Search results for: E. coli expression

Authors: Paula Buldres, Jorge Toledo

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Objectives: This study aimed to characterize potential probiotic strains isolated from canine breast milk for use in dogs with enteropathies. Methodology: Six canine breast milk strains, one canine colostrum strain, and one control porcine breast milk strain were characterized. According to its functional properties of resistance to acids, different concentrations of bile salts, and pancreatin, its presumptive properties of safety and inhibitory effect on pathogens, non-cytotoxic characteristics, and adhesion to the intestine. The immunomodulatory effect of formulations with better probiotic characterization in vitro and in vivo was also analyzed. Results: Two strains characterized as potential probiotics were obtained, which corresponded to the canine strains (TUCO-16 and TUCO-17), presenting resistance to acidic pH, bile salts, and pancreatin, as well as an inhibitory effect on pathogenic Escherichia coli, Salmonella sp., and Clostridium perfringens. Strains TUCO-16 and TUCO-17 induced a significant increase in the expression of TNF-α and IL-8 in canine macrophages, respectively. Expression analyses of pattern recognition receptors in DH82 cells suggest that TUCO-16 and TUCO-17 might increase the TLR2 expression marker, and porcine strain (TUCO-4) increases the NOD2 expression marker. Based on the count obtained and the encapsulation yield, the best formulations correspond to FOS-Inulin for the TUCO-17 and TUCO-4 strains; Maltodextrin-Inulin for TUCO-16. All the strains are non-cytotoxic. The strain that showed the highest adhesion to intestinal epithelial cells was TUCO-17 with the FOS-Inulin formulation. On the other hand, the probiotics decreased the expression of pro-inflammatory markers in vivo, both in the intestine and in the spleen of mice. Conclusion: The combination of these three strains under study (TUCO-16, TUCO-17, and TUCO-4) would cover the probiotic properties in formulation and immunomodulation of all the markers under study.

Keywords: probiotics, gastrointestinal infec, dog, probiotic formulation, immunomodulatory probiotics

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Authors: Fariha Akhter Chowdhury, Sabrina Mahboob, Anamika Saha, Afrin Jahan, Mohammad Nurul Islam

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Urinary tract infection (UTI) is frequently experienced by the female population where the prevalence increases with aging. Escherichia coli, one of the most common UTI causing organisms, retains glutathione defense mechanism that aids the organism to withstand the harsh physiological environment of urinary tract, host oxidative immune response and even to affect antibiotic-mediated cell death and the emergence of resistance. In this study, we aimed to investigate the glutathione reductase activity of uropathogenic E. coli (UPEC) by observing the reduced glutathione (GSH) level alteration under stressful condition. Urine samples of 58 patients with UTI were collected. Upon isolation and identification, 88% of the samples presented E. coli as UTI causing organism among which randomly selected isolates (n=9), obtained from urine samples of female patients, were considered for this study. E. coli isolates were grown under normal and stressful conditions where H₂O₂ was used as the stress-inducing agent. GSH level estimation of the isolates in both conditions was carried out based on the colorimetric measurement of 5,5'-dithio-bis (2-nitrobenzoic acid) (DTNB) and GSH reaction product using microplate reader assay. The GSH level of isolated E. coli sampled from adult patients decreased under stress compared to normal condition (p = 0.011). On the other hand, GSH production increased markedly in samples that were collected from elderly subjects (p = 0.024). A significant partial correlation between age and change of GSH level was found as well (p = 0.007). This study may help to reveal ways for better understanding of E. coli pathogenesis of UTI prevalence in elderly patients.

Keywords: Escherichia coli, glutathione reductase activity, oxidative stress, reduced glutathione (GSH), urinary tract infection (UTI)

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Authors: Fatemeh Moradian, Razieh Ghorbani, Poria Biparva

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Growing resistance of microorganisms to potent antibiotics has renewed a great interest towards investigating bactericidal properties of nanoparticles and their Nano composites as an alternative. The use of metal nanoparticles to combat bacterial infections is one of the most wide spread applications of nanotechnology in the field of antibacterial. Nanomaterials have unique properties compared to their bulk counterparts. In this report, we demonstrate the antimicrobial activity of zerovalent Iron(ZVI) and Ag(silver) nanoparticles against Gram-negative bacteria E.coli(DH5α) and Pseudomonas aeruginosa. At first ZVI and Ag nanoparticles were synthesized by chemical reduction method and using scanning electron microscopy (SEM) the nanoparticle size determined. Different concentrations of Ag and ZVI nanoparticles were added to bacteria on nutrient agar medium. Minimum inhibitory concentration (MIC) of Ag and Fe nanoparticles for P. aeruginosa were 5µM and 1µg as well as for E.coli were 6µM. and 10 µg, respectively. Among the two nanoparticles, ZVI showed that the greatest antimicrobial activity against E.coli and Ag nanoparticle on P.aeruginosa. Results suggested that the bactericidal effect of metal nanoparticles has been attributed to their small size as well as high surface to volume ratio and NPs could be used as an effective antibacterial material.

Keywords: bactericidal properties, MIC, nanoparticle, SEM

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Authors: Zohreh Rashmei

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Introduction: Plasma inactivation is one of the emerging technologies in biomedical field and has been applied to the inactivation of microorganisms in water. The inactivation effect has been attributed to the presence of active plasma species, i.e. OH, O, O3, H2O2, UV and electric fields, generated by the discharge of plasma. Material and Method: To evaluate germicidal effects of plasma, the electric spark discharge device was used. After the effect of the plasma samples were collected for culture medium agar plate count. In addition to biological experiments, the concentration of hydrogen peroxide was also measured. Results: The results showed that Plasma is able to inactivate a high concentration of E. coli. After a short period of plasma radiation on the surface of water, the amount log8 reduced the microbial load. Starting plasma radiation on the surface of the water, the measurements show of production and increasing the amount of hydrogen peroxide in water. So that at the end of the experiment, the concentration of hydrogen peroxide to about 100 mg / l increased. Conclusion: Increasing the concentration of hydrogen peroxide is directly related to the reduction of microbial load. The results of E. coli culture in media containing certain concentrations of H2O2 showed that E. coli can not to grow in a medium containing more than 2/5 mg/l of H2O2. Surely we can say that the main cause of killing bacteria is a molecule of H2O2.

Keywords: plasma, hydrogen peroxide, disinfection, E. coli

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Authors: Ali Mohagheghzadeh, Samad Vaez Badiegard, Bita Shomali

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Nowadays, with the increase in population, the need for protein sources is increasing. Different bacteria can cause food poisoning while most of the symptoms of food poisoning are similar to those of gastrointestinal infections. As a result, the diagnosis of bacteria and viruses causing food poisoning would not be possible without a stool culture. Cases of food poisoning are often accompanied by gastrointestinal disorders such as diarrhea, vomit, and gastrointestinal stomach cramps. Thus, providing enough food, taking into account health issues has always been a concern of authorities. Since E. coli bacterium is one of the important indicators of food hygiene and quality, producing food without being contaminated by this bacterium is desired in the food industry. This study aimed at assessing the E. coli contamination of poultry meat produced in slaughterhouses. Samples were taken from critical areas of slaughterhouses, namely the feather picking area, viscera and carcass evacuation area the area after cooling chillers. The results showed that 60% of contamination occurs in feather picking area. Among antiseptic and detergent materials, the highest reduction belongs to Epimax.

Keywords: slaughterhouse, E. coli, Epimax, contamination

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Authors: Lotem Sarid, Meirav Trebicz-Geffen, Serge Ankri

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Queuosine (Q) is a naturally occurring modified nucleoside that occurs in the first position of transfer RNA anticodons such as Asp, Asn, His, and Tyr. As eukaryotes lack pathways to synthesize queuine, the nucleobase of queuosine, they must obtain it from their diet or gut microbiota. Our previous work investigated the effects of queuine on the physiology of the eukaryotic parasite Entamoeba histolytica and defined the enzyme EhTGT responsible for its incorporation into tRNA. To our best knowledge, it is unknown how E. histolytica salvages Q from gut bacteria. We used N-acryloyl-3-aminophenylboronic acid (APB) PAGE analysis to demonstrate that E. histolytica trophozoites can salvage queuine from Q or E. coli K12 but not from the modified E. coli QueC strain, which cannot produce queuine. Next, we examined the role of EhDUF2419, a protein with homology to DNA glycosylase, as a queuine salvage enzyme in E. histolytica. When EhDUF2419 expression is silenced, it inhibits Q's conversion to queuine, resulting in a decrease in Q-tRNA levels. We also observed that Q protects control trophozoites from oxidative stress (OS), but not siEhDUF2419 trophozoites. Overall, our data reveal that EhDUF2419 is central for the salvaging of queuine from bacteria and for the resistance of the parasite to OS.

Keywords: entamoeba histolytica, epitranscriptomics, gut microbiota, queuine, queuosine, response to oxidative stress, tRNA modification.

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Authors: Karl Kingsley

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Many mechanisms are involved in the control of cellular differentiation and growth, which are often dysregulated in many cancers. Many distinct pathways are involved in these mechanisms of control, including deoxyribonuclease (DNA) methyltransferase and histone deacetylase (HDAC) activation that controls both genetic and epigenetic modifications and micro ribonucleic acid (RNA) expression. Less is known about the expression of DNA methyltransferase (DNMT) and HDAC in oral cancers and the effect on microRNA expression. The primary objective of this study was to evaluate the expression of DNMT and HDAC family members in oral cancer and the concomitant expression of cancer-associated microRNAs. Using commercially available oral cancers, including squamous cell carcinoma (SCC)-4, SCC-9, SCC-15, and SCC-25, RNA was extracted and screened for DNMT, HDAC, and microRNA expression using highly-specific primers and quantitative polymerase chain reaction (qPCR). These data revealed low or absent expression of DNMT-1, which is associated with cellular differentiation but increased expression of DNMT-3a and DNMT-3b in all SCC cell lines compared with normal non-cancerous cell controls. In addition, no expression of HDAC1 and HDAC2 expression was found among the normal, non-cancerous cells but was highly expressed in each of the SCC cell lines examined. Differential expression of oncogenic and cancer-associated microRNAs was also observed among the SCC cell lines, including miR-21, miR-133, miR-149, miR-155, miR-365, and miR-720. These findings also appeared to vary according to observed growth rates among these cells. These data may be the first to demonstrate the expression and association between HDAC and DNMT3 family members among oral cancers. In addition, the differential expression of these epigenetic modifiers may be associated with the expression of specific microRNAs in these cancers, which have not previously been observed to the best of the author's knowledge. In addition, some associations and relationships may exist between the expression of these biomarkers and the rates of growth and proliferation, which may suggest that these expression patterns might represent potentially useful biomarkers to determine tumor aggressiveness and other phenotypic behaviors among oral cancers.

Keywords: oral cancer, DNA methyltransferase, histone deacetylase, microRNA

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Authors: Alouache Souhila, Messai Yamina, Torres Carmen, Bakour Rabah

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Objective: It has often been reported an association between antibiotic resistance and virulence. However, resistance to quinolones seems to be an exception, it tends instead to be associated with an attenuation of virulence, particularly in clinical strains. The purpose of this study was to evaluate the potential virulence of 28 quinolone-resistant E. coli strains recovered from water at the inflow (n=16) and outflow (n=12) of an urban wastewater treatment plant (WWTP). Methods: E. coli isolates were selected on Tergitol-7 agar supplemented with 2µg/ml of ciprofloxacin, they were screened by PCR for 11 virulence genes related to Extraintestinal pathogenic E. coli (ExPEC): papC, papG, afa/draBC, sfa/foc, kpsMTII, iutA, iroN, hlyF, ompT, iss and traT. The phylogenetic groups were determined by PCR and clonal relationship was evaluated by ERIC-PCR. Results: Genotyping by ERIC-PCR showed 7 and 12 DNA profiles among strains of wastewater (inflow) and treated water (outflow), respectively. Strains were assigned to the following phylogenetic groups: B2 (n = 1, 3.5%), D (n = 3, 10.7%), B1 (n = 10, 35.7%.) and A (n = 14, 50%). A total of 8 virulence-associated genes were detected, traT (n=19, 67.8%), iroN (n= 16, 57 .1%), hlyF (n=15, 53 .5%), ompT (n=15, 53 .5%), iss (n=14, 50%), iutA (n=9, 32.1%) , sfa/foc (n=7, 25%) and kpsMTII (n=2, 7.1%). Combination of virulence factors allowed to define 16 virulence profiles. The pathotype APEC was observed in 17.8% (D=1, B1=4) and human ExPEC in 7% (B2=1, D=1) of strains. Conclusion: The study showed that quinolone-resistant E. coli strains isolated from wastewater and treated water in WWTP harbored virulence genes with the presence of APEC and human ExPEC strains.

Keywords: E. coli, quinolone-resistance, virulence, WWTP

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Authors: Eleni Poulou-Sidiropoulou, Charalampos N. Bompas, Martina Samiotaki, Alexios Vlamis-Gardikas

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The glutaredoxin (Grx) and thioredoxin (Trx) systems keep the intracellular environment reduced in almost all organisms. In Escherichia coli (E. coli), the Grx system relies on NADPH+ to reduce GSH reductase (GR), the latter reducing oxidized diglutathione to glutathione (GSH) which in turn reduces cytosolic Grxs, the electron donors for different intracellular substrates. In the Trx system, GR and GSH are replaced by Trx reductase (TrxR). Three of the Grxs of E. coli (Grx1, 2, 3) are reduced by GSH, while Grx4 is likely reduced by TrxR. Trx1 and Grx1 from E. coli may reduce ribonucleotide reductase Ia to ensure a constant supply of deoxyribonucleotides for the synthesis of DNA. The role of the other three Grxs is relatively unknown, especially for Grx2 that may amount up to 1 % of total cellular protein in the stationary phase of growth. The protein is known as a potent antioxidant, but no specific functions have been attributed to it. Herein, affinity chromatography of cellular extracts on immobilized Grx2, followed by MS analysis of the resulting eluates, was employed to identify protein ligands that could provide insights into the biological role of Grx2. Ionic, strong non-covalent, and covalent (disulfide) interactions with relevant proteins were detected. As a means of verification, the identified ligands were subjected to in silico docking with monothiol Grx2. In other experiments, protein extracts from E. coli cells lacking the gene for Grx2 (grxB) were compared to those of wild type. Taken together, the two approaches suggest that Grx2 is involved in protein synthesis, nucleotide metabolism, DNA damage repair, stress responses, and various metabolic processes. Grx2 appears as a versatile protein that may participate in a wide range of biological pathways beyond its known general antioxidant function.

Keywords: Escherichia coli, glutaredoxin 2, interactome, thiol-disulfide oxidoreductase

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Authors: Reshaa Alruwaili

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The primary aim of this study was to examine the hypothesis presented by Self Determination theory which suggests that autonomy impacts positively the expression of love. Other hypotheses were also examined which suggest that other variables explain the expression of love, including: dyadic adjustment (dyadic consensus, dyadic satisfaction and dyadic cohesion), couple satisfaction, age, gender, the length of marriage, number of children and attachment styles. The participants were Saudi couples, which provided the opportunity to consider the influence of Saudi culture on the expression of love. A questionnaire was employed to obtain measures of all the relevant variables, including a measure of expression of love that was built from 27 items, constituting verbal, physical and caring features, and a measure of autonomy based on three features: authorship, interest-taking and susceptibility. Data were collected from both members of 34 Saudi couples. Descriptive analysis of both expression of love and autonomy was conducted. Correlation and regression were used to assess the relationships between expression of love and autonomy and other variables. Results indicated that Saudi couples who most often express their love tend to be more than somewhat autonomous. Not much difference was found between husbands and wives in expressing love, although wives were slightly more autonomous than husbands. Expression of love was enhanced by the autonomy of the participants to a greater extent when dyadic satisfaction was controlled, since the latter was negatively correlated with autonomy and had no effect on the expression of love. Basic psychological needs, dyadic consensus and dismissive-avoidant attachment improve the expression of love, while it is decreased by the number of children.

Keywords: autonomy, determination theory, expression of love, dyadic adjustment

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Authors: Yakup Ulusu, Faruk Ozel, Numan Eczacioglu, Abdurrahman Ozen, Sabriye Acikgoz

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GFP discovered in the mid-1970s, has been used as a marker after replicated genetic study by scientists. In biotechnology, cell, molecular biology, the GFP gene is frequently used as a reporter of expression. In modified forms, it has been used to make biosensors. Many animals have been created that express GFP as an evidence that a gene can be expressed throughout a given organism. Proteins labeled with GFP identified locations are determined. And so, cell connections can be monitored, gene expression can be reported, protein-protein interactions can be observed and signals that create events can be detected. Additionally, monitoring GFP is noninvasive; it can be detected by under UV-light because of simply generating fluorescence. Moreover, GFP is a relatively small and inert molecule, that does not seem to treat any biological processes of interest. The synthesis of GFP has some steps like, to construct the plasmid system, transformation in E. coli, production and purification of protein. GFP carrying plasmid vector pBAD–GFPuv was digested using two different restriction endonuclease enzymes (NheI and Eco RI) and DNA fragment of GFP was gel purified before cloning. The GFP-encoding DNA fragment was ligated into pET28a plasmid using NheI and Eco RI restriction sites. The final plasmid was named pETGFP and DNA sequencing of this plasmid indicated that the hexa histidine-tagged GFP was correctly inserted. Histidine-tagged GFP was expressed in an Escherichia coli BL21 DE3 (pLysE) strain. The strain was transformed with pETGFP plasmid and grown on LuiraBertoni (LB) plates with kanamycin and chloramphenicol selection. E. coli cells were grown up to an optical density (OD 600) of 0.8 and induced by the addition of a final concentration of 1mM isopropyl-thiogalactopyranoside (IPTG) and then grown for additional 4 h. The amino-terminal hexa-histidine-tag facilitated purification of the GFP by using a His Bind affinity chromatography resin (Novagen). Purity of GFP protein was analyzed by a 12 % sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). The concentration of protein was determined by UV absorption at 280 nm (Varian Cary 50 Scan UV/VIS spectrophotometer). Synthesis of GFP-Polymer composite nanofibers was produced by using GFP solution (10mg/mL) and polymer precursor Polyvinylpyrrolidone, (PVP, Mw=1300000) as starting materials and template, respectively. For the fabrication of nanofibers with the different fiber diameter; a sol–gel solution comprising of 0.40, 0.60 and 0.80 g PVP (depending upon the desired fiber diameter) and 100 mg GFP in 10 mL water: ethanol (3:2) mixtures were prepared and then the solution was covered on collecting plate via electro spinning at 10 kV with a feed-rate of 0.25 mL h-1 using Spellman electro spinning system. Results show that GFP-based nano-fiber can be used plenty of biomedical applications such as bio-imaging, bio-mechanic, bio-material and tissue engineering.

Keywords: biomaterial, GFP, nano-fibers, protein expression

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Authors: Young Ah Kim, Hyunsoo Kim, Eun-Jeong Yoon, Young Hee Seo, Kyungwon Lee

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Extended-spectrum β-lactamase (ESBL)-producing Escherichia coli from food animals are considered as a reservoir for transmission of ESBL genes to human. The aim of this study is to assess the prevalence and molecular epidemiology of ESBL-producing E. coli colonization in pigs, farm workers, and farm environments to elucidate the transmission of multidrug-resistant clones from animal to human. Nineteen pig farms were enrolled across the country in Korea from August to December 2017. ESBL-producing E. coli isolates were detected in 190 pigs, 38 farm workers, and 112 sites of farm environments using ChromID ESBL (bioMerieux, Marcy l'Etoile, France), directly (stool or perirectal swab) or after enrichment (sewage). Antimicrobial susceptibility tests were done with disk diffusion methods and blaTEM, blaSHV, and blaCTX-M were detected with PCR and sequencing. The genomes of the four CTX-M-55-producing E. coli isolates from various sources in one farm were entirely sequenced to assess the relatedness of the strains. Whole genome sequencing (WGS) was performed with PacBio RS II system (Pacific Biosciences, Menlo Park, CA, USA). ESBL genotypes were 85 CTX-M-1 group (one CTX-M-3, 23 CTX-M-15, one CTX-M-28, 59 CTX-M-55, one CTX-M-69) and 60 CTX-M-9 group (41 CTX-M-14, one CTX-M-17, one CTX-M-27, 13 CTX-M-65, 4 CTX-M-102) in total 145 isolates. The rectal colonization rates were 53.2% (101/190) in pigs and 39.5% (15/38) in farm workers. In WGS, sequence types (STs) were determined as ST69 (E. coli PJFH115 isolate from a human carrier), ST457 (two E. coli isolates PJFE101 recovered from a fence and PJFA1104 from a pig) and ST5899 (E. coli PJFA173 isolate from the other pig). The four plasmids encoding CTX-M-55 (88,456 to 149, 674 base pair), whether it belonged to IncFIB or IncFIC-IncFIB type, shared IncF backbone furnishing the conjugal elements, suggesting of genes originated from same ancestor. In conclusion, the prevalence of ESBL-producing E. coli in swine farms was surprisingly high, and many of them shared common ESBL genotypes of clinical isolates such as CTX-M-14, 15, and 55 in Korea. It could spread by horizontal transfer between isolates from different reservoirs (human-animal-environment).

Keywords: Escherichia coli, extended-spectrum β-lactamase, prevalence, whole genome sequencing

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Authors: Debjani Ghosh, Goutam Chowdhury, Prosenjit Samanta, Asish Kumar Mukhopadhyay

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Acute diarrhoea caused by diarrheagenic Escherichia coli (DEC) is one of the major public health problem in developing countries, mainly in Asia and Africa. DEC consists of six pathogroups, but the majority of the cases were associated with the three pathogropus, enterotoxigenic E. coli (ETEC), enteroaggregative E. coli (EAEC), and enteropathogenic E. coli (EPEC). Hence, we studied the prevalence and antimicrobial resistance of these three major DEC pathogroups in hospitalized diarrheal patients in Kolkata, India, during 2012-2019 with a large sample size. 8,891 stool samples were processed, and 7.8% of them was identified as DEC infection screened by multiplex PCR, in which ETEC was most common (47.7%) followed by EAEC (38.4%) and EPEC (13.9%). Clinical patient history suggested that children <5 years of age were mostly affected with ETEC and EAEC, whereas people within >5-14 years of age were significantly associated with EPEC and ETEC infections. Antibiogram profile showed a high prevalence of multidrug resistant (MDR) isolates among DEC (56.9%), in which 9% were resistant to antibiotics of six different antimicrobial classes. Screening of the antibiotic resistance conferring genes in DEC showed the presence of blaCTX-M (30.2%) in highest number followed by blaTEM (27.5%), tetB (18%), sul2 (12.6%), strA (11.8%), aadA1 (9.8%), blaOXA-1 (9%), dfrA1 (1.6%) and blaSHV (1.2%) which indicates the existence of mobile genetic elements in those isolates. Therefore, the presence of MDR DEC strains in higher number alarms the public health authorities to take preventive measures before the upsurge of the DEC caused diarrhea cases in near future.

Keywords: diarrheagenic escherichia coli, ETEC, EAEC, EPEC

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Authors: Cheng-Yen Lu, Chin-Yuan Hsu

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Ambient temperature reduction (ATR) can extend the lifespan of an annual fish (Nothobranchius rachovii), but the underlying mechanism is unknown. In this study, the expression, concentration, and activity of cellular-degraded molecules were evaluated in the muscle of N. rachovii reared under high (30 °C), moderate (25 °C), and low (20 °C) ambient temperatures by biochemical techniques. The results showed that (i) the activity of the 20S proteasome, the expression of microtubule-associated protein 1 light chain 3-II (LC3-II), the expression of lysosome-associated membrane protein type 2a (Lamp 2a), and lysosome activity increased with ATR; (ii) the expression of the 70 kD heat shock cognate protein (Hsc 70) decreased with ATR; (iii) the expression of the 20S proteasome, the expression of lysosome-associated membrane protein type 1 (Lamp 1), the expression of molecular target of rapamycin (mTOR), the expression of phosphorylated mTOR (p-mTOR), and the p-mTOR/mTOR ratio did not change with ATR. These findings indicated that ATR activated the activity of proteasome, macroautophagy, and chaperone-mediated autophagy. Taken together these data reveal that ATR likely activates cellular degradation activity to extend the lifespan of N. rachovii.

Keywords: ambient temperature reduction, autophagy, degradation activity, lifespan, proteasome

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Authors: Mazoun Nasser Rashid Al Kharusi, Nada Al Siyabi

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Objectives: The main aim of this study is to Find the rate of susceptibility of ESBL E. coli causing UTI to oral antibiotics. Secondary objectives: Prevalence of ESBL E. coli from community urine samples, identify the best empirical oral antibiotics with the least resistance rate for UTI and identify alternative oral antibiotics for testing and utilization. Methods: This study is a retrospective descriptive study of the last three years in five major hospitals in Oman (Khowla Hospital, AN’Nahdha Hospital, Rustaq Hospital, Nizwa Hospital, and Ibri Hospital) equipped with a microbiologist. Inclusion criteria include all eligible outpatient urine culture isolates, excluding isolates from admitted patients with hospital-acquired urinary tract infections. Data was collected through the MOH database. The MOH hospitals are using different types of testing, automated methods like Vitek2 and manual methods. Vitek2 machine uses the principle of the fluorogenic method for organism identification and a turbidimetric method for susceptibility testing. The manual method is done by double disc diffusion for identifying ESBL and the disc diffusion method is for antibiotic susceptibility. All laboratories follow the clinical laboratory science institute (CLSI) guidelines. Analysis was done by SPSS statistical package. Results: Total urine cultures were (23048). E. coli grew in (11637) 49.6% of the urine, whereas (2199) 18.8% of those were confirmed as ESBL. As expected, the resistance rate to amoxicillin and cefuroxime is 100%. Moreover, the susceptibility of those ESBL-producing E. coli to nitrofurantoin, trimethoprim+sulfamethoxazole, ciprofloxacin and amoxicillin-clavulanate is progressing over the years; however, still low. ESBL E. coli was predominating in the female gender and those aged 66-74 years old throughout all the years. Other oral antibiotic options need to be explored and tested so that we add to the pool of oral antibiotics for ESBL E. coli causing UTI in the community. Conclusion: High rate of ESBL E. coli in urine from the community. The high resistance rates to oral antibiotics highlight the need for alternative treatment options for UTIs caused by these bacteria. Further research is needed to identify new and effective treatments for UTIs caused by ESBL-E. Coli.

Keywords: UTI, ESBL, oral antibiotics, E. coli, susceptibility

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Authors: Charalampos N. Bompas, Eleni Poulou-Sidiropoulou, Martina Samiotaki, Alexios Vlamis-Gardikas

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Ιn all living organisms, antioxidant defenses are orchestrated by the thioredoxin (Trx) and glutaredoxin (Grx) systems. The Trx system of Escherichia coli (E. coli) is comprised of Trx1 and Trx2, both reduced by thioredoxin reductase (TrxR). The Grx system consists of four Grxs (Grx1, Grx2, Grx3, and Grx4), all reduced by glutathione (GSH) except for Grx4, which is reduced by TrxR. Under normal conditions, the GSH reductase of the Grx system keeps GSH at its reduced state. NADPH+ provides the electrons for all reductions in the Trx and Grx systems. Although the role of the E. coli Trx system is widely known, the function of the Grx system reflects the main property of Grx1, which is the reduction of ribonucleotide reductase Ia (RRIa). E. coli Grx3 (encoded by grxC) may also reduce RRIa in vitro but with slow kinetics. The molecule may account for up to 0.4% of total soluble protein and has been the subject of extensive structural studies. Its biological function, however, remains unknown. Herein, affinity chromatography with monothiol Grx3 serving as bait was used to detect the interactions of Grx3 with other proteins. Different types of interactions were identified (covalent, weak, and strong non-covalent) that suggested novel functions for Grx3. In silico approaches were employed to validate selected interactions. In addition, total protein extracts from the null mutant for grxC and the wild-type strain were compared. The overall findings suggest that Grx3 is involved in various metabolic processes, protein synthesis, and stress responses, expanding the recognized functions of Grx3 beyond the possible reduction of RRIa.

Keywords: escherichia coli, glutaredoxin 3, interactome, thiol-disulfide oxidoreductase

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Authors: Sanaa Rashed Abdallah, Yasser F. Hassan

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The paper will introduce an approach where a rough sets, gene expression programming and rough neural networks are used cooperatively for learning and classification support. The Objective of gene expression programming rough neural networks (GEP-RNN) approach is to obtain new classified data with minimum error in training and testing process. Starting point of gene expression programming rough neural networks (GEP-RNN) approach is an information system and the output from this approach is a structure of rough neural networks which is including the weights and thresholds with minimum classification error.

Keywords: rough sets, gene expression programming, rough neural networks, classification

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Authors: B. M. Y. I. Basnayake, G. G. T. Nisansala, P. I. J. B. Wijewickrama, U. S. Weerathunga, K. W. M. Y. D. Gunasekara, N. K. Jayasekera, A. W. Kalupahana, R. S. Kalupahana, A. Silva- Fletcher, K. S. A. Kottawatta

Abstract:

Antimicrobial resistance (AMR) has attracted significant attention worldwide as an emerging threat to public health. Understanding the role of livestock and wildlife with the shared environment in the maintenance and transmission of AMR is of utmost importance due to its interactions with humans for combating the issue in one health approach. This study aims to investigate the extent of AMR distribution among wild animals, livestock, and environment cohabiting in a rural ecosystem in Sri Lanka: Hambegamuwa. One square km area at Hambegamuwa was mapped using GPS as the sampling area. The study was conducted for a period of five months from November 2020. Voided fecal samples were collected from 130 wild animals, 123 livestock: buffalo, cattle, chicken, and turkey, with 36 soil and 30 water samples associated with livestock and wildlife. From the samples, Escherichia coli (E. coli) was isolated, and their AMR profiles were investigated for 12 antimicrobials using the disk diffusion method following the CLSI standard. Seventy percent (91/130) of wild animals, 93% (115/123) of livestock, 89% (32/36) of soil, and 63% (19/30) of water samples were positive for E. coli. Maximum of two E. coli from each sample to a total of 467 were tested for the sensitivity of which 157, 208, 62, and 40 were from wild animals, livestock, soil, and water, respectively. The highest resistance in E. coli from livestock (13.9%) and wild animals (13.3%) was for ampicillin, followed by streptomycin. Apart from that, E. coli from livestock and wild animals revealed resistance mainly against tetracycline, cefotaxime, trimethoprim/ sulfamethoxazole, and nalidixic acid at levels less than 10%. Ten cefotaxime resistant E. coli were reported from wild animals, including four elephants, two land monitors, a pigeon, a spotted dove, and a monkey which was a significant finding. E. coli from soil samples reflected resistance primarily against ampicillin, streptomycin, and tetracycline at levels less than in livestock/wildlife. Two water samples had cefotaxime resistant E. coli as the only resistant isolates out of 30 water samples tested. Of the total E. coli isolates, 6.4% (30/467) was multi-drug resistant (MDR) which included 18, 9, and 3 isolates from livestock, wild animals, and soil, respectively. Among 18 livestock MDRs, the highest (13/ 18) was from poultry. Nine wild animal MDRs were from spotted dove, pigeon, land monitor, and elephant. Based on CLSI standard criteria, 60 E. coli isolates, of which 40, 16, and 4 from livestock, wild animal, and environment, respectively, were screened for Extended Spectrum β-Lactamase (ESBL) producers. Despite being a rural ecosystem, AMR and MDR are prevalent even at low levels. E. coli from livestock, wild animals, and the environment reflected a similar spectrum of AMR where ampicillin, streptomycin, tetracycline, and cefotaxime being the predominant antimicrobials of resistance. Wild animals may have acquired AMR via direct contact with livestock or via the environment, as antimicrobials are rarely used in wild animals. A source attribution study including the effects of the natural environment to study AMR can be proposed as this less contaminated rural ecosystem alarms the presence of AMR.

Keywords: AMR, Escherichia coli, livestock, wildlife

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Authors: Satoshi Furukawa, Satomu Morita, Lisa Wingenfeld, Katsuji Nishi, Masahito Hitosugi

Abstract:

We studied the expression of hypoxia-related antigens (e.g., cold-inducible antigens and apoptotic antigens) in the myocardium and the cerebellumthat were obtained from individuals after death by carbon monoxide or hypothermia. The immunohistochemistry results revealed that expression of cold-inducible RNA binding protein (CIRBP) and RNA-binding protein 3 (RBM3) may be associated with hpyothermic and the hypoxic conditions. The expression of CIRBP and RBM3 in the myocardium was different from their expression in the cerebellum, especially in the Purkinje cells. The results indicate that agonal duration influences antigen expression. In the hypothermic condition, the myocardium uses more ATP since the force of the excitation-contraction coupling of the myocardium increases by more than 400% when the experimental temperature is reduced from 35°C to 25°C. The results obtained in this study indicate that physicians should pay attention to the myocardium when cooling the patient’s body to protect the brain.

Keywords: carbon monoxide death, cerebellum, CIRBP, hypothermic death, myocardium, RBM3

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Authors: Malalage Mudara Peiris

Abstract:

Objectives of the study are: the biosynthesis of silver nanoparticles (AgNPs) using Escherichia coli, Acinetobacter baumannii and Staphylococcus aureus, characterization of silver nanoparticles and determination of antimicrobial activity against E. coli, P. aeruginosa, S. aureus, MRSA, and C. Albicans. Methods: E. coli (ATCC 25922), A. baumanii (clinical strain), S. aureus (clinical strain) cultured in nutrient broth medium were used for biosynthesis of AgNPs. Culture conditions (AgNO3 concentration, pH, incubation time and temperature) were optimized. Characterization of synthesized NPs was done by UV-Visible spectroscopy. The antimicrobial activity of the synthesized NPs was studied using the good diffusion assay against E. coli, S. aureus, MRSA (Methicillin-resistant Staphylococcus aureus), P. aeruginosa and C. Albicans. Results: All the selected bacteria produced silver nanoparticles at alkaline pH above 0.3 g/L AgNO3 concentration. The optimum reaction temperature was 60oC. According to the UV-Visible spectroscopy, the maximum absorbance was found to be around 420 - 430 nm indicating the presence of AgNPs. According to the good diffusion results, AgNPs produced by S. aureus resulted in the larger zone of inhibition (ZOI) against the selected pathogens, while AgNPs produced by E. coli showed comparatively smaller ZOI. In general, biosynthesized AgNPs were highly effective against gram-negative bacteria compared to gram-positive bacterial and fungal species. Conclusions: Green AgNPs produced by each bacterium show antimicrobial activity against the selected pathogens. AgNPs produced by S. aureus are the most effective NPs among tested AgNPs, while AgNPs produced by E. coli are the least effective. Further characterization of NPs is required to study the physical properties of silver NPs.

Keywords: green nanotechnology, silver nanoparticles, bacteria, antimicrobial activity

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Authors: Nadia Obaed, Lexi Frankel, Amalia Ardeljan, Denis Nigel, Anniki Witter, Omar Rashid

Abstract:

Breast cancer is the most common cancer among women, with a lifetime risk of one in eight of all women in the United States. Although breast cancer is prevalent throughout the world, the uneven distribution in incidence and mortality rates is shaped by the variation in population structure, environment, genetics and known lifestyle risk factors. Furthermore, the bacterial profile in healthy and cancerous breast tissue differs with a higher relative abundance of bacteria capable of causing DNA damage in breast cancer patients. Previous bacterial infections may change the composition of the microbiome and partially account for the environmental factors promoting breast cancer. One study found that higher amounts of Staphylococcus, Bacillus, and Enterobacteriaceae, of which Escherichia coli (E. coli) is a part, were present in breast tumor tissue. Based on E. coli’s ability to damage DNA, it is hypothesized that there is an increased risk of breast cancer associated with previous E. coli infection. Therefore, the purpose of this study was to evaluate the correlation between E. coli infection and the incidence of breast cancer. Holy Cross Health, Fort Lauderdale, provided access to the Health Insurance Portability and Accountability (HIPAA) compliant national database for the purpose of academic research. International Classification of Disease 9th and 10th Codes (ICD-9, ICD-10) was then used to conduct a retrospective analysis using data from January 2010 to December 2019. All breast cancer diagnoses and all patients infected versus not infected with E. coli that underwent typical E. coli treatment were investigated. The obtained data were matched for age, Charlson Comorbidity Score (CCI score), and antibiotic treatment. Standard statistical methods were applied to determine statistical significance and an odds ratio was used to estimate the relative risk. A total of 81286 patients were identified and analyzed from the initial query and then reduced to 31894 antibiotic-specific treated patients in both the infected and control group, respectively. The incidence of breast cancer was 2.51% and present in 2043 patients in the E. coli group compared to 5.996% and present in 4874 patients in the control group. The incidence of breast cancer was 3.84% and present in 1223 patients in the treated E. coli group compared to 6.38% and present in 2034 patients in the treated control group. The decreased incidence of breast cancer in the E. coli and treated E. coli groups was statistically significant with a p-value of 2.2x10-16 and 2.264x10-16, respectively. The odds ratio in the E. coli and treated E. coli groups was 0.784 and 0.787 with a 95% confidence interval, respectively (0.756-0.813; 0.743-0.833). The current study shows a statistically significant decrease in breast cancer incidence in association with previous Escherichia coli infection. Researching the relationship between single bacterial species is important as only up to 10% of breast cancer risk is attributable to genetics, while the contribution of environmental factors including previous infections potentially accounts for a majority of the preventable risk. Further evaluation is recommended to assess the potential and mechanism of E. coli in decreasing the risk of breast cancer.

Keywords: breast cancer, escherichia coli, incidence, infection, microbiome, risk

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Authors: Rohita Gupta, G. S. Brah, R. Verma, C. S. Mukhopadhayay

Abstract:

Although chicken strains show differences in susceptibility to a number of diseases, the underlying immunological basis is yet to be elucidated. In the present study, heterophils were subjected to LPS stimulation and total RNA extraction, further differential gene expression was studied in broiler, layer and indigenous Aseel strain by Real Time RT-PCR at different time periods before and after induction. The expression of the 14 AvBDs and chTLR 1, 2, 3, 4, 5, 7, 15 and 21 was detectable in heterophils. The expression level of most of the AvBDs significantly increased (P<0.05) 3 hours post in vitro lipopolysaccharide challenge. Higher expression level and stronger activation of most AvBDs, NFkB-1 and IRF-3 in heterophils was observed with the stimulation of LPS in layer compared to broiler, and in Aseel compared to both layer and broiler. This investigation will allow more refined interpretation of immuno-genetic basis of the variable disease resistance/susceptibility in divergent stock of chicken including indigenous breed. Moreover, this study will be helpful in formulation of strategy for isolation of antimicrobial peptides from heterophils.

Keywords: differential expression, heterophils, cytokines, defensin, TLR

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Authors: Rui Liu, Pengyu Cui, Nan Jiang

Abstract:

At present, there is much research on the application of centralized management and cross-integration application of basic geographic information. However, the idea of information integration and sharing between land, sea, and air navigation targets is not deeply applied into the research of navigation information service, especially in the information expression. Targeting at this problem, the paper carries out works about the expression pattern of navigation electronic map for ground vehicles under air and land united navigation mechanism. At first, with the support from multi-source information fusion of GIS vector data, RS data, GPS data, etc., an air and land united information expression pattern is designed aiming at specific navigation task of emergency rescue in the earthquake. And then, the characteristics and specifications of the united expression of air and land navigation information under the constraints of map load are summarized and transferred into expression rules in the rule bank. At last, the related navigation experiment is implemented to evaluate the effect of the expression pattern. The experiment selects evaluation factors of the navigation task accomplishment time and the navigation error rate as the main index, and make comparisons with the traditional single information expression pattern. To sum up, the research improved the theory of navigation electronic map and laid a certain foundation for the design and realization of united navigation system in the aspect of real-time navigation information delivery.

Keywords: navigation electronic map, united navigation, multi-element expression pattern, multi-source information fusion

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Authors: Ala Rakhshandeh, Zahra Ashkar, Solmaz Dehghani Dolatabadi, Hossein Bayat

Abstract:

The present study has been conducted to investigate the role of family emotional climate and emotional expression style in the academic well-being of students with military parents. Children, including 280 female students of Shahriar police officers, were selected by random sampling method, and they have been investigated through Alfred B. Hillburn's family emotional climate questionnaire (1964), King and Ammon's emotional expression questionnaire (1990), and Pitrinen, Sweeney, and Falto's academic well-being questionnaire (2014). The data were analyzed using statistical methods of correlation coefficient and stepwise multiple regression under the SPSS23 program. The results reveal that the variables of family emotional climate and emotional expression can explain 36.4% of the variance in academic well-being. This finding reveals that with an increase of standard deviation on the scores of family emotional climate and emotional expression, 0.513 and 0.155 standard deviations are added to the scores of academic well-being, respectively. The emotional climate of the family has a superior distinctive role in predicting the educational well-being of female students. Thus, the emotional climate of the family and the style of emotional expression play a meaningful role in the academic well-being of students with the military parent.

Keywords: emotional climate, family, emotional expression style, academic well-being

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Authors: Sana Mohammad Jafar, Hossaini Seyahi Zohreh

Abstract:

Infections and food intoxication caused by microbial contamination of food is of major issues in different countries, and diseases caused by the consumption of contaminated food included a large percentage of the country's health problems. Since traditional cheese for cultural reasons, good taste and smell in many parts of the area still has the important place in people's food basket, transmission of pathogenic bacteria could be at risk human health through the consumption of this food. In this study selected randomly 100 samples of 250 grams of traditional cheeses supplied in the city Behbahan market and adjacent to the ice was transferred to the laboratory and microbiological tests were performed immediately. According to the results, from 100 samples tested traditional cheese, 94 samples (94% of samples) were contaminated with coliforms, which of this number 75 samples (75% of samples) the contamination rate was higher than the limit (more than 100 cfu/g). Of the total samples, 36 samples (36% of samples) were contaminated with fecal coliform which of this number 30 samples (30% of samples) were contaminated with Escherichia.coli bacteria. Based on the results of agglutination test,no samples was found positive as pathogenic Escherichia.coli.

Keywords: determination, traditional cheese, Behbahan, Escherichia coli

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Authors: J. Shiue, I. H. Chen, S. W. Y. Chiu, Y. L. Wang

Abstract:

In this work, we present a nondestructive method to characterize bacteria in a TEM system. Unlike the conventional TEM specimen preparation method, which needs to thin the specimen in a destructive way, or spread the samples on a tiny millimeter sized carbon grid, our method is easy to operate without the need of sample pretreatment. With a specially designed transparent chip that allows the electron beam to pass through, and a custom made chip holder to fit into a standard TEM sample holder, the bacteria specimen can be easily prepared on the chip without any pretreatment, and then be observed under TEM. The centimeter-sized chip is covered with Au nanoparticles in the surface as the markers which allow the bacteria to be observed easily on the chip. We demonstrate the success of our method by using E. coli as an example, and show that high-resolution TEM images of E. coli can be obtained with the method presented. Some E. coli morphology characteristics imaged using this method are also presented.

Keywords: bacteria, chip, nanoparticles, TEM

Procedia PDF Downloads 289

Authors: Amit Chaudhary, B. S. Yadav, P. K. Maurya, A. M., S. Srivastava, S. Singh, A. Mani

Abstract:

Cold shock protein A (CspA) is major cold inducible protein present in Escherichia coli. The protein is involved in stabilizing secondary structure of RNA by working as chaperone during cold temperature. Two RNA binding motifs play key role in the stabilizing activity. This study aimed to investigate implications of low temperature on structure and RNA binding activity of E. coli CspA. Molecular dynamics simulations were performed to compare the stability of the protein at 37°C and 10 °C. The protein was mutated at RNA binding motifs and docked with RNA to assess the stability of both complexes. Results suggest that CspA as well as CspA-RNA complex is more stable at low temperature. It was also confirmed that RNP1 and RNP2 play key role in RNA binding.

Keywords: CspA, homology modelling, mutation, molecular dynamics simulation

Procedia PDF Downloads 347

Authors: Rohita Gupta, G. S. Brah, R. Verma, C. S. Mukhopadhayay

Abstract:

Although chicken strains show differences in susceptibility to a number of diseases, the underlying immunological basis is yet to be elucidated. In the present study, heterophils were subjected to LPS stimulation and total RNA extraction, further differential gene expression was studied in broiler, layer and indigenous Aseel strain by Real Time RT-PCR at different time periods before and after induction. The expression of the 14 AvBDs and chTLR 1, 2, 3, 4, 5, 7, 15 and 21 was detectable in heterophils. The expression level of most of the AvBDs significantly increased (P<0.05) 3 hours post in vitro lipopolysaccharide challenge. Higher expression level and stronger activation of most AvBDs, NFkB-1 and IRF-3 in heterophils was observed, with the stimulation of LPS in layer compared to broiler, and in Aseel compared to both layer and broiler. This investigation will allow more refined interpretation of immuno-genetic basis of the variable disease resistance/susceptibility in divergent stock of chicken including indigenous breed. Moreover this study will be helpful in formulation of strategy for isolation of antimicrobial peptides from heterophils.

Keywords: differential expression, heterophils, cytokines, defensin, TLR

Procedia PDF Downloads 584

Authors: Azhar Jabareen, Mahmoud Huleihel

Abstract:

BRCA-1 is a multifunctional tumor suppressor, whose expression is activated by the estrogen (E2)-liganded ERα receptor. The activated ERα is a transcriptional factor which activates various genes either by direct binding to the DNA at E2-responsive elements (EREs) and indirectly associated with a range of alternative non-ERE elements. Interference with BRCA-1 expression and/or functions leads to high risk of breast or/and ovarian cancer. Our lab investigated the involvement of Human T-cell leukemia Virus Type 1 (HTLV-1) in breast cancer, since HTLV-1 Tax was found to strongly inhibit BRCA-1 expression. In addition, long exposure of 12-O-tetradecanoylphorbol-13-acetate (TPA), which is one of the stress-inducing agents activated the HTLV-1 promoter. So here the involvement of TPA in breast cancer had been examined by testing the effect of TPA on BRCA-1 and ERE expression. The results showed that TPA activated both BRCA-1 and ERE expression. In the 12 hours TPA activated the tow promoters more than others time, and after 24 hours the level of the tow promoters was decreased. Tax inhibited BRCA-1 expression but did not succeed to inhibit the effect of TPA. Then the activation of the two promoters was not through ERα pathway because TPA had no effect on ERα binding to the two promoters of the BRCA-1 and ERE. Also, the activation was not via nuclear factor kappa B (NF-κB) pathway because when the inhibitory of NF-κB had been added to the TPA, it still activated the tow promoters. However, it seems that 53BP1 may be involved in TPA activation of these promoters because ectopic high expression of 53BP1 significantly reduced the TPA activity. In addition, in the presence of Bisindolylmaleimide-I (BI)- the inhibitor of Protein Kinase C (PKC)- there was no activation for the two promoters, so the PKC is agonized BRCA-1 and ERE activation.

Keywords: BRCA-1, ERE, HTLV-1, TPA

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Authors: Rufai Yakubu And Suleiman Kabiru

Abstract:

Triterpenoids are a diverse class of secondary metabolites with potential antimicrobial properties. In this study, the crude extracts from ethyl acetate was obtained with ultrasonic extraction method. Using a combined chromatographic separation method to isolate squalene (1) stigmasterol (2), stigmasta-5,22-diene-3-ol acetate (3), γ-sitosterol (4), lupeol (5), taraxasterol (6), and betulinic acid (7) from ethyl acetate extracts. Ethyl acetate crude extracts and isolated compounds were both screened for antimicrobial activity and minimum inhibitory concentration (MIC). For ethyl acetate crude extracts with concentrations of (1.5, 0.75, 0.35, & 0.168 mg/mL) indicated marginal antibacterial activity with a range of 17, 20 and 14 mm zone of inhibition for Staphylococcus aureus, Escherichia coli and Candida albicans and lower minimum inhibitory concentrations ranges from 18.75 µg/ml to 150 µg/mL. Butulinic acid showed the highest activity against E. coli and C. albicans at 15 mm and 15 mm followed by Lupeol against S. aureus, E. coli and C. albicans at 13, 12, 12 mm. Moreso, no antimicrobial activity for both S. aureus and C. albicans with squalene except for E. coli which showed activity at 11 mm with 300 µg/mL (MIC). Thus, abundant triterpenoids in Deinbollia pinnata will be another centered area for antimicrobial drug discovery.

Keywords: triterpenoid, antimicrobial potentials, deinbollia pinnata, aerial parts

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