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1.
Neural Regeneration Research ; 18(1):38-46, 2023.
Article in English | EMBASE | ID: covidwho-2313974

ABSTRACT

Obesity is associated with several diseases, including mental health. Adipose tissue is distributed around the internal organs, acting in the regulation of metabolism by storing and releasing fatty acids and adipokine in the tissues. Excessive nutritional intake results in hypertrophy and proliferation of adipocytes, leading to local hypoxia in adipose tissue and changes in these adipokine releases. This leads to the recruitment of immune cells to adipose tissue and the release of pro-inflammatory cytokines. The presence of high levels of free fatty acids and inflammatory molecules interfere with intracellular insulin signaling, which can generate a neuroinflammatory process. In this review, we provide an up-to-date discussion of how excessive obesity can lead to possible cognitive dysfunction. We also address the idea that obesity-associated systemic inflammation leads to neuroinflammation in the brain, particularly the hypothalamus and hippocampus, and that this is partially responsible for these negative cognitive outcomes. In addition, we discuss some clinical models and animal studies for obesity and clarify the mechanism of action of anti-obesity drugs in the central nervous system.Copyright © 2023 Wolters Kluwer Medknow Publications. All rights reserved.

2.
European Journal of Medicinal Chemistry Reports ; 6 (no pagination), 2022.
Article in English | EMBASE | ID: covidwho-2303478

ABSTRACT

Globally cancer is the second leading cause of death;a drug that can cure cancer with the utmost negligible side effects is still a distant goal. Due to increasing antibiotic resistance, microbial infection remains a grave global health security threat. The ongoing coronavirus pandemic increased the risk of microbial and fungal infection. A new series of 3-(4-methyl-2-arylthiazol-5-yl)-5-aryl-1,2,4-oxadiazole (7a-t) have been synthesized. The structure of synthesized compounds was confirmed by the spectrometric analysis. The newly synthesized compounds were screened for cytotoxic activity against breast cell lines MCF-7 and MDA-MB-231. Against the MCF-7 cell line compounds 7f, 7 g and 7n showed excellent activity with GI50 0.6 muM to <100 nM concentration. Compound 7b showed good activity against MDA-MB-231 cell line with GI50 47 muM. The active derivatives 7b, 7e, 7f, 7 g and 7n were further evaluated for cytotoxicity against the epithelial cell line derived from the human embryonic kidney (HEK 293) and were found nontoxic. The thiazolyl-1,2,4-oxadiazole derivatives were also screened to evaluate theirs in vitro antimicrobial potential against Escherichia coli (NCIM 2574), Proteus mirabilis (NCIM 2388), Bacillus subtilis (NCIM 2063), Staphylococcus albus (NCIM 2178), Candida albicans (NCIM 3100) and Aspergillus niger (ATCC 504). Amongst the 7a-t derivatives, six compounds 7a, 7d, 7f, 7n, 7o, 7r showed good antifungal activity against C. albicans and eight compounds 7c, 7d, 7 g, 7h, 7i, 7k, 7l and 7o showed good activity against A. niger. The potential cytotoxic and antifungal activity suggested that the thiazolyl-1,2,4-oxadiazole derivatives could assist in the development of lead compounds for the treatment of cancer and microbial infections.Copyright © 2022 The Authors

3.
J Biomol Struct Dyn ; 40(13): 5748-5758, 2022 08.
Article in English | MEDLINE | ID: covidwho-2248784

ABSTRACT

The COVID-19 being a preconized global pandemic by the World Health Organization needs persuasive immediate research for possible medications. The present study was carried out with a specific aim to computationally evaluate and identify compounds derived from Bacillus species as the plausible inhibitors against 3-chymotrypsin-like main protease (3CLpro) or main protease (MPro), which is a key enzyme in the life-cycle of coronavirus. The compounds were isolated from the crude extracts of Bacillus species. Among the isolated compounds, novel inhibitory leads were identified using in silico techniques. Molecular docking revealed that stigmasterol (-8.3 kcal/mol), chondrillasterol (-7.9 kcal/mol) and hexadecnoic acid (-6.9 kcal/mol)) among others bind in the substrate-binding pocket and also interacted with the catalytic dyad of the 3-CLpro. Further evaluation using 50 ns molecular dynamic simulation and MMPB-GBSA indicated that among the top three docking hits, hexadecanoic acid was found to be the most promising anti-COVID-19 lead against the main protease. Hexadecanoic acid might serve as a potent anti-SARS-CoV-2 compound to combat COVID-19, however, in vitro and in vivo validation and optimization is needed.Communicated by Ramaswamy H. Sarma.


Subject(s)
Bacillus , COVID-19 Drug Treatment , Bacillus/metabolism , Humans , Molecular Docking Simulation , Molecular Dynamics Simulation , Palmitic Acid , Protease Inhibitors/chemistry , Protease Inhibitors/pharmacology
4.
Comput Struct Biotechnol J ; 21: 1550-1556, 2023.
Article in English | MEDLINE | ID: covidwho-2236645

ABSTRACT

Bacillus subtilis spores are considered to be efficient and useful vehicles for the surface display and delivery of heterologous proteins. In this study, we prepared recombinant spores with the receptor binding domain (RBD) of the SARS-CoV-2 spike glycoprotein displayed on their surface in fusion with the CotZ or CotY spore coat proteins as a possible tool for the development of an oral vaccine against the SARS-CoV-2 virus. The RBD was attached to the N-terminus or C-terminus of the coat proteins. We also directly adsorbed non-recombinantly produced RBD to the spore surface. SDS-PAGE, western blot and fluorescence microscopy were used to analyze RBD surface expression on purified spores. Results obtained from both display systems, recombinant and non-recombinant, demonstrated that RBD was present on the spore surfaces.

5.
Foods ; 12(2)2023 Jan 09.
Article in English | MEDLINE | ID: covidwho-2199952

ABSTRACT

Bacillus subtilis natto is used in the production of natto, a traditional fermented soy food, and has beneficial immunomodulatory effects in humans. Single-stranded RNA (ssRNA) viruses, including influenza and coronavirus, often cause global pandemics. We proposed a human cell culture model mimicking ssRNA viral infection and investigated the ability of B. subtilis natto to induce antiviral effects in the model. The gene expressions were analyzed using quantitative real-time reverse transcription PCR. M1-phenotype macrophages derived from THP-1 cells strongly express the Toll-like receptor 8 (76.2-hold), CD80 (64.2-hold), and CCR7 (45.7-hold) mRNA compared to M0 macrophages. One µg/mL of resiquimod (RSQ)-stimulation induced the expression of IRF3 (1.9-hold), CXCL10 (14.5-hold), IFNß1 (3.5-hold), ISG20 (4.4-hold), and MxA (1.7-hold) mRNA in the M1-phenotype macrophages. Based on these results, the RSQ-stimulated M1-phenotype macrophages were used as a cell culture model mimicking ssRNA viral infection. Moreover, the B. subtilis natto XF36 strain induced the expression of genes associated with antiviral activities (IFNß1, IFNλ1, ISG20, and RNase L) and anti-inflammatory activities (IL-10) in the cell culture model. Thus, it is suggested that the XF36 suppresses viral infections and excessive inflammation by inducing the expression of genes involved in antiviral and anti-inflammatory activities.

6.
FEBS Open Bio ; 12:171, 2022.
Article in English | EMBASE | ID: covidwho-1976644

ABSTRACT

Antimicrobial agents are demanded to treat infectious diseases, one of the leading causes of death worldwide. Longstanding thoughtless use of antibiotics has led to the development and spread of drug-resistant microorganisms. This problem has become especially acute within the Covid-19 pandemic. Antimicrobial peptides (AMPs) are considered as effective therapeutic agents that are less prone to the development of bacterial resistance than antibiotics. In the present study, we identified antimicrobial non-toxic peptides from the leech metagenome protein database using a gradient boosting approach (CatBoost). The antimicrobial activity of the peptides was tested against Grampositive and Gram-negative bacteria (Bacillus subtilis, Staphylococcus aureus, Staphylococcus haemolyticus, Escherichia coli, Mycobacterium smegmatis). One of the analyzed peptides, peptide HMAMP-2, showed the best antimicrobial activity against the tested bacteria at low concentrations. Viability and hemolysis evaluation of cells incubated with peptides demonstrated peptide HMAMP-2 did not affect cell viability at high concentrations. It was experimentally shown that the peptide killed pathogens due to the membranolytic mechanism of action. According to nuclear magnetic resonance analysis, the peptide adopted an a-helical structure in a membrane environment. In addition, we demonstrated that peptide Hm-AMP2 binds to LPS, which supported the immunomodulatory properties of the peptide. Thus, peptide Hm-AMP2 is characterized as a promising non-hemolytic and non-toxic antimicrobial compound. Peptide Hm-AMP2 is likely involved in the regulation of the inflammatory response of the innate immune system, which should be studied further. The developed algorithm for the identification of antimicrobial peptides with a reduced toxic effect can be used for the rational design of effective non-toxic antibacterial agents.

7.
Front Immunol ; 13: 925922, 2022.
Article in English | MEDLINE | ID: covidwho-1933699

ABSTRACT

Although feline coronavirus (FCoV) infection is extremely common in cats, there are currently few effective treatments. A peptide derived from the heptad repeat 2 (HR2) domain of the coronavirus (CoV) spike protein has shown effective for inhibition of various human and animal CoVs in vitro, but further use of FCoV-HR2 in vivo has been limited by lack of practical delivery vectors and small animal infection model. To overcome these technical challenges, we first constructed a recombinant Bacillus subtilis (rBSCotB-HR2P) expressing spore coat protein B (CotB) fused to an HR2-derived peptide (HR2P) from a serotype II feline enteric CoV (FECV). Immunogenic capacity was evaluated in mice after intragastric or intranasal administration, showing that recombinant spores could trigger strong specific cellular and humoral immune responses. Furthermore, we developed a novel mouse model for FECV infection by transduction with its primary receptor (feline aminopeptidase N) using an E1/E3-deleted adenovirus type 5 vector. This model can be used to study the antiviral immune response and evaluate vaccines or drugs, and is an applicable choice to replace cats for the study of FECV. Oral administration of rBSCotB-HR2P in this mouse model effectively protected against FECV challenge and significantly reduced pathology in the digestive tract. Owing to its safety, low cost, and probiotic features, rBSCotB-HR2P is a promising oral vaccine candidate for use against FECV/FCoV infection in cats.


Subject(s)
Coronavirus Infections , Coronavirus, Feline , Animals , Bacillus subtilis/genetics , CD13 Antigens/metabolism , Cats , Coronavirus, Feline/genetics , Coronavirus, Feline/metabolism , Disease Models, Animal , Humans , Immunity , Mice , Peptides/metabolism , Spike Glycoprotein, Coronavirus/metabolism , Spores, Bacterial/genetics
8.
Vaccines (Basel) ; 10(7)2022 Jun 24.
Article in English | MEDLINE | ID: covidwho-1911718

ABSTRACT

The coronavirus diseases 2019 (COVID-19) pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infections have threatened the world for more than 2 years. Multiple vaccine candidates have been developed and approved for emergency use by specific markets, but multiple doses are required to maintain the antibody level. Preliminary safety and immunogenicity data about an oral dose vaccine candidate using recombinant Bacillus subtilis in healthy adults were reported previously from an investigator-initiated trial in Hong Kong. Additional data are required in order to demonstrate the safety and efficacy of the candidate as a heterologous booster in vaccinated recipients. In an ongoing, placebo-controlled, observer-blinded, fixed dose, investigator-initiated trial conducted in the Macau, we randomly assigned healthy adults, 21 to 62 years of age to receive either placebo or a Bacillus subtilis oral dose vaccine candidate, which expressed the spike protein receptor binding domain of SARS-CoV-2 on the spore surface. The primary outcome was safety (e.g., local and systemic reactions and adverse events); immunogenicity was a secondary outcome. For both the active vaccine and placebo, participants received three courses in three consecutive days. A total of 16 participants underwent randomization: 9 participants received vaccine and 7 received placebo. No observable local or systemic side-effect was reported. In both younger and older adults receiving placebo, the neutralizing antibody levels were gradually declining, whereas the participants receiving the antibody booster showed an increase in neutralizing antibody level.

9.
Vet Sci ; 9(5)2022 Apr 25.
Article in English | MEDLINE | ID: covidwho-1892994

ABSTRACT

Bacillus subtilis (B. subtilis), a probiotic bacterium and feeding additive, is widely used for heterologous antigen expression and protective immunisation. Porcine epidemic diarrhoea virus (PEDV) invades swine via mucosal tissue. To enhance the mucosal immune response to PEDV, we modified B. subtilis to express a PEDV antigen and used it as a mucosal vaccine delivery system. Initially, we constructed a recombinant B. subtilis strain (B.s-RCL) that expressed the PEDV spike protein and L-Lectin-ß-GF, with the goal of inducing mucosal secretory immunoglobulin A (sIgA) and anti-PEDV serum immunoglobulin G (IgG) production, as well as to increase the number of microfold cells (M cells). Following the oral administration of B.s-RCL to mice, the small intestinal PEDV-specific sIgA expression levels significantly increased, as well as the increased number of B.s-RCL adhered to M cells. Moreover, we found that mice administered B.s-RCL exhibited markedly higher percentages of CD4+ and CD8+ T cells in the mesenteric lymph nodes and spleen compared to the control mice. Furthermore, we found that intestinal mucosa sIgA and serum anti-PEDV IgG levels were higher in mice orally immunised with B.s-RCL, suggesting that the mice could be more resistant to PEDV. In this study, we developed a novel oral vaccine to prevent porcine diarrhoea epidemics.

10.
J Infect Chemother ; 28(8): 1212-1215, 2022 Aug.
Article in English | MEDLINE | ID: covidwho-1864584

ABSTRACT

A 70-year-old woman, who started on hemodialysis 7 months before for end-stage renal disease due to diabetic nephropathy and was diagnosed with symptomatic multiple myeloma 1 month before, was admitted to our hospital with critical coronavirus disease 2019 and treated with long-term immunosuppressive therapy such as steroids and tocilizumab. During treatment, Bacillus subtilis was detected in the blood cultures. We could not exclude the association of natto (fermented soybeans) with B. subtilis var. natto, which the patient had been eating every day from 8 days after admission. She was prohibited from eating natto and treated with vancomycin. Later, B. subtilis detected in the blood culture was identified as B. subtilis var. natto, which was identical with those contained in the natto that the patient consumed daily using a next-generation sequencer. Gut dysbiosis due to old age, malignant tumor, diabetes mellitus, end-stage renal disease, and intestinal inflammation caused by severe acute respiratory syndrome coronavirus 2 increased intestinal permeability and the risk of bacterial translocation, causing B. subtilis var. natto bacteremia. Therefore, careful consideration might be given to the intake of fermented foods containing live bacteria in patients with severe immunocompromised conditions.


Subject(s)
Bacteremia , COVID-19 Drug Treatment , COVID-19 , Kidney Failure, Chronic , Multiple Myeloma , Soy Foods , Aged , Bacillus subtilis , Bacteremia/drug therapy , COVID-19/complications , Eating , Female , Humans , Kidney Failure, Chronic/complications , Kidney Failure, Chronic/therapy , Multiple Myeloma/complications , Multiple Myeloma/drug therapy , Renal Dialysis , Soy Foods/microbiology
11.
Biochemical and Cellular Archives ; 21(2):1-2, 2021.
Article in English | EMBASE | ID: covidwho-1812557
12.
Research Journal of Pharmacy and Technology ; 15(1):127-136, 2022.
Article in English | EMBASE | ID: covidwho-1744018

ABSTRACT

The diversity in Jordan’s flora due to its geographical areas make is well noted in the scientific literature. The challenge of disease and death caused by infectious diseases like viruses and bacteria, and as infectious diseases evolve and pathogens develop resistance to existing pharmaceuticals, the search for new novel leads, possibly with different modes of action, against bacterial and viral diseases has intensified in recent years. The intent of this review is to provide prevalent information on the antibacterial and antiviral potential in medicinal plants in Jordan, mode of action, type of viruses and bacteria, and phytochemical contents. It has been demonstrated by several studies presented in this review that medicinal plants in Jordan are rich in phytochemicals and possess antiviral and antibacterial properties.

13.
8th Colombian Congress and International Conference on Air Quality and Public Health, CASAP 2021 ; 2021.
Article in Spanish | Scopus | ID: covidwho-1746121

ABSTRACT

This study was carried out in order to determine the presence of potentially pathogenic microorganisms in the air of a home located in the municipality of Tausa and its possible relationship with ARIs (Acute Respiratory Infections), for which a microbiological analysis was carried out in order to identify bacteria that are possibly causing respiratory illnesses in the household. The sampling consisted of the use of a suction equipment (MAS100 Eco), during different time bands and spaces inside and outside the house. Later the respective analysis was carried out in the laboratory and 6 different mesophilic microorganisms were identified, which were: Salmonella tiphy, Bacillus subtilis, Acinetobacter baumannii, Escherichia coli, Staphylococcus aureus, and Kingella kingae. In addition, each of these bacteria were individually analyzed to understand the dynamics of the pollutant load in the home. Finally, the identified mesophilic microorganisms correspond to bacteria with some degree of pathogenicity and/or health effects, in the same way the morbidity data from the Tausa medical center were analyzed. Here we found that the population under 13 years old is the most affected by ARI, and that the bacteria present more easily affect this type of population, generating a wider perspective on the possibilities of having more patients diagnosed with ARI, as found in the home. The data presented here were affected and biased due to the health emergency caused by COVID19. © 2021 IEEE.

14.
Biocell ; 46(SUPPL 1):180, 2022.
Article in English | EMBASE | ID: covidwho-1675776

ABSTRACT

The SARS-CoV-2 coronavirus, which causes respiratory syndrome COVID-19, has a protein nucleocapsid that envelops the viral ssRNA. The main protein of the nucleocapsid is the Np protein, which presents limited homology with nucleoproteins of other coronaviruses and therefore turns out to be an attractive antigen for the development of specific anti-Np antibodies. These antibodies can be used for the development of diagnostic systems that allow the detection of the viral antigen in infected individuals from saliva samples. In this context, our group has developed a labelling system called FasTAG®, which allows the immobilization of recombinant proteins on the surface of Gram+ formaldehyde inactivated bacteria. In this system, the recombinant proteins expressed in heterologous systems are fused to the C-terminal domain of S-Layer proteins of Lactobacillus sp. Then, the intrinsic affinity this domain possesses for the membranes of Gram+ bacteria is used for the immobilization of the recombinant proteins of interest. In this way, it is possible to purify specific antibodies against an antigen of interest. Based on the above, the objective of this work was to evaluate the functionality of the FasTAG® system to purify specific anti-Np antibodies. For this, the recombinant protein Np-FasTAG® was incubated for 12 hours at 4 °C with a matrix made up of B. subtilis inactivated with 3% formaldehyde. Next, for the optimization of the protein fixation process to the matrix, a compound factorial design was carried out, the variables of which were: formaldehyde concentration (0.5-1.5-2.5% v/v) and time of incubation (15-30-45 minutes). The optimal condition was determined as the one that minimizes the detachment of the Np protein and maximizes the detachment of the specific antibodies. Turning out to be the optimal condition for the elaboration of the affinity matrix 2.5% v/v of formaldehyde and 15 minutes. Then, in order to evaluate the application of the affinity matrix in the purification of specific antibodies, it was incubated for 1 hour with polyclonal antibodies obtained from chicken egg yolks and the serum of goats immunized with the Np antigen. Next, to study the elution conditions of the antibodies, a compound factorial design was performed using variables: pH, time, and SDS concentration. The best elution condition was obtained for pH 10.5 and 15 minutes. Subsequently, the purified antibodies were evaluated by SDS-PAGE and ELISA. As a result, it was possible to purify 3.5 μg of anti-Np IgG and 3.1 μg of anti-Np IgY per mg of resin. Finally, the set of experiments carried out here demonstrate the potential and functionality of this system for the purification of specific anti-Np antibodies and their use for diagnostic purposes.

15.
Avicenna J Med Biotechnol ; 14(1): 70-78, 2022.
Article in English | MEDLINE | ID: covidwho-1649919

ABSTRACT

Background: Lipopeptides are potential microbial metabolites that are abandoned with broad spectrum biopharmaceutical properties ranging from antimicrobial, antiviral and anticancer, etc. Clinical studies are not much explored beyond the experimental methods to understand drug mechanisms on target proteins at the molecular level for large molecules. Due to the less available studies on potential target proteins of lipopeptide based drugs, their potential inhibitory role for more obvious treatment on disease have not been explored in the direction of lead optimization. However, Computational approaches need to be utilized to explore drug discovery aspects on lipopeptide based drugs, which are time saving and cost-effective techniques. Methods: Here a ligand-based drug discovery approach is coupled with reverse pharmacophore-mapping for the prediction of potential targets for antiviral (SARS-nCoV-2) and anticancer lipopeptides. Web-based servers PharmMapper and Swiss Target Prediction are used for the identification of target proteins for lipopeptides surfactin and iturin produced by Bacillus subtilis. Results: The studies have given the insight to treat the diseases with next-generation large molecule therapeutics. Results also indicate the affinity for Angiotensin-Converting Enzymes (ACE) and proteases as the potential viral targets for these categories of peptide therapeutics. A target protein for the Human Papilloma Virus (HPV) has also been mapped. Conclusion: The work will further help in exploring computer-aided drug designing of novel compounds with greater efficiency where the structure of the target proteins and lead compounds are known.

16.
Vaccines (Basel) ; 10(1)2021 Dec 21.
Article in English | MEDLINE | ID: covidwho-1580373

ABSTRACT

Various types of vaccines, such as mRNA, adenovirus, and inactivated virus by injection, have been developed to prevent SARS-CoV-2 infection. Although some of them have already been approved under the COVID-19 pandemic, various drawbacks, including severe side effects and the requirement for sub-zero temperature storage, may hinder their applications. Bacillus subtilis (B. subtilis) is generally recognized as a safe and endotoxin-free Gram-positive bacterium that has been extensively employed as a host for the expression of recombinant proteins. Its dormant spores are extraordinarily resistant to the harsh environment in the gastrointestinal tract. This feature makes it an ideal carrier for oral administration in resisting this acidic environment and for release in the intestine. In this study, an engineered B. subtilis spore expressing the SARS-CoV-2 spike protein receptor binding domain (sRBD) on the spore surface was developed. In a pilot test, no adverse health event was observed in either mice or healthy human volunteers after three oral courses of B. subtilis spores. Significant increases in neutralizing antibody against sRBD, in both mice and human volunteers, after oral administration were also found. These findings may enable the further clinical developments of B. subtilis spores as an oral vaccine candidate against COVID-19 in the future.

17.
Biochem Biophys Res Commun ; 570: 21-25, 2021 09 17.
Article in English | MEDLINE | ID: covidwho-1309162

ABSTRACT

Natto, a traditional Japanese fermented soybean food, is well known to be nutritious and beneficial for health. In this study, we examined whether natto impairs infection by viruses, such as severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) as well as bovine herpesvirus 1 (BHV-1). Interestingly, our results show that both SARS-CoV-2 and BHV-1 treated with a natto extract were fully inhibited infection to the cells. We also found that the glycoprotein D of BHV-1 was shown to be degraded by Western blot analysis and that a recombinant SARS-CoV-2 receptor-binding domain (RBD) was proteolytically degraded when incubated with the natto extract. In addition, RBD protein carrying a point mutation (UK variant N501Y) was also degraded by the natto extract. When the natto extract was heated at 100 °C for 10 min, the ability of both SARS-CoV-2 and BHV-1 to infect to the cells was restored. Consistent with the results of the heat inactivation, a serine protease inhibitor inhibited anti-BHV-1 activity caused by the natto extract. Thus, our findings provide the first evidence that the natto extract contains a protease(s) that inhibits viral infection through the proteolysis of the viral proteins.


Subject(s)
COVID-19 Drug Treatment , Plant Extracts/pharmacology , SARS-CoV-2/drug effects , Soy Foods , Soybeans/chemistry , Animals , COVID-19/metabolism , COVID-19/pathology , COVID-19/virology , Cattle , Cells, Cultured , Chlorocebus aethiops , Herpesviridae Infections/drug therapy , Herpesviridae Infections/metabolism , Herpesviridae Infections/pathology , Herpesviridae Infections/virology , Herpesvirus 1, Bovine/drug effects , Herpesvirus 1, Bovine/isolation & purification , Herpesvirus 1, Bovine/pathogenicity , Humans , Plant Extracts/chemistry , SARS-CoV-2/isolation & purification , SARS-CoV-2/pathogenicity , Viral Proteins/antagonists & inhibitors , Viral Proteins/metabolism
18.
Bioresour Bioprocess ; 8(1): 58, 2021.
Article in English | MEDLINE | ID: covidwho-1299325

ABSTRACT

Cell-free protein synthesis (CFPS) systems have become an ideal choice for pathway prototyping, protein production, and biosensing, due to their high controllability, tolerance, stability, and ability to produce proteins in a short time. At present, the widely used CFPS systems are mainly based on Escherichia coli strain. Bacillus subtilis, Corynebacterium glutamate, and Vibrio natriegens are potential chassis cells for many biotechnological applications with their respective characteristics. Therefore, to expand the platform of the CFPS systems and options for protein production, four prokaryotes, E. coli, B. subtilis, C. glutamate, and V. natriegens were selected as host organisms to construct the CFPS systems and be compared. Moreover, the process parameters of the CFPS system were optimized, including the codon usage, plasmid synthesis competent cell selection, plasmid concentration, ribosomal binding site (RBS), and CFPS system reagent components. By optimizing and comparing the main influencing factors of different CFPS systems, the systems can be optimized directly for the most influential factors to further improve the protein yield of the systems. In addition, to demonstrate the applicability of the CFPS systems, it was proved that the four CFPS systems all had the potential to produce therapeutic proteins, and they could produce the receptor-binding domain (RBD) protein of SARS-CoV-2 with functional activity. They not only could expand the potential options for in vitro protein production, but also could increase the application range of the system by expanding the cell-free protein synthesis platform. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s40643-021-00413-2.

19.
Appl Environ Microbiol ; 87(3)2021 01 15.
Article in English | MEDLINE | ID: covidwho-1035279

ABSTRACT

Airborne disinfection of high-containment facilities before maintenance or between animal studies is crucial. Commercial spore carriers (CSC) coated with 106 spores of Geobacillus stearothermophilus are often used to assess the efficacy of disinfection. We used quantitative carrier testing (QCT) procedures to compare the sensitivity of CSC with that of surrogates for nonenveloped and enveloped viruses, including severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), mycobacteria, and spores, to an aerosolized mixture of peroxyacetic acid and hydrogen peroxide (aPAA-HP). We then used the QCT methodology to determine relevant process parameters to develop and validate effective disinfection protocols (≥4-log10 reduction) in various large and complex facilities. Our results demonstrate that aPAA-HP is a highly efficient procedure for airborne room disinfection. Relevant process parameters such as temperature and relative humidity can be wirelessly monitored. Furthermore, we found striking differences in inactivation efficacies against some of the tested microorganisms. Overall, we conclude that dry fogging a mixture of aPAA-HP is highly effective against a broad range of microorganisms as well as material compatible with relevant concentrations. Furthermore, CSC are artificial bioindicators with lower resistance and thus should not be used for validating airborne disinfection when microorganisms other than viruses have to be inactivated.IMPORTANCE Airborne disinfection is not only of crucial importance for the safe operation of laboratories and animal rooms where infectious agents are handled but also can be used in public health emergencies such as the current severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic. We show that dry fogging an aerosolized mixture of peroxyacetic acid and hydrogen peroxide (aPAA-HP) is highly microbicidal, efficient, fast, robust, environmentally neutral, and a suitable airborne disinfection method. In addition, the low concentration of dispersed disinfectant, particularly for enveloped viral pathogens such as SARS-CoV-2, entails high material compatibility. For these reasons and due to the relative simplicity of the procedure, it is an ideal disinfection method for hospital wards, ambulances, public conveyances, and indoor community areas. Thus, we conclude that this method is an excellent choice for control of the current SARS-CoV-2 pandemic.


Subject(s)
COVID-19/prevention & control , Disinfectants/pharmacology , Disinfection/methods , Mycobacterium/drug effects , SARS-CoV-2/drug effects , Spores, Bacterial/drug effects , Aerosols , Cell Line , Decontamination/methods , Geobacillus stearothermophilus/drug effects , Hydrogen Peroxide , Particle Size , Peracetic Acid , Steam
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