Potential of Nanoparticles Chitosan for Delivery pcDNA3.1-tat

The development of Jembrana disease vaccine is importance to prevent the loss of Bali cattle industry in Indonesia. This study aims to prepare a Jembrana DNA vaccine. The data Tat protein sequences gained from NCBI and the consensus process has been finished by the MultAlign program, and then Cloning of the pcDNA3.1-tat has been successfully performed on E. coli DH5α and confirmed by PCR, restriction analysis and sequencing. The propagated plasmids were prepared as DNA-chitosan complex and physiochemical characterized using Particle Size Analyzer. Complex with a 1:2 (wt/wt) ratio of DNA and chitosan have a mean diameter of 268.5 nm and zeta potential +25.1 mV and the value of Cytotoxicity Assay 80-90% as compared to the untreated cells that used as negative control, so it can be concluded that nanoparticles chitosan has good potential as a carrier agent for pcDNA3.1-tat.

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Potential of Nanoparticles Chitosan for Delivery pcDNA3.1-SB3-HBcAg

BIO Web of Conferences ◽

10.1051/bioconf/20214107003 ◽

2021 ◽

Vol 41 ◽

pp. 07003

Author(s):

Lalu Unsunnidhal ◽

Raudatul Jannah ◽

Abdul Haris ◽

Agus Supinganto ◽

Asmarani Kusumawati

Keyword(s):

Hepatitis B ◽

Restriction Analysis ◽

Negative Control ◽

Core Antigen ◽

Base Sequence ◽

E Coli ◽

Particle Size Analyzer ◽

B Virus ◽

Good Potential ◽

Hepatitis B Core Antigen

Hepatitis B virus (HBV) is a DNA virus that causes hepatitis in humans. This study aims to prepare a Hepatitis DNA vaccine. he optimized base sequence of the SB3-HBcAg gene was derived from the nucleotide base sequence of the Hepatitis B core antigen B3 HBcAg subgenotype, and then Cloning of the pcDNA3.1-SB3-HBcAg has been successfully performed on E. coli DH5α and confirmed by PCR, restriction analysis and sequencing. The propagated plasmids were prepared as DNA-chitosan complex and physiochemical characterized using Particle Size Analyzer. Complex with a 4:1 (wt/wt) ratio of DNA with 0.04% concentration and chitosan have a mean diameter of 231.7 nm and zeta potential +12.3 mV and the value of Cytotoxicity Assay 80-90% as compared to the untreated cells that used as negative control, so it can be concluded that nanoparticles chitosan has good potential as a carrier agent for pcDNA3.1-SB3-HBcAg.

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The Order in Which Antibiotics are Administered Affects the Fitness Costs of Adherent-Invasive E. coli (AIEC) Strains

10.31219/osf.io/46fae ◽

2019 ◽

Author(s):

Jordan B Gregg

Keyword(s):

Crohn’S Disease ◽

Crohn's Disease ◽

Combination Therapy ◽

Agar Plate ◽

Sequential Therapy ◽

Negative Control ◽

Clinical Settings ◽

Fitness Costs ◽

Inflammatory Gene Expression ◽

E Coli

AIEC-LF82 is a strain of bacteria that is surmised to have a role in causing IBD and Crohn’s disease by activating pro-inflammatory gene expression in organisms. Using antibiotics via combination therapy has been a technique used in clinical settings in an attempt to treat the strains, however, the attempts have not been that effective nor efficient in terms of completely halting the growth and colonization of AIEC to treat IBD and Crohn's disease patients. Research has shown that regarding hindering or preventing the colonization bacterial colonies, sequential therapy tends to be more effective and time-efficient than combination therapy, with fewer adverse effects. To test if this is also the case with the AIEC-LF82 strain of bacteria, I first tested AIEC’s response to combination therapy using the Penicillin-Streptomycin, Kanamycin-Chloramphenicol, antimicrobial peptide (AMP), Kanamycin, SPE phase and LB agar plates, all of which were experimental plates other than the LB agar plate that acted as the negative control. I then tested AIEC-LF82’s response to sequential therapy using the LB+ Kan + Spe, LB + AMP + Spe, LB+ Kan/Cam + Spe, LB + P/S + Spe, LB + P/S + Kan and LB + P/S + AMP and one LB agar plate acting as the negative control. The only differences between sets a and b were the order in which antibiotics were administered in the six aforementioned treatment sets. Ultimately, I found that set b of sequential therapy, strong-weak antibiotic treatments, was the most effective treatment but that set a regarding sequential therapy was actually the least effective of all of the treatments. In conclusion, using strong-weak sequential antibiotic therapy treatments appears to be a potentially promising option to treat patients suffering from Crohn's disease and IBD.

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POS0384 A NOVEL MECHANISM LINKING MUCOSAL BACTERIA WITH AUTOANTIBODY RESPONSES IN RA: ACETYLATED BACTERIAL LYSATE AS A MODEL ANTIGEN

Annals of the Rheumatic Diseases ◽

10.1136/annrheumdis-2021-eular.1709 ◽

2021 ◽

Vol 80 (Suppl 1) ◽

pp. 422.1-422

Author(s):

M. Volkov ◽

A. S. B. Kampstra ◽

K. van Schie ◽

J. Kwekkeboom ◽

T. Huizinga ◽

...

Keyword(s):

B Cells ◽

Cell Activation ◽

Positive Control ◽

Negative Control ◽

B Cell Activation ◽

Human Fibrinogen ◽

Post Translational Modifications ◽

E Coli ◽

Bacterial Proteins ◽

Citrullinated Proteins

Background:Rheumatoid arthritis (RA) is characterized by autoantibodies against post-translationally modified proteins (AMPA) such as citrullinated, carbamylated and acetylated proteins. Importantly, these antibodies are highly multireactive, as they often recognize more than one of these post-translational modifications. Despite extensive research, the antigens inducing the breach of tolerance remain unknown, although microbial antigens are often suspected. Various bacteria are known to be capable of acetylation, therefore, it is intriguing to know what mechanisms can underlie the breach of tolerance towards acetylated proteins and development of anti-acetylated protein antibodies (AAPA).Objectives:To investigate whether acetylated proteins of bacterial origin (1) are recognized by human derived AMPA and AMPA expressing B cells; and (2) can induce AMPA development when used to immunize mice.Methods:Acetylated E. coli proteins were acquired with two separate methods (Figure 1A): by culturing E. coli in a condition promoting auto-acetylation (intrinsically acetylated bacterial proteins, IABP), or by directly acetylating lysate-derived proteins via a chemical reaction (extrinsically acetylated BP, EABP). Acetylated ovalbumin (AcOVA) served as positive control for AAPA induction in mice, non-acetylated BP (NABP) and phosphate buffer saline (PBS) served as negative control. Mice were immunized with these proteins and the resulting antibody response was studied by ELISA. Furthermore, EABP/IABP/NABP were investigated for recognition by human-derived AAPA with ELISA and AAPA-expressing B cells with spleen tyrosine kinase (Syk) phosphorylation assay; acetylated human fibrinogen and native fibrinogen served as positive and negative control.Results:Repetitive immunization of mice with EABP resulted in an AMPA response recognizing acetylated, carbamylated and citrullinated proteins. AMPA titers in these mice exceeded the titers in the positive control mice immunized with AcOVA and were substantially higher than in the NABP-immunized mice (Figure 1B). Human-derived monoclonal AAPA recognized EABP and IABP (not shown). B cell activation (measured by Syk phosphorylation) assay indicated that AAPA expressing B cells recognized EABP and (to a lesser extent) IABP, but not NABP (Figure 1C).Conclusion:Acetylated bacterial proteins are potent antigens that can induce cross-reactive AMPA responses in mice and they are recognized by human AAPA. This suggests that acetylated bacterial proteins could possibly be involved in the breach of tolerance in RA.Acknowledgements:We thank Dr. Can Araman and Prof. Chunaram Choudhary for their advice regarding optimization of bacterial auto-acetylation.Disclosure of Interests:None declared

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Evaluation of antipyretic activity of hydroalcoholic extract of Corchorus depressus Linn. in Escherichia coli–induced pyretic rabbits

European Journal of Inflammation ◽

10.1177/2058739218792958 ◽

2018 ◽

Vol 16 ◽

pp. 205873921879295

Author(s):

Saeed Ahmad ◽

Muhammad Akram ◽

Syed Muhammad Ali Shah ◽

Sabira Sultana

Keyword(s):

Escherichia Coli ◽

Positive Control ◽

Negative Control ◽

The Body ◽

Hydroalcoholic Extract ◽

Antipyretic Effect ◽

E Coli ◽

Antipyretic Activity ◽

Experimental Treatments

This study was conducted to investigate the antipyretic effect of the hydroalcoholic extract of Corchorus depressus Linn. against Escherichia coli ( E. coli)-induced pyrexia in rabbits. Hydroalcohalic extracts of C. depressus were given orally at 25, 50, and 100 mg/kg for antipyretic affect in E. coli-induced fever in rabbits. The animals were divided into five groups of five each. Among these five groups, three received various doses of experimental treatments, whereas the fourth one served as positive control and received paracetamol. The fifth group of animals served as negative control and received no treatment. The body temperature of the rabbits was measured rectally over a period of 5 h. C. depressus exhibited better effects at dose rate of 25, 50, and 100 mg/kg. The hydroalcoholic extract of C. depressus has significant antipyretic effect. These results lend support to the popular use of C. depressus in traditional medicine as a remedy for pyrexia and suggest that the characterization of the principles for such activity deserves further investigation.

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Mapping of HIV-1 Tat protein sequences required for binding to Tar RNA

Virology ◽

10.1016/0042-6822(91)90598-6 ◽

1991 ◽

Vol 182 (2) ◽

pp. 570-577 ◽

Cited By ~ 14

Author(s):

James Kamine ◽

Paul Loewenstein ◽

Maurice Green

Keyword(s):

Protein Sequences ◽

Tat Protein ◽

Tar Rna ◽

Hiv 1

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miR-215 Targeting Novel Genes EREG, NIPAL1 and PTPRU Regulates the Resistance to E.coli F18 in Piglets

Genes ◽

10.3390/genes11091053 ◽

2020 ◽

Vol 11 (9) ◽

pp. 1053

Author(s):

Chao-Hui Dai ◽

Fang Wang ◽

Shi-Qin Wang ◽

Zheng-Chang Wu ◽

Sheng-Long Wu ◽

...

Keyword(s):

Target Genes ◽

Core Promoter ◽

Negative Control ◽

Enzyme Linked Immunosorbent Assay ◽

Nucleotide Polymorphisms ◽

Weaned Piglets ◽

Promoter Regions ◽

E Coli ◽

Rnai Technology ◽

Genes Expression

Previous research has revealed that miR-215 might be an important miRNA regulating weaned piglets’ resistance to Escherichia coli (E. coli) F18. In this study, target genes of miR-215 were identified by RNA-seq, bioinformatics analysis and dual luciferase detection. The relationship between target genes and E. coli infection was explored by RNAi technology, combined with E. coli stimulation and enzyme linked immunosorbent assay (ELISA) detection. Molecular regulating mechanisms of target genes expression were analyzed by methylation detection of promoter regions and dual luciferase activity assay of single nucleotide polymorphisms (SNPs) in core promoter regions. The results showed that miR-215 could target EREG, NIPAL1 and PTPRU genes. Expression levels of three genes in porcine intestinal epithelial cells (IPEC-J2) in the RNAi group were significantly lower than those in the negative control pGMLV vector (pGMLV-NC) group after E. coli F18 stimulation, while cytokines levels of TNF-α and IL-1β in the RNAi group were significantly higher than in the pGMLV-NC group. Variant sites in the promoter region of three genes could affect their promoter activities. These results suggested that miR-215 could regulate weaned piglets’ resistance to E. coli F18 by targeting EREG, NIPAL1 and PTPRU genes. This study is the first to annotate new biological functions of EREG, NIPAL1 and PTPRU genes in pigs, and provides a new experimental basis and reference for the research of piglets disease-resistance breeding.

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Influence of treatment and refrigeration time on antimicrobial activity of goat and sheep colostrum

Journal of Dairy Research ◽

10.1017/s0022029919000657 ◽

2019 ◽

Vol 86 (4) ◽

pp. 450-453

Author(s):

Maria D. Ruiz-Diaz ◽

Anastasio Argüello ◽

Daniel Padilla ◽

Bernadette Earley ◽

Noemi Castro

Keyword(s):

Antimicrobial Activity ◽

Petri Dish ◽

Negative Control ◽

E Coli ◽

Research Communication ◽

Different Temperatures ◽

Effect Of Treatment

AbstractThe aim of the studies presented in this research communication was to compare species of origin (goat and sheep) and the effect of treatment (pasteurization at 56, 63 and 72 °C, skimming and curding) and refrigeration time on colostrum antimicrobial activity (AnAc). Two experiments were performed. In experiment 1, twenty-four first milking colostrum samples were obtained (12 goats, 12 sheep) and an aliquot of each sample was subjected to 6 different treatments, control (untreated), pasteurization at 56, 63 and 72 °C, skimming and curding. Colostrum AnAc was tested directly against E. coli using disks in a Petri dish and Enrofloxacin (antibiotic) and saline serum as positive and negative control, respectively. Species had no effect (P > 0.05) on colostrum AnAc, and neither did pasteurization at different temperatures or skimming. However, curding showed the lowest colostrum AnAc (P < 0.05) in both species. In the second experiment, four treatments were assayed, control, pasteurization at 56 and 63 °C and skimming. An aliquot of twelve goat colostrum samples were refrigerated after treatments for 10 d at 4 °C. Colostrum AnAc was measured at 0, 2, 4, 6, 8, and 10 d. A reduction in colostrum AnAc was observed due to refrigeration time. The results suggest that if farmers use frozen colostrum for neonates, the process of curding colostrum or refrigeration at 4 °C longer than 4 d is not recommended.

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Effect of different phytases on the performance, nutrient retention and tibia composition in broiler chickens

Archives Animal Breeding ◽

10.7482/0003-9438-56-104 ◽

2013 ◽

Vol 56 (1) ◽

pp. 1028-1038 ◽

Cited By ~ 2

Author(s):

A. Ptak ◽

D. Józefiak ◽

B. Kierończyk ◽

M. Rawski ◽

K. Żyła ◽

...

Keyword(s):

Broiler Chickens ◽

Nutrient Retention ◽

Nutrient Utilization ◽

Positive Control ◽

Negative Control ◽

Metabolizable Energy ◽

Feed Conversion ◽

E Coli ◽

Performance Response ◽

Apparent Metabolizable Energy

Abstract. The effect of different phytases on the performance response, nutrient utilization and tibia characteristics of chickens was investigated. The five pelleted diets were the following: positive control (PC) with added monocalcium phosphate; negative control (NC) formulated with equivalency values of phytase for Ca and digestible P; and three further diets where different phytases were individually added to the NC diet at 500 FTU/kg. The phytases were derived either from Aspergillus (phytase I), or E. coli (phytases II and III). Compared to PC, the performance parameters, as well as apparent metabolizable energy (AMEn), mineral retention, bone breaking force and tibia mineral content were suppressed by the reduction of dietary Ca and digestible P. All phytases enhanced the overall body weight gains and feed conversion ratio in comparison with NC, but none outperformed PC. Only phytase II improved AMEn compared to NC and PC group. However only phytase I outperformed NC group in terms of mineral retention and P retention was higher than phytase II and III. No significant differences were observed in fat digestibility and N retention. Bone strength among phytases did not differ and all improved this parameter compared to the NC diet. However, even though all phytases enhanced tibia minerals content, the improvement was less pronounced with phytase III. Moreover, the differences in all analysed tibia minerals between phytase III and II were significant suggesting that even among 6-phytases derived from and expressed in the same organism, different efficacy or mode of action can occur.

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Protective Effect of Curcumin on Carbapenem-Resistant Escherichia coli–Induced Lung Injury in Rats

International Surgery ◽

10.9738/intsurg-d-15-00256.1 ◽

2016 ◽

Vol 101 (7-8) ◽

pp. 304-312 ◽

Cited By ~ 1

Author(s):

Cagla Bali ◽

Nejat Altintas ◽

Ozlem Ozmete ◽

Ibrahim Gelincik ◽

Hakan Yabanoglu ◽

...

Keyword(s):

Escherichia Coli ◽

Lung Injury ◽

Intraperitoneal Injection ◽

Negative Control Group ◽

Positive Control ◽

Negative Control ◽

Positive Control Group ◽

Control Group ◽

E Coli ◽

Carbapenem Resistant

Curcumin has remarkable anti-inflammatory and antioxidant properties. The aim of this study was to investigate the protective effects of curcumin on a rat model of carbapenem-resistant Escherichia coli–induced acute lung injury (ALI). Thirty-two rats were randomly allocated to 4 groups to induce an ALI: negative control group (rats not infected with E coli with no antibiotic treatment), positive control group (rats infected with E coli with no antibiotic treatment), imipenem group (rats infected with E coli that received intraperitoneal injection of imipenem), and the imipenem+curcumin group (rats infected with E coli that received intraperitoneal injection of imipenem and were fed on curcumin).The rats were killed, and lung tissues samples were harvested for biochemical analyses and histopathologic examination. Total antioxidant status (TAS), total oxidant status (TOS), tumor necrosis factor α (TNFα), and interleukin-6 (IL6) were measured. TOS increased in the positive control group (P < 0.001) and decreased in the imipenem and imipenem+curcumin groups (P < 0.001 and P < 0.001, respectively). TAS decreased in the positive control group (P = 0.005). Imipenem treatment did not increase TAS, but the imipenem+curcumin group increased TAS (P = 0.014). TNFα and IL6 increased in the positive control group compared with the negative control group (P < 0.001 and P = 0.010, respectively). Imipenem decreased TNFα (P < 0.001), but did not decrease IL6 (P = 0.418). Imipenem+curcumin decreased TNFα (P < 0.001); this decrease was more pronounced compared with the imipenem group (P = 0.008). IL6 decreased in the curcumin group compared with the positive control group (P = 0.011). Curcumin combined with imipenem can be an alternative therapeutic agent to overcome the resistance of E coli strains.

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Benzoic Acid Combined with Essential Oils Can Be an Alternative to the Use of Antibiotic Growth Promoters for Piglets Challenged with E. coli F4

Animals ◽

10.3390/ani10111978 ◽

2020 ◽

Vol 10 (11) ◽

pp. 1978

Author(s):

Leticia Mendonça Rodrigues ◽

Túlio Otávio de Araújo Lima Neto ◽

Cesar Augusto Pospissil Garbossa ◽

Claudia Cassimira da Silva Martins ◽

Dino Garcez ◽

...

Keyword(s):

Benzoic Acid ◽

Essential Oils ◽

Cellular Proliferation ◽

Block Design ◽

Average Daily Gain ◽

Negative Control ◽

Growth Promoter ◽

Final Body Weight ◽

Total Period ◽

E Coli

Benzoic acid (BA) and essential oils (EOs) (thymol, 2-methoxyphenol, eugenol, piperine, and curcumin) are being studied to minimize the impairment of gastrointestinal functions in weanling piglets. This study evaluates the effects of combining BA and EO on the performance and intestinal health of piglets challenged with E. coli F4 (1 mL, 106 CFU/mL). In total, 270 weaned piglets were used in a randomized block design with six treatments: positive control, with 40 mg/kg colistin (PC); negative control, without the growth promoter (NC); negative control +5 g/kg benzoic acid (BA); negative control +2 g/kg combination of BA+EO (BA+EO2); negative control +3 g/kg combination of BA+EO (BA+EO3); negative control +4 g/kg combination of BA+EO (BA+EO4). BA+EO3 presented a greater average daily gain (ADG) (p = 0.0013) and better feed-to-gain ratio (p = 0.0138), compared to NC, from 21 to 35 days age. For the total period, BA, BA+EO3, and BA+EO4 were similar to PC and superior to NC for ADG (p = 0.0002) and final body weight (BW) (p = 0.0002). No difference (p > 0.05) was verified for diarrhea, microbial population, production of volatile fatty acids, pH, weight of organs, cellular proliferation, and cholecystokinin count. NC and BA+EO4 resulted in a higher villus height in the jejunum (p = 0.0120) compared to BA+EO3. The use of BA or the combination of BA and EO at 3 g/kg provides improved performance, aside from being an economically viable alternative to replace colistin.

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