Search results for: isolated

Authors: María Reveles-Miranda, Daniella Pacheco-Catalán

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Sierra Papacal commissary is located north of Merida, Yucatan, México, where the indigenous Maya population predominates. Due to its location, the region has an elevation of fewer than 4.5 meters above sea level, with a high risk of flooding associated with storms and hurricanes and a high vulnerability of infrastructure and housing in the presence of strong gusts of wind. In environmental contingencies, the challenge is providing an autonomous electrical supply using renewable energy sources that cover vulnerable populations' health, food, and water pumping needs. To address this challenge, a hybrid energy storage module is proposed for the emergency photovoltaic (PV) system of the community shelter in Sierra Papacal, Yucatán, which combines high-energy-density batteries and high-power-density supercapacitors (SC) in a single module, providing a quick response to energy demand, reducing the thermal stress on batteries and extending their useful life. Incorporating SC in energy storage modules can provide fast response times to power variations and balanced energy extraction, ensuring a more extended period of electrical supply to vulnerable populations during contingencies. The implemented control strategy increases the module's overall performance by ensuring the optimal use of devices and balanced energy exploitation. The operation of the module with the control algorithm is validated with MATLAB/Simulink® and experimental tests.

Keywords: batteries, community shelter, environmental contingencies, hybrid energy storage, isolated photovoltaic system, supercapacitors

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Authors: Matthew Omoniyi Adebola, Jude E Amadi

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Ground nut (Arachis hypogaea) is one of the most popular commercial crops in Nigeria. Its suc-cessful production has been drastically affected by early leaf spot disease caused by Mycosphae-rella arachidis Deighton. In vitro control of the pathogen by six medicinal plants (Entada afri-cana, Vitex doniana, Lawsonia inermis, Azadirachta indica, Acalypha hispida and Nuaclea lati-folia) was assessed in this study. The extracts of the plants were prepared using cold and hot wa-ter and alcohol. The pathogen was isolated from ground nut infected with early leaf spot disease. The results revealed a great significant difference (P<0.05) in yield of extracts between cold water, hot water, and alcohol extracts. A significant difference (P<0.05) was observed in percentage concentrations of the various phytochemical constituents present in the extracts. Flavonoids per-centage concentration was the highest (0.68 - 1.95%) followed by saponnin(0.09-1.53%) in N. latifolia extracts. Steroiods had the least percentage concentrations (0.00- 0.09%)followed by terpenoids(0.02–0.71%) and proanthocyannin (0.05 – 0.86%). N. latifolia extracts produced the highest percentage concentrations (0.07–1.95%) of all the phytochemicals followed by A. indi-ca(0.05–1.64%)and least concentrations were obtained in A. hispidia(0.09 – 0.87%)and V. do-niana (0.00–0.88%). The extracts inhibited spore germination and growth of M. arachidis. The inhibition by alcohol extracts was high and significantly different (P>0.05) from cold and hot water extracts. Alcohol extract of L. inermis gave 100% spore germination inhibition followed by N. latifolia and A.indica with 97.75% and 85.60% inhibition respectively. Therefore, field trials of these six medicinal plants on the control of early leaf spot disease of ground nut are rec-ommended.

Keywords: groundnut, phytochemicals, medicinal plants, extracts, inhibition

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Authors: Leila Najafian

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Sarcoplasmic protein hydrolysates (SPHs) and myofibrillar protein hydrolysates (MPHs) from patin (Pangasius sutchi) were produced using two types of proteases: Papain and Alcalase. 1,1-diphenyl-2-picrylhydrazyl (DPPH), 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) diammonium salt (ABTS) radical scavenging activities and metal chelating activity assays for antioxidant activities were carried out on the SPHs and MPHs. The hydrolysates were isolated and purified by ultrafiltration, gel filtration and reverse phase high-performance liquid chromatography (RP-HPLC) and liquid chromatography with tandem mass spectrometry detection (LC-MS/MS) was used in identifying peptide sequences. The results showed that when the DH of MPHs increased, the protein solubility increased, while the highest amount of the protein solubility of SPHs was after 60 min incubation. The effect of DH on antioxidant activities of SPHs and MPHs was investigated. Among the hydrolysates, papain-MPH and Alcalase-SPH, which had the highest antioxidant activities, were purified. The potent fractions obtained from RP-HPLC of sarcoplasmic (SI 3 fraction) and myofibrillar (MI 4 fraction) hydrolysates showed the highest DPPH radical scavenging activity. The FVNQPYLLYSVHMK peptide for MPH and the LVVDIPAALQHA peptide for SPH exhibited the highest antioxidant activity. The presence of hydrophobic and hydrophilic amino acids, namely leucine (L), valine (V), phenylalanine (F), histidine (H) and proline (P), in the peptide sequences of SPH and MPH are believed to contribute to high antioxidant activity. Hence, SPH and MPH from patin have the potential as a natural functional ingredient in food and pharmaceutical industry.

Keywords: patin (Pangasius sutchi), protein hydrolysates, antioxidative peptides, mass spectrometry

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Authors: Khalda Amr, Noha Eltaweel, Sherif Ismail, Hala Raslan

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Introduction: Osteoarthritis is the most common form of arthritis that involves the wearing away of the cartilage that caps the bones in the joints. While rheumatoid arthritis is an autoimmune disease in which the immune system attacks the joints, beginning with the lining of joints. In this study, we aimed to study the top deregulated miRNAs that might be the cause of pathogenesis in both diseases. Methods: Eight cases were recruited in this study: 4 rheumatoid arthritis (RA), 2 osteoarthritis (OA) patients, as well as 2 healthy controls. Total RNA was isolated from plasma to be subjected to miRNA profiling by NGS. Sequencing libraries were constructed and generated using the NEBNextR UltraTM small RNA Sample Prep Kit for Illumina R (NEB, USA), according to the manufacturer’s instructions. The quality of samples were checked using fastqc and multiQC. Results were compared RA vs Controls and OA vs. Controls. Target gene prediction and functional annotation of the deregulated miRNAs were done using Mienturnet. The top deregulated miRNAs in each disease were selected for further validation using qRT-PCR. Results: The average number of sequencing reads per sample exceeded 2.2 million, of which approximately 57% were mapped to the human reference genome. The top DEMs in RA vs controls were miR-6724-5p, miR-1469, miR-194-3p (up), miR-1468-5p, miR-486-3p (down). In comparison, the top DEMs in OA vs controls were miR-1908-3p, miR-122b-3p, miR-3960 (up), miR-1468-5p, miR-15b-3p (down). The functional enrichment of the selected top deregulated miRNAs revealed the highly enriched KEGG pathways and GO terms. Six of the deregulated miRNAs (miR-15b, -128, -194, -328, -542 and -3180) had multiple target genes in the RA pathway, so they are more likely to affect the RA pathogenesis. Conclusion: Six of our studied deregulated miRNAs (miR-15b, -128, -194, -328, -542 and -3180) might be highly involved in the disease pathogenesis. Further functional studies are crucial to assess their functions and actual target genes.

Keywords: next generation sequencing, mirnas, rheumatoid arthritis, osteoarthritis

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Authors: Sayyed-javad Ziaolhagh, Ali-Reza Saadatifar

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Introduction: Athletes frequently perform anabolic steroid resistance exercise, but the effects of medical doses and abuse along with resistance exercise on structural damage to the Pancreases and also jujube extract are unknown. The aim of this study was to investigate the effects of resistance training on body weight and hip fractures induced by boldenone injection in male rats. Materials and methods: In this experimental study, 30 male Wistar rats aged 8-12 weeks (weight 202±9.34 g) were randomly divided into five groups: control, boldenone, extract of iujuba+boldenone, boldenone+resistance training and boldenone+resistance training +extract of jujuba. The resistance training program included climbing the ladder for 8 weeks, 3 days a week, 1 session training in a day and each session consisted of the 3 sets and 5 repetitions. Injection was conducted in depth in the hamstring once a week on an appointed day. After anesthesia, autopsy was performed, and the cardiac tissue was isolated. Results: Results showed that boldenone caused tissue damage, congestion, and nuclei unclear and diffuse. In the group "resistance + Boldenone," The Pancreases tissue showed a high degree of hyperemia, and the muscle cells were somewhat abnormal. In boldenone + jujube, the appearance of the tissue was normal, and the rejuvenating effect was visible. Conclusion: Boldenone appears to cause structural damage to the Pancreases tissue. Strength training with Jujube Extract can reduce part of the pancreatic system disorders (necrosis and inflammation) caused by anabolic steroid use.

Keywords: boldenone, Jujube extract, pancreases tissue, resistance training

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Authors: Anyaphat Srithanasuwan, Noppason Pangprasit, Montira Intanon, Phongsakorn Chuammitri, Witaya Suriyasathaporn, Ynte H. Schukken

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Streptococcus uberis is one of the most common mastitis-causing pathogens, with a wide range of intramammary infection (IMI) durations and pathogenicity. This study aimed to compare shared or unique virulence factor gene clusters distinguishing persistent and transient strains of S. uberis. A total of 139 S. uberis strains were isolated from three small-holder dairy herds with a high prevalence of S. uberis mastitis. The duration of IMI was used to categorize bacteria into two groups: transient and persistent strains with an IMI duration of less than 1 month and longer than 2 months, respectively. Six representative S. uberis strains, three from each group (transience and persistence) were selected for analysis. All transient strains exhibited multi-locus sequence types (MLST), indicating a highly diverse population of transient S. uberis. In contrast, MLST of persistent strains was available in an online database (pubMLST). Identification of virulence genes was performed using whole-genome sequencing (WGS) data. Differences in genomic size and number of virulent genes were found. For example, the BCA gene or alpha-c protein and the gene associated with capsule formation (hasAB), found in persistent strains, are important for attachment and invasion, as well as the evasion of the antimicrobial mechanisms and survival persistence, respectively. These findings suggest a genetic-level difference between the two strain types. Consequently, a comprehensive study of 139 S. uberis isolates will be conducted to perform an in-depth genetic assessment through WGS analysis on an Illumina platform.

Keywords: Streptococcus Uberis, mastitis, whole genome sequence, intramammary infection, persistent S. Uberis, transient s. Uberis

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Authors: Sreeja Shaw, Prosenjit Samanta, Asish Kumar Mukhopadhyay

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Cholera is still a global public health burden and is caused by Vibrio cholerae O1 and O139 serogroups. Evidence from recent outbreaks in Haiti and Yemen suggested that circulating V. cholerae O1 El Tor variant strains are continuously changing to cause more ruinous outbreaks worldwide, and most of them have emerged from the Indian subcontinents. Therefore, we studied the changing virulence characteristics along with the antibiotic resistance profile of V. cholerae O1strains isolated from seasonal outbreaks in three cholera endemic regions during 2018, Gujarat and Maharashtra in Western India (87 strains), and to compare those features with the isolates of West Bengal in Eastern India (48 strains) collected during the same period. All the strains from Western India were of Ogawa serotype, polymyxin B-sensitive, hemolytic, and contained a large fragment deletion in VSP-II genomic region similar with Yemen outbreak strains and carried more virulent Haitian genetic alleles of major virulence associated genes ctxB, tcpA, and rtxA. Conversely, 14.6% (7/48) of the strains from Eastern India were belong to the Inaba serotype, polymyxin B-resistant, non-hemolytic, harbored intact VSP-II region, classical ctxB, Haitian tcpA, and El Tor rtxA alleles. Interestingly, resistance to tetracycline and chloramphenicol was seen in isolates from both regions, which are not very common among V. cholerae O1 isolates in India. Therefore, this study indicated West Bengal as a diverse region where two different types of El Tor variant hypervirulent strains are co-existed, probably competing for their better environmental survival, which may result in severe irrepressible disease outcome in the future.

Keywords: cholera, vibrio cholerae, polymyxin B, Non-hemolytic, ctxB, tcpA, rtxA, VSP-II

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Authors: Elif Coskun Daggecen, Seyma Dokucu, Yekta Gezginc, Ismail Akyol

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Fermented dairy food quality is mainly determined by the sensory perception and influenced by many factors. Today, starter cultures for fermented foods are being developed to have a constant quality in these foods. Streptococcus thermophilus is one of the main species of most a starter cultures of yogurt fermentation. This species produces lactate by lactose fermentation from pyruvate. On the other hand, a small amount of pyruvate can alternatively be converted to various typical yoghurt flavor compounds such as diacetyl, acetoin, acetaldehyde, or acetic acid, for which the activity of three genes are shown to be especially important; ldh, nox and als. Up to date, commercially produced yoghurts have not yet met the desired aromatic properties that Turkish consumers find in traditional homemade yoghurts. Therefore, it is important to select starters carrying favorable metabolic characteristics from natural isolates. In this study, 30 strains of Str. Thermophilus were isolated from traditional Turkish yoghurts obtained from different regions of the country. In these strains, transcriptional levels of ldh, nox and als genes were determined via a newly developed qPCR protocol, which is a more reliable and precision method for analyzing the quantitative and qualitative expression of specific genes in different experimental conditions or in different organisms compared to conventional analytical methods. Additionally, the metabolite production potentials of the isolates were measured. Of all the strains examined, 60% were found to carry the metabolite production potential and the gene activity which appeared to be suitable to be used as a starter culture. Probable starter cultures were determined according to real-time PCR results.

Keywords: gene expression, RT-PCR, starter culture, Streptococcus thermophilus

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Authors: Hosam A. Shawky, Amr A. Abdel Fatah, Moustafa M. S. Abo ElFad, Abdel Hameed M. El-Aassar

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Water desalination projects based on reverse osmosis technology are being introduced in Egypt to combat drinking water shortage in remote areas. Reverse osmosis (RO) desalination is a pressure driven process. This paper focuses on the design of an integrated brackish water and seawater RO desalination and solar Photovoltaic (PV) technology. A small Mobile PV driven RO desalination plant prototype without batteries is designed and tested. Solar-driven reverse osmosis desalination can potentially break the dependence of conventional desalination on fossil fuels, reduce operational costs, and improve environmental sustainability. Moreover, the innovative features incorporated in the newly designed PV-RO plant prototype are focusing on improving the cost effectiveness of producing drinkable water in remote areas. This is achieved by maximizing energy yield through an integrated automatic single axis PV tracking system with programmed tilting angle adjustment. An autonomous cleaning system for PV modules is adopted for maximizing energy generation efficiency. RO plant components are selected so as to produce 4-5 m3/day of potable water. A basic criterion in the design of this PV-RO prototype is to produce a minimum amount of fresh water by running the plant during peak sun hours. Mobility of the system will provide potable water to isolated villages and population as well as ability to provide good drinking water to different number of people from any source that is not drinkable.

Keywords: design, reverse osmosis, photovoltaic, energy, desalination, Egypt

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Authors: Rozalina Keremedchieva, Ivan Svinyarov, Milen G. Bogdanov

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In continuation of a research project on the application of ionic liquids (ILs) as an alternative to the conventional organic solvents used in the recovery of value added chemicals of industrial interest1-3 we developed a procedure for back extraction and isolation in pure form of the biologically active alkaloid glaucine from IL-based aqueous solutions. One of the approaches applied was the formation of two-phase systems (IL-ATPS) by the addition of kosmotropic salts to the plant extract. The ability of the salts (Na2CO3, MgSO4, (NH4)2SO4, NaH2PO4) to induce the formation of two-phase systems and the influence of pH value on the partition coefficients of glaucine was comprehensively studied. As a result, it was found that the target alkaloid is preferably partitioned into the IL-rich phase regardless of the pH value of the medium and thus shows the inapplicability of the approach used for the isolation of the target compound from the ionic liquid. However, the results obtained can be used as a platform for the development of an analytical method for the quantitative determination of low concentrations of glaucine in biological samples. We further examined the ability of a series of organic solvents such as diethyl ether, Tert-butylmethyl ether, ethyl acetate, butyl acetate, toluene, chloroform, dichloromethane to recover glaucine form raw IL-based aqueous extracts. Optimal conditions for quantitative extraction of glaucine into chloroform were found from which, after removal of the solvent and subsequent recrystallization from ethanol, the target compound was isolated in a high purity as a hydrobromide salt – The form in which it entrance as an active ingredient in various medicines.

Keywords: natural products, ionic liquids, solid-liquid extraction, liquid-liquid extraction

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Authors: Ramasamy Tamizhselvi, Leema George, Madhav Bhatia

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We have earlier shown that mouse pancreatic acinar cells produce hydrogen sulfide (H2S) and play a role in the pathogenesis of acute pancreatitis. This study is to determine the effect of H2S on TLR4 mediated innate immune signaling in acute pancreatitis via substance P (SP). Male Swiss mice were treated with hourly intraperitoneal injection of caerulein (50μg/kg) for 10 hour. DL-propargylglycine (PAG) (100 mg/kg i.p.), an inhibitor of H2S formation was administered 1h after the induction of acute pancreatitis. Pancreatic acinar cells from male Swiss mice were incubated with or without caerulein (10–7 M for 60 min) and CP-96345 (NK1R inhibitor). To better understand the effect of H2S in inflammation, acinar cells were stimulated with caerulein after addition of H2S donor, NaHS. In addition, caerulein treated pancreatic acinar cells were pretreated with PAG (30 µM), for 1h. H2S inhibitor, PAG, eliminated TLR4, IRAK4, TRAF6 and NF-kB levels in an in vitro and in vivo model of caerulein-induced acute pancreatitis. PPTA gene deletion reduced TLR4, MyD88, IRAK4, TRAF6, adhesion molecules and NF-kB in caerulein treated pancreatic acinar cells whereas administration of NaHS resulted in further rise in TLR4 and NF-kB levels in caerulein treated pancreatic acinar cells. In addition, acini isolated from mice and treated with PPTA gene receptor NK1R antagonist CP96345 did not exhibit further increase in TLR4, IRAK4, TRAF6, adhesion molecules and NF-kB levels after NaHS pretreatment. The present findings show for the first time that in acute pancreatitis, H2S up-regulates TLR4 pathway and NF-kB via substance P.

Keywords: preprotachykinin-A gene, H2S, TLR4, acute pancreatitis

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Authors: Arfan Ali, Idrees Ahmad Nasir

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The critical evaluation of tolerance in tomato plants against the induced saline soil were assessed by transcript analysis of genes coding for products potentially involved in stress tolerance. A reverse transcriptase PCR experiment was performed with Hsp90-1, MT2, and GR1like protein genes using RNA isolated from different tissues of tomato plants. Four strains of Bacillus magisterium were inoculated with 100 Mm & 200 Mm concentrations of salt. Eleven treatments each ten replica pots were installed in green house experiment and the parameters taken into account were morphological (length, weight, number of leaves, leaf surface area), chemical (anthocyanin, chlorophyll-a, chlorophyll-b, carotenoids) and biological (gene expression). Results bare a response i.e. highest response of MT2 like gene was at 24 hpi and the highest levels of GR1 like protein transcript accumulation were detected at 36 hpi. The chemical and morphological parameters at diverse salt concentrations bequeath superlative response amongst strains which candidly flank on Zm7 and Zm4. Therefore, Bacillus magisterium Zm7 strains and somehow Zm4 strain can be used in saline condition to make plants tolerant. The overall performance of strains Zm7, Zm6, and Zm4 was found better for all studied traits under salt stress conditions. Significant correlations among traits root length, shoot length, number of leaves, leaf surface area, carotenoids, anthocyanin, chlorophyll-a and chlorophyll-b were found and suggested that the salt tolerance in tomato may be improved through the use of PGPR strains.

Keywords: Bacillus magisterium, gene expression glutathione reductase, metallothionein, PGPR, Rhizobacteria, saline

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Authors: Reine Suci Wulandari, Rosa Suryantini

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Albizia (Paraserianthes falcataria) is a woody plant species that has a high economic value and multifunctional. Albizia is important timber, medicinal plants and can also be used as a plant to rehabilitate critical lands. The demand value of Albizia is increased so that the large quantities and high quality of seeds are required. In vitro propagation techniques are seed propagation that can produce more seeds and quality in a short time. In vitro cultures require growth regulators that can be obtained from biological agents such as endophytic fungi. Endophytic fungi are micro fungi that colonize live plant tissue without producing symptoms or other negative effects on host plants and increase plant growth. The purposes of this research were to isolate and identify endophytic fungi isolated from the root of Albizia and to study the effect of endophytic fungus on the growth of Albizia in vitro. The methods were root isolation, endophytic fungal identification, and inoculation of endophytic fungi to Albizia plants in vitro. Endophytic fungus isolates were grown on PDA media before being inoculated with Albizia sprouts. Incubation is done for 4 (four) weeks. The observed growth parameters were live explant percentage, percentage of explant shoot, and percentage of explant rooted. The results of the research showed that 6 (six) endophytic fungal isolates obtained from the root of Albizia, namely Aspergillus sp., Verticillium sp, Penicillium sp., Trichoderma sp., Fusarium sp., and Acremonium sp. Statistical analysis found that Trichoderma sp. and Fusarium sp. affect in vitro growth of Albizia. Endophytic fungi from the results of this research were potential as plant growth promoting. It can be applied to increase productivity either through increased plant growth and increased endurance of Albizia seedlings to pests and diseases.

Keywords: Albizia, endophytic fungi, propagation, in vitro

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Authors: Lukas Ded

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Studying the tissue structure and histogenesis in the natural, 3D context is challenging but highly beneficial process. Contrary to classical approach of the physical tissue sectioning and subsequent imaging, it enables to study the relationships of individual cellular and histological structures in their native context. Recent developments in the tissue clearing approaches and microscopic volume imaging/data processing enable the application of these methods also in the areas of developmental and reproductive biology. Here, using the CLARITY tissue procedure and 3D confocal volume imaging we optimized the protocol for clearing, staining and imaging of the mice seminiferous tubules isolated from the testes without cardiac perfusion procedure. Our approach enables the high magnification and fine resolution axial imaging of the whole diameter of the seminiferous tubules with possible unlimited lateral length imaging. Hence, the large continuous pieces of the seminiferous tubule can be scanned and digitally reconstructed for the study of the single tubule seminiferous stages using nuclear dyes. Furthermore, the application of the antibodies and various molecular dyes can be used for molecular labeling of individual cellular and subcellular structures and resulting 3D images can highly increase our understanding of the spatiotemporal aspects of the seminiferous tubules development and sperm ultrastructure formation. Finally, our newly developed algorithms for 3D data processing enable the massive parallel processing of the large amount of individual cell and tissue fluorescent signatures and building the robust spermatogenic models under physiological and pathological conditions.

Keywords: CLARITY, spermatogenesis, testis, tissue clearing, volume imaging

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Authors: M. D. Asemoloye, S. G. Jonathan, A. Rafiq, O. J. Olawuyi, D. O. Adejoye

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Calmodulin is one of the intracellular calcium proteins that regulates large spectrum of enzymes and cellular functions including metabolism of cyclic nucleotides and glycogen. The potentials of calmodulin gene in fungi necessitates their genetic response and their strong cassette of enzyme secretions for pesticide degradation. Therefore, this study was carried out to investigate the ‘Transcriptomic’ response of calmodulin encoding genes in Talaromyces fungi in response to 2, 2-dichlorovinyl dimethyl phosphate (DDVP or Dichlorvos) an organophosphate pesticide and γ-Hexachlorocyclohexane (Lindane) an organochlorine pesticide. Fungi strains isolated from rhizosphere from grasses rhizosphere in pesticide polluted sites were subjected to percentage incidence test. Two most frequent fungi were further characterized using ITS gene amplification (ITS1 and ITS4 combinations), they were thereafter subjected to In-vitro DDVP and lindane tolerance tests at different concentrations. They were also screened for presence and expression of calmodulin gene (caM) using RT-PCR technique. The two Talaromyces strains had the highest incidence of 50-72% in pesticide polluted site, they were both identified as Talaromyces astroroseus asemoG and Talaromyces purpurogenum asemoN submitted in NCBI gene-bank with accession numbers KY488464 and KY488468 respectively. T. astroroseus KY488464 tolerated DDVP (1.23±0.023 cm) and lindane (1.11±0.018 cm) at 25 % concentration while T. purpurogenum KY488468 tolerated DDVP (1.33±0.061 cm) and lindane (1.54±0.077 cm) at this concentration. Calmodulin gene was detected in both strains, but RT-PCR expression of caM gene revealed at 900-1000 bp showed an under-expression of caM in T. astrorosues KY488464 but overexpressed in T. purpurogenum KY488464. Thus, the calmodulin gene response of these fungal strains to both pesticides could be considered in monitoring the potentials of fungal strains to pesticide tolerance and bioremediation of pesticide in polluted soil.

Keywords: Calmodulin gene, pesticide, RT-PCR, talaromyces, tolerance

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Authors: Gloria Pinilla, Jose Manuel Baena, Patricia Gálvez-Martín, Juan Antonio Marchad

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Cartilage is a dense connective tissue with limited self-repair properties. Currently, the therapeutic use of autologous or allogenic chondrocytes makes up an alternative therapy to the pharmacological treatment. The design of a bioprinted 3D cartilage with chondrocytes and biodegradable biomaterials offers a new therapeutic alternative able of bridging the limitations of current therapies in the field. We have developed an enhanced printing processes-Injection Volume Filling (IVF) to increase the viability and survival of the cells when working with high-temperature thermoplastics without the limitation of the scaffold geometry in contact with cells. We have demonstrated the viability of the printing process using chondrocytes for cartilage regeneration. This development will accelerate the clinical uptake of the technology and overcomes the current limitation when using thermoplastics as scaffolds. An alginate-based hydrogel combined with human chondrocytes (isolated from osteoarthritis patients) was formulated as bioink-A and the polylactic acid as bioink-B. The bioprinting process was carried out with the REGEMAT V1 bioprinter (Regemat 3D, Granada-Spain) through a IVF. The printing capacity of the bioprinting plus the viability and cell proliferation of bioprinted chondrociytes was evaluated after five weeks by confocal microscopy and Alamar Blue Assay (Biorad). Results showed that the IVF process does not decrease the cell viability of the chondrocytes during the printing process as the cells do not have contact with the thermoplastic at elevated temperatures. The viability and cellular proliferation of the bioprinted artificial 3D cartilage increased after 5 weeks. In conclusion, this study demonstrates the potential use of Regemat V1 for 3D bioprinting of cartilage and the viability of bioprinted chondrocytes in the scaffolds for application in regenerative medicine.

Keywords: cartilage regeneration, bioprinting, bioink, scaffold, chondrocyte

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Authors: Pampita Chakraborty, Sukumar Mukherjee

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The study was done to determine the microbiological profile and changing pattern of the pathogens causing UTI in the ICU patients. All the patients admitted to the ICU with urinary catheter insertion for more than 48hours were included in the study. Urine samples were collected in a sterile container with aseptic precaution using disposable syringe and was processed as per standards. Antimicrobial susceptibility test was done by Disc Diffusion method as per CLSI guidelines. A total of 100 urine samples were collected from ICU patients, out of which 30% showed significant bacterial growth and 7% showed growth of candida spp. Prevalence of UTI was more in female (73%) than male (27.%). Gram-negative bacilli 26(86.67%) were more common in our study followed by gram-positive cocci 4(13.33%). The most common uropathogens isolated were Escherichia coli 14 (46.67%), followed by Klebsiella spp 7(23.33%), Staphylococcus aureus 4(13.33%), Acinetobacter spp 3(10%), Enterococcus faecalis 1(3.33%) and Pseudomonas aeruginosa 1(3.33%). Most of the Gram-negative bacilli were sensitive to amikacin (80%) and nitrofurantoin (80%), where as all gram-positive organisms were sensitive to Vancomycin. A large number ESBL producers were also observed in this study. The study finding showed that E.coli is the predominant pathogen and has increasing resistance pattern to the commonly used antibiotics. The study proposes that the adherence to antibiotic policy is the key ingredients for successful outcome in ICU patients and also emphasizes that repeated evaluation of microbial characteristics and continuous surveillance of resistant bacteria is required for selection of appropriate antibiotic therapy.

Keywords: antimicrobial sensitivity, intensive care unit, nosocomial infection, urinary tract infection

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Authors: B. Samuel Raj, Solomon R. D. Jebakumar

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Microbial fuel cells (MFCs) are an emerging and promising method for achieving sustainable energy since they can remove contaminated organic matter and simultaneously generate electricity. Our approach was driven in three different ways like Bacterial fuel cell, Soil Microbial fuel cell (Soil MFC) and Plant Microbial fuel cell (Plant MFC). Bacterial MFC: Sulphate reducing bacteria (SRB) were isolated and identified as the efficient electricigens which is able to produce ±2.5V (689mW/m2) and it has sustainable activity for 120 days. Experimental data with different MFC revealed that high electricity production harvested continuously for 90 days 1.45V (381mW/m2), 1.98V (456mW/m2) respectively. Biofilm formation was confirmed on the surface of the anode by high content screening (HCS) and scanning electron Microscopic analysis (SEM). Soil MFC: Soil MFC was constructed with low cost and standard Mudwatt soil MFC was purchased from keegotech (USA). Vermicompost soil (V1) produce high energy (± 3.5V for ± 400 days) compared to Agricultural soil (A1) (± 2V for ± 150 days). Biofilm formation was confirmed by HCS and SEM analysis. This finding provides a method for extracting energy from organic matter, but also suggests a strategy for promoting the bioremediation of organic contaminants in subsurface environments. Our Soil MFC were able to run successfully a 3.5V fan and three LED continuously for 150 days. Plant MFC: Amaranthus candatus (P1) and Triticum aestivium (P2) were used in Plant MFC to confirm the electricity production from plant associated microbes, four uniform size of Plant MFC were constructed and checked for energy production. P2 produce high energy (± 3.2V for 40 days) with harvesting interval of two times and P1 produces moderate energy without harvesting interval (±1.5V for 24 days). P2 is able run 3.5V fan continuously for 10days whereas P1 needs optimization of growth conditions to produce high energy.

Keywords: microbial fuel cell, biofilm, soil microbial fuel cell, plant microbial fuel cell

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Authors: Parminder Kaur, Chandrajit B. Majumder

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The unrestrained usage of pesticides to meet the burgeoning demand of enhanced crop productivity has led to the serious contamination of both terrestrial and aquatic ecosystem. The remediation of mixture of pesticides is a challenging affair regarding inadvertent mixture of pesticides from agricultural lands treated with various compounds. Global concerns about the excessive use of pesticides have driven the need to develop more effective and safer alternatives for their remediation. We focused our work on the microbial degradation of a mixture of three Type II-pyrethroids, namely Cypermethrin, Cyhalothrin and Deltamethrin commonly applied for both agricultural and domestic purposes. The fungal strains (Fusarium strain 8-11P and Fusarium sp. zzz1124) had previously been isolated from agricultural soils and their ability to biotransform this amalgam was studied. In brief, the experiment was conducted in two growth systems (added carbon and carbon-free) enriched with variable concentrations of pyrethroids between 100 to 300 mgL⁻¹. Parameter optimization (pH, temperature, concentration and time) was done using a central composite design matrix of Response Surface Methodology (RSM). At concentrations below 200 mgL⁻¹, complete removal was observed; however, degradation of 95.6%/97.4 and 92.27%/95.65% (in carbon-free/added carbon) was observed for 250 and 300 mgL⁻¹ respectively. The consortium has been shown to degrade the pyrethroid mixture (300 mg L⁻¹) within 120 h. After 5 day incubation, the residual pyrethroids concentration in unsterilized soil were much lower than in sterilized soil, indicating that microbial degradation predominates in pyrethroids elimination with the half-life (t₁/₂) of 1.6 d and R² ranging from 0.992-0.999. Overall, these results showed that microbial consortia might be more efficient than single degrader strains. The findings will complement our current understanding of the bioremediation of mixture of Type II pyrethroids with microbial consortia and potentially heighten the importance for considering bioremediation as an effective alternative for the remediation of such pollutants.

Keywords: bioremediation, fungi, pyrethroids, soil

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Authors: Fatma Koksal Çakirlar, Si̇nem Ozdemir, Selcan Akyol, Revazi̇ye Gulesen, Murat Gunaydin, Nevri̇ye Gonullu, Belkis Levent, Nuri̇ Kiraz

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Vibrio cholerae O1 and O139 are the causative agent of epidemic or pandemic cholera. V. cholerae O1 is generally accepted as a non-invasive enterotoxigenic organism causing gastroenteritis of various severities. Non-O1 V. cholerae can cause small outbreaks of diarrhea due to consumption of contaminated food and water. Particularly, the patients with achlorydria have a risk for vibrio infections. There are numerous case reports of bacteremia caused by vibrio in patients with predisposing conditions like cirrhosis, nephrotic syndrome, diabetes, hematologic malignancy, gastrectomy, and AIDS. We described in this study the first case of nontoxigenic, non-01/non-O139 V. cholerae isolated from the blood culture of a 77-year-old female patient with hipertension, diabetes, coronary artery disease, gout and about 9 years ago migrated breast cancer history. The patient with complaints of shortness of breath, fever and malaise admitted to our emergency clinic were evaluated. There was no diarrhea or abdominal symptoms in the patient. No growth in her urine culture, but blood culture (BACTEC 9120 system, Becton Dickinson, USA) was positive for non-01/non-O139 V. cholerae that was identified by conventional methods and Phoenix automated system (BD Diagnostic Systems, Sparks, MD). It does not secrete the cholera toxin. The agglutination test was negative with polyvalent O1 antisera and O139 antiserum. Empirically ceftriaxone was administered to the patient and she was discharged with improvement in general condition. In this study we report bacteremia by non-01/non-O139 V. cholerae that is rare in the worldwide and first in Turkey.

Keywords: bacteremia, blood culture, immunocompromised patient, Non-O1 vibrio cholerae

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Authors: Y. Hajjam, I. T. Alami, S. M. Udupa, S. Cherkaoui

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Faba bean (Vicia faba L.) is an important food legume crop in Morocco. It is mainly used as human food and feed for animals. Faba bean also plays an important role in cereal-based cropping systems, when rotated with cereals it improves soil fertility by fixing N2 in root nodules mediated by Rhizobium. Both faba bean and its biological nitrogen fixation symbiotic bacterium Rhizobium are affected by different stresses such as: salinity, drought, pH, heavy metal, and the uptake of inorganic phosphate compounds. Therefore, the aim of the present study was to evaluate the phenotypic diversity among the faba bean rhizobial isolates and to select the tolerant strains that can fix N2 under environmental constraints for inoculation particularly for affected soils, in order to enhance the productivity of faba bean and to improve soil fertility. Result have shown that 62% of isolates were fast growing with the ability of producing acids compounds , while 38% of isolates are slow growing with production of alkalins. Moreover, 42.5% of these isolates were able to solubilize inorganic phosphate Ca3(PO4)2 and the index of solubilization was ranged from 2.1 to 3.0. The resistance to extreme pH, temperature, water stress heavy metals and antibiotics lead us to classify rhizobial isolates into different clusters. Finally, the authentication test under greenhouse conditions showed that 55% of the rhizobial isolates could induce nodule formation on faba bean (Vicia faba L.) under greenhouse experiment. This phenotypic characterization may contribute to improve legumes and non legumes crops especially in affected soils and also to increase agronomic yield in the dry areas.

Keywords: rhizobia, vicia faba, phenotypic characterization, nodule formation, environmental constraints

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Authors: Sarra Bouhallel, Sara Legbedj, Rima Messaoud, Sofia Saffarbatti

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In the context of global waste management and sustainable resource utilization, millets emerge as a vital yet underutilized cereal resource. Despite their exceptional nutritional profile and resilience to harsh environmental conditions, their potential remains largely untapped. This study aims to contribute to the valorization of seven Algerian pearl millet landraces (Pennisetum glaucum (L.) R. Br) from the southern region by focusing on the characterization of their starches. Utilizing both conventional wet milling, incorporating sodium azide as a microbial growth inhibitor, and a novel green technology—Ultrasound-assisted isolation, we explore avenues for enhancing the functional properties of these starches. Analysis of key functional properties such as swelling power and water solubility index reveals significant enhancements, particularly during heat treatment near the gelatinization temperature [70 - 80 °C]. Furthermore, our investigation into the influence of pre-treatment methods on isolated starches highlights the potential of Ultrasound-assisted isolation in reducing absorbency and water solubility compared to conventional methods. Through rigorous data analysis using SPSS software (Version 23), we ascertain the efficiency of Ultrasound-assisted isolation, underscoring its promising role in the valorization of pearl millet waste. This research not only sheds light on the functional properties of pearl millet starch but also underscores the imperative of sustainable waste management in harnessing the full potential of underutilized cereal resources.

Keywords: isolation, solubility, starch, swelling, ultrasound

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Authors: Sophie Mills, Constantinos Aristotelous, Leila L. Touil, Richard C. W. James

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Objectives: Historical studies of the anatomy of the spinal accessory nerve (SAN) have reported conflicting results regarding its relationship with the internal jugular vein (IJV). A literature review was undertaken to establish the prevalence of anatomical variations of the SAN encountered during routine neck dissection surgery in order to increase awareness and reduce morbidity associated with iatrogenic SAN injury. Materials and Methods: The largest systematic review to date was performed using PRISMA-ScR guidelines, which yielded nine articles following the application of inclusion and exclusion criteria. A case report is also included, which demonstrates the rare anatomical relationship of the SAN traversing a fenestrated IJV, seen for the first time in the senior author’s career. Results: The mean number of dissections per study was 119, of which 55.6% (n=5) studies were performed on cadaver subjects, and 44.4% (n=4) were surgical dissections. Incidences of the SAN lateral to the IJV and medial to the IJV ranged from 38.9%-95.7% and 2.8%-57.4%, respectively. Over half of the studies reported incidences of the SAN traversing the IJV in 0.9%-2.8% of dissections. One study reported an isolated variant of the SAN dividing around the IJV with a prevalence of 0.5%. Conclusion: At the level of the posterior belly of the digastric muscle, the surgeon can anticipate the identification of the SAN lateral to the IJV in approximately three-quarters of cases, whilst around one-quarter are estimated to be medial. A mean of 1.6% of SANs traverses a fenestration of the vein. It is essential for surgeons to be aware of these anatomical variations and their prevalence to prevent injury to vital structures during surgery.

Keywords: anatomical variant, internal jugular vein, neck dissection, spinal accessory nerve

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Authors: Bedilu Tafesse

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Conservation of crop genetic resources presents a challenge of identifying specific determinants driving maintenance of diversity at farm and agroecosystems. The objectives of this study were to identify socioeconomic, market and agroecological determinants of farmers’ maintenance of wheat diversity at the household level and derive implications for policies in designing on-farm conservation programs. We assess wheat diversity at farm level using household survey data. A household decision making model is conceptualized using microeconomic theory to assess and identify factors influencing on-farm rice diversity. The model is then tested econometrically by using various factors affecting farmers’ variety choice and diversity decisions. The findings show that household-specific socioeconomic, agroecological and market factors are important in determining on-farm wheat diversity. The significant variables in explaining richness and evenness of wheat diversity include distance to the nearest market, subsistence ratio, modern variety sold, land types and adult labour working in agriculture. The statistical signs of the factors determining wheat diversity are consistent in explaining the richness, dominance and evenness among rice varieties. Finally, the study implies that the cost-effective means of promoting and sustaining on-farm conservation programmes is to target them in market isolated geographic locations of high crop diversity where farm households have more heterogeneity of agroecological conditions and more active family adult labour working on-farm.

Keywords: diversity indices, dominance, evenness, on-farm conservation, wheat diversity, richness

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Authors: Lok Bahadur Shrestha, Narayan Raj Bhattarai, Basudha Khanal

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Introduction: Coagulase-negative staphylococci (CoNS) are the normal commensal of human skin and mucous membranes. The study was carried out to study the prevalence of CoNS among clinical isolates, to characterize them up to species level and to compare the three conventional methods for detection of biofilm formation. Objectives: to characterize the clinically significant coagulase-negative staphylococci up to species level, to compare the three phenotypic methods for the detection of biofilm formation and to study the antimicrobial susceptibility pattern of the isolates. Methods: CoNS isolates were obtained from various clinical samples during the period of 1 year. Characterization up to species level was done using biochemical test and study of biofilm formation was done by tube adherence, congo red agar, and tissue culture plate method. Results: Among 71 CoNS isolates, seven species were identified. S. epidermidis was the most common species followed by S. saprophyticus, S. haemolyticus. Antimicrobial susceptibility pattern of CoNS documented resistance of 90% to ampicillin. Resistance to cefoxitin and ceftriaxone was observed in 55% of the isolates. We detected biofilm formation in 71.8% of isolates. The sensitivity of tube adherence method was 82% while that of congo red agar method was 78%. Conclusion: Among 71 CoNS isolated, S. epidermidis was the most common isolates followed by S. saprophyticus and S. haemolyticus. Biofilm formation was detected in 71.8% of the isolates. All of the methods were effective at detecting biofilm-producing CoNS strains. Biofilm former strains are more resistant to antibiotics as compared to biofilm non-formers.

Keywords: CoNS, congo red agar, bloodstream infections, foreign body-related infections, tissue culture plate

Procedia PDF Downloads 198

Authors: Ingrid R. Avanzi, Marcela dos P. G. Baltazar, Louise H. Gracioso, Luciana J. Gimenes, Bruno Karolski, Elen A. Perpetuo, Claudio Auguto Oller do Nascimento

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In this study was developed a natural process using a biological system for the uptake of Copper and possible removal of copper from wastewater by dead biomass of the strain Burkholderia cenocepacia. Dead and live biomass of Burkholderia cenocepacia was used to analyze the equilibrium and kinetics of copper biosorption by this strain in function of the pH. Living biomass exhibited the highest biosorption capacity of copper, 50 mg g−1, which was achieved within 5 hours of contact, at pH 7.0, temperature of 30°C, and agitation speed of 150 rpm. The dead biomass of Burkholderia cenocepacia may be considered an efficiently bioprocess, being fast and low-cost to production of copper and also a probably nano-adsorbent of this metal ion in wastewater in bioremediation process. In this study was developed a natural process using a biological system for the uptake of Copper and possible removal of copper from wastewater by dead biomass of the strain Burkholderia cenocepacia. Dead and live biomass of Burkholderia cenocepacia was used to analyze the equilibrium and kinetics of copper biosorption by this strain in function of the pH. Living biomass exhibited the highest biosorption capacity of copper, 50 mg g−1, which was achieved within 5 hours of contact, at pH 7.0, temperature of 30°C, and agitation speed of 150 rpm. The dead biomass of Burkholderia cenocepacia may be considered an efficiently bioprocess, being fast and low-cost to production of copper and also a probably nano-adsorbent of this metal ion in wastewater in bioremediation process.

Keywords: biosorption, dead biomass, biotechnology, copper recovery

Procedia PDF Downloads 337

Authors: Astorga-Trejo Rebeca, Fonseca-Peralta Héctor Manuel, Beltrán-Arredondo Laura Ivonne, Castro-Martínez Claudia

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The use of biomass as feedstock for the production of fuels and other chemicals of interest is an ever-growing accepted option in the way to the development of biorefinery complexes; in the Mexican state of Sinaloa, two million tons of residues from corn crops are produced every year, most of which can be converted to bioethanol and other products through biotechnological conversion using yeast and other microorganisms. Therefore, the objective of this work was to take advantage of corn stover and evaluate its potential as a substrate for the production of second-generation bioethanol (2G), enzymes, and xylitol. To produce bioethanol 2G, an acid-alkaline pretreatment was carried out prior to saccharification and fermentation. The microorganisms used for the production of enzymes, as well as for the production of xylitol, were isolated and characterized in our workgroup. Statistical analysis was performed using Design Expert version 11.0. The results showed that it is possible to obtain 2G bioethanol employing corn stover as a carbon source and Saccharomyces cerevisiae ItVer01 and Candida intermedia CBE002 with yields of 0.42 g and 0.31 g, respectively. It was also shown that C. intermedia has the ability to produce xylitol with a good yield (0.46 g/g). On the other hand, qualitative and quantitative studies showed that the native strains of Fusarium equiseti (0.4 IU/mL - xylanase), Bacillus velezensis (1.2 IU/mL – xylanase and 0.4 UI/mL - amylase) and Penicillium funiculosum (1.5 IU / mL - cellulases) have the capacity to produce xylanases, amylases or cellulases using corn stover as raw material. This study allowed us to demonstrate that it is possible to use corn stover as a carbon source, a low-cost raw material with high availability in our country, to obtain bioproducts of industrial interest, using processes that are more environmentally friendly and sustainable. It is necessary to continue the optimization of each bioprocess.

Keywords: biomass, corn stover, biorefinery, bioethanol 2G, enzymes, xylitol

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Authors: Adil Basuki Ahza, Michael Liong, Subarna Suryatman

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Healthy chips now dominating the snack market shelves. More than 80% processed snack foods in the market are chips. This research takes the advantages of twin extrusion technology to produce two types of product, i.e. directly expanded and intermediate ready-to-fry or microwavable chips. To improve the functional quality, the cereal-tuber based mix was enriched with antioxidant rich mix of temurui, celery, carrot and isolated soy protein (ISP) powder. Objectives of this research were to find best composite cassava-sorghum ratio, i.e. 60:40, 70:30 and 80:20, to optimize processing conditions of extrusion and study the microstructural, physical and sensorial characteristics of the final products. Optimization was firstly done by applying metering section of extruder barrel temperatures of 120, 130 and 140 °C with screw speeds of 150, 160 and 170 rpm to produce direct expanded product. The intermediate product was extruded in 100 °C and 100 rpm screw speed with feed moisture content of 35, 40 and 45%. The directly expanded products were analyzed for color, hardness, density, microstructure, and organoleptic properties. The results showed that interaction of ratio of cassava-sorghum and cooking methods affected the product's color, hardness, and bulk density (p<0.05). Extrusion processing conditions also significantly affected product's microstructure (p<0.05). The direct expanded snacks of 80:20 cassava-sorghum ratio and fried expanded one 70:30 and 80:20 ratio shown the best organoleptic score (slightly liked) while baking the intermediate product with microwave were resulted sensorial not acceptable quality chips.

Keywords: cassava-sorghum composite, extrusion, microstructure, physical characteristics

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Authors: Prittesh Patel, Rushabh Shah, Krishnamurthy Ramar, Vakulbhushan Bhaskar

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The present research work aims at finding genetic differences in the genomes of sugarcane red rot isolates Colletotrichum falcatum Went using Random Amplified Polymorphic DNA (RAPD) and interspersed simple sequence repeat (ISSR) molecular markers. Ten isolates of C. falcatum isolated from different red rot infected sugarcane cultivars stalk were used in present study. The amplified bands were scored across the lanes obtained in 15 RAPD primes and 21 ISSR primes successfully. The data were analysed using NTSYSpc 2.2 software. The results showed 80.6% and 68.07% polymorphism in RPAD and ISSR analysis respectively. Based on the RAPD analysis, ten genotypes were grouped into two major clusters at a cut-off value of 0.75. Geographically distant C. falcatum isolate cfGAN from south Gujarat had a level of similarity with Coimbatore isolate cf8436 presented on separate clade of bootstrapped dendrograms. First and second cluster consisted of five and three isolates respectively, indicating the close relation among them. The 21 ISSR primers produced 119 distinct and scorable loci in that 38 were monomorphic. The number of scorable loci for each primer varied from 2 (ISSR822) to 8 (ISSR807, ISSR823 and ISSR15) with an average of 5.66 loci per primer. Primer ISSR835 amplified the highest number of bands (57), while only 16 bands were obtained by primers ISSR822. Four primers namely ISSR830, ISSR845, ISSR4 and ISSR15 showed the highest value of percentage of polymorphism (100%). The results indicated that both of the marker systems RAPD and ISSR, individually can be effectively used in determination of genetic relationship among C falcatum accessions collected from different parts of south Gujarat.

Keywords: Colletotrichum falcatum, ISSR, RAPD, Red Rot

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Authors: Shashank Gupta, Iteash Agarwal, Vijayalaxmi Mogiligidda, Rajesh Kumar Krishnan, Sruthi Chennuri, Deepika Aggarwal, Anwesha Hoodati, Sheroy Cooper, Ranjan, Mohammad Bilal Sheik, Bhavya K. M., Manasa Hegde, M. Naveen Krishna, Amit Kumar Chauhan, Mallikarjun Korrapati, Sumit Singh, J. B. Singh, Sunil Sud, Sunil Gupta, Sidhartha Pant, Sankar, Neha Agrawal, Ashish Ranjan, Piyush Mohapatra, Roopak T., Arsh Ahmad, Nanjunda M., Dilip Singh

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Major research interests on quantum key distribution (QKD) are primarily focussed on increasing 1. point-to-point transmission distance (1000 Km), 2. secure key rate (Mbps), 3. security of quantum layer (device-independence). It is great to push the boundaries on these fronts, but these isolated approaches are neither scalable nor cost-effective due to the requirements of specialised hardware and different infrastructure. Current and future QKD network requires addressing different sets of challenges apart from distance, key rate, and quantum security. In this regard, we present ChaQra -a sub-quantum network with core features as 1) Crypto agility (integration in the already deployed telecommunication fibres), 2) Software defined networking (SDN paradigm for routing different nodes), 3) reliability (addressing denial-of-service with hybrid quantum safe cryptography), 4) upgradability (modules upgradation based on scientific and technological advancements), 5) Beyond QKD (using QKD network for distributed computing, multi-party computation etc). Our results demonstrate a clear path to create and accelerate quantum secure Indian subcontinent under the national quantum mission.

Keywords: quantum network, quantum key distribution, quantum security, quantum information

Procedia PDF Downloads 56