The ecosystems of certain abandoned mines contain arsenic-resistant bacteria capable of performing detoxification when an ars gene is present in the bacterial genome. The ars gene has already been isolated from Pseudomonas putida and identified as a member of the membrane transport regulatory deoxyribonucleic acid family. The arsenite-oxidizing bacterial strains isolated in the present study were found to grow in the presence of 66.7 mM sodium arsenate($V;\;Na_2HAsO_4{\cdot}7H_2O$), yet experienced inhibited growth when the sodium arsenite($III;\;NaAsO_2$) concentration was higher than 26 mM. Batch experiment results showed that Pseudomonas putida strain OS-5 completely oxidized 1 mM of As(III) to As(V) within 35 h. An arsB gene encoding a membrane transport regulatory protein was observed in arsenite-oxidizing Pseudomonas putida strain OS-5, whereas arsB, arsH, and arrA were detected in strain OS-19, arsD and arsB were isolated from strain RW-18, and arsR, arsD, and arsB were found in E. coli strain OS-80. The leader gene of arsR, -arsD, was observed in a weak acid position. Thus, for bacteria exposed to weak acidity, the ars system may cause changes to the ecosystems of As-contaminated mines. Accordingly, the present results suggest that arsR, arsD, arsAB, arsA, arsB, arsC, arsH, arrA, arrB, aoxA, aoxB, aoxC, aoxD, aroA, and aroB may be useful for arsenite-oxidizing bacteria in abandoned arsenic-contaminated mines.
Naturally immobilized tannase (tannin acyl hydrolase, E.C. 3.1.1.20) has many advantages, as it avoids the expensive and laborious operation of isolation, purification, and immobilization, plus it is highly stable in adverse pH and temperature. However, in the case of cell-associated enzymes, since the enzyme is associated with the biomass, separation of the pure biomass is necessary. However, tannic acid, a known inducer of tannase, forms insoluble complexes with media proteins, making it difficult to separate pure biomass. Therefore, this study optimizes the production of cell-associated tannase using a "protein-tannin complex" free media. An exploratory study was first conducted in shake-flasks to select the inducer, carbon source, and nitrogen sources. As a result it was found that gallic acid induces tannase synthesis, a tryptose broth gives higher biomass, and lactose supplementation is beneficial. The medium was then optimized using response surface methodology based on the full factorial central composite design in a 3 l bioreactor. A $2^3$ factorial design augmented by 7 axial points (${\alpha}$ = 1.682) and 2 replicates at the center point was implemented in 17 experiments. A mathematical model was also developed to show the effect of each medium component and their interactions on the production of cell-associated tannase. The validity of the proposed model was verified, and the optimized medium was shown to produce maximum cell-associated tannase activity of 9.65 U/l, which is 93.8% higher than the activity in the basal medium, after 12 h at pH 5.0, $30^{\circ}C$. The optimum medium consists of 38 g/l lactose, 50 g/l tryptose, and 2.8 g/l gallic acid.
This experiment was carried out to develope the production of specific yolk antibody from laying hens immunized with antigens from Salmonella enteritidis. Antigenic protein isolated from the flagella of Salmonella enteritidis, determined by SDS-PAGE, was pure and has a molecular mass of approximately 54.6 kDa. It was observed that the antibody titers both in egg yolk and serum were performed at 2 weeks after immunization with flagella antigen to the laying hen. And the level was increased gradually to 6 weeks after immunization. At the time of 6 weeks, the antibody titer of yolk showed higher than that of serum. According to the results of specificity test(ELISA), the yolk antibody did not react with different bacterial strains(S. choleraesuis, ETEC Kl2:K99, K88,987P), but reacted only with S. enteritidis strain. The contents of immunoglobulin(IgY) in an egg yolk was 106mg approximately. By the isolation procedure of IgY from the egg yolk, 88.3 percent of IgY content was recovered in this study.
Antimicrobial activity of Plantain(Plantago asiatica L.) was investigated. Methanol extract of dried Plantain was fractionated to hexane, chloroform, ethylacetate, butanol and aqueous fraction. Ethylacetate fraction among these fractions showed the highest inhibitory effect on the microorganisms such as B. subtilis, E. coli, and V. parahaemolyticus at 500 $\mu\textrm{g}$/disc. Ethylacetate fraction was further fractionated into 8 fractions by silica gel column and thin layer chromatography(TLC). The results showed that ethylacetate fractions No. 2 and 3 had the highest anti-microbial activity. They were mixed again, re-separated, and seven fractions were obtained. Among them, No.4 and 6 fraction had the highest inhibitory effect on the microorganisms, which were then separated into four fractions. In the 3rd fractionation, No.4 fraction was identified as hexadecanoic acid by HPLC, $^1$H-NMR and GC-MS.
Jeong, Yeon Gyeom;Park, Bo Mi;Kim, Min Ju;Park, Jin Il;Jung, Yeoun Joong;Oh, Eun Gyoung
Korean Journal of Fisheries and Aquatic Sciences
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v.54
no.4
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pp.388-396
/
2021
Antimicrobial resistance patterns of Escherichia coli were investigated. Strains were isolated from 310 shellfish, 36 crustaceans, and 12 fish collected off the West Coast of Korea from April 2019 to October 2020. Two hundred and ninety-five E. coli strains were isolated from shellfish, 100 from crustaceans, and 54 from fish. Strains isolated from shellfish showed the highest resistance to ampicillin (27.5%), whereas those from crustaceans were resistant to sulfisoxazole (30.0%) and those from fish were resistant to ampicillin (59.3%) and sulfisoxazole (59.3%). Ceftazidime resistance was observed in strains isolated from short neck and hard clams, whereas gentamicin resistance was observed in strains from fish. Multi-drug resistance was observed in 56 strains (48.7%) isolated from shellfish, 11 (28.2%) from crustaceans and 27 (73.0%) from fish. Depending on the source of isolation, the strains showed specific antimicrobial resistance tendency. Strains isolated from shellfish showed 12 different multi-drug resistance patterns, whereas those from crustaceans showed high resistance (59%) to a single antimicrobial agent and those from fish showed a broad trend of multi-drug resistance to more than eight antimicrobials.
Unsunnidhal, Lalu;Wasito, Raden;Setyawan, Erif Maha Nugraha;Warsani, Ziana;Kusumawati, Asmarani
Journal of Veterinary Science
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v.22
no.6
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pp.76.1-76.15
/
2021
Background: The development of a vaccine for Jembrana disease is needed to prevent losses in Indonesia's Bali cattle industry. A DNA vaccine model (pEGFP-C1-tat) that requires a functional delivery system will be developed. Polylactic-co-glycolic acid (PLGA) may have potential as a delivery system for the vaccine model. Objectives: This study aims to evaluate the in vitro potential of PLGA as a delivery system for pEGFP-C1-tat. Methods: Consensus and codon optimization for the tat gene was completed using a bioinformatic method, and the product was inserted into a pEGFP-C1 vector. Cloning of the pEGFP-C1-tat was successfully performed, and polymerase chain reaction (PCR) and restriction analysis confirmed DNA isolation. PLGA-pEGFP-C1-tat solutions were prepared for encapsulated formulation testing, physicochemical characterization, stability testing with DNase I, and cytotoxicity testing. The PLGA-pEGFP-C1-tat solutions were transfected in HeLa cells, and gene expression was observed by fluorescent microscopy and real-time PCR. Results: The successful acquisition of transformant bacteria was confirmed by PCR. The PLGA:DNA:polyvinyl alcohol ratio formulation with optimal encapsulation was 4%:0.5%:2%, physicochemical characterization of PLGA revealed a polydispersity index value of 0.246, a particle size of 925 nm, and a zeta potential value of -2.31 mV. PLGA succeeded in protecting pEGFP-C1-tat from enzymatic degradation, and the percentage viability from the cytotoxicity test of PLGA-pEGFP-C1-tat was 98.03%. The PLGA-pEGFP-C1-tat demonstrated luminescence of the EGFP-tat fusion protein and mRNA transcription was detected. Conclusions: PLGA has good potential as a delivery system for pEGFP-C1-tat.
This case report describes a technique in which endodontic treatment and permanent indirect restoration were completed in the same clinical appointment with the aid of a computer-aided design/computer-aided manufacturing (CAD/CAM) system. Two patients were diagnosed with irreversible pulpitis of the mandibular first molar. After access preparation, root canals were located, irrigation was performed until bleeding ceased, and the coronal tooth structure was prepared for indirect restoration. Then, utilizing an interim 3-mm build-up of the endodontic access cavity, a hemi-arch digital scan was performed with an intraoral scanner. Subsequent to digital scanning, restoration design was performed simultaneously with the endodontic procedure. The root canals were shaped using the Race system under irrigation with 2.5% sodium hypochlorite followed by root canal filling. The pulp chamber was subsequently filled with a 3-mm-thick composite resin restoration mimicking the interim build-up previously utilized to facilitate block milling in the CAD/CAM system. Clinical try-in of the permanent onlay restoration was followed by acid etching, application of a 5th generation adhesive, and cementation of the indirect restoration. Once the restoration was cemented, rubber dam isolation was removed, followed by occlusal adjustment and polishing. After 2 years of follow-up, the restorations were esthetically and functionally satisfactory, without complications.
The Great Sebkha of Oran is a closed depression located in northwestern of Algeria. Despite the ranking of this sebkha among the wetlands of global importance by Ramsar Convention in 2002, no studies on the fungal community in this area have been carried out. In our study, samples were collected from two different regions. The first region is characterized by halophilic vegetation and cereal crops and the second by a total absence of vegetation. The isolated strains were identified morphologically then by molecular analysis. The biotechnological interest of the strains was evaluated by testing their ability to grow at different concentration of NaCl and to produce extracellular enzymes (i.e., lipase, amylase, protease, and cellulase) on solid medium. The results showed that the soil of sebkha is alkaline, with the exception of the soil of cereal crops that is neutral, and extremely saline. In this work, the species Gymnoascus halophilus, Trichoderma gamsii, the two phytopathogenic fungi, Fusarium brachygibbosum and Penicillium allii, and the teleomorphic form of P. longicatenatum observed for the first time in this species, were isolated for the first time in Algeria. The halotolerance test revealed that the majority of the isolated are halotolerant. Wallemia sp. and two strains of G. halophilus are the only obligate halophilic strains. All strains are capable to secrete at least one of the four tested enzymes. The most interesting species presenting the highest enzymatic index were Aspergillus sp. strain A4, Chaetomium sp. strain H1, P. vinaceum, G. halophilus, Wallemia sp. and Ustilago cynodontis.
Kim, Ji-Yul;Woo, E-Eum;Ha, Lee Su;Ki, Dae-Won;Lee, In-Kyoung;Yun, Bong-Sik
Mycobiology
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v.47
no.2
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pp.256-260
/
2019
Neuraminidase (NA) cleaves the glycosidic bond linkages of sialic acids to release the mature virions from infected cells and has been an attractive therapeutic target for anti-influenza agents. In our ongoing investigation of NA inhibitors in mushroom extracts, we found that the extract the fruiting body of Glaziella splendens potently inhibited neuraminidase. The fruiting bodies of G. splendens were extracted and partitioned successively with hexane, ethyl acetate, and butanol. The ethyl acetate soluble-layer was subjected to silica gel and Sephadex LH-20 column chromatographies, and MPLC to obtain five compounds (1-5). Their structures were determined by spectroscopic methods. NA inhibitory activity of these compounds was evaluated using NAs from recombinant rvH1N1, H3N2, and H5N1 influenza A viruses. One compound (1) was elucidated as a new azaphilone derivative, and four compounds (2-5) were identified as entonaemin A, comazaphilone D, rubiginosin A, and entonaemin B, respectively. Compounds 3 and 4 showed considerable inhibitory activity against three types of neuraminidases with the $IC_{50}$ values of 30.9, 41.8, and $35.7{\mu}M$ for 3 and 46.5, 50.4, and $29.9{\mu}M$ for 4, respectively. This study reveals that the fruiting bodies of G. splendens possess azaphilone derivatives with the NA inhibitory activity. This is the first report on the isolation of neuraminidase inhibitors from the fruiting bodies of G. splendens.
For the last decades, criticism on Frankenstein has tried to make a link between Victor's Creature and Rousseaurean "man in a state of nature." Like the Rousseaurean savage in a state of animal, the monster has only basic instincts least needed for his survival, i.e. self-preservation, but turns into a civilized man after learning language. Most critics argue that, despite the monster's acquisition of language, his failure in entry into a cultural and linguistic community is the outcome of a lack of sympathy for him by others, which displays the stark existence of epistemological barriers between them. That is to say, the monster imagines his being the same as others in the pre-linguistic stage but, in the linguistic stage, he realizes that he is different from others. Interpreting the Rousseaurean idea of language, which appears in his writings, as much more focused on emotion than many critics think, I read the dispute between Victor and his Creature as a variation of parent-offspring conflict. Shelley criticizes Rousseau's parental negligence in putting his children into a foundling hospital and leaving them dying there. The monster's revenge on uncaring Victor parallels the likely retaliation Rousseau's displaced children would perform against Rousseau, which Shelley imaginatively reproduces in her novel. The conflict between the monster and Victor is due to a disrupted attachment between parent and child in terms of Darwinian developmental psychology. Affective asynchrony between parent and child, which refers to a state of lack of mutual favorable feelings, accounts for numerous dysfunctional families. This paper shifts a focus from a semiotics-oriented perspective on the monster's social isolation to a Darwinian perspective, drawing attention to emotional problems transpiring in familial interactions. In doing so, it finds that language is a means of communicating one's internal emotions to others along with other means such as facial expressions and body movements. It also demonstrates that how to promote emotional well-being in either familial or social relationships entirely depends on the way in which one employs language that can entail either pleasure or anger on hearers' part.
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