Search results for: candidate gene of type-2 diabetes

Authors: Donludee Jaisut, Norihisa Kato, Thanutchaporn Kumrungsee, Kiyoshi Kawai, Somkiat Prachayawarakorn, Patchalee Tungtrakul

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Jasmine Rice is a principle food of Thai people. However, glycemic index of jasmine rice is in high level, risk of type II diabetes after consuming. Burdock root is a good source of non-starch polysaccharides such as inulin. Inulin acts as prebiotic and helps reduce blood-sugar level. The purpose of this research was to reduce digestion rate of jasmine rice by coating burdock root extract on rice surface, using top-spay fluidized bed coating technique. Coating experiments were performed by spraying burdock root solution onto Jasmine rice kernels (Khao Dawk Mali-105; KDML), which had an initial moisture content of 11.6% wet basis, suspended in the fluidized bed. The experimental conditions were: solution spray rates of 31.7 mL/min, atomization pressure of 1.5 bar, spray time of 10 min, time of drying after spraying of 30 s, superficial air velocity of 3.2 m/s and drying temperatures of 60°C. The coated rice quality was evaluated in terms of the moisture content, texture, whiteness and digestion rate. The results showed that initial and final moisture contents of samples were the same in concentration 8% (v/v) and 10% (v/v). The texture was insignificantly changed from that of uncoated sample. The whiteness values were varied on concentration of burdock root extract. Coated samples were slower digested.

Keywords: burdock root, digestion, drying, rice

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Authors: Sadhana Shukla, Pushplata Singh, Nidhi Didwania

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Arbuscular mycorrhizal (AM) fungi are a highly advantageous and versatile group of fungi that significantly contribute to the formation of soil organic matter by creating a demand for plant carbon (C) and distributing it through below-ground hyphal biomass, regardless of their substantial contribution in enhancing net primary productivity and accumulating additional photosynthetic fixed C in the soil. The genetic role of AM fungi in carbon cycling is largely unexplored. In our study, we propose that AM fungi significantly interact with the soil, particularly: the provision of photosynthates by plants. We have studied the expression of AM fungi genes involved in CO₂ sequestration during host-plant interaction was investigated by qPCR studies. We selected Rhizophagus proliferus (AM fungi) and Oryza sativa (Rice) (inoculated with or without 200ppg AMF inoculums per plant) and investigated the effect of AM fungi on soil organic carbon (SOC) and rice growth under field conditions. Results thus provided faster SOC turnover, 35% increased nutrient uptake in plants and pronounced hyphal biomass of AM fungi which enhanced soil carbon storage by 15% in comparison to uninoculated plants. This study will offer a foundation for delving into various carbon-soil studies while also advancing our comprehension of the relationship between AM fungi and the sustainability of agricultural ecosystems.

Keywords: arbuscular mycorrhizal (AM) fungi, carbon sequestration, gene expression, soil health, plant development.

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Authors: Ashit Syngle, Nidhi Garg, Pawan Krishan

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Objective: Bone marrow derived stem cells, endothelial progenitor cells (EPCs), protect against atherosclerotic vascular damage. However, EPCs are depleted in AS and contribute to the enhanced cardiovascular risk. Statins have a protective effect in CAD and diabetes by enhancing the proliferation, migration and survival of EPCs. Therapeutic potential of augmenting EPCs to treat the heightened cardiovascular risk of AS has not yet been exploited. We aimed to investigate the effect of rosuvastatin on EPCs population and inflammation in AS. Methods: 30 AS patients were randomized to receive 6 months of treatment with rosuvastatin (10 mg/day, n=15) and placebo (n=15) as an adjunct to existing stable anti-rheumatic drugs. EPCs (CD34+/CD133+) were quantified by Flow Cytometry. Inflammatory measures (BASDAI, BASFI, CRP and ESR), pro-inflammatory cytokines (TNF-α, IL-6 and IL-1) and lipids were measured at baseline and after treatment. Results: At baseline, inflammatory measures and pro-inflammatory cytokines were elevated and EPCs depleted among both groups. EPCs increased significantly (p < 0.01) after treatment with rosuvastatin. At 6 months, BASDAI, BASFI, ESR, CRP, TNF-α, and IL-6 improved significantly in rosuvastatin group. Significant negative correlation was observed between EPCs and BASDAI, CRP and IL-6 after rosuvastatin treatment. Conclusion: First study to show that rosuvastatin augments EPCs population in AS. This defines a novel mechanism of rosuvastatin treatment in AS: the augmentation of EPCs with improvement in proinflammatory cytokines and inflammatory disease activity. The augmentation of EPCs by rosuvastatin may provide a novel strategy to prevent cardiovascular events in AS.

Keywords: ankylosing spondylitis, Endothelial Progenitor Cells, inflammation, pro-inflammatory cytokines, rosuvastatin

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Authors: Hoda Mahgoub, Abeer Hanafy

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Linagliptin (LNG) belongs to dipeptidyl-peptidase-4 (DPP-4) inhibitor class. DPP-4 inhibitors represent a new therapeutic approach for the treatment of type 2 diabetes in adults. The aim of this work was to develop and validate an accurate and reproducible HPLC method for the determination of LNG with high sensitivity in rat plasma. The method involved separation of both LNG and pindolol (internal standard) at ambient temperature on a Zorbax Eclipse XDB C18 column and a mobile phase composed of 75% methanol: 25% formic acid 0.1% pH 4.1 at a flow rate of 1.0 mL.min-1. UV detection was performed at 254nm. The method was validated in compliance with ICH guidelines and found to be linear in the range of 5–1000ng.mL-1. The limit of quantification (LOQ) was found to be 5ng.mL-1 based on 100µL of plasma. The variations for intra- and inter-assay precision were less than 10%, and the accuracy values were ranged between 93.3% and 102.5%. The extraction recovery (R%) was more than 83%. The method involved a single extraction step of a very small plasma volume (100µL). The assay was successfully applied to an in-vivo pharmacokinetic study of LNG in rats that were administered a single oral dose of 10mg.kg-1 LNG. The maximum concentration (Cmax) was found to be 927.5 ± 23.9ng.mL-1. The area under the plasma concentration-time curve (AUC0-72) was 18285.02 ± 605.76h.ng.mL-1. In conclusion, the good accuracy and low LOQ of the bioanalytical HPLC method were suitable for monitoring the full pharmacokinetic profile of LNG in rats. The main advantages of the method were the sensitivity, small sample volume, single-step extraction procedure and the short time of analysis.

Keywords: HPLC, linagliptin, pharmacokinetic study, rat plasma

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Authors: Chia-Jung Li, Kuan-Hao Tsui

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As women age, the quality of their oocytes deteriorates irreversibly, leading to reduced fertility. To better understand the role of Ferroptosis-related genes in ovarian aging, we employed a multi-omics analysis approach, including spatial transcriptomics, single-cell RNA sequencing, human ovarian pathology, and clinical biopsies. Our study identified excess lipid peroxide accumulation in aging germ cells, metal ion accumulation via oxidative reduction, and the interaction between ferroptosis and cellular energy metabolism. We used multi-histological prediction of ferroptosis key genes to evaluate 75 patients with ovarian aging insufficiency and then analyzed changes in hub genes after supplementing with DHEA, Ubiquinol CoQ10, and Cleo-20 T3 for two months. Our results demonstrated a significant increase in TFRC, GPX4, NCOA4, and SLC3A2, which were consistent with our multi-component prediction. We theorized that these supplements increase the mitochondrial tricarboxylic acid cycle (TCA) or electron transport chain (ETC), thereby increasing antioxidant enzyme GPX4 levels and reducing lipid peroxide accumulation and ferroptosis. Overall, our findings suggest that supplementation intervention significantly improves IVF outcomes in senescent cells by enhancing metal ion and energy metabolism and enhancing oocyte quality in aging women.

Keywords: multi-omics, nutrients, ferroptosis, ovarian aging

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Authors: Modi Alsubaie, Willem Kuyken, Rebecca Abbott, Barnaby Dunn, Chris Dickens, Tina Keil, William Henley

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Background: Recently, there has been an increased interest in studying the effects of mindfulness-based interventions for people with psychological and physical problems. However, the mechanisms of action in these interventions that lead to beneficial physical and psychological outcomes have yet to be clearly identified. Purpose: The aim of this paper is to review, systematically, the evidence to date on the mechanisms of action in mindfulness interventions in populations with physical and/or psychological conditions. Method: Searches of seven databases (PsycINFO, Medline (Ovid), Cochrane Central Register of Controlled Trials, EMBASE, CINAHL, AMED, ClinicalTrials.gov) were undertaken in June 2014 and July 2015. We evaluated to what extent the studies we identified met the criteria suggested by Kazdin for establishing mechanisms of action within a psychological treatment (2007, 2009). Results: We identified four trials examining mechanisms of mindfulness interventions in those with comorbid psychological and physical health problems and 14 in those with psychological conditions. These studies examined a diverse range of potential mechanisms, including mindfulness and rumination. Of these candidate mechanisms, the most consistent finding was that greater self-reported change in mindfulness mediated superior clinical outcomes. However, very few studies fully met the Kazdin criteria for examining treatment mechanisms. Conclusion: There was evidence that global changes in mindfulness are linked to better outcomes. This evidence pertained more to interventions targeting psychological rather than physical health conditions. While there is promising evidence that MBCT/MBSR intervention effects are mediated by hypothesised mechanisms, there is a lack of methodological rigour in the field of testing mechanisms of action for both MBCT and MBSR, which precludes definitive conclusions.

Keywords: MBCT, MBSR, mechanisms, physical conditions, psychological conditions, systematic review

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Authors: Yen-Hui Chen, Terry Walker

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As the world experiences an energy crisis, investing in sustainable energy resources is a pressing mission for many countries. Microalgae-derived biodiesel has attracted intensive attention as an important biofuel, and microalgae Chlorella protothecoides lipid is recognized as a renewable source for microalgae-derived biodiesel production. Supercritical carbon dioxide (SC-CO₂) is a promising green solvent that may potentially substitute the use of organic solvents for lipid extraction; however, the efficiency of SC-CO₂ extraction may be affected by many variables, including temperature, pressure and extraction time individually or in combination. In this study, response surface methodology (RSM) was used to optimize the process parameters, including temperature, pressure and extraction time, on C. protothecoides lipid yield by SC-CO₂ extraction. A second order polynomial model provided a good fit (R-square value of 0.94) for the C. protothecoides lipid yield. The linear and quadratic terms of temperature, pressure and extraction time—as well as the interaction between temperature and pressure—showed significant effects on lipid yield during extraction. The optimal lipid yield from the model was predicted as the temperature of 59 °C, the pressure of 350.7 bar and the extraction time 2.8 hours. Under these conditions, the experimental lipid yield (25%) was close to the predicted value. The principal fatty acid methyl esters (FAME) of C. protothecoides lipid-derived biodiesel were oleic acid methyl ester (60.1%), linoleic acid methyl ester (18.6%) and palmitic acid methyl ester (11.4%), which made up more than 90% of the total FAMEs. In summary, this study indicated that RSM was useful to characterize the optimization the SC-CO₂ extraction process of C. protothecoides lipid yield, and the second-order polynomial model could be used for predicting and describing the lipid yield very well. In addition, C. protothecoides lipid, extracted by SC-CO₂, was suggested as a potential candidate for microalgae-derived biodiesel production.

Keywords: Chlorella protothecoides, microalgal lipids, response surface methodology, supercritical carbon dioxide extraction

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Authors: Wijayanada Rupasinghe, A. H. Dinithi Jayasekara

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In a modern democracy, new media can be used by governments to involve citizens in decision-making, and by civil society to engage people in specific issues. However new media can also be used to broaden political participation by helping citizens to communicate with their representatives and with each other. Arguably this political communication is most important during election campaigns when political parties and candidates seek to mobilize citizens and persuade them to vote for a given party or candidate. The new media must be used by Parliaments, Parliamentarians, governments and political parties as they are highly effective tools to involve and inform citizens in public policymaking and in the formation of governments. But all these groups must develop strategies to deal with a wide array of both positive and negative effects of these rapidly growing media.New media has begun to take precedent over other communication outlets in part because of its heightened accessibility and usability. Using personal website can empower the public in a way that is far faster, cheaper and more pervasive than other forms of communication. They encourage pluralism, reach young people more than other media and encourage greater participation, accountability and transparency. This research discusses the impact politicians’ personal websites has over their overall electability and likability and explores the integration of website is an essential campaign tactic on both the local and national level. This research examined the impact of having personal website have over the way constituents view politicians. This research examined how politicians can use their website in the most effective fashion and incorporate these new media outlets as essential campaign tools and tactics. A mixed-method approach using content analysis. Content analysis selected thirty websites in sri Lankan politicians. Research revealed that politician’s new media usage significantly influenced and enriched the experience an individual has with the public figure.

Keywords: election campaign ministers, new media, parliament, politicians websites

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Authors: Ava Faridi, Pouya Faridi, Aleksandr Kakinen, Ibrahim Javed, Thomas P. Davis, Pu Chun Ke

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Human islet amyloid polypeptide (IAPP) is a hormone associated with glycemic control and type 2 diabetes. Biophysically, the chirality of IAPP fibrils has been little explored with respect to the aggregation and toxicity of the peptide. Biochemically, it remains unclear as for how protein expression in pancreatic beta cells may be altered by cell exposure to the peptide, and how such changes may be mitigated by nanoparticle inhibitors for IAPP aggregation. In this study, we first demonstrated the elimination of the IAPP nucleation phase and shortening of its elongation phase by silica nanoribbons. This accelerated IAPP fibrillization translated to reduced toxicity, especially for the right-handed silica nanoribbons, as revealed by cell viability, helium ion microscopy, as well as zebrafish embryo survival, developmental and behavioral assays. We then examined the proteomes of βTC6 pancreatic beta cells exposed to the three main aggregation states of monomeric, oligomeric and amyloid fibrillar IAPP, and compared that with cellular protein expression modulated by graphene quantum dots (GQDs). A total of 29 proteins were significantly regulated by different forms of IAPP, and the majority of these proteins were nucleotide-binding proteins. A regulatory capacity of GQDs against aberrant protein expression was confirmed. These studies have demonstrated the great potential of employing nanomaterials targeting the mesoscopic enantioselectivity and protein expression dysregulation in pancreatic beta cells.

Keywords: graphene quantum dots, IAPP, silica nanoribbons, protein expression, toxicity

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Authors: Nadia A. Mohamed, Zakaria El-Khayat, Wagdy K. B. Khalil, Mehrez E. El-Naggar

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Drug nephrotoxicity is still a problem for patients who have taken drugs for elongated periods or permanently. Ultrasound-assisted sol−gel method was used to prepare hollow structured poroussilica nanoemulsion loaded with sodium salicylate as a model drug. The work was extended to achieve the target of the current work via investigating the protective role of this nanoemulsion model as anti-inflammatory drug or ginger for its antioxidant effect against cisplatin-induced nephrotoxicity in male albino rats. The results clarify that the nanoemulsion model was synthesized using ultrasonic assisted with small size and well stabilization as proved by TEM and DLS analysis. Additionally, blood urea nitrogen (BUN), Serum creatinine (SC) and Urinary total protein (UTP) were increased, and the level of creatinine clearance (Crcl) was decreased. All those were met with disorders in oxidative stress and downregulation in the expression of the nephrin gene. Also, histopathological changes of the kidney tissue were observed. These changes back to normal by treatment with silica nanoparticles loaded sodium salicylate (Si-Sc-NPs), ginger or both. Conclusions oil/water nanoemulsion of (Si-Sc NPs) and ginger showed a protective and promising preventive strategy against nephrotoxicity due to their antioxidant and anti-inflammatory effects, and that offers a new approach in attenuating drug induced nephrotoxicity.

Keywords: sodium salicylate nanoencapsulation, nephrin mRNA, drug nephrotoxicity, cisplatin, experimental rats

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Authors: H. Sakamoto, J. Tochimoto, S. Kurosawa, M. Suzuki, S. Oguri

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Salt stress adversely affects plant growth at various stages of development including seed germination, seedling establishment, vegetative growth and finally reproduction. Because of their immobile nature, plants have evolved mechanisms to sense and respond to salt stress. Seed dormancy is an adaptive trait that enables seed germination to coincide with favorable environmental conditions. We identified a novel locus of Arabidopsis, designated SHG1 (salt hypersensitive germination 1), whose disruption leads to reduced germination rate under moderate salt stress conditions. SHG1 encodes a transmembrane protein with an ankyrin repeat motif that has been implicated in diverse cellular processes such as signal transduction. The SGH1-disrupted Arabidopsis mutant died at the cotyledon stage when sown on salt-containing medium, although wild type plants could form true leaves under the same conditions. On the other hand, this mutant showed similar phenotypes to wild type plants when sown on medium without salt and transferred to salt-containing medium at the vegetative stage. These results suggested that SHG1 played indispensable role in the seed germination and seedling establishment under moderate salt stress conditions. SHG1 may be involved in the release of seed dormancy.

Keywords: germination, ankyrin repeat, arabidopsis, salt tolerance

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Authors: Nicolae Bold, Daniel Nijloveanu

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The idea of cropping-system is a method used by farmers. It is an environmentally-friendly method, protecting the natural resources (soil, water, air, nutritive substances) and increase the production at the same time, taking into account some crop particularities. The combination of this powerful method with the concepts of genetic algorithms results into a possibility of generating sequences of crops in order to form a rotation. The usage of this type of algorithms has been efficient in solving problems related to optimization and their polynomial complexity allows them to be used at solving more difficult and various problems. In our case, the optimization consists in finding the most profitable rotation of cultures. One of the expected results is to optimize the usage of the resources, in order to minimize the costs and maximize the profit. In order to achieve these goals, a genetic algorithm was designed. This algorithm ensures the finding of several optimized solutions of cropping-systems possibilities which have the highest profit and, thus, which minimize the costs. The algorithm uses genetic-based methods (mutation, crossover) and structures (genes, chromosomes). A cropping-system possibility will be considered a chromosome and a crop within the rotation is a gene within a chromosome. Results about the efficiency of this method will be presented in a special section. The implementation of this method would bring benefits into the activity of the farmers by giving them hints and helping them to use the resources efficiently.

Keywords: chromosomes, cropping, genetic algorithm, genes

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Authors: Monika Szymańska-Czerwińsk, Kinga Zaręba-Marchewka, Krzysztof Niemczuk

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Chlamydiaceae are Gram-negative bacteria distributed worldwide in animals and humans. One of them is Chlamydia gallinacea recently discovered. Available data show that C. gallinacea is dominant chlamydial agent found in poultry in European and Asian countries. The aim of the studies was screening of poultry flocks in order to evaluate frequency of C. gallinacea shedding and genetic diversity. Sampling was conducted in different regions of Poland in 2019-2020. Overall, 1466 cloacal/oral swabs were collected in duplicate from 146 apparently healthy poultry flocks including chickens, turkeys, ducks, geese and quails. Dry swabs were used for DNA extraction. DNA extracts were screened using a Chlamydiaceae 23S rRNA real-time PCR assay. To identify Chlamydia species, specific real-time PCR assays were performed. Furthermore, selected samples were used for sequencing based on ompA gene fragments and variable domains (VD1-2, VD3-4). In total, 10.3% of the tested flocks were Chlamydiaceae-positive (15/146 farms). The presence of Chlamydiaceae was confirmed mainly in chickens (13/92 farms) but also in turkey (1/19 farms) and goose (1/26 farms) flocks. Eleven flocks were identified as C. gallinacea-positive while four flocks remained unclassified. Phylogenetic analysis revealed at least 16 genetic variants of C. gallinacea. Research showed that Chlamydiaceae occur in a poultry flock in Poland. The strains of C. gallinacea as dominant species show genetic variability.

Keywords: C. gallinacea, emerging agent, poultry, real-time PCR

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Authors: M. Medrano, M. D. M. S., L. Younes, M. D.

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Introduction: Incidence of tibial hemimelia is 1:1,000,000. Due to pre-existing case studies and literature, there is now a better understanding of the genetics, etiology and pathoanatomy of tibial hemimelia, but an underlying cause is generally unknown. This presentation aims to discuss a rare, congenital lower limb deficiency observed in a patient in order to identify potential prenatal risk factors and future considerations for the patient’s well-being. Observation: A newborn female child, born full term via spontaneous vaginal delivery after induction of labor to unaffected and non-consanguineous parents. The prenatal course was notable for limited and disjointed prenatal care as well as maternal tobacco and marijuana use, anemia of pregnancy, and inadequate weight gain. Prenatal imaging showed lower extremity deformity with the inability to visualize tibia and bilateral clubfeet in the setting of Intrauterine Growth Restriction (IUGR). The patient presented with right equino varus deformity of the foot and right knee joint deformity. Radiological imaging showed the absence of the right tibia and varus angulation of the right foot with dislocation of the tibiotalar joint. Normal femur with lateral and mild anterior displacement of a wide fibula (Weber Type VIIa). Due to the absence of the patient’s tibia and knee extensor mechanism, the patient was not a candidate for reconstructive surgery and ultimately underwent successful right knee disarticulation. Discussion and Conclusion: By utilizing a retrospective chart review of this case, possible risk factors in prenatal care may be identified and add to existing knowledge on etiology. Hopefully, a cause can be clearly identified in the future and, thus, addressed in the prenatal period. In addition, we can investigate the patient’s well-being and adjustment post-operatively to support outpatient management of an uncommon anomaly.

Keywords: Tibial hemimelia, prenatal care, pediatric orthopedics, congenital deformity

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Authors: Mansour F. Hussein, Mohammed A. Alshaikh, Riyadh S. Al-Jumaah, A. GarelNabi, I. Al-Khalifa, Osama B. Mohammed

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Serum samples from 489 male and female camels were tested for antibodies against C. burnetii using indirect enzyme-linked immunosorbent assay (ELISA). Antibodies to C. burnetii were recorded in sera of 252 (51.64%) camels. Significant differences in prevalence were found between male and female camels, juvenile and adult camels, different ecotypes and different sampling locations. 307 camels were simultaneously tested for C. burnetii antibodies by ELISA and indirect immunofluorescence (IFA). Close agreement was found between the results of the two tests. A high prevalence of C. burnetii antibodies was also recorded in milk samples tested by ELISA. Clinical samples from serologically positive camels were subjected to PCR analysis using primers which amplify the repetitive transposon-like and transposase gene regions of C. burnetii. Positive DNA amplification was obtained from both regions, with highest shedding of C. burnetii in faecal samples (27.59%) followed, in descending order, by urine (23.81%), blood (15.85%) and milk (6.5%). The present results indicate that camels are a major reservoir of C. burnetii in Saudi Arabia. The high prevalence of infection in camels, the poor sanitary standards under which the animals are kept and the consumption of raw camel milk indicate that camels could also be a major source of transmission of Q fever to humans in Saudi Arabia.

Keywords: Arabian camel, Camelus dromedarius, Coxiella brunetii, ELISA, immunofluoresence, PCR

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Authors: Valizadeh Gever Bita, Joel Caren, Louisa Huand, Yu-Cheng Zhu, Esmaeil Amiri

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Within a honey bee colony, queen is the sole reproducer of fertilized eggs, while queens are safeguarded by worker bees, trophallactic behavior and food sharing activities could expose them to agrochemicals. Here, we assessed the effects of three widely used pesticides—Acephate, Bifenthrin, and Chlorantraniliprole— on worker bees, to investigate indirect effects on the physiology and reproductive traits of queens as well as the eggs they produce. Using RT-qPCR we measured the expression of several detoxification and immune genes in adult worker bees, queens, and freshly laid eggs after pesticide exposure. These analyses aimed to elucidate the physiological changes in queens and potential transgenerational effects. While no significant changes in reproductive traits were observed following Chlorantraniliprole and Bifenthrin exposure, Acephate caused adverse effects on egg size, egg-laying activity, and queen weight. The expression of detoxification, immune and antioxidant-related genes in workers, queens and freshly laid eggs changed over time in response to these pesticides. The results of this investigation revealed that pesticides can cause negative impact on queen physiology and reproduction indirectly through their effects on exposed worker bees. These effects can potentially extend to the next generation of honey bees.

Keywords: apis mellifera, egg laying, detoxification enzymes, gene expression, honey bee queen

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Authors: Rosetta Moshirian Farahi, Ahya Abdi Ali, Sara Gharavi

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The major mechanism of Pseudomonas aeruginosa resistance to fluoroquinolones is the alteration of target enzymes, type II and IV topoisomerases due to mutations in the quinolone resistance-determining regions (QRDR) of the gyrA and parC genes coding A subunits of these enzymes. 37 isolates from patients with burn wounds and 20 isolates from blood, urine and sputum specimen were selected to evaluate mutations involved in antibiotic resistance and were subsequently verified for their resistance to ciprofloxacin. QRDRs regions of gyrA and parC were amplified by polymerase chain reaction (PCR) and were subsequently sequenced. 90% of isolates with MIC≥8 µg/ml to ciprofloxacin had a mutation in gyrA gene in which threonine at position 83 changed to isoleucine. 87.5% of isolates had mutation in parC, Serine 87 changed. 75% had Ser87Leu and 12.5% possessed Serin87Trp. Various silent mutations were also detected such as Val103Val, Ala118Ala, Ala136Ala, His132His in gyrA and Ala115Ala in parC. The data indicates that the common mutation in gyrA is Thr83Ile and in parC is Ser87Leu/Trp. No individual parC mutation was observed while mutations in gyrA and parC occurred simultaneously and appears to be the main reason of high-level resistance to fluoroquinolones in patients with burn wounds and urine infection. The vast majority of P.aeruginosa isolates had mutation in parC which can play a crucial role in increased resistance of these isolates. This is a report of parC mutations from resistant P. aeruginosa isolates from Iran, Tehran.

Keywords: P. aeruginosa, fluoroquinolones, gyrA, parC, antibiotic resistance

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Authors: Amirhossein Moghanian, Morteza Elsa, Mehrnaz Aminitabar

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Modified bioactive glass has been considered as a promising multifunctional candidate in bone repair and regeneration due to its attractive properties. The present study mainly aims to evaluate how the individual substitution of lithium (L-BG) and magnesium (M-BG) for calcium can affect the in vitro bioactivity of sol-gel derived substituted 58S bioactive glass (BG); and to present one composition in both of the 60SiO2–(36-x)CaO–4P₂O₅–(x)Li₂O and 60SiO₂ –(36-x)CaO–4P₂O₅–(x)MgO quaternary systems (where x= 0, 5, 10 mol.%) with improved biocompatibility, enhanced alkaline phosphatase (ALP) activity, and the most efficient antibacterial activity against methicillin-resistant staphylococcus aureus bacteria. To address these aims, and study the effect of CaO/Li₂O and CaO/MgO substitution up to 10 mol % in 58S-BGs, the samples were characterized by X-ray diffraction, Fourier transform infrared spectroscopy, inductively coupled plasma atomic emission spectrometry and scanning electron microscopy after immersion in simulated body fluid up to 14 days. Results indicated that substitution of either CaO/ Li₂O and CaO/ MgO had a retarding effect on in vitro hydroxyapatite (HA) formation due to the lower supersaturation degree for nucleation of HA compared with 58s-BG. Meanwhile, magnesium had a more pronounced effect. The 3-(4,5dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and alkaline phosphatase (ALP) assays showed that both substitutions of CaO/ Li₂O and CaO/ MgO up to 5mol % in 58s-BGs led to increased biocompatibility and stimulated proliferation of the pre-osteoblast MC3T3 cells with respect to the control. On the other hand, the substitution of either Li or Mg for Ca in the 58s BG composition resulted in improved bactericidal efficiency against MRSA bacteria. Taken together, sample 58s-BG with 5 mol % CaO/Li₂O substitution (BG-5L) was considered as a multifunctional biomaterial in bone repair/regeneration with improved biocompatibility, enhanced ALP activity as well as significant antibacterial activity against methicillin-resistant staphylococcus aureus (MRSA) bacteria.

Keywords: alkaline, alkaline earth, bioactivity, biomedical applications, sol-gel processes

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Authors: Muhammad Jawad Saleem, Faisal Hafeez, Muhammad Arshad, Afifa Naeem, Ayesha Iftekhar

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Bacillus thuringiensis is a gram-positive spore-forming bacterium that belongs to the Bacillus cereus group of Bacilli and it produces ICP (insecticidal crystal protein) Cry toxins or Cysts toxins. Spores are produced as parasporal crystalline inclusions bodies (also known as endotoxins) at the onset of sporulation during the stationary growth phase. During vegetative growth that does not form crystals and is called vegetative insecticidal proteins (VIP) and secreted an insecticidal protein (SIP). Bacillus thuringiensis (Bt) is important for pest management either in the form of insecticides or through incorporated in the gene of the crop. Bioassays were conducted on the F2 generation of 1st instar larvae of H. armigera by the diet incorporation method to determine the susceptibility to Bt Cry toxins (Cry1Ac, Cry2Ab, Cry2A). The median lethal concentration (LC₅₀) of Cry1Ac, Cry2Ab, Cry2A ranged from 0.11 to 1.06 µg/ml and moult inhibitory concentration (MIC₅₀) of Cry1Ac, Cry2Ab, Cry2A ranged from 0.05 to 0.25 µg/ml. Cry1Ac was found most toxic to 1st instar larvae of H. armigera as compared to other Bt Cry toxins (Cry1Ac, Cry2Ab, Cry2A). The experimental results are important to policy-makers and technology providers to develop strategies for the exploitation of transgenic Bt cotton varieties as a component of integrated pest management.

Keywords: Bt toxin, Cry1Ac, Cry2Ab, Cry2A, susceptibility, Helicoverpa armigera

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Authors: Aniket Kumar, Jacob Peedicayil

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Major depressive disorder (MDD) is a common and important psychiatric disorder. Several clinical features of MDD suggest an epigenetic basis for its pathogenesis. Since epigenetics (heritable changes in gene expression not involving changes in DNA sequence) may underlie the pathogenesis of MDD, epigenetic drugs such as DNA methyltransferase inhibitors (DNMTi) and histone deactylase inhibitors (HDACi) may be useful for treating MDD. The available literature indexed in Pubmed on preclinical drug trials of epigenetic drugs for the treatment of MDD was investigated. The search terms we used were ‘depression’ or ‘depressive’ and ‘HDACi’ or ‘DNMTi’. Among epigenetic drugs, it was found that there were 3 preclinical trials using HDACi and 3 using DNMTi for the treatment of MDD. All the trials were conducted on rodents (mice or rats). The animal models of depression that were used were: learned helplessness-induced animal model, forced swim test, open field test, and the tail suspension test. One study used a genetic rat model of depression (the Flinders Sensitive Line). The HDACi that were tested were: sodium butyrate, compound 60 (Cpd-60), and valproic acid. The DNMTi that were tested were: 5-azacytidine and decitabine. Among the three preclinical trials using HDACi, all showed an antidepressant effect in animal models of depression. Among the 3 preclinical trials using DNMTi also, all showed an antidepressant effect in animal models of depression. Thus, epigenetic drugs, namely, HDACi and DNMTi, may prove to be useful in the treatment of MDD and merit further investigation for the treatment of this disorder.

Keywords: DNA methylation, drug discovery, epigenetics, major depressive disorder

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Authors: Fatemeh Kourkinejad Gharaei, Tahereh Safari, Zahra Saebinasab

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Metabolic syndrome (MetS) is a set of risk factors associated with cardiovascular diseases, atherosclerosis, and type 2 diabetes. Nonalcoholic fatty liver disease (NAFLD) is the most important liver disorder in metabolic syndrome. High fructose consumption increases the risk of NAFLD. Eugenol shows anti-thrombotic, insulin-sensitive, fat-reducing effects. This study was designed to investigate the protective role of eugenol in NAFLD caused by metabolic syndrome. Methods: Thirty male Wistar rats were randomly divided into five groups; group 1, drinking water intake animals; group 2, fructose, group 3, fructose+eugenol solvent; group 4, fructose+ eugenol 50mg/kg and group 5, fructose+ eugenol 100mg/kg. At the end of the experiment, after 12 hours of fasting and under anesthesia, blood samples were taken for measurement of fast blood glucose (FBS), SGOT, AGPT, LDL, HDL, cholesterol, triglyceride. Results: FBG significantly increased in group 2 compared to group 1 (p < 0.001); however, it significantly decreased in groups 4 and 5 compared to group 2 (p < 0.05). SGOT and SGPT levels significantly increased in group 2 compared to drinking water alone (p < 0.001). However, SGOT and SGPT levels significantly decreased in groups 4 and 5. MDA and LTDS significantly increased in group 2 compared with drinking water alone (p < 0.01), while MDA and LTDS decreased in 4 and 5 groups compared to group 2 (p < 0.05), which confirms the pathology results related to the liver damage. Conclusion: Eugenol has protective effects on the liver and fat accumulation in liver cells.

Keywords: eugenol, fructose, metabolic syndrome, nonalcoholic fatty liver disease

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Authors: Hansraj Riteesh Bookun, Emily Maree Stevens, Jarryd Leigh Solomon, Anthony Chan

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Objective: This was a retrospective cross-sectional study evaluating the safety and efficacy of femoral endarterectomy using the eversion technique with transection as opposed to the conventional endarterectomy technique with either vein or synthetic patch arterioplasty. Methods: Between 2010 to mid 2017, 19 patients with mean age of 75.4 years, underwent eversion femoral endarterectomy with transection by a single surgeon. There were 13 males (68.4%), and the comorbid burden was as follows: ischaemic heart disease (53.3%), diabetes (43.8%), stage 4 kidney impairment (13.3%) and current or ex-smoking (73.3%). The indications were claudication (45.5%), rest pain (18.2%) and tissue loss (36.3%). Results: The technical success rate was 100%. One patient required a blood transfusion following bleeding from intraoperative losses. Two patients required blood transfusions from low post operative haemogloblin concentrations – one of them in the context of myelodysplastic syndrome. There were no unexpected returns to theatre. The mean length of stay was 11.5 days with two patients having inpatient stays of 36 and 50 days respectively due to the need for rehabilitation. There was one death unrelated to the operation. Conclusion: The eversion technique with transection is safe and effective with low complication rates and a normally expected length of stay. It poses the advantage of not requiring a synthetic patch. This technique features minimal extraneous dissection as there is no need to harvest vein for a patch. Additionally, future endovascular interventions can be performed by puncturing the native vessel. There is no change to the femoral bifurcation anatomy after this technique. We posit that this is a useful adjunct to the surgeon’s panoply of vascular surgical techniques.

Keywords: endarterectomy, eversion, femoral, vascular

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Authors: Kita R. Ashman, Anthony R. Rendall, Matthew R. E. Symonds, Desley A. Whisson

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Forest cover is decreasing globally, chiefly due to the conversion of forest to agricultural landscapes. In contrast, the area under plantation forestry is increasing significantly. For wildlife occupying landscapes where native forest is the dominant land cover, plantations generally represent a lower value habitat; however, plantations established on land formerly used for pasture may benefit wildlife by providing temporary forest habitat and increasing connectivity. This study investigates the influence of landscape, site, and climatic factors on koala population density in far south-west Victoria where there has been extensive plantation establishment. We conducted koala surveys and habitat characteristic assessments at 72 sites across three habitat types: plantation, native vegetation blocks, and native vegetation strips. We employed a hierarchical modeling framework for estimating abundance and constructed candidate multinomial N-mixture models to identify factors influencing the abundance of koalas. We detected higher mean koala density in plantation sites (0.85 per ha) than in either native block (0.68 per ha) or native strip sites (0.66 per ha). We found five covariates of koala density and using these variables, we spatially modeled koala abundance and discuss factors that are key in determining large-scale distribution and density of koala populations. We provide a distribution map that can be used to identify high priority areas for population management as well as the habitat of high conservation significance for koalas. This information facilitates the linkage of ecological theory with the on-ground implementation of management actions and may guide conservation planning and resource management actions to consider overall landscape configuration as well as the spatial arrangement of plantations adjacent to the remnant forest.

Keywords: abundance modelling, arboreal mammals plantations, wildlife conservation

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Authors: Adewole Tomiwa Adetunji, Francis Bayo Lewu, Richard Mundembe

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Plants require nitrogen (N) to support desired production levels. There is a need for better understanding of N transport mechanism in order to improve N assimilation by plant root. Nitrogen is available to plants in the form of nitrate or ammonium, which are transported into the cell with the aid of various transport proteins. Ammonium transporters (AMTs) play a role in the uptake of ammonium, the form in which N is preferentially absorbed by plants. Solanum tuberosum AMT1 (StAMT1) was amplified, sequenced and characterized using molecular biology and bioinformatics methods. Nucleotide database sequences were used to design 976 base pairs AMT1-specific primers which include forward primer 5’- GCCATCGCCGCCGCCGG-3’ and reverse primer 5’-GGGTCAGATCCATACCCGC-3’. These primers were used to amplify the Solanum tuberosum AMT1 internal regions. Nucleotide sequencing, alignment and phylogenetic analysis assigned StAMT1 to the AMT1 family due to the clade and high similarity it shared with other plant AMT1 genes. The deduced amino acid sequences showed that StAMT1 is 92%, 83% and 76% similar to Solanum lycopersicum LeAMT1.1, Lotus japonicus LjAMT1.1, and Solanum lycopersicum LeAMT1.2 respectively. StAMT1 fragments were shown to correspond to the 5th-10th trans-membrane domains. Residue StAMT1 D15 is predicted to be essential for ammonium transport, while mutations of StAMT1 S76A may further enhance ammonium transport.

Keywords: ammonium transporter, bioinformatics, nitrogen, primers, Solanum tuberosum

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Authors: Asma Naseer Cheema, Attya Bhatti, Jabar Ali, John Peter

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Background: High cholesterol levels, endothelial dysfunction, inefficient coagulation cascade and hyper inflammatory response all are the basis of coronary artery disease (CAD). Several studies are carried out to see the genetic influence of these factors on disease outcome. Objective: The objective of our study was to see the association of 10 putative loci with coronary artery disease in our population. Materials & Methods: We screened our population for 10 putative loci of CAD showing significant association (p < 5x10-8) with candidate genes (regulating the cholesterol metabolism, endothelial function, coagulation cascade and inflammatory response of body). Hardy-Weinberg equilibrium and linkage disequilibrium in cases and controls s were estimated separately. Approximately 5-10 ng of dried DNA in 384 well plate format was used to genotype each sample on the Sequenom iPLEX assay at University of Pittsburgh Genomics and Proteomics Core Laboratories. It was built on single-base primer extension with the MALDI-TOF MS detection possessing high sensitivity and specificity. The SNPs were genotyped through Taqman assay. Hardy Weinberg test was applied. The 10 SNPs were selected as genetic markers for this study (rs579459, rs1561198, rs2954029, rs1122608, rs17114036, rs9515203, rs10947789, rs7173743, rs2895811, rs2075650). Results: Mean age of the patient was 52 ± 11 years. Blood pressure and positive family history was found a significant risk factor for CAD. None of the selected SNPs showed significant association with coronary artery disease in our population (p>0.05). Conclusion: rs579459, rs1561198, rs2954029, rs1122608, rs17114036, rs9515203, rs10947789, rs7173743, rs2895811, rs2075650 are not significant genetic markers for CAD in our population.

Keywords: CAD, genetic markers, loci, risk factors

Procedia PDF Downloads 371

Authors: C. Bolakis, I. S. Karanasiou, D. Grbovic, G. Karunasiri, N. Uzunoglu

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A new approach for efficient detection of THz radiation in biomedical imaging applications is proposed. A double-layered absorber consisting of a 32 nm thick aluminum (Al) metallic layer, located on a glass medium (SiO2) of 1 mm thickness, was fabricated and used to design a fine-tuned absorber through a theoretical and finite element modeling process. The results indicate that the proposed low-cost, double-layered absorber can be tuned based on the metal layer sheet resistance and the thickness of various glass media taking advantage of the diversity of the absorption of the metal films in the desired THz domain (6 to 10 THz). It was found that the composite absorber could absorb up to 86% (a percentage exceeding the 50%, previously shown to be the highest achievable when using single thin metal layer) and reflect less than 1% of the incident THz power. This approach will enable monitoring of the transmission coefficient (THz transmission ‘’fingerprint’’) of the biosample with high accuracy, while also making the proposed double-layered absorber a good candidate for a microbolometer pixel’s active element. Based on the aforementioned promising results, a more sophisticated and effective double-layered absorber is under development. The glass medium has been substituted by diluted poly-si and the results were twofold: An absorption factor of 96% was reached and high TCR properties acquired. In addition, a generalization of these results and properties over the active frequency spectrum was achieved. Specifically, through the development of a theoretical equation having as input any arbitrary frequency in the IR spectrum (0.3 to 405.4 THz) and as output the appropriate thickness of the poly-si medium, the double-layered absorber retains the ability to absorb the 96% and reflects less than 1% of the incident power. As a result, through that post-optimization process and the spread spectrum frequency adjustment, the microbolometer detector efficiency could be further improved.

Keywords: bio-imaging, fine-tuned absorber, fingerprint, microbolometer

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Authors: Atena Sadat Hosseini, Mohammadhossein Habibi

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Acute myeloid leukemia (AML) is the most typically diagnosed leukemia. In older adults, AML imposes a dismal outcome. AML originates with a dominant mutation, then adds collaborative, transformative mutations leading to myeloid transformation and clinical/biological heterogeneity. Several chemotherapeutic drugs are used for this cancer. These drugs are naturally associated with several side effects, and finding a more accurate molecular mechanism of these drugs can have a significant impact on the selection and better candidate of drugs for treatment. In this study, we evaluated bortezomibin myeloma cells using bioinformatics analysis and evaluation of RNA-Seq data. Then investigated the molecular pathways proteins- proteins interactions associated with this chemotherapy drug. A total of 658upregulated genes and 548 downregulated genes were sorted.AUF1 (hnRNP D0) binds and destabilizes mRNA, degradation of GLI2 by the proteasome, the role of GTSE1 in G2/M progression after G2 checkpoint, TCF dependent signaling in response to WNT demonstrated in upregulated genes. Besides insulin resistance, AKT phosphorylates targets in the nucleus, cytosine methylation, Longevity regulating pathway, and Signal Transduction of S1P Receptor were related to low expression genes. With respect to this results, HIST2H2AA3, RP11-96O20.4, ZED9, PRDX1, and DOK2, according to node degrees and betweenness elements candidates from upregulated genes. in the opposite side, PYURF, NRSN1, FGF23, UPK3BL, and STAG3 were a prominent role in downregulated genes. Sum up, Using in silico analysis in the present study, we conducted a precise study ofbortezomib molecular mechanisms in myeloma cells. so that we could take further evaluation to discovermolecular cancer therapy. Naturally, more additional experimental and clinical procedures are needed in this survey.

Keywords: myeloma cells, acute myeloid leukemia, bioinformatics analysis, bortezomib

Procedia PDF Downloads 93

Authors: Ananthakrishna L. R., Ramesh D., Kumar Wodeyar, Kotresh A. M., Gururaj P. M.

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Malnad Gidda is the indigenous recognized cattle breed of Shivamogga District of Karnataka state, India is known for its disease resistance to many of the infectious diseases. There are 25 LRR (Leucine Rich Repeats) identified in bovine (Bos indicus) TLR9. The amino acid sequence of LRR is deduced to nucleotide sequence in BLASTx bioinformatic online tools. LRR2 to LRR10 are involved in pathogen recognition and binding in human TLR9 which showed a higher degree of nucleotide variations with respect to disease resistance to various pathogens. Hence, primers were designed to amplify the flanking sequences of LRR2 to LRR10, to discover the nucleotide variations if any, in Malnad Gidda breed of Cattle which is associated with disease resistance. The DNA isolated from peripheral blood mononuclear cells of ten Malnad Gidda cattle. A desired and specific amplification product of 0.8 kb was obtained at an annealing temperature of 56.6ᵒC. All the PCR products were sequenced on both sides by gene-specific primers. The sequences were compared with TLR9 sequence of cross breed cattle obtained from NCBI data bank. The sequence analysis between Malnad Gidda and crossbreed cattle revealed no nucleotide variations in the region LRR2 to LRR9 which shows the conserved in pathogen binding domain (LRR) of TLR9.

Keywords: leucine rich repeats, Malnad Gidda, cross breed, TLR9

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Authors: Miguel García-Ferrús, Ana González, María A. Ferrús

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The most demanded organic foods worldwide are those that are consumed fresh, such as fruits and vegetables. However, there is a knowledge gap about some aspects of organic food microbiological quality and safety. Organic fruits and vegetables are more exposed to pathogenic microorganisms due to surface contact with natural fertilizers such as animal manure, wastes and vermicompost used during farming. It has been suggested that some emergent pathogens, such as Helicobacter pylori or Arcobacter spp., could reach humans through the consumption of raw or minimally processed vegetables. Therefore, the objective of this work was to study the contamination of organic fresh green leafy vegetables by Arcobacter spp. and Helicobacter pylori. For this purpose, a total of 24 vegetable samples, 13 lettuce and 11 spinach were acquired from 10 different ecological supermarkets and greengroceries and analyzed by culture and PCR. Arcobacter spp. was detected in 5 samples (20%) by PCR, 4 spinach and one lettuce. One spinach sample was found to be also positive by culture. For H. pylori, the H. pylori VacA gene-specific band was detected in 12 vegetable samples (50%), 10 lettuces and 2 spinach. Isolation in the selective medium did not yield any positive result, possibly because of low contamination levels together with the presence of the organism in its viable but non-culturable form. Results showed significant levels of H. pylori and Arcobacter contamination in organic vegetables that are generally consumed raw, which seems to confirm that these foods can act as transmission vehicles to humans.

Keywords: Arcobacter sp., Helicobacter pylori, Organic Vegetables, Polymerase Chain Reaction (PCR)

Procedia PDF Downloads 164

Authors: Rueyhung Weng, Chia-En Huang, Jung-Chi-Liao

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The transition zone (TZ) serves as a diffusion barrier to regulate the ins and outs of the proteins recruited to the primary cilia. TCTN2 is one of the TZ proteins and its mutation causes Joubert syndrome, a serious multi-organ disease. Despite its important medical relevance, the functions of TCTN2 remain elusive. Here we created a TCTN2 gene deleted retinal pigment epithelial cells (RPE1) using CRISPR/Cas9-based genome editing technique and used this knockout line to reveal roles of TCTN2. TCTN2 knockout RPE1 cells displayed a significantly reduced ciliogenesis or a shortened primary cilium length in the cilium-remaining population. Intraflagellar transport protein IFT88 aberrantly accumulated at the tip of TCTN2 deficient cells. Guanine nucleotide exchange factor Arl13B was mostly absent from the ciliary compartment, with a small population localizing at the ciliary tip. The deficient TZ was corroborated with the mislocalization of two other TZ proteins TMEM67 and MKS1. In addition, TCTN2 deficiency induced TZ impairment led to the suppression of Sonic hedgehog signaling in response to Smoothened (Smo) agonist. Together, depletion of TCTN2 destabilizes other TZ proteins and considerably alters the localization of key transport and signaling-associated proteins, including IFT88, Arl13B, and Smo.

Keywords: CRISPR/Cas9, primary cilia, Sonic hedgehog signaling, transition zone

Procedia PDF Downloads 351