scholarly journals Creation of recombinant Chlamydomonas reinhardtii strains expressing codon optimized vp28 gene from white spot symdrome virus

2018 ◽  
Vol 40 (1) ◽  
pp. 92-99
Author(s):  
Nguyen Minh Huong ◽  
Ha Thi Thu ◽  
Nguyen Thi Hoa ◽  
Dinh Duy Khang ◽  
Dang Diem Hong ◽  
...  
Keyword(s):  
Chlamydomonas Reinhardtii ◽  
White Spot ◽  
Future Research ◽  
Natural Food ◽  
Rt Pcr ◽  
Edible Vaccines ◽  
Colony Pcr ◽  
E Coli ◽  
Pharmaceutical Proteins ◽  
Vp28 Gene

White spot syndrome virus (WSSV) is the leading cause of shrimp mortality in farms all over the world. In Vietnam, for the last five to ten years, WSSV has always been among the top causes of diseases and loss in our shrimp aquaculture. VP28 and VP26 are two capsid proteins of WSSV that commonly used as biomarkers for diagnosis and target antigens for vaccine against WSSV. Recombinant VP28 (rVP28) has been studied and expressed in various expression systems including E. coli, yeast and baculovirus. rVP28 expressed in these systems showed effective protection against WSSV in shrimps, though there remains limitation on expression efficiency and safety. Recently, green microalgae Chlamydomonas reinhardtii has been widely used to express pharmaceutical proteins and edible vaccines for aquaculture thanks to its advantages as a safe and efficient host. C. reinhardtii is also used as nutrious natural food for shrimps due to its benefits towards shrimp health and growth. In this study, the codons of vp28 gene was adapted and chemically synthesized, and transformed into the nucleus genome of C. reinhardtii using electroporation. The presence of a codon optimized vp28 gene in C. reinhardtii genome was confirmed by colony PCR and sequencing; and its expression level was examined by RT-PCR. These results proved our success in creating transgenic C. reinhardtiii expressing rVP28 and set the foundation for our future research on edible vaccine against WSSV for shrimp.

An Updated Review on COVID-19

2020 ◽  
Vol 20 ◽  
Author(s):  
Mahmoud Ahmed Ebada ◽  
Ahmed Wadaa Allah ◽  
Eshak Bahbah ◽  
Ahmed Negida
Keyword(s):  
Screening Tool ◽  
Distress Syndrome ◽  
Reproduction Number ◽  
Case Fatality ◽  
Current Data ◽  
Future Research ◽  
Rt Pcr ◽  
Viral Structure ◽  
Discharge Criteria ◽  
Serology Testing

: Coronavirus Disease (COVID-19) pandemic has affected more than seven million individuals in 213 countries worldwide with a basic reproduction number ranging from 1.5 to 3.5 and an estimated case fatality rate ranging from 2% to 7%. A substantial proportion of COVID-19 patients are asymptomatic; however, symptomatic cases might present with fever, cough, and dyspnoea or severe symptoms up to acute respiratory distress syndrome. Currently, RNA RT-PCR is the screening tool, while bilateral chest CT is the confirmatory clinical diagnostic test. Several drugs have been repurposed to treat COVID-19, including chloroquine or hydroxychloroquine with or without azithromycin, lopinavir/ritonavir combination, remdesivir, favipiravir, tocilizumab, and EIDD-1931. Recently, Remdesivir gained FDA emergency approval based on promising early findings from the interim analysis of 1063 patients. The recently developed serology testing for SARSCoV-2 antibodies opened the door to evaluate the actual burden of the disease and to determine the rate of the population who have been previously infected (or developed immunity). This review article summarizes current data on the COVID-19 pandemic starting from the early outbreak, viral structure and origin, pathogenesis, diagnosis, treatment, discharge criteria, and future research.

scholarly journals Microarray and RT-PCR screening for white spot syndrome virus immediate-early genes in cycloheximide-treated shrimp

Virology ◽  
2005 ◽  
Vol 334 (2) ◽  
pp. 327-341 ◽  
Author(s):  
Wang-Jing Liu ◽  
Yun-Shiang Chang ◽  
Chung-Hsiung Wang ◽  
Guang-Hsiung Kou ◽  
Chu-Fang Lo
Keyword(s):  
White Spot Syndrome Virus ◽  
Immediate Early Genes ◽  
Immediate Early ◽  
White Spot ◽  
Rt Pcr ◽  
Pcr Screening ◽  
Early Genes ◽  
White Spot Syndrome

scholarly journals A New Clone Sweeps Clean: the Enigmatic Emergence of Escherichia coli Sequence Type 131

2014 ◽  
Vol 58 (9) ◽  
pp. 4997-5004 ◽  
Author(s):  
Ritu Banerjee ◽  
James R. Johnson
Keyword(s):  
Escherichia Coli ◽  
Sequence Type ◽  
Rapid Expansion ◽  
Future Research ◽  
Content Type ◽  
Extended Spectrum Beta Lactamase ◽  
E Coli ◽  
Phylogenetic Studies ◽  
Ecological Success

ABSTRACTEscherichia colisequence type 131 (ST131) is an extensively antimicrobial-resistantE. coliclonal group that has spread explosively throughout the world. Recent molecular epidemiologic and whole-genome phylogenetic studies have elucidated the fine clonal structure of ST131, which comprises multiple ST131 subclones with distinctive resistance profiles, including the (nested) H30, H30-R, and H30-Rx subclones. The most prevalent ST131 subclone, H30, arose from a single common fluoroquinolone (FQ)-susceptible ancestor containing allele 30 offimH(type 1 fimbrial adhesin gene). An early H30 subclone member acquired FQ resistance and launched the rapid expansion of the resulting FQ-resistant subclone, H30-R. Subsequently, a member of H30-R acquired the CTX-M-15 extended-spectrum beta-lactamase and launched the rapid expansion of the CTX-M-15-containing subclone within H30-R, H30-Rx. Clonal expansion clearly is now the dominant mechanism for the rising prevalence of both FQ resistance and CTX-M-15 production in ST131 and inE. coligenerally. Reasons for the successful dissemination and expansion of the key ST131 subclones remain undefined but may include increased transmissibility, greater ability to colonize and/or persist in the intestine or urinary tract, enhanced virulence, and more-extensive antimicrobial resistance compared to otherE. coli. Here we discuss the epidemiology and molecular phylogeny of ST131 and its key subclones, possible mechanisms for their ecological success, implications of their widespread dissemination, and future research needs.

scholarly journals Експериментальне зараження поросят вірусом епідемічної діареї свиней

2017 ◽  
Vol 19 (77) ◽  
pp. 208-213
Author(s):  
D. Masiuk ◽  
A. Sosnitskiy ◽  
A. Kokarev ◽  
S. Koliada
Keyword(s):  
Real Time ◽  
The Body ◽  
Control Group ◽  
Rt Pcr ◽  
Internal Organs ◽  
Real Time System ◽  
E Coli ◽  
Neonatal Piglets ◽  
And Control ◽  
Experimental Group

There were infected neonatal piglets in the first days of their lives PED virus suspension derived from pigs previously PED patients. Diagnosis for PED in piglets donor virus PED was inserted complex method for clinical and epizootic performance and confirmed the identification PEDV by PCR-RT using the test system «EZ-RED/TGE/PDCoV MPX 1.0 Real time RT-PCR» company Tetracore (USA) Thermocyclers CFX 96 Real-Time System company BIO RAD (USA). Homogenate small intestine of pigs PEDV donor, prepared in a blender for PCR in a thick band of 18 animal carcasses, frozen at -18 °C without cryopreservation and kept 359 days. Before infecting pigs and strip defrost by RT-PCR identified the concentration of the virus genome equivalents (GE) without establishing viable virions quantitative pathogen. For Sample 20 selected analog neonatal piglets, divided them into 3 experimental groups (group 1 – 5 piglets, group 2 – 5 piglets and group 3 – 7 piglets) and one control (3 piglets). Research pigs infected per os virus-containing suspension with a concentration PEDV 1.03×106 GE/cm3. The dose for infection first group was 6 cm3 (6.18×106 GE/cm3), for the second – 5 cm3 (5,15 × 106 GE/cm3), for the third – 4 cm3 (4.12 GE×106/cm3) homogenate. The fourth group – control (not infected). All the pigs were in identical conditions that fully meet the physiological needs of the body. Of the 17 infected pigs only 2 was infected PEDV. PED was confirmed by laboratory methods. In bacteriological examination of internal organs of pigs that came out of a research experiment and control group were diagnosed colibacteriosis. In the control group was isolated from heart and intestinal non-pathogenic for white mice E. coli. From pigs 1 and 2 research groups has been allocated to white mice nonpathogenic E. coli, is set colibacteriosis; 2 experimental group found in one pig hemolytic E. coli; 3 experimental group from the internal organs of pigs in conjunction with non-pathogenic for mice intestinal former cane isolated Klesiella spp., is diagnosed with mixed infection (E. coli, Klesiella spp.). From the intestine of experimental and control pigs do not identified beneficial microflora – aerococcus, lactobacteria, bifidobacteria and cultured putrefactive anaerobic spore facultative and non spore microflora.

scholarly journals ANTIBACTERIAL AND ANTIOXIDANT ACTIVITIES OF COCOA POD THAT ASSOCIATED IN MALTODEXTRIN IN VARIOUS CONCENTRATION

2018 ◽  
Vol 5 (2) ◽  
pp. 123
Author(s):  
Asriani Hasanuddin ◽  
Chairil Anwar ◽  
Marhawati Mappatoba ◽  
Hafsah Hafsah
Keyword(s):  
Antimicrobial Activity ◽  
Antioxidant Activity ◽  
Antioxidant Activities ◽  
Diffusion Method ◽  
Natural Food ◽  
Skin Extract ◽  
E Coli ◽  
Dpph Method ◽  
Antioxidant And Antimicrobial Activity ◽  
Peel Extract

Cocoa pod extract ((Theobroma cacao L.) has antioxidant and antimicrobial activity that has the potential as a natural food preservative. However, in its use the cocoa fruit skin extract has a disadvantage because the short shelf time and its application to food are limited, efforts are needed to prevent damage and extend shelf life, one of the efforts that can be done is by encapsulating the extract.This study aims to determine the antibacterial activity and antioxidant encapsulation of cocoa peel extract, this study begins with the extraction of cocoa pods with ethanol solvent by comparing cocoa pods : solvent 1: 4 The skin of cacao cocoa fruit used is yellow harvested cocoa fruit, then chopped and dried to form flour.The sample is extracted by maceration with ethanol solvent Antioxidant test is done by DPPH method, while antibacterial test is carried out by the well diffusion method. This study used a completely randomized design method (CRD) with 5 treatments using a maltodextrin concentration of 20% (M1); 30% (M2); 40% (M3); 50% (M4) and 60% (M5). The results showed that the treatment gave the highest yield in the treatment of 60% maltodextrin concentration (M5), while the highest antioxidant activity was obtained in the treatment of 20% maltodextrin (M1) with IC50 75.98 µg / mL and the treatment with the lowest antioxidant activity was obtained at treatment of 60% maltodextrin concentration (M5) with IC50 value 114.89 µg / mL. While for the antimicrobial activity also obtained with the same results, namely treatment of 20% (M1) obtained a higher inhibition diameter compared to treatment at 30%; 40%; 50% and 60% for all types of bacteria. The inhibition diameter in the treatment of the concentration of maltodextrin 20% (M1) for E. coli bacteria is between 4.12 mm - 10.95 mm, Salmonella sp is 2.85 mm - 8 , 25 mm and for Staphylococcusaureus of 5.15 mm - 13.90 mm and the lowest inhibition diameter was obtained in the treatment of 60% maltodextrin concentration (M5) for E. coli bacteria of between 2.0 mm - 4.79 mm, Salmonella sp of 1.15 mm - 4.35 mm and for Staphylococcusaureusat 2.76 mm - 5.17 mm.This study concluded that the encapsulation of cocoa peel extract using 20% maltodextrin had the highest antioxidant and antimicrobial activity when compared with other treatments namely 30% concentration; 40%; 50% and 60% but for the treatment of 20% and 30% there is no difference. Ethanol extract of cocoa pods can be made in the form of encapsulates which are very likely to be used as natural preservatives.

scholarly journals Response to BNT162b2 mRNA COVID-19 vaccine among healthcare workers in Italy: a 3-month follow-up

2021 ◽  
Author(s):  
Domenico Ponticelli ◽  
Fabiana Madotto ◽  
Sara Conti ◽  
Ippazio C. Antonazzo ◽  
Andrea Vitale ◽  
...  
Keyword(s):  
Healthcare Workers ◽  
Future Research ◽  
Nasopharyngeal Swab ◽  
Special Focus ◽  
Rt Pcr ◽  
Vaccine Response ◽  
Serology Testing ◽  
Effectiveness Data

AbstractThis study investigated the response to BNT162b2 mRNA COVID-19 vaccine among healthcare workers (HCWs) in an Italian teaching hospital. 444 participants were surveyed with either multiple RT-PCR assays for detection of SARS-CoV-2 nucleic acid in nasopharyngeal swabs or serology testing for the research of virus-specific immunoglobulins. Adverse events following immunization (AEFI) were reported. Two weeks after the first dose anti-SARS-CoV-2 antibodies exceeded reactivity cut-off in 82.5% the participants. Four HCWs tested positive at nasopharyngeal swab after 3 months. More than three-quarters reported AEFIs. Our findings offer an insight regarding the vaccine response after 3 months from its administration, with a special focus on effectiveness data, as well as the type and number of AEFIs complained by HCW recipients. The presented study may serve as reference for future research which will be necessary to explore the long-term safety of this vaccine, especially in population at high risk for infection, such as HCWs.

scholarly journals First Report of Cucurbit aphid-borne yellows virus in Spain

Plant Disease ◽  
2004 ◽  
Vol 88 (8) ◽  
pp. 907-907 ◽  
Author(s):  
M. Juarez ◽  
V. Truniger ◽  
M. A. Aranda
Keyword(s):  
Nucleotide Sequence ◽  
Nucleotide Sequence Identity ◽  
Rt Pcr ◽  
Sequence Comparisons ◽  
E Coli ◽  
Sequence Identity ◽  
Tissue Print ◽  
Sigma Chemical ◽  
Beet Pseudo Yellows Virus ◽  
Cucumis Melo L

In late spring 2003, field-grown melon plants (Cucumis melo L.) showing bright yellowing of older leaves were observed near Valladolises in Campo de Cartagena, Murcia, Spain. Symptoms resembled those caused by viruses of the genus Crinivirus (family Closteroviridae), but absence or very low populations of whiteflies were observed. However, diseased foci showed clear indications of heavy aphid infestations. Later, during the fall of 2003, squash plants (Cucurbita pepo L.) grown in open fields in the same area showed similar symptoms. Tissue print hybridizations to detect Cucurbit yellow stunting disorder virus (CYSDV) and Beet pseudo yellows virus (BPYV) in symptomatic samples were negative. CYSDV and BPYV are two yellowing-inducing criniviruses previously described in Spain. In contrast, standard double-antibody sandwich enzyme-linked immunosorbent assays (DAS-ELISA) with antiserum against Cucurbit aphid-borne yellows virus (CABYV; genus Polerovirus, family Luteoviridae) that was kindly provided by H. Lecoq (INRA-Montfavet Cedex, France) were consistently positive. Definitive confirmation of CABYV associated with symptomatic samples was obtained by performing reverse-transcription polymerase chain reaction (RT-PCR) analyses for the CABYV coat protein gene. Total RNA extracts (TRI reagent; Sigma Chemical, St. Louis, MO) were obtained from symptomatic and asymptomatic leaf samples and RT-PCR reactions were carried out using the primers 5′-GAATACGGTCGCGGCTAGAAATC-3′ (CE9) and 5′-CTATTTCGGGTTCTGGACCTGGC-3′ (CE10) based on the CABYV sequence published by Guilley et al. (2). A single DNA product of approximately 600 bp was obtained only from symptomatic samples. Amplified DNA fragments from two independent samples (samples 36-2 and 37-5) were cloned in E. coli and sequenced (GenBank Accession Nos. AY529653 and AY529654). Sequence comparisons showed a 95% nucleotide sequence identity between the two sequences. A 97% and 94% nucleotide sequence identity was found among 36-2 and 37-5, respectively and the CABYV sequence published by Guilley et al. (2). CABYV seems to be widespread throughout the Mediterranean Basin (1,3) but to our knowledge, it has not previously been described in Spain. Additionally, our data suggest that significant genetic variability might be present in the Spanish CABYV populations. References: (1) Y. Abou-Jawdah et al. Crop Prot. 19:217, 2000. (2) H. Guilley et al. Virology 202:1012, 1994. (3) H. Lecoq et al. Plant Pathol. 41:749, 1992.

scholarly journals OmpC regulation differs between ST131 and non-ST131 Escherichia coli clinical isolates and involves differential expression of the small RNA MicC

2020 ◽  
Vol 75 (5) ◽  
pp. 1151-1158
Author(s):  
Corey S Suelter ◽  
Nancy D Hanson
Keyword(s):  
Escherichia Coli ◽  
Half Life ◽  
Selective Advantage ◽  
Resistance Mechanisms ◽  
Northern Blotting ◽  
Rt Pcr ◽  
E Coli ◽  
Protein Levels ◽  
Porin Proteins ◽  
Mrna Half Life

Abstract Background Virulence genes and the expression of resistance mechanisms undoubtedly play a role in the successful spread of the pandemic clone Escherichia coli ST131. Porin down-regulation is a chromosomal mechanism associated with antibiotic resistance. Translation of porin proteins can be impacted by modifications in mRNA half-life and the interaction among small RNAs (sRNAs), the porin transcript and the sRNA chaperone Hfq. Modifications in the translatability of porin proteins could impact the fitness and therefore the success of E. coli ST131 isolates in the presence of antibiotic. Objectives To identify differences in the translatability of OmpC and OmpF porins for different STs of E. coli by comparing steady-state RNA levels, mRNA half-life, regulatory sRNA expression and protein production. Methods RNA expression was evaluated using real-time RT–PCR and OmpC mRNA half-life by northern blotting. OmpC, OmpF and Hfq protein levels were evaluated by immunoblotting. Results Differences between ST131 and non-ST131 isolates included: (i) the level of OmpC RNA and protein produced with mRNA expression higher for ST131 but OmpC protein levels lower compared with non-ST131 isolates; (ii) OmpC mRNA half-life (21–30 min for ST131 isolates compared with <2–23 min for non-ST131 isolates); and (iii) levels of the sRNA MicC (2- to 120-fold for ST131 isolates compared with −4- to 70-fold for non-ST131 isolates). Conclusions Mechanisms involved in the translatability of porin proteins differed among different STs of E. coli. These differences could provide a selective advantage to ST131 E. coli when confronted with an antibiotic-rich environment.

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