0, 1, 2, 3 and 4 kg B ha⁻¹, and 5, 10, 15 and 20 kg Zn ha⁻¹, combined B and Zn application at 1 kg B and 5 kg Zn ha⁻¹, 2 kg B and 10 kg Zn ha⁻¹. Colemanite ore were used as source of B and zinc sulfate for Zn. The second season wheat crop was planted in the same plots after the interval period of 30 days and during this time gap soil was fallow. Boron and Zn application significantly enhanced the plant height, number of tillers, Grains panicle⁻¹ seed index fewer empty grains panicle⁻¹ and yield of rice crop at all defined levels as compared to control. The highest yield (10.00 tons/ha) was recorded at 2 Kg B, 10 Kg Zn ha⁻¹ rates. Boron and Zn concentration in grain and straw significantly increased. The application of B also improved the nutrition status of rice as B, protein and total carbohydrates content of grain augmented. The analysis of soil samples collected after harvest of rice crop showed that the B and Zn content in post-harvest soil samples was high in colemanite and zinc sulfate applied plots. The residual B and Zn were also effectual for the second season wheat crop, as the growth parameters plant height, number of tillers, earhead length, weight 1000 grains, B and Zn content of grain significantly improved. The highest wheat grain yield (4.23 tons/ha) was recorded at the residual rates of 2 kg B and 10 kg Zn ha⁻¹ than the other treatments. This study showed that one application of B and Zn can increase crop yields for at least two consecutive seasons and the mineral colemanite can confidently be used as source of B for rice crop because very small quantities of these nutrients are consumed by first season crop and remaining amount was present in soil which were used by second season wheat crop for healthy growth. Consequently, there is no need to apply these micronutrients to the following crop when it is applied on the previous one.

Keywords: residual boron, zinc, rice, wheat

Procedia PDF Downloads 156

Authors: D. Nagalakshmi, S. Parashuramulu K. Sadasiva Rao, G. Aruna, L. Vikram

Abstract:

The zinc (Zn) is the second most abundant trace element in mammals and birds, forming structural component of over 300 enzymes, playing an important role in anti-oxidant defense, immune response and reproduction. Organic trace minerals are more readily absorbed from the digestive tract and more biologically available compared with its inorganic salt. Thus, the present study was undertaken on 60 adult female Sprague Dawley rats (275±2.04 g) for experimental duration of 12 weeks to investigate the effect of dietary Zn supplementation from various organic sources on immunity, reproduction, oxidative defense mechanism and blood biochemical profile. The rats were randomly allotted to 30 replicates (2 per replicate) which were in turn randomly allotted to 5 dietary treatments varying in Zn source i.e., one inorganic source (Zn carbonate) and 4 organic sources (Zn-proteinate, Zn-propionate, Zn-amino acid complex and Zn-methionine) so as to supply NRC recommended Zn concentration (12 ppm Zn). Supplementation of organic Zn had no effect on various haematological and serum biochemical constituents compared to inorganic Zn fed rats. The TBARS and protein carbonyls concentration in liver indicative of oxidative stress was comparable between various organic and inorganic groups. The glutathione reductase activity in haemolysate (P<0.05) and reduced glutathione concentration in liver (P<0.01) was higher when fed organic Zn and RBC catalase activity was higher (P<0.01) on Zn methionine compared to other organic sources tested and the inorganic source. The humoral immune response assessed as antibody titres against sheep RBC was higher (P<0.05) when fed organic sources of zinc compared to inorganic source. The cell mediated immune response expressed as delayed type hypersensitivity reaction was higher (P<0.05) in rats fed Zn propionate with no effect of other organic Zn sources. The serum progesterone concentration was higher (P<0.05) in rats fed organic Zn sources compared to inorganic zinc. The data on ovarian folliculogenesis indicated that organic Zn supplementation increased (P<0.05) the number of graafian follicles and corpus luteum with no effect on primary, secondary and tertiary follicle number. The study indicated that rats fed organic sources of Zn had higher antioxidant enzyme activities, immune response and serum progesterone concentration with higher number of mature follicles. Though the effect of feeding various organic sources were comparable, rats fed zinc methionine had higher antioxidant activity and cell mediated immune response was higher in rats on Zn propionate.

Keywords: organic zinc, immune, rats, reproductive

Procedia PDF Downloads 287

Authors: M. M. Essa, S. Subash, M. Akbar, S. Al-Adawi, A. Al-Asmi, G. J. Guillemein

Abstract:

Objective: Transgenic (tg) mice which contain an amyloid precursor protein (APP) gene mutation, develop extracellular amyloid beta (Aβ) deposition in the brain, and severe memory and behavioural deficits with age. These mice serve as an important animal model for testing the efficacy of novel drug candidates for the treatment and management of symptoms of Alzheimer's disease (AD). Several reports have suggested that oxidative stress is the underlying cause of Aβ neurotoxicity in AD. Pomegranates contain very high levels of antioxidants and several medicinal properties that may be useful for improving the quality of life in AD patients. In this study, we investigated the effect of dietary supplementation of Omani pomegranate extract on the memory, anxiety and learning skills along with inflammation in an AD mouse model containing the double Swedish APP mutation (APPsw/Tg2576). Methods: The experimental groups of APP-transgenic mice from the age of 4 months were fed custom-mix diets (pellets) containing 4% pomegranate. We assessed spatial memory and learning ability, psychomotor coordination, and anxiety-related behavior in Tg and wild-type mice at the age of 4-5 months and 18-19 months using the Morris water maze test, rota rod test, elevated plus maze test, and open field test. Further, inflammatory parameters also analysed. Results: APPsw/Tg2576 mice that were fed a standard chow diet without pomegranates showed significant memory deficits, increased anxiety-related behavior, and severe impairment in spatial learning ability, position discrimination learning ability and motor coordination along with increased inflammation compared to the wild type mice on the same diet, at the age of 18-19 months In contrast, APPsw/Tg2576 mice that were fed a diet containing 4% pomegranates showed a significant improvements in memory, learning, locomotor function, and anxiety with reduced inflammatory markers compared to APPsw/Tg2576 mice fed the standard chow diet. Conclusion: Our results suggest that dietary supplementation with pomegranates may slow the progression of cognitive and behavioural impairments in AD. The exact mechanism is still unclear and further extensive research needed.

Keywords: Alzheimer's disease, pomegranates, oman, cognitive decline, memory loss, anxiety, inflammation

Procedia PDF Downloads 530

Authors: Ieva Bruzauskaite, Jovile Raudoniute, Karina Poliakovaite, Danguole Zabulyte, Daiva Bironaite, Ruta Aldonyte

Abstract:

Introduction: Air pollution is a growing global problem which has been shown to be responsible for various adverse health outcomes. Immunotoxicity, such as dysregulated inflammation, has been proposed as one of the main mechanisms in air pollution-associated diseases. Chronic obstructive pulmonary disease (COPD) is among major morbidity and mortality causes worldwide and is characterized by persistent airflow limitation caused by the small airways disease (obstructive bronchiolitis) and irreversible parenchymal destruction (emphysema). Exact pathways explaining the air pollution induced and mediated disease states are still not clear. However, modern societies understand dangers of polluted air, seek to mitigate such effects and are in need for reliable biomarkers of air pollution. We hypothesise that post-translational modifications of structural proteins, e.g. citrullination, might be a good candidate biomarker. Thus, we have designed this study, where mice were exposed to diesel exhaust and the ongoing protein modifications and inflammation in lungs and other tissues were assessed. Materials And Methods: To assess the effects of diesel exhaust a in vivo study was designed. Mice (n=10) were subjected to everyday 2-hour exposure to diesel exhaust for 14 days. Control mice were treated the same way without diesel exhaust. The effects within lung and other tissues were assessed by immunohistochemistry of formalin-fixed and paraffin-embedded tissues. Levels of inflammation and citrullination related markers were investigated. Levels of parenchymal damage were also measured. Results: In vivo study corroborates our own data from in vitro and reveals diesel exhaust initiated inflammatory shift and modulation of lung peptidyl arginine deiminase 4 (PAD4), citrullination associated enzyme, levels. In addition, high levels of citrulline were observed in exposed lung tissue sections co-localising with increased parenchymal destruction. Conclusions: Subacute exposure to diesel exhaust renders mice lungs inflammatory and modifies certain structural proteins. Such structural changes of proteins may pave a pathways to lost/gain function of affected molecules and also propagate autoimmune processes within the lung and systemically.

Keywords: air pollution, citrullination, in vivo, lungs

Procedia PDF Downloads 157

Authors: Samane Eftekhari Ranjbar

Abstract:

Diabetes mellitus, a metabolic disorder characterized by elevated blood glucose levels, ranks among the leading causes of adult mortality. This study investigates the impact of an eight-week high-intensity interval training (HIIT) program combined with sodium nitrite supplementation on TNF- α, MURF1, and PI3K in a type 2 diabetes rodent model. Elevated TNF-α levels have been associated with insulin resistance, while MURF1 and PI3K play roles in muscle atrophy and insulin signaling pathways, respectively. In this experimental study, 15 eight-week-old rats from the Sara Laboratory Center in Tabriz were assigned to one of five groups: healthy control, diabetic control, diabetic with sodium nitrite supplementation, diabetic with eight weeks of intermittent exercise, and diabetic with eight weeks of interval training plus sodium nitrite supplementation. The HIIT protocol was designed to span eight weeks, with five weekly sessions at specified intensities and durations. Sodium nitrite, known for its vasodilatory and cytoprotective properties, was administered via injection. The findings revealed that the HIIT program and sodium nitrite supplementation influenced the examined biomarkers. ANOVA test outcomes indicated statistically significant differences in TNF- α (P=0.001), MURF1 (P=0.001), and PI3K (P=0.001) concentrations among the various groups. The healthy control group exhibited substantially decreased TNF- α, and MURF1 levels, as well as elevated PI3K levels compared to the diabetic control group. The exercise group, in conjunction with sodium nitrite supplementation, demonstrated a significant rise in PI3K levels (P=0.001) and a decline in TNF- α levels (P=0.018) relative to the diabetic control group. These results suggest that the combined intervention may help improve insulin sensitivity and reduce inflammation. However, MURF1 levels, which are related to muscle atrophy, showed no significant difference (P=0.24). In conclusion, in type 2 diabetic rats, an eight-week high-intensity interval training program with sodium nitrite supplementation does not affect MURF1 levels but does influence PI3K and TNF- α levels. This combination may hold potential for improving insulin sensitivity and reducing inflammation in type 2 diabetes patients, warranting further investigation and potential translation to human clinical trials.

Keywords: high-intensity interval training, sodium nitrate supplementation, type 2 diabetes, tumor necrosis factor-alpha, phosphatidylinositol-3-kinase, muscle RING-finger protein-1

Procedia PDF Downloads 90

Authors: Flóra Karsai-Rektenwald, Khongorzul Odgerel, Vanda Villányi, Zoltán Gábor Tóth, Zsófia Bánfalvi

Abstract:

GIGANTEA (GI) is a plant-specific, circadian clock-regulated, nuclear protein involved in diverse processes from flowering to stress responses. In the obligate short-day tuberising Andigenum Group potatoes, GI is indirectly involved in determination of the time of tuber initiation. The goal of our study was to get information on the function of GI in the day-length independent tuberising commercial potato cultivar ‘Désirée’, a tetraploid plant carrying two GI genes, one on chromosome 4 (GI.04) and another one on chromosome 12 (GI.12). Functional analysis of the two GI genes was attempted by targeted mutagenesis using the CRISPR-Cas9 system. Two sets of mutants were generated. The mutations were mapped at nucleotide level and the plants grown in a greenhouse. GI is located in the nucleus and interacts with at least five proteins. Three out of them, two photoreceptors and FKF1, bind on GI close to the nuclear localisation (NLS) signal to the so called LOV domain. In Andigenum Group potatoes, FKF1 interacts not only with GI but form a triplex with CDF1, a positive regulator of tuberisation, and transport it to proteasomes, where CDF1 is degraded. Three GI.04 and three GI.12 null mutants were selected from the first set of mutagenesis. Although, the deletions did not reach the NLS and LOV domain in any of the six mutants all GI. 04 and two GI.12 mutants were shorter than the control suggesting that both GIs are involved in vegetative growth regulation and the deleted region might be important in terms of conformation or stability of the proteins. From the second set of mutagenesis, three null mutants carrying mutations in one of the two GI genes and three mutants carrying mutations in both GI genes were selected for detailed analysis. Deletions in the GI mutants of this set disrupted the NLS and extended to the LOV domain. Nevertheless, none of the single GI gene mutations influenced the time of tuberisation or the tuber number and yield, whereas one of the GI.04 and all GI.12 mutants were shorter than the ‘Désirée’ control. Furthermore, all GI.12 mutants showed early senescence. The early senescence of mutants carrying mutations in both GI genes was even more pronounced, resulting in substantial yield loss in one of the double mutants. These results raise the possibility that the two GI genes can substitute each other in term of tuberisation or they are not involved in it, the yield loss is due to the early death of the plants. To distinguish between the two possibilities yeast two-hybrid experiments were initiated to detect the interaction between the GI proteins and FKF1 and between FKF1 and CDF1 originated from ‘Désirée’.

Keywords: gene editing, tuberisation, senescence, Solanum tuberosum

Procedia PDF Downloads 13

Authors: Aidin Foroutan, David S. Wishart, Leluo L. Guan, Carolyn Fitzsimmons

Abstract:

Maximizing efficiency and growth potential of beef cattle requires not only genetic selection (i.e. residual feed intake (RFI)) but also adequate nutrition throughout all stages of growth and development. Nutrient restriction during gestation has been shown to negatively affect post-natal growth and development as well as fertility of the offspring. This, when combined with RFI may affect progeny traits. This study aims to investigate the impact of selection for divergent genetic potential for RFI and maternal nutrition during early- to mid-gestation, on bull calf traits such as fertility and muscle development using multiple ‘omics’ approaches. Comparisons were made between High-diet vs. Low-diet and between High-RFI vs. Low-RFI animals. An epigenetics experiment on semen samples identified 891 biomarkers associated with growth and development. A gene expression study on Longissimus thoracis muscle, semimembranosus muscle, liver, and testis identified 4 genes associated with muscle development and immunity of which Myocyte enhancer factor 2A [MEF2A; induces myogenesis and control muscle differentiation] was the only differentially expressed gene identified in all four tissues. An initial metabolomics experiment on serum samples using nuclear magnetic resonance (NMR) identified 4 metabolite biomarkers related to energy and protein metabolism. Once all the biomarkers are identified, bioinformatics approaches will be used to create a database covering all the ‘omics’ data collected from this project. This database will be broadened by adding other information obtained from relevant literature reviews. Association analyses with these data sets will be performed to reveal key biological pathways affected by RFI and maternal nutrition. Through these association studies between the genome and metabolome, it is expected that candidate biomarker genes and metabolites for feed efficiency, fertility, and/or muscle development are identified. If these gene/metabolite biomarkers are validated in a larger animal population, they could potentially be used in breeding programs to select superior animals. It is also expected that this work will lead to the development of an online tool that could be used to predict future traits of interest in an animal given its measurable ‘omics’ traits.

Keywords: biomarker, maternal nutrition, omics, residual feed intake

Procedia PDF Downloads 193

Authors: Sultan Çağırıcı, Arsida Bajrami, Beyza Aslan, Hacı Ali Erdoğan, Nejla Sözer Topçular, Dilek Bozkurt, Vildan Yayla

Abstract:

Objective: Corticobasal syndrome (CBS) is phenotypically characterized by asymmetric rigidity, apraxia, alien-limb phenomenon, cortical sensory loss, dystonia and myoclonus. The underlying pathologies consists of corticobasal degeneration (CBD), progressive supra nuclear palsy, Alzheimer's, Creutzfeldt-Jakob and frontotemporal degeneration. CBD is a degenerative disease with clinical symptoms related to the prominent involvement of cerebral cortex and basal ganglia. CBD is a pathological diagnosis and antemortem clinical diagnosis may change many times. In this paper, we described the clinical features and discussed a cases diagnosed with symmetric CBS because of its rarity. Case: Seventy-five-year-old woman presented with a three years history of difficulty in speaking and reading. Involuntary hand jerks and slowness of movement also had began in the last six months. In the neurological examination the patient was alert but not fully oriented. The speech was non-fluent, word finding difficulties were present. Bilateral limited upgaze, bradimimia, bilateral positive cogwheel' rigidity but prominent in the right side, postural tremor and negative myoclonus during action on the left side were detected. Receptive language was normal but expressive language and repetition were impaired. Acalculia, alexia, agraphia and apraxia were also present. CSF findings were unremarkable except for elevated protein level (75 mg/dL). MRI revealed bilateral symmetric cortical atrophy prominent in the frontoparietal region. PET showed hypometabolism in the left caudate nucleus. Conclusion: The increase of data related to neurodegenerative disorders associated with dementia, movement disorders and other findings results in an expanded range of diagnosis and transitions between clinical diagnosis. When considered the age of onset, clinical symptoms, imaging findings and prognosis of this patient, clinical diagnosis was CBS and pathologic diagnosis as probable CBD. Imaging of CBD usually consist of typical asymmetry between hemispheres. Still few cases with clinical appearance of CBD may show symmetrical cortical cerebral atrophy. It is presented this case who was diagnosed with CBD although we found symmetrical cortical cerebral atrophy in MRI.

Keywords: symmetric cortical atrophy, corticobasal degeneration, corticobasal syndrome

Procedia PDF Downloads 459

Authors: Alexandra C. Blaga, Dan Caşcaval, Alexandra Tucaliuc, Madalina Poştaru, Anca I. Galaction

Abstract:

Amino acids are valuable chemical products used in in human foods, in animal feed additives and in the pharmaceutical field. Recently, there has been a noticeable rise of amino acids utilization throughout the world to include their use as raw materials in the production of various industrial chemicals: oil gelating agents (amino acid-based surfactants) to recover effluent oil in seas and rivers and poly(amino acids), which are attracting attention for biodegradable plastics manufacture. The amino acids can be obtained by biosynthesis or from protein hydrolysis, but their separation from the obtained mixtures can be challenging. In the last decades there has been a continuous interest in developing processes that will improve the selectivity and yield of downstream processing steps. The liquid-liquid extraction of amino acids (dissociated at any pH-value of the aqueous solutions) is possible only by using the reactive extraction technique, mainly with extractants of organophosphoric acid derivatives, high molecular weight amines and crown-ethers. The purpose of this study was to analyse the separation of nine amino acids of acidic character (l-aspartic acid, l-glutamic acid), basic character (l-histidine, l-lysine, l-arginine) and neutral character (l-glycine, l-tryptophan, l-cysteine, l-alanine) by reactive extraction with di-(2-ethylhexyl)phosphoric acid (D2EHPA) dissolved in butyl acetate. The results showed that the separation yield is controlled by the pH value of the aqueous phase: the reactive extraction of amino acids with D2EHPA is possible only if the amino acids exist in aqueous solution in their cationic forms (pH of aqueous phase below the isoeletric point). The studies for individual amino acids indicated the possibility of selectively separate different groups of amino acids with similar acidic properties as a function of aqueous solution pH-value: the maximum yields are reached for a pH domain of 2–3, then strongly decreasing with the pH increase. Thus, for acidic and neutral amino acids, the extraction becomes impossible at the isolelectric point (pHi) and for basic amino acids at a pH value lower than pHi, as a result of the carboxylic group dissociation. From the results obtained for the separation from the mixture of the nine amino acids, at different pH, it can be observed that all amino acids are extracted with different yields, for a pH domain of 1.5–3. Over this interval, the extract contains only the amino acids with neutral and basic character. For pH 5–6, only the neutral amino acids are extracted and for pH > 6 the extraction becomes impossible. Using this technique, the total separation of the following amino acids groups has been performed: neutral amino acids at pH 5–5.5, basic amino acids and l-cysteine at pH 4–4.5, l-histidine at pH 3–3.5 and acidic amino acids at pH 2–2.5.

Keywords: amino acids, di-(2-ethylhexyl) phosphoric acid, reactive extraction, selective extraction

Procedia PDF Downloads 431

Authors: Ahmad Alsheikh Kaddour

Abstract:

Herbal medicine has been a long-standing phenomenon in Arab countries since ancient times because of its breadth and moderate temperament. Therefore, it possesses a vast natural and economic wealth of medicinal and aromatic herbs. This prompted ancient Egyptians and Arabs to discover and exploit them. The economic importance of the plant is not only from medicinal uses; it is a plant of high economic value for its various uses, especially in food, cosmetic and aromatic industries. It is also an ornamental plant and soil stabilization. The main objective of this research is to study the chemical changes that occur in the plant during the growth period, as well as the production of plant buds, which were previously considered unwanted plants. The research was carried out in the period 2021-2022 in the valley of Al-Shaflah (common caper), located in Qumhana village, 7 km north of Hama Governorate, Syria. The results of the research showed a change in the percentage of chemical components in the plant parts. The ratio of protein content and the percentage of fatty substances in fruits and the ratio of oil in the seeds until the period of harvesting of these plant parts improved, but the percentage of essential oils decreased with the progress of the plant growth, while the Glycosides content where improved with the plant aging. The production of buds is small, with dimensions as 0.5×0.5 cm, which is preferred for commercial markets, harvested every 2-3 days in quantities ranging from 0.4 to 0.5 kg in one cut/shrubs with 3 years’ age as average for the years 2021-2022. The monthly production of a shrub is between 4-5 kg per month. The productive period is 4 months approximately. This means that the seasonal production of one plant is 16-20 kg and the production of 16-20 tons per year with a plant density of 1,000 shrubs per hectare, which is the optimum rate of cultivation in the unit of mass, given the price of a kg of these buds is equivalent to 1 US $; however, this means that the annual output value of the locally produced hectare ranges from 16,000 US $ to 20,000 US $ for farmers. The results showed that it is possible to transform the cultivation of this plant from traditional random to typical areas cultivation, with a plant density of 1,000-1,100 plants per hectare according to the type of soil to obtain production of medicinal and nutritious buds, as well as, the need to pay attention to this national wealth and invest in the optimal manner, which leads to the acquisition of hard currency through export to support the national income.

Keywords: common caper, medicinal plants, propagation, medical, economic importance

Procedia PDF Downloads 73

Authors: Kangana Sengar, Sanjay Deb, Ramesh Dawar

Abstract:

Introduction: Ki-67 can be a useful marker in determining proliferative activity in patients with multiple myeloma (MM). However, using Ki-67 alone results in the erroneous inclusion of non-myeloma cells leading to false high counts. We have used Dual IHC (immunohistochemistry) staining with Ki-67 and CD138 to enhance specificity in assessing proliferative activity of bone marrow plasma cells. Aims and objectives: To estimate the proportion of proliferating (Ki-67 expressing) plasma cells in patients with MM and correlation of Ki-67 with other known prognostic parameters. Materials and Methods: Fifty FFPE (formalin fixed paraffin embedded) blocks of trephine biopsies of cases diagnosed as MM from 2010 to 2015 are subjected to H & E staining and Dual IHC staining for CD 138 and Ki-67. H & E staining is done to evaluate various histological parameters like percentage of plasma cells, pattern of infiltration (nodular, interstitial, mixed and diffuse), routine parameters of marrow cellularity and hematopoiesis. Clinical data is collected from patient records from Medical Record Department. Each of CD138 expressing cells (cytoplasmic, red) are scored as proliferating plasma cells (containing a brown Ki¬67 nucleus) or non¬proliferating plasma cells (containing a blue, counter-stained, Ki-¬67 negative nucleus). Ki-67 is measured as percentage positivity with a maximum score of hundred percent and lowest of zero percent. The intensity of staining is not relevant. Results: Statistically significant correlation of Ki-67 in D-S Stage (Durie & Salmon Stage) I vs. III (p=0.026) and ISS (International Staging System) Stage I vs. III (p=0.019), β2m (p=0.029) and percentage of plasma cells (p < 0.001) is seen. No statistically significant correlation is seen between Ki-67 and hemoglobin, platelet count, total leukocyte count, total protein, albumin, S. calcium, S. creatinine, S. LDH, blood urea and pattern of infiltration. Conclusion: Ki-67 index correlated with other known prognostic parameters. However, it is not determined routinely in patients with MM due to little information available regarding its relevance and paucity of studies done to correlate with other known prognostic factors in MM patients. To the best of our knowledge, this is the first study in India using Dual IHC staining for Ki-67 and CD138 in MM patients. Routine determination of Ki-67 will help to identify patients who may benefit with more aggressive therapy. Recommendation: In this study follow up of patients is not included, and the sample size is small. Studying with larger sample size and long follow up is advocated to prognosticate Ki-67 as a marker of survival in patients with multiple myeloma.

Keywords: bone marrow, dual IHC, Ki-67, multiple myeloma

Procedia PDF Downloads 159

Authors: Ellana Boada, Eric Santos-Clotas, Alba Cabrera-Codony, Maria Martin, Lluis Baneras, Frederic Gich

Abstract:

Volatile methylsiloxanes (VMS) are a group of manmade silicone compounds widely used in household and industrial applications that end up on the biogas produced through the anaerobic digestion of organic matter in landfills and wastewater treatment plants. The presence of VMS during the biogas energy conversion can cause damage on the engines, reducing the efficiency of this renewable energy source. Non regenerative adsorption onto activated carbon is the most widely used technology to remove siloxanes from biogas, while new trends point out that biotechnology offers a low-cost and environmentally friendly alternative to conventional technologies. The first objective of this research was to enrich, isolate and identify bacterial species able to grow using siloxane molecules as a sole carbon source: anoxic wastewater sludge was used as initial inoculum in liquid anoxic enrichments, adding D4 (as representative siloxane compound) previously adsorbed on activated carbon. After several months of acclimatization, liquid enrichments were plated onto solid media containing D4 and thirty-four bacterial isolates were obtained. 16S rRNA gene sequencing allowed the identification of strains belonging to the following species: Ciceribacter lividus, Alicycliphilus denitrificans, Pseudomonas aeruginosa and Pseudomonas citronellolis which are described to be capable to degrade toxic volatile organic compounds. Kinetic assays with 8 representative strains revealed higher cell growth in the presence of D4 compared to the control. Our second objective was to characterize the community composition and diversity of the microbial community present in the enrichments and to elucidate whether the isolated strains were representative members of the community or not. DNA samples were extracted, the 16S rRNA gene was amplified (515F & 806R primer pair), and the microbiome analyzed from sequences obtained with a MiSeq PE250 platform. Results showed that the retrieved isolates only represented a minor fraction of the microorganisms present in the enrichment samples, which were represented by Alpha, Beta, and Gamma proteobacteria as dominant groups in the category class thus suggesting that other microbial species and/or consortia may be important for D4 biodegradation. These results highlight the need of additional protocols for the isolation of relevant D4 degraders. Currently, we are developing molecular tools targeting key genes involved in siloxane biodegradation to identify and quantify the capacity of the isolates to metabolize D4 in batch cultures supplied with a synthetic gas stream of air containing 60 mg m⁻³ of D4 together with other volatile organic compounds found in the biogas mixture (i.e. toluene, hexane and limonene). The isolates were used as inoculum in a biotrickling filter containing lava rocks and activated carbon to assess their capacity for siloxane removal. Preliminary results of biotrickling filter performance showed 35% of siloxane biodegradation in a contact time of 14 minutes, denoting that biological siloxane removal is a promising technology for biogas upgrading.

Keywords: bacterial cultivation, biogas upgrading, microbiome, siloxanes

Procedia PDF Downloads 260

Authors: Tatyane Lopes de Freitas, Antonio Diogo S. Vieira, Susana Marta Isay Saad, Maria Ines Genovese

Abstract:

The fruit processing industries generate a large volume of residues to produce juices, pulps, and jams. These residues, or by-products, consisting of peels, seeds, and pulps, are routinely discarded. Fruits are rich in bioactive compounds, including polyphenols, which have positive effects on health. Dry residues from two fruits, acerola (M. emarginata D. C.) and orange (C. sinensis), were characterized in relation to contents of ascorbic acid, minerals, total dietary fibers, moisture, ash, lipids, proteins, and carbohydrates, and also high performance liquid chromatographic profile of flavonoids, total polyphenols and proanthocyanidins contents, and antioxidant capacity by three different methods (Ferric reducing antioxidant power assay-FRAP, Oxygen Radical Absorbance Capacity-ORAC, 1,1-diphenyl-2-picrylhydrazil (DPPH) radical scavenging activity). Acerola by-products presented the highest acid ascorbic content (605 mg/100 g), and better antioxidant capacity than orange by-products. The dry residues from acerola demonstrated high contents of proanthocyanidins (617 µg CE/g) and total polyphenols (2525 mg gallic acid equivalents - GAE/100 g). Both presented high total dietary fiber (above 60%) and protein contents (acerola: 10.4%; orange: 9.9%), and reduced fat content (acerola: 1.6%; orange: 2.6%). Both residues showed high levels of potassium, calcium, and magnesium, and were considered sources of these minerals. With acerola by-product, four formulations of probiotics fermented milks were produced: F0 (without the addition of acerola residue (AR)), F2 (2% AR), F5 (5% AR) and F10 (10% AR). The physicochemical characteristics of the fermented milks throughout of storage were investigated, as well as the impact of in vitro simulated gastrointestinal conditions on flavonoids and probiotics. The microorganisms analyzed maintained their populations around 8 log CFU/g during storage. After the gastric phase of the simulated digestion, the populations decreased, and after the enteric phase, no colonies were detected. On the other hand, the flavonoids increased after the gastric phase, maintaining or suffering small decrease after enteric phase. Acerola by-products powder is a valuable ingredient to be used in functional foods because is rich in vitamin C, fibers and flavonoids. These flavonoids appear to be highly resistant to the acids and salts of digestion.

Keywords: acerola, orange, by-products, fermented milk

Procedia PDF Downloads 133

Authors: Sharat Chandra, Zilong Wang, Ru-Rong Ji, Andrey Bortsov

Abstract:

Chronic pain (CP) therapeutic approaches have limited efficacy. As a result, doctors are prescribing opioids for chronic pain, leading to opioid overuse, abuse, and addiction epidemic. Therefore, the development of effective and safe CP drugs remains an unmet medical need. Voltage-gated sodium (Nav) channels act as cardiovascular and neurological disorder’s molecular targets. Nav channels selective inhibitors are hard to design because there are nine closely-related isoforms (Nav1.1-1.9) that share the protein sequence segments. We are targeting the Nav1.7 found in the peripheral nervous system and engaged in the perception of pain. The objective of this project was to screen a 1.5 million compound library for identification of inhibitors for Nav1.7 with analgesic effect. In this study, we designed a protocol for identification of isoform-selective inhibitors of Nav1.7, by utilizing the prior information on isoform-selective antagonists. First, a similarity search was performed; then the identified hits were docked into a binding site on the fourth voltage-sensor domain (VSD4) of Nav1.7. We used the FTrees tool for similarity searching and library generation; the generated library was docked in the VSD4 domain binding site using FlexX and compounds were shortlisted using a FlexX score and SeeSAR hyde scoring. Finally, the top 25 compounds were tested with molecular dynamics simulation (MDS). We reduced our list to 9 compounds based on the MDS root mean square deviation plot and obtained them from a vendor for in vitro and in vivo validation. Whole-cell patch-clamp recordings in HEK-293 cells and dorsal root ganglion neurons were conducted. We used patch pipettes to record transient Na⁺ currents. One of the compounds reduced the peak sodium currents in Nav1.7-HEK-293 stable cell line in a dose-dependent manner, with IC50 values at 0.74 µM. In summary, our computer-aided analgesic discovery approach allowed us to develop pre-clinical analgesic candidate with significant reduction of time and cost.

Keywords: chronic pain, voltage-gated sodium channel, isoform-selective antagonist, similarity search, virtual screening, analgesics development

Procedia PDF Downloads 125

Authors: Ali Hajimohammadi, Mohammad Salahlou, Saeed Nazifi, Abbas Rowshan-Ghasrodashti, Ahmadreza Mirzaei

Abstract:

Several studies have shown that the liver is vulnerable to detrimental alterations characterized by functional and morphological changes during the transitional period. Probiotic supplementation is pivotal in enhancing metabolic functions by orchestrating a harmonious equilibrium within the intestinal microbial community, thereby averting gastrointestinal infections and elevating the overall well-being, growth, and performance of ruminants. This study was designed to evaluate the impact of probiotic and yeast cell wall extract (YCW) supplementation on metabolic and hepatic functions during the transitional period. A cohort of forty dry cows was randomly divided into four groups, namely the probiotic group (Pr) receiving a basal diet combined with a blend of Bacillus subtilis, Bacillus lechiniformis, Streptococcus Thermophilis, and Enterococcus faecium; the YCW group receiving a basal diet enriched with Saccharomyces cerevisiae; the probiotic and yeast cell wall extract group (P & Y) receiving a basal diet supplemented with a mixture of probiotic and yeast cell wall extract; and the control group adhering to the basal diet. The intervention was initiated 21 days before calving and persisted until 28 days post-calving, except for the control group. The study entailed the collection of blood samples at four sampling times, encompassing 21 days preceding calving, seven days before calving, seven days post-calving, and four weeks post-calving. Multiple biochemical parameters were assessed, including urea, blood urea nitrogen (BUN), Gamma-glutamyl transferase (GGT), total bilirubin (TB), albumin, total protein (TP), globulin, glucose, triglyceride, cholesterol, and liver functionality index. The results showed that the Pr group exhibited reduced average levels of urea, BUN, triglycerides, and GGT compared to the control group (P > 0.05). Similarly, the YCW group demonstrated lowered average BUN, TB, and cholesterol levels than the control group (P > 0.05). Notably, the liver functionality index exhibited a discernible trend towards elevation in the Pr group compared to the control group (P = 0.06) and the P & Y group (P = 0.007). In essence, the supplementation of YCW and probiotics is associated with advantageous effects on metabolic parameters and liver functionality.

Keywords: dairy cattle, probiotic, yeast extract supplement, liver functionality index, transition period

Procedia PDF Downloads 8

Authors: Haris Setiawan, Phongsakorn Chuammitri, Korawan Sringarm, Montira Intanon, Anucha Sathanawongs

Abstract:

Capacitation is a prerequisite to achieving sperm competency to penetrate the oocyte naturally occurring in vivo throughout the female reproductive tract and entangling secretory fluid and epithelial cells. One of the crucial compounds in the oviductal fluid which promotes capacitation is albumin, secreted in major concentrations. However, the difficulties in the collection and the inconsistency of the oviductal fluid composition throughout the estrous cycle have replaced its function with serum-based albumins such as bovine serum albumin (BSA). BSA has been primarily involved and evidenced for their stabilizing effect to maintain the acrosome intact during the capacitation process, modulate hyperactivation, and elevate the number of sperm bound to zona pellucida. Contrary to its benefits, the use of blood-derived products in the culture system is not sustainable and increases the risk of disease transmissions, such as Creutzfeldt-Jakob disease (CJD) and bovine spongiform encephalopathy (BSE). Moreover, it has been asserted that this substance is an aeroallergen that produces allergies and respiratory problems. In an effort to identify an alternative sustainable and non-toxic albumin source, the present work evaluated sperm reactions to a capacitation medium containing albumin derived from the flesh of the snakehead fish (Channa striata). Before examining the ability of this non-serum albumin to promote capacitation in bovine sperm, the presence of albumin was detected using bromocresol purple (BCP) at the level of 25% from snakehead fish extract. Following the SDS-PAGE and densitometric analysis, two major bands at 40 kDa and 47 kDa consisting of 57% and 16% of total protein loaded were detected as the potential albumin-related bands. Significant differences were observed in all kinematic parameters upon incubation in the capacitation medium. Moreover, consistently higher values were shown for the kinematic parameters related to hyperactivation, such as amplitude lateral head (ALH), velocity curve linear (VCL), and linearity (LIN) when sperm were treated with 3 mg/mL of snakehead fish albumin among other treatments. Likewise, substantial differences of higher acrosome intact presented in sperm upon incubation with various concentrations of snakehead fish albumin for 90 minutes, indicating that this level of snakehead fish albumin can be used to replace the bovine serum albumin. However, further study is highly required to purify the albumin from snakehead fish extract for more reliable findings.

Keywords: capacitation promoter, snakehead fish, non-serum albumin, bovine sperm

Procedia PDF Downloads 115