Search results for: bacteria isolation

Authors: O. I. Adeniran, M. A. Mogale

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Advanced glycation end-products (AGEs) have been implicated in the development and progression of vascular complications of diabetes mellitus and other age-related disease such as Alzheimer’s disease, heart diseases, stroke and limb amputation. The aim of the study was to determine the anti-glycation activity and AGE-cross-linking breaking ability of Sclerocarya birrea stem-bark extracts (SBSBETs). Hexane, ethyl acetate, methanol and water extracts of Sclerocarya birrea stem-bark and standard inhibitor, aminoguanidine (AG) were incubated with bovine serum albumin (BSA)-fructose mixture for 20 and 40 days. The amounts of total immunogenic AGEs (TIAGEs), fluorescent AGEs (FAGEs) and carboxymethyl lysine (CML) formed were determined and the percentage anti-glycation activity of each plant extract calculated. The ability of SBSBETs to break fructose-derived BSA-AGE-collagen cross-links was also investigated. All SBSBETs under investigation demonstrated less anti-glycation activity against TIAGE, FAGEs and CML than AG after 20 days incubation. After 40 days incubation, ethyl acetate, methanol and water SBSBETs demonstrated lower anti-glycation activity against TIAGEs than AG but exerted higher anti-glycation activity than AG against FAGEs. All SBSBETs except water demonstrated lower anti-glycation activity than AG against CML. With regard to the ability of SBSBETs to breakdown fructose-derived AGEs cross-links, the polar SBSBETs demonstrated higher ability to break AGE-cross-links than the non-polar ones. The results of this study may lead to the isolation of bio-active phyto-chemicals from SBSBETs that may be used for the prevention of vascular complication of diabetes.

Keywords: advanced glycation end-products, anti-glycation, cross-link breaking, Sclerocarrya birrea

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Authors: Yemane Tadesse Gebreslassie, Henok Gidey Gebretnsae

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Nanotechnology has emerged as a highly promising field of research with wide-ranging applications across various scientific disciplines. In recent years, tin oxide has garnered significant attention due to its intriguing properties, particularly when synthesized in the nanoscale range. While numerous physical and chemical methods exist for producing tin oxide nanoparticles, these approaches tend to be costly, energy-intensive, and involve the use of toxic chemicals. Given the growing concerns regarding human health and environmental impact, there has been a shift towards developing cost-effective and environmentally friendly processes for tin oxide nanoparticle synthesis. Green synthesis methods utilizing biological entities such as plant extracts, bacteria, and natural biomolecules have shown promise in successfully producing tin oxide nanoparticles. However, scaling up the production to an industrial level using green synthesis approaches remains challenging due to the complexity of biological substrates, which hinders the elucidation of reaction mechanisms and formation processes. Thus, this review aims to provide an overview of the various sources of biological entities and methodologies employed in the green synthesis of tin oxide nanoparticles, as well as their impact on nanoparticle properties. Furthermore, this research delves into the strides made in comprehending the mechanisms behind the formation of nanoparticles as documented in existing literature. It also sheds light on the array of analytical techniques employed to investigate and elucidate the characteristics of these minuscule particles.

Keywords: nanotechnology, tin oxide, green synthesis, formation mechanisms

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Authors: Anita Vidacs, Csaba Vagvolgyi, Judit Krisch

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The attachment of microorganisms to different surfaces and the development of biofilms can lead to outbreaks of food-borne diseases and economic losses due to perished food. In food processing environments, bacterial communities are generally formed by mixed cultures of different species. Plants are sources of several antimicrobial substances that may be potential candidates for the development of new disinfectants. We aimed to investigate cinnamon (Cinnamomum zeylanicum), marjoram (Origanum majorana), and thyme (Thymus vulgaris). Essential oils and their major components (cinnamaldehyde, terpinene-4-ol, and thymol) on four-species biofilms of E. coli, L. monocytogenes, P. putida, and S. aureus. Experiments had three parts: (i) determination of minimum bactericide concentration and the killing time with microdilution methods; (ii) elimination of the four-species 24– and 168-hours old biofilm from stainless steel, polypropylene, tile and wood surfaces; and (iii) comparing the disinfectant effect with industrial used per-acetic based sanitizer (HC-DPE). E. coli and P. putida were more resistant to investigated essential oils and their main components in biofilm, than L. monocytogenes and S. aureus. These Gram-negative bacteria were detected on the surfaces, where the natural based disinfectant had not total biofilm elimination effect. Most promoted solutions were the cinnamon essential oil and the terpinene-4-ol that could eradicate the biofilm from stainless steel, polypropylene and even from tile, too. They have a better disinfectant effect than HC-DPE. These natural agents can be used as alternative solutions in the battle against bacterial biofilms.

Keywords: biofilm, essential oils, surfaces, terpinene-4-ol

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Authors: R. S. Komartin, B. Balanuca, R. Stan

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the last decades, studies about the use of polymeric materials of plant origin, considering environmental concerns, have captured the interest of researchers because these represent an alternative to petroleum-derived materials. Vegetable oils are one of the preferred alternatives for petroleum-based raw materials having long aliphatic chains similar to hydrocarbons which means that can be processed using conventional chemistry. Epoxidized vegetable oils (EVO) are among the most interesting products derived from oil both for their high reactivity (epoxy group) and for the potential to react with compounds from various classes. As in the case of epoxy resins starting from petrochemical raw materials, those obtained from EVO can be crosslinked with different agents to build polymeric networks and can also be reinforced with various additives to improve their thermal and mechanical performances. Among the multitude of known EVO, the most common in industrial practice are epoxidized linseed oils (ELO) and epoxidized soybean oils (ESO), the first with an iodine index over 180, the second having a lower iodine index but being cheaper. On the other hand, lignin (Ln) is the second natural organic material as a spread, whose use has long been hampered because of the high costs associated with its isolation and purification. In this context, our goal was to obtain new composite materials with satisfactory intermediate properties in terms of stiffness and elasticity using the characteristics of ELO and Ln and choosing the proper curing procedure. In the present study linseed oil (LO) epoxidation was performed using peracetic acid generated in situ. The obtained bio-based epoxy resin derived from linseed oil was used further to produce the new composites byloading Ln in various mass ratios. The resulted ELO-Ln blends were subjected to a dual-curing protocol, namely photochemical and thermal. The new ELO-Ln composites were investigated by FTIR spectrometry, thermal stability, water affinity, and morphology. The positive effect of lignin regarding the thermal stability of the composites could be proved. The results highlight again the still largely unexplored potential of lignin in industrial applications.

Keywords: composite materials, dual curing, epoxidized linseed oil, lignin

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Authors: Zhanara Suleimenova, Raushan Blieva, Aigerim Zhakipbekova, Inkar Tapenbayeva, Zhanar Narmuratova

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Green processes are based on innovative technologies that do not negatively affect the environment. Industrial enzymes originated from biological systems can effectively contribute to sustainable development through being isolated from microorganisms which are fermented using primarily renewable resources. Many widespread microorganisms secrete a significant amount of biocatalysts into the environment, which greatly facilitates the task of their isolation and purification. The ability to control the enzyme production through the regulation of their biosynthesis and the selection of nutrient media and cultivation conditions allows not only to increase the yield of enzymes but also to obtain enzymes with certain properties. In this regard, large potentialities are embedded in immobilized cells. Enzyme production technology in a secreted active form enabling industrial application on an economically feasible scale has been developed. This method is based on the immobilization of enzyme producers on a solid career. Immobilizing has a range of advantages: decreasing the price of the final product, absence of foreign substances, controlled process of enzyme-genesis, the ability of various enzymes' simultaneous production, etc. Design of proposed equipment gives the opportunity to increase the activity of immobilized cell culture filtrate comparing to free cells, growing in periodic culture conditions. Such technology allows giving a 10-times raise in culture productivity, to prolong the process of fungi cultivation and periods of active culture liquid generation. Also, it gives the way to improve the quality of filtrates (to make them more clear) and exclude time-consuming processes of recharging fermentative vials, that require manual removing of mycelium.

Keywords: industrial enzymes, immobilization, submerged fermentation, microscopic fungi

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Authors: Louise Elizabeth Bolton

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Statement of the problem: There are an estimated 328 million cases of COPD worldwide. It is likely to become the third biggest cause of death by 2030. The impact of living with palliative care needs arising from COPD disrupts an individual’s existential situation. Understandings of individuals' existential situations within COPD are limited within the research literature and are rarely addressed within clinical practice, yet existential suffering has been linked to poor health-related quality of life for those living with other chronic conditions. The purpose of this integrative review is to provide a synthesis of existing evidence on existential suffering for those living with palliative care needs arising from COPD. Methods: This is an integrative review undertaken in accordance with PRISMA guidelines. Nine electronic databases were searched from April 2019 to January 2021. Thirty-five empirical research papers of both qualitative and quantitative methodologies, alongside systematic literature reviews, were included. Data analysis was undertaken using an integrative thematic analysis approach. Findings: Identified themes of existential suffering when living with palliative care needs arising from COPD are as follows: Liminality, Lamented Life, Loss of Personal Liberty, Life Meaning and Existential isolation. The absence of life meaning and purpose was of most importance to patients. Conclusion and Significance: This integrative review provides a synthesis of international evidence upon the presence of existential suffering. It is present and of significant impact within the daily lives of those living with palliative care needs arising from COPD. The absence of life meaning has the most significant impact, requiring further exploration of both its physical and psychological impact. Rediscovery of life meaning diminishes feelings of worthlessness and hopelessness in daily life and facilitates feelings of inner peace. For those with COPD living with such a relentless symptom burden, a positive existential situation is desirable.

Keywords: palliative care, COPD, existential suffering, end of life care

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Authors: Suwattana Visetnan, Premruethai Supungul, Sureerat Tang, Ikuo Hirono, Anchalee Tassanakajon, Vichien Rimphanitchayakit

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In animals, infection by Gram-negative bacteria and certain viruses activates the Imd signaling pathway wherein the a NF-κB transcription factor, Relish, is a key regulatory protein for the synthesis of antimicrobial proteins. Infection by yellow head virus (YHV) activates the Imd pathway. To investigate the expression of genes involved in YHV infection and under the influence of PmRelish regulation, RNA interference and suppression subtractive hybridization (SSH) are employed. The genes in forward library expressed in shrimp after YHV infection and under the activity of PmRelish were obtained by subtracting the cDNAs from YHV-infected and PmRelish-knockdown shrimp with cDNAs from YHV-infected shrimp. Opposite subtraction gave a reverse library whereby an alternative set of genes under YHV infection and no PmRelish expression was obtained. Sequencing of 252 and 99 cDNA clones from the respective forward and reverse libraries were done and annotated through blast search against the GenBank sequences. Genes involved in defense and homeostasis were abundant in both libraries, 31% and 23% in the forward and reverse libraries, respectively. They were predominantly antimicrobial proteins, proteinases and proteinase inhibitors. The expression of antimicrobial protein genes, ALFPm3, crustinPm1, penaeidin3 and penaeidin5 were tested under PmRelish silencing and Gram-negative bacterium V. harveyi infection. Together with the results previously reported, the expression of penaeidin5 and also penaeidin3 but not ALFPm3 and crustinPm1 were under the regulation of PmRelish in the Imd pathway.

Keywords: relish, yellow head virus, penaeus monodon, antimicrobial proteins

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Authors: Lazic Marina, Sugden Scott, Sharma Kanta Hem, Sauvageau Dominic, Stein Lisa

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Methane is a highly potent greenhouse gas mostly released through anthropogenic activities. Methane represents a low-cost and sustainable feedstock used for the biological production of value-added compounds by bacteria known as methanotrophs. In addition to methane, these organisms can utilize methanol, another cheap carbon source that is a common industrial by-product. Alphaproteobacteria methanotrophs can utilize both methane and methanol to produce the biopolymer polyhydroxybutyrate. The goal of this study was to examine the effect of methanol on polyhydroxybutyrate production in Methylocystis sp. Rockwell and to identify the optimal methane: methanol ratio that will improve PHB without reducing biomass production. Three methane: methanol ratios (4, 2.5., and 0.5) and three nitrogen source (ammonium or nitrate) concentrations (10 mM, 1 mM, and 0.1 mM) were combined to generate 18 growing conditions (9 per carbon source). The production of polyhydroxybutyrate and biomass was analyzed at the end of growth. Overall, the methane: methanol ratios that promoted polyhydroxybutyrate synthesis without reducing biomass were 4 and 2.5 and the optimal nitrogen concentration was 1 mM for both ammonium and nitrate. The physiological mechanism behind the beneficial effect of combining methane and methanol as carbon sources remain to be discovered. One possibility is that methanol has a dual role as a carbon source at lower concentrations and as a stringent response trigger at higher concentrations. Nevertheless, the beneficial effect of methanol and optimal nitrogen concentration for PHB production was confirmed, providing a basis for future physiological analysis and conditions for process scale-up.

Keywords: methane, methanol, methanotrophs, polyhydroxybutyrate, methylocystis sp. rockwell, single carbon bioconversions

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Authors: Y. V. S. Annapurna, V. V. Lakshmi

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Urinary tract infection (UTI) is one of the most common infectious diseases at the community level with a high rate of morbidity . This is further augmented by increase in the number of resistant and multi resistant strains of bacteria particularly by those producing Extended spectrum of beta lactamases. The present study was aimed at analysis of antibiograms of E.coli and Klebsiella sp causing urinary tract infections. Between November 2011 and April 2013, a total of 1120 urine samples were analyzed,. Antibiotic sensitivity testing was done with 542(48%) isolates of E.coli and 446(39%) of Klebsiella sp using the standard disc diffusion method against eleven commonly used antibiotics .Organisms showed high susceptibility to Amikacin and Netilimicin and low susceptibility to Cephalosporins. MAR index was calculated for the multidrug resistant strains. Maximum MAR index detected among the isolates was 0.9. Phenotypic identification for ESBL production was confirmed by double disk synergy test (DDST) according to CLSI guidelines. Plasmid profile of the isolates was carried out using alkaline hydrolysis method. Agarose-gel electrophoresis showed presence of high-molecular weight plasmid DNA among the ESBL strains. This study emphasizes the importance of indiscriminate use of antibiotics which if discontinued, in turn would prevent further development of bacterial drug resistance. For this, a proper knowledge of susceptibility pattern of uropathogens is necessary before prescribing empirical antibiotic therapy and it should be made mandatory.

Keywords: escherichia coli, extended spectrum of beta lactamase, Klebsiella spp, Uropathogens

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Authors: Atefeh Esmailnejad, Mohammad Abbaszadeh Hasiri

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Coxiella burnetii is a gram-negative obligatory intracellular bacterium that causes Q fever, a significant zoonotic disease. Sheep, cattle, and goats are the most commonly reported reservoirs for the bacteria, but infected cats and dogs have also been implicated in the transmission of the disease to human. The aim of present study was to investigate the presence of antibodies against Coxiella burnetii among companion dogs in Fars province, South of Iran. A total of 181 blood samples were collected from asymptomatic dogs, mostly referred to Veterinary Hospital of Shiraz University for regular vaccination. The IgG antibody detection against Coxiella burnetii was made by indirect Enzyme-linked Immunosorbent Assay (ELISA), employing phase I and II Coxiella burnetii antigens. A logistic regression model was developed to analyze multiple risk factors associated with seropositivity. An overall seropositivity of 7.7% (n=14) was observed. Prevalence was significantly higher in adult dogs above five years (18.18 %) compared with dogs between 1 and five years (7.86 %) and less than one year (6.17%) (P=0.043). Prevalence was also higher in male dogs (11.21 %) than in female (2.7 %) (P=0.035). There were no significant differences in the prevalence of positive cases and breed, type of housing, type of food and exposure to other farm animals (P>0.05). The results of this study showed the presence of Coxiella burnetii infection among the companion dogs population in Fars province. To our knowledge, this is the first study regarding Q fever in dogs carried out in Iran. In areas like Iran, where human cases of Q fever are not common or remain unreported, the public health implications of Q fever seroprevalence in dogs are quite significant.

Keywords: Coxiella burnetii, dog, Iran, Q fever

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Authors: Siti Helmyati, Euis Nurdiyawati, Joko Susilo, Endri Yuliati, Siti Fadhilatun Nashriyah, Kurnia Widyastuti

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Background: Iron fortification is one of the most effective and sustainable strategies to overcome anemia. It contradictively, has negative effect on gut microbiota balance. Pathogenic bacteria required iron for their growth. The iron source have greatly affect iron absorption in the intestine. Probiotic can inhibit the growth of pathogen. Lactobacillus plantarum Dad 13, Indonesian local isolate provides many benefits for health while fructo-oligosaccharides (FOS) provides selective substrates for probiotics’ growth. Objective: To determine the effect of iron fortification (NaFeEDTA and FeSO4) on antibacterial activity of synbiotic fermented milk. Methods: The antibacterial activity test was performed using the disc diffusion method. Paper discs were soaked in three kinds of synbiotic fermented milk, which are: 1) fortified with NaFeEDTA, 2) FeSO4 and 3) control. Escherichia coli was inoculated on nutrient agar medium. The ability of inhibition was shown by the formation of clear zone around the paper disc and measured in diameter (mm). Results: Synbiotic fermented milk fortified with iron (either NaFeEDTA or FeSO4) had antibacterial activity against Escherichia coli with diameter of clear zone were 6.53 mm and 12.3 mm, respectively (p<0.05). Compared to control (10.73 mm), synbiotic fermented milk fortified with FeSO4 had similar antibacterial activity (p>0.05). Conclusions: In vitro, synbiotic fermented milk fortified with NaFeEDTA and FeSO4 had different antibacterial activity against Escherichia coli. Iron fortification compound affected the antibacterial activity of synbiotic fermented milk.

Keywords: lactobacillus plantarum Dad 13, FOS, NaFeEDTA, FeSO4, antibacterial activity

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Authors: Neha Bhadauria, Indu Gaur, Shilpi Shilpi, Susmita Goswami, Prabir K. Paul

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The aerial surface of plants colonized by Human Enteric Pathogens ()has been implicated in outbreaks of enteric diseases in humans. Practice of organic farming primarily using animal dung as manure and sewage water for irrigation are the most significant source of enteric pathogens on the surface of leaves, fruits and vegetables. The present work aims to have an insight into the molecular mechanism of interaction of Human Enteric Pathogens or their metabolites with cell wall receptors in plants. Tomato plants grown under aseptic conditions at 12 hours L/D photoperiod, 25±1°C and 75% RH were inoculated individually with S. fonticola and K. pneumonia. The leaves from treated plants were sampled after 24 and 48 hours of incubation. The cell wall and cytoplasmic proteins were extracted and isocratically separated on 1D SDS-PAGE. The sampled leaves were also subjected to formaldehyde treatment prior to isolation of cytoplasmic proteins to study protein-protein interactions induced by Human Enteric Pathogens. Protein bands extracted from the gel were subjected to MALDI-TOF-TOF MS analysis. The foremost interaction of Human Enteric Pathogens on the plant surface was found to be cell wall bound receptors which possibly set ups a wave a critical protein-protein interaction in cytoplasm. The study revealed the expression and suppression of specific cytoplasmic and cell wall-bound proteins, some of them being important components of signaling pathways. The results also demonstrated HEP induced rearrangement of signaling pathways which possibly are crucial for adaptation of these pathogens to plant surface. At the end of the study, it can be concluded that controlling the over-expression or suppression of these specific proteins rearrange the signaling pathway thus reduces the outbreaks of food-borne illness.

Keywords: cytoplasmic protein, cell wall-bound protein, Human Enteric Pathogen (HEP), protein-protein interaction

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Authors: Ana Paola Echavarria, Mariuxi Medina, Haydelba D'Armas, Carmita Jaramillo, Diana San Martin

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The use of medicinal plants is one of the oldest and most extended medical therapies that goes back to prehistoric times, and nowadays, they are also used in the preparation of phytopharmaceuticals with options to cure diseases. The test for the determination of fungi was carried out in the Pharmacy Pilot Plant (treatment of the leaves of the plant species) and the Microbiology Laboratory (determination of fungi of the plant species, using growth medium called Sabouraud agar plus the vegetal sample), of the Academic Unit of Chemical Sciences and Health, of the Universidad Tecnica de Machala. Subsequently, colony counting was performed, both macroscopic, which is determined in the growth medium of the seeding, and microscopic, to identify the germinative forms using blue lactophenol. The procedure was repeated in duplicate to replicate the results data. The determination of the total fungal content of the following plant species was evaluated: Cymbopogon citratus (lemon verbena), Melissa officinalis (lemon balm), Taraxacum officinale (dandelion), Artemisia absinthium (absinthe), Piper carpunya (guaviduca), Moringa oleifera (moringa), Coriandrum sativum (coriander), Momordica charantia (achochilla), Borago officinalis (borage), Aloysia citriodora (cedron), Ambrosia artemisifolia (altamisa) and Ageratum conyzoides (mastrante). The results obtained showed that all the samples of the twelve plant species studied developed filamentous fungi, with great variability of them, within the permissible limits and contemplated by the Ecuadorian Institute of Normalization (INEN), being suitable as raw material for its use in the preparation of nutraceuticals and medicinal products or phytodrugs; with the exception of A. conyzoides (mastranto) which is the only species that exceeds the regulation in the average of dilutions.

Keywords: colonies, fungi, medicinal plants, microbiological quality, Sabouraud agar

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Authors: Areej Javaid, Nishat Fatima, Mehwish Naseer

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There is a tremendous use of biofertilizers in agriculture to increase crop productivity. Pakistan spends a huge amount on the purchase of synthetic fertilizers every year. The use of natural compounds to harness crop productivity is the major area of interest nowadays due to being safe for human health and the environment as well. Legumes have the intrinsic quality to enrich the nutrient status of soil because of the presence of nitrogen fixation bacteria on nodules. This research determined the effect of biofertilizer on nodulation attributes and yield of the pea plant. Seeds of pea varieties were treated with a slurry of biofertilizer prepared in a 10% sugar solution just before seed sowing. The impact of biofertilizer on different parameters of growth, yield and nodulation was observed. Analysis of variance showed that plant height, days to flowering, number of nodes, days to first pod, root length and plant height exhibited significant genetic variation. All the yield parameters, including the number of pods per plant, number of seeds per pod, seed fresh and dry weight showed significant results under treatment. Among nodulation parameters, nodule number responded positively to biofertilizer treatment. Genotypes 2001-40 showed better performance followed by 2001-20 and LINA-PAK in all the parameters, whereas 2001-40 and 2001-20 performed well in nodulation and yield parameters. Consequently, seed treatment with biofertilizer before sowing is recommended to obtain higher crop yield.

Keywords: biological nitrogen fixation, correlation analysis, quantitative inheritance, varietal responses

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Authors: Gordana Ćetković, Sanja Podunavac-Kuzmanović, Jasna Čanadanović-Brunet, Vesna Tumbas Šaponjac, Vanja Šeregelj, Jelena Vulić, Slađana Stajčić

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The color of food is one of the decisive factors for consumers. Potential toxicity of artificial food colorants has led to the consumers' preference for natural products over products with artificial colors. Natural pigments have many bioactive functions, such as antioxidant, provitamin and many other. Having this in mind, the acceptability of natural colorants by the consumers is much higher. Being present in all photosynthetic plant tissues carotenoids are probably most widespread pigments in nature. Carrot (Daucus carota) is a good source of functional food components. Carrot is especially rich in carotenoids, mainly α- and β-carotene and lutein. For this study, carrot was extracted using classical extraction with hexane and ethyl acetate, as well as supercritical CO₂ extraction. The extraction efficiency was evaluated by estimation of carotenoid yield determined spectrophotometrically. Classical extraction using hexane (18.27 mg β-carotene/100 g DM) was the most efficient method for isolation of carotenoids, compared to ethyl acetate classical extraction (15.73 mg β-carotene/100 g DM) and supercritical CO₂ extraction (0.19 mg β-carotene/100 g DM). Three carrot extracts were tested in terms of antioxidant activity using DPPH and reducing power assay as well. Surprisingly, ethyl acetate extract had the best antioxidant activity on DPPH radicals (AADPPH=120.07 μmol TE/100 g) while hexane extract showed the best reducing power (RP=1494.97 μmol TE/100 g). Hexane extract was chosen as the most potent source of carotenoids and was encapsulated in whey protein by freeze-drying. Carotenoid encapsulation efficiency was found to be high (89.33%). Based on our results it can be concluded that carotenoids from carrot can be efficiently extracted using hexane and classical extraction method. This extract has the potential to be applied in encapsulated form due to high encapsulation efficiency and coloring capacity. Therefore it can be used for dietary supplements development and food fortification.

Keywords: carotenoids, carrot, extraction, encapsulation

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Authors: Antonietta Maoloni, Federica Cardinali, Vesna Milanović, Andrea Osimani, Ilario Ferrocino, Maria Rita Corvaglia, Luca Cocolin, Lucia Aquilanti

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Table olives (Olea europaea L.) are among the most important fermented vegetables all over the world, while sea fennel (Crithmum maritimum L.) is an emerging food crop with interesting nutritional and sensory traits. Both of them are characterized by the presence of several bioactive compounds with potential beneficial health effects, thus representing two valuable substrates for the manufacture of innovative vegetable-based preserves. Given these premises, the present study was aimed at exploring the co-fermentation of table olives and sea fennel to produce new high-value preserves. Spanish style or Greek style processing method and the use of a multiple strain starter were explored. The preserves were evaluated for their microbial dynamics and key sensory traits. During the fermentation, a progressive pH reduction was observed. Mesophilic lactobacilli, mesophilic lactococci, and yeasts were the main microbial groups at the end of the fermentation, whereas Enterobacteriaceae decreased during fermentation. An evolution of the microbiota was revealed by metataxonomic analysis, with Lactiplantibacillus plantarum dominating in the late stage of fermentation, irrespective of processing method and use of the starter. Greek style preserves resulted in more crunchy and less fibrous than Spanish style one and were preferred by trained panelists.

Keywords: lactic acid bacteria, Lactiplantibacillus plantarum, metataxonomy, panel test, rock samphire

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Authors: Asma Lamin, Anna H. Kaksonen, Ivan Cole, Paul White, Xiao-Bo Chen

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Microbiologically influenced corrosion (MIC) is a process in which microorganisms initiate, facilitate, or accelerate the electrochemical corrosion reactions of metallic components. Several reports documented that MIC accounts for about 20 to 40 % of the total cost of corrosion. Biofilm formation due to the presence of microorganisms on the surface of metal components is known to play a vital role in MIC, which can lead to severe consequences in various environmental and industrial settings. Quorum sensing (QS) system plays a major role in regulating biofilm formation and control the expression of some microbial enzymes. QS is a communication mechanism between microorganisms that involves the regulation of gene expression as a response to the microbial cell density within an environment. This process is employed by both Gram-positive and Gram-negative bacteria to regulate different physiological functions. QS involves production, detection, and responses to signalling chemicals, known as auto-inducers. QS controls specific processes important for the microbial community, such as biofilm formation, virulence factor expression, production of secondary metabolites and stress adaptation mechanisms. The use of QS inhibitors (QSIs) has been proposed as a possible solution to biofilm related challenges in many different applications. Although QSIs have demonstrated some strength in tackling biofouling, QSI-based strategies to control microbially influenced corrosion have not been thoroughly investigated. As such, our research aims to target the QS mechanisms as a strategy for mitigating MIC on metal surfaces in engineered systems.

Keywords: quorum sensing, quorum quenching, biofilm, biocorrosion

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Authors: Duaa Mughal, Syeda Areeba Nadeem, Shakil Ahmed, Ishtiaq Ahmed Khan

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The identification of porcine DNA in Halal food items is critical to ensuring compliance with dietary restrictions and religious beliefs. In Islam, Porcine is prohibited as clearly mentioned in Quran (Surah Al-Baqrah, Ayat 173). The purpose of this study was to compare two DNA extraction procedures for detecting 0.001% of porcine DNA in processed Halal food sample mixtures containing chicken, camel, veal, turkey and goat meat using the TaqMan Real-Time PCR technology. In this research, two different commercial kit protocols were compared. The processed sample mixtures were prepared by spiking known concentration of porcine DNA to non-porcine food matrices. Afterwards, TaqMan Real-Time PCR technique was used to target a particular porcine gene from the extracted DNA samples, which was quantified after extraction. The results of the amplification were evaluated for sensitivity, specificity, and reproducibility. The results of the study demonstrated that two DNA extraction techniques can detect 0.01% of porcine DNA in mixture of Halal food samples. However, as compared to the alternative approach, Eurofins| GeneScan GeneSpin DNA Isolation kit showed more effective sensitivity and specificity. Furthermore, the commercial kit-based approach showed great repeatability with minimal variance across repeats. Quantification of DNA was done by using fluorometric assay. In conclusion, the comparison of DNA extraction methods for detecting porcine DNA in Halal food sample mixes using the TaqMan Real-Time PCR technology reveals that the commercial kit-based approach outperforms the other methods in terms of sensitivity, specificity, and repeatability. This research helps to promote the development of reliable and standardized techniques for detecting porcine DNA in Halal food items, religious conformity and assuring nutritional.

Keywords: real time PCR (qPCR), DNA extraction, porcine DNA, halal food authentication, religious conformity

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Authors: Sharique Ahmed, Khushtar Anwar Salman

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Alpha-1-antitrypsin (AAT) is a major plasma serine protease inhibitor (SERPIN). Hereditary AAT deficiency is one of the common diseases in some part of the world. AAT is mainly produced in the liver and functions to protect the lung against proteolytic damage (e.g., from neutrophil elastase) acting as the major inhibitor for neutrophil elastase. α (1)-Antitrypsin (AAT) deficiency is an under recognized genetic condition that affects approximately 1 in 2,000 to 1 in 5,000 individuals and predisposes to liver disease and early-onset emphysema. Not only does α-1-antitrypsin deficiency lead to disabling syndrome of pulmonary emphysema, there are other disorders too which include ANCA (antineutrophilic cytoplasmic antibody) positive Wegener's granulomatosis, diffuse bronchiectasis, necrotizing panniculitis in α-1-antitrypsin phenotype (S), idiopathic pulmonary fibrosis and steroid dependent asthma. Augmentation therapy with alpha-1 antitrypsin (AAT) from human plasma has been available for specific treatment of emphysema due to AAT deficiency. Apart from this several observations have also suggested a role for endogenous suppressors of HIV-1, alpha-1 antitrypsin (AAT) has been identified to be one of those. In view of its varied important role in humans, serum from a mammalian source was chosen for the isolation and purification. Studies were performed on the homogeneous fraction. This study suggests that the buffalo serum α-1-antritrypsin has characteristics close to ovine, dog, horse and more importantly to human α-1-antritrypsin in terms of its hydrodynamic properties such as molecular weight, carbohydrate content, etc. The similarities in the hydrodynamic properties of buffalo serum α-1-antitrypsin with other sources of mammalian α-1-antitrypsin mean that it can be further studied and be a potential source for "augmentation therapy", as well as a source of AAT replacement therapy to raise serum levels above the protective threshold. Other parameters like the amino acid sequence, the effect of denaturants, and the thermolability or thermostability of the inhibitor will be the interesting basis of future studies on buffalo serum alpha-1 antitrypsin (AAT).

Keywords: α-1-antitrypsin, augmentation therapy , hydrodynamic properties, serine protease inhibitor

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Authors: Nisha James, Shubha Ranganathan

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Sex trafficking is a type of human trafficking involving prostitution of individuals for sexual exploitation. The stigma and social isolation they face in the society often makes it difficult for them to become rehabilitated from trafficking, due to which many of them continue in prostitution for years after being sex trafficked. Victims are subjected to violations of their fundamental human rights, deprived of basic medical facilities and undergo long-term abuse. This paper focuses on the role of Non-Governmental Organizations (NGOs) in the rescue and rehabilitation of victims of sex trafficking. Semi-structured interviews were conducted with 26 survivors of sex trafficking, five sex workers and 14 non-community staff members of a project running NGO in the city of Chennai in South India. Chennai has a number of NGOs that are involved in HIV/AIDS awareness and prevention programs. In many cases, rehabilitation of sex trafficking victims is also a mandate of these NGOs. This particular NGO was also involved in development activities towards the eradication of HIV/AIDS. For instance, they were engaged in inculcating safe sex practices among high-risk groups such as sex workers or in fighting for sex worker rights. The study found that the NGO’s role in combatting sex trafficking is overrun by the way it approaches these issue related to HIV/AIDS. Further, their activities are dependent solely on funding. Given that gradually, international funding for HIV/AIDS has slowly been withdrawn, there have been problems such as reduction in the salary of the project staff, the outreach workers and peer educators, many of whom were survivors of sex trafficking who have been able to survive on their wages instead of continuing in prostitution. Therefore, till date, the project funding has helped in making them aware of the health and social consequences of continuing in prostitution, and in supporting them socioeconomically, but the lack of funding may also lead the NGO workers into a state of unemployment, poverty and eventually into being re-trafficked. The study concludes by pointing to the need for disengaging anti-trafficking efforts from the HIV/AIDS related programs.

Keywords: non-governmental organization role, non-governmental organization staff, sex trafficking survivors, sex workers

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Authors: Alejandro Arnaldo Barroso Martinez

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Unlike sex, which is constituted by anatomic-physiological differences, gender is a social construction. Our thoughts and behaviors as females and males are not etched in stone by our biology but rather from how society expects us to think and behave based on our sex assignment in the womb. Social expectations, values, and roles are taken on by individuals and shape the ways considered acceptable and linked to our bodies, feelings, and interpersonal relationships. Furthermore, these evolve into dire consequences for those who do not meet these disciplinary, economic, and cultural standards. Then, the social learning of gender identity implies the individual’s psychological sense of being, and it might be highly linked to a sense of life and suicide attempts. As a result, suicide has been considered a gender issue with differences in the rates and means used by men and women worldwide. Nevertheless, there has been a misunderstanding of the meaning of being male or female in a particular context and how it becomes a risk process for suicide attempts. For this reason, the general objective of the current research is to explain how this process occurs in Cuba. From a Critical Sociology and Social Psychology, a qualitative methodology was developed through six case studies and qualitative in-depth interviews. The analysis is focused on the sequence and interplay between two dimensions of meaning: signifiers and voices. Findings show that the risk process of suicide attempts in Cuba means some patriarchal beliefs and practices as part of informal educational models and some positivist practices in mental health attention. Findings also show that community relations create a sense of belonging, and it is a protection against suicide attempts in Cuba. Those frames of signifiers and voices explain in both males and females but differently when and how they are suffering from isolation, violence, the normalization of emotional awareness, and emotional distress expression. Suicide prevention programs should take gender learning into account as a cultural process.

Keywords: social constructions, gender identity, meanings, suicide attempt

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Authors: Manish Madappa, Madhavi Gayathri Raman

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The primary vehicle of human communication is language. A breakdown occurring in any aspect of communication may lead to frustration and isolation among the learners and the teachers. Over seven percent of the population in the world currently experience limitations and those children who exhibit a deviant/deficient language acquisition curve even when being in a language rich environment as their peers may be at risk of having a language disorder or language impairment. The difficulty may be in the word level [vocabulary/word knowledge] and/or the sentence level [syntax/morphology) Children with SLI appear to be developing normally in all aspects except for their receptive and/or expressive language skills. Thus, it is utmost importance to identify children with or at risk of SLI so that an early intervention can foster language and social growth, provide the best possible learning environment with special support for language to be explicitly taught and a step in providing continuous and ongoing support. The present study looks at Kannada English bilingual children and works towards identifying children at risk of “specific language impairment”. The study was conducted through an exploratory study which systematically enquired into the narratives of young Kannada-English bilinguals and to investigate the data for story structure in their narrative formulations. Oral narrative offers a rich source of data about a child’s language use in a relatively natural context. The fundamental objective is to ensure comparability and to be more universal and thus allows for the evaluation narrative text competence. The data was collected from 10 class three students at a primary school in Mysore, Karnataka and analyzed for macrostructure component reflecting the goal directed behavior of a protagonist who is motivated to carry out some kind of action with the intention of attaining a goal. The results show that the children exhibiting a deviation of -1.25 SD are at risk of SLI. Two learners were identified to be at risk of Specific Language Impairment with a standard deviation of more the 1.25 below the mean score.

Keywords: bilingual, oral narratives, SLI, macrostructure

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Authors: Nacéra Benoussaid, Lehalali Meriem, Benchabane Messaoud

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Our work focuses on the production of compound antibiotic effect of volatile nature namely hydrogen cyanide and the production and identification of molecules phénazinique by some strains of fluorescent Pseudomonas spp isolated from the rhizosphere of some trees for a possible use as bio pesticides antifungal effect and/or antibiotic. We tested the production of hydrogen cyanide of 21 strains of Pseudomonas spp. fluorescent among them 19 strains (90, 47%) showed a positive cyanogenesis.The antagonism test executed in vitro showed that Pseudomonas strains have a higher anti fungal effect relative to their antibacterial effect with diameters of inhibition zones up to 3, 9 cm recorded by the strain F48 against Coleosporiumsp compared with recorded results against bacteria with a maximum inhibition of 1, 26 cm among this antagonistic strain.Three strains were selected by testing for producing phénazines namely PI9, BB9 and F20. The effect of the antimicrobial activity was performed on different culture media (GN, King B, ISP2 and PDA). The results of our study allowed us to retain the King B medium as ideal medium for the production of secondary metabolite. The produced phenazinique compounds was extracted from various organic solvents, and after the results of antibiographie against germs - targets, the extracts of ethyl acetate gave the best results compared to dichloromethane and hexane.The Analysis of these compounds of antibiotic phenazinique effect within layer chromatography (CCM) and high performance liquid chromatography( HPLC) indicate that both strains PI9 and F20 are productive of phenazine-1-carboxylic acid (PCA). The BB9 strain is suspected to be productive of another phenazinique compound.

Keywords: Pseudomonas ssp. fluorescents, antagonism in vitro, secondary metabolite, phenazines, biopesticide.

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Authors: Shengxin Yu

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Earthquakes can cause varying degrees of damage to building and bridge structures. Traditional laminated natural rubber bearings (NRB) exhibit inadequate energy dissipation and restraint, particularly under near-fault ground motions, resulting in excessive displacements in the superstructure. This paper presents a composite natural rubber bearing (NFUD-NRB) incorporating two types of shape memory alloy (SMA) U-shaped dampers (UD). The bearing exhibits adjustable features, predominantly characterized by partial self-centering and multi-level energy dissipation, facilitated by nickel-titanium-based SMA (NiTi-SMA) and iron-based SMA (Fe-SMA) UDs. The hysteresis characteristics of NFUD-NRB can be tailored by manipulating the configuration of NiTi-SMA and Fe-SMA UDs. Firstly, the proposed bearing's geometric configuration and working principle are introduced. The rationality of the modeling strategy for the bearing is validated through existing experimental results. Parameterized numerical simulations are subsequently performed to investigate the partially self-centering behavior of NFUD-NRB. The findings indicate that NFUD-NRB can attain the anticipated nonlinear behavior and deliver adequate energy dissipation. Finally, the impact of NFUD-NRB on improving the seismic resilience of highway bridges is examined using the OpenSees software, with particular emphasis on the seismic performance of NFUD-NRB under near-fault ground motions. System-level analysis reveals that bridge systems equipped with NFUD-NRBs exhibit satisfactory residual deformations and higher energy dissipation than those equipped with traditional NRBs. Moreover, NFUD-NRB markedly mitigates the detrimental impacts of near-fault ground motions on the main structure of bridges.

Keywords: partially self-centering behavior, energy dissipation, natural rubber bearing, shape memory alloy, U-shaped damper, numerical investigation, near-fault ground motion

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Authors: M. Gomaa Abdoelatef, Robert M. Field, Lee, Yong-Kwan

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Growing human populations have placed increased demands on water supplies and a heightened interest in desalination infrastructure. Key elements of the economics of desalination projects are thermal and electrical inputs. With growing concerns over the use of fossil fuels to (indirectly) supply these inputs, coupling of desalination with nuclear power production represents a significant opportunity. Individually, nuclear and desalination technologies have a long history and are relatively mature. For desalination, Reverse Osmosis (RO) has the lowest energy inputs. However, the economically driven output quality of the water produced using RO, which uses only electrical inputs, is lower than the output water quality from thermal desalination plants. Therefore, modern desalination projects consider that RO should be coupled with thermal desalination technologies (MSF, MED, or MED-TVC) with attendant steam inputs to permit blending to produce various qualities of water. A large nuclear facility is well positioned to dispatch large quantities of both electrical and thermal power. This paper considers the supply of thermal energy to a large desalination facility to examine heat balance impact on the nuclear steam cycle. The APR1400 nuclear plant is selected as prototypical from both a capacity and turbine cycle heat balance perspective to examine steam supply and the impact on electrical output. Extraction points and quantities of steam are considered parametrically along with various types of thermal desalination technologies to form the basis for further evaluations of economically optimal approaches to the interface of nuclear power production with desalination projects. In our study, the thermodynamic evaluation will be executed by DE-TOP which is the IAEA desalination program, it is approved to be capable of analyzing power generation systems coupled to desalination systems through various steam extraction positions, taking into consideration the isolation loop between the APR-1400 and the thermal desalination plant for safety concern.

Keywords: APR-1400, desalination, DE-TOP, IAEA, MSF, MED, MED-TVC, RO

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Authors: Ji-Won Park, Jong-Wha Kim

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Triploid L. lancifolium have a wide geographic distribution. By contrast, diploid L. lancifolium have limited distributions in the islands and coastal regions of the South and West Korean Peninsula and northern Tsushima Island, Japan. L. lancifolium diploids and triploids are not sympatrically distributed with other lily species or ploidy lines in West Sea and South Sea Islands of the Korean Peninsula. This observation raises the following questions: 'Why have autotriploid L. lancifolium never been observed in those isolated islands?', 'What mechanism excludes the occurrence of autotriploids, if they arise?'. To determine the occurrence and survival of triploid plants in natural diploid populations of tiger lily (Lilium lancifolium), ploidy analysis was conducted on natural open-pollinated seeds produced from plants grown on isolated islands, and on hybrid seeds produced by artificial crossing between plant populations originating on different Korean islands. Normal seeds were classified into five grades depending on the ratio of embryo/endosperm lengths, including 5/5, 4/5, 3/5, 2/5, and 1/5. Triploids were not observed among seedlings produced from natural open pollinations on isolated islands. Triploids were detected only in seedlings of underdeveloped seed grades(3/5 and 2/5) from artificial crosses between populations from different isolated islands. The triploid occurrence frequency was calculated as 0.0 for natural open-pollinated seedlings and 0.000582 for artificial crosses(6 triploids from 10,303 seedlings). Triploids were produced from crosses between isolated populations located at least 70 km apart; no triploids were detected in inter-population crosses of plants originating on the same islands. Triploid seedlings have very low viability in soil. We analyzed factors affecting triploid occurrence and survival in natural diploid populations of L. lancifolium. The results suggest that triploids originate from fertilization between plants that are genetically isolated due to geographical isolation and/or genotypic differences.

Keywords: Lilium lancifolium, autotriploid, natural population, genetic distance, 2n female gamete

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Authors: Raymond Ezenweani, Jeffrey Ogbebor

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The menace of aquatic pollution has become increasingly of great concern and the effects of this pollution as a result of anthropogenic activities cannot be over emphasized. Phycoremediation is the application of algal remediation technology in the removal of harmful products from the environment. Harmful products also known as pollutants are usually introduced into the environment through variety of processes such as industrial discharge, agricultural runoff, flooding, and acid rain. This work has to do with the capability of algae in the efficient removal of different pollutants, ranging from hydrocarbons, eutrophication, agricultural chemicals and wastes, heavy metals, foul smell from septic tanks or dumps through different processes such as bioconversion, biosorption, bioabsorption and biodecomposition. Algae are capable of bioconversion of environmentally persistent compounds to degradable compounds and also capable of putting harmful bacteria growth into check in waste water remediation. Numerous algal organisms such as Nannochloropsis spp, Chlorella spp, Tetraselmis spp, Shpaerocystics spp, cyanobacteria and different macroalgae have been tested by different researchers in laboratory scale and shown to have 100% efficiency in environmental remediation. Algae as a result of their photosynthetic capacity are also efficient in air cleansing and management of global warming by sequestering carbon iv oxide in air and converting it into organic carbon, thereby making food available for the other organisms in the higher trophic level of the aquatic food chain. Algae play major role in the sustenance of the aquatic ecosystem by their virtue of being photosynthetic. They are the primary producers and their role in environmental sustainability is remarkable.

Keywords: Algae , Pollutant, ., Phycoremediation, Aquatic, Sustainability

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Authors: Zonglin Yang, Tatsuya Akiyama, Kerry S. Williamson, Michael J. Franklin, Thiruvarangan Ramaraj

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Pseudomonas aeruginosa is an opportunistic pathogen that forms surface-associated microbial communities (biofilms) on artificial implant devices and on human tissue. Biofilm infections are difficult to treat with antibiotics, in part, because the bacteria in biofilms are physiologically heterogeneous. One measure of biological heterogeneity in a population of cells is to quantify the cellular concentrations of ribosomes, which can be probed with fluorescently labeled nucleic acids. The fluorescent signal intensity following fluorescence in situ hybridization (FISH) analysis correlates to the cellular level of ribosomes. The goals here are to provide computationally and statistically robust approaches to automatically quantify cellular heterogeneity in biofilms from a large library of epifluorescent microscopy FISH images. In this work, the initial steps were developed toward these goals by developing an automated biofilm detection approach for use with FISH images. The approach allows rapid identification of biofilm regions from FISH images that are counterstained with fluorescent dyes. This methodology provides advances over other computational methods, allowing subtraction of spurious signals and non-biological fluorescent substrata. This method will be a robust and user-friendly approach which will enable users to semi-automatically detect biofilm boundaries and extract intensity values from fluorescent images for quantitative analysis of biofilm heterogeneity.

Keywords: image informatics, Pseudomonas aeruginosa, biofilm, FISH, computer vision, data visualization

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Authors: Fatemeh Zeinali Sehrig

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Background: miRNAs play an important role in the post-transcriptional regulation of genes, including genes involved in DNA methylation (DNMTs), and are also important regulators of oncogenic pathways. The study of microRNAs and DNMTs in breast cancer allows the development of targeted treatments and early detection of this cancer. Methods and Materials: Clinical Patients and Samples: Institutional guidelines, including ethical approval and informed consent, were followed by the Ethics Committee (Ethics code: IR.IAU.TABRIZ.REC.1401.063) of Tabriz Azad University, Tabriz, Iran. In this study, tissues of 100 patients with breast cancer and tissues of 100 healthy women were collected from Noor Nejat Hospital in Tabriz. The basic characteristics of the patients with breast cancer included: 1)Tumor grade(Grade 3 = 5%, Grade 2 = 87.5%, Grade 1 = 7.5%), 2)Lymph node(Yes = 87.5%, No = 12.5%), 3)Family cancer history(Yes = 47.5%, No = 41.3%, Unknown = 11.2%), 4) Abortion history(Yes = 36.2%).In silico methods (data gathering, process, and build networks): Gene Expression Omnibus (GEO), a high-throughput genomic database, was queried for miRNAs expression profiles in breast cancer. For Experimental protocol Tissue Processing, Total RNA isolation, complementary DNA(cDNA) synthesis, and quantitative real time PCR (QRT-PCR) analysis were performed. Results: In the present study, we found significant (p.value<0.05) changes in the expression level of miRNAs and DNMTs in patients with breast cancer. In bioinformatics studies, the GEO microarray data set, similar to qPCR results, showed a decreased expression of miRNAs and increased expression of DNMTs in breast cancer. Conclusion: According to the results of the present study, which showed a decrease in the expression of miRNAs and DNMTs in breast cancer, it can be said that these genes can be used as important diagnostic and therapeutic biomarkers in breast cancer.

Keywords: gene expression omnibus, microarray dataset, breast cancer, miRNA, DNMT (DNA methyltransferases)

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Authors: L. A. Bosnea, T. Moschakis, C. Biliaderis

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Two probiotic strains of Lactobacillus paracasei subsp. paracasei (E6) and Lactobacillus paraplantarum (B1), isolated from traditional Greek dairy products, were microencapsulated by complex coacervation using whey protein isolate (WPI, 3% w/v) and gum arabic (GA, 3% w/v) solutions mixed at different polymer ratio (1:1, 2:1 and 4:1). The effect of total biopolymer concentration on cell viability was assessed using WPI and GA solutions of 1, 3 and 6% w/v at a constant ratio of 2:1. Also, several parameters were examined for optimization of the microcapsule formation, such as inoculum concentration and the effect of ionic strength. The viability of the bacterial cells during heat treatment and under simulated gut conditions was also evaluated. Among the different WPI/GA weight ratios tested (1:1, 2:1, and 4:1), the highest survival rate was observed for the coacervate structures made with the ratio of 2:1. The protection efficiency at low pH values is influenced by both concentration and the ratio of the added biopolymers. Moreover, the inoculum concentration seems to affect the efficiency of microcapsules to entrap the bacterial cells since an optimum level was noted at less than 8 log cfu/ml. Generally, entrapment of lactobacilli in the complex coacervate structure enhanced the viability of the microorganisms when exposed to a low pH environment (pH 2.0). Both encapsulated strains retained high viability in simulated gastric juice (>73%), especially in comparison with non-encapsulated (free) cells (<19%). The encapsulated lactobacilli also exhibited enhanced viability after 10–30 min of heat treatment (65oC) as well as at different NaCl concentrations (pH 4.0). Overall, the results of this study suggest that complex coacervation with WPI/GA has a potential to deliver live probiotics in low pH food systems and fermented dairy products; the complexes can dissolve at pH 7.0 (gut environment), releasing the microbial cells.

Keywords: probiotic, complex coacervation, whey, encapsulation

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