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New Biotechnology Dec 2022We have developed a novel urea-inducible recombinant protein production system by exploiting the Proteus mirabilis urease ureR-ureD promoter region and the ureR...
We have developed a novel urea-inducible recombinant protein production system by exploiting the Proteus mirabilis urease ureR-ureD promoter region and the ureR AraC-family transcriptional regulator. Experiments using the expression of β-galactosidase and green fluorescent protein (GFP) showed that promoter activity is tightly regulated and that varying the concentration of urea can give up to 100-fold induction. Production of proteins of biopharmaceutical interest has been demonstrated, including human growth hormone (hGH), a single chain antibody fragment (scFv) against interleukin-1β and a potential Neisserial vaccine candidate (BamA). Expression levels can be fine-tuned by temperature and different urea concentrations, and can be induced with readily available garden fertilisers and even urine. As urea is an inexpensive, stable inducer, a urea-induced expression system has the potential to considerably reduce the costs of large-scale recombinant protein production.
Topics: Humans; Urea; Bacterial Proteins; Escherichia coli; Proteus mirabilis; Recombinant Proteins; Bacterial Outer Membrane Proteins; Escherichia coli Proteins
PubMed: 36273806
DOI: 10.1016/j.nbt.2022.10.003 -
Journal of the American Association For... May 2022The exclusion of opportunistic pathogens is important for protecting animal health and ensuring desired research outcomes in highly immunodeficient mice. has been...
The exclusion of opportunistic pathogens is important for protecting animal health and ensuring desired research outcomes in highly immunodeficient mice. has been associated with gastrointestinal tract lesions, septicemia, pyelonephritis, splenomegaly, and hepatitis and can influence select mouse models. To inform health-surveillance practices after we experienced difficulty in excluding from our mouse colony, we aimed to determine the likelihood of detecting -positive immunocompromised (SRG), immunovague (), and immunocompetent (CD1) colony mice through culture and PCR testing; to evaluate transmission via 2 sentinel-based approaches (direct contact and indirect dirty-bedding transfer); and to further characterize associated pathology. We hypothesized that immunocompromised mice would be better detectors and transmitters of . Multiple logistic regression models were used for analysis and included PCR copy number, repeated testing, age, sex, and antibiotic-treated (trimethoprim-sulfamethoxazole) diet as covariates. Repeated testing over 10 wk showed that -colonized immunocompromised colony mice were 95 times more likely than immunocompetent mice to test positive by culture and 30 times more likely by PCR assay. Sentinel mice were 15 times more likely to test positive by PCR assay for when exposed by direct contact compared with dirty bedding and 18 times more likely to test positive when exposed to positive immunocompromised as compared with immunocompetent colony mice. After 10 wk of exposure, 3.8% of dirty-bedding sentinel PCR tests were positive, as compared with 30.7% of contact sentinels. Only immunocompromised mice on antibiotic diet (37.5%) developed lesions of the urogenital tract and abdominal cavity consistent with known pathology of . Our findings suggest that PCR testing of dirty-bedding sentinels alone is not sufficient for the detection of in mouse colonies. Direct-contact sentinels and testing of colony mice-especially if immunocompromised-with adjunct culture may facilitate successful bioexclusion.
Topics: Animals; Anti-Bacterial Agents; Bedding and Linens; Housing, Animal; Mice; Proteus mirabilis
PubMed: 35277210
DOI: 10.30802/AALAS-JAALAS-21-000104 -
Archives of Razi Institute Dec 2022belongs to the family Enterobacteriaceae and is capable of transforming in shape from rod to elongated and swarming motility by flagella. It is an opportunity for...
belongs to the family Enterobacteriaceae and is capable of transforming in shape from rod to elongated and swarming motility by flagella. It is an opportunity for bacteria and can cause different clinical diseases. Therefore, this study aimed to assay and detect a sequence of genes that encode for antibiotic resistance in multidrug resistance clinical isolates of , including , , qnrA, , and secondly to investigate the relationship in the phylogenetic tree among these genes in Iraq comparison with global strains in NCBI. The study included the identifying of 500 clinical samples depending on morphological and biochemical tests and confirming diagnosis by the VITEK-2 Compact system. The confirmed isolates of were 95 clinical isolates (19%). Antibiotic susceptibility test of all these isolates was done using twelve antibiotics tested using Amoxicillin, Aztreonam, Imipenem, Cefoxitin, Amikacin, Ceftazidem, Ciprofloxacin, Nalidixic acid, Gentamicin, Sulphamethazol-trimethoprim, Cefotaxime, Amoxicillin-clavulanic acid. The results showed that multidrug resistance isolates contained the genes in different levels as follow gene (90%), gene (80%) , gene (100%), gene (80%). These genes were sequenced and detected phylogenetic relationships among these genes and global genes were documented in NCBI. The results showed that some Iraqi isolates contain genetic variation compared to global strains. Therefore, this variation was detected and registered in NCBI of all five antibiotic resistance genes mentioned above and accepted under accession numbers of gene (LC613168.1), gene (LC613166.1), gene (LC613169.1), gene (LC613170.1).
Topics: Anti-Bacterial Agents; Drug Resistance, Multiple, Bacterial; Phylogeny; Proteus Infections; Proteus mirabilis
PubMed: 37274906
DOI: 10.22092/ARI.2022.358489.2230 -
Clinical Microbiology and Infection :... Sep 2023To analyse carbapenemases in Proteus mirabilis and assess the performance of carbapenemase detection assays.
OBJECTIVES
To analyse carbapenemases in Proteus mirabilis and assess the performance of carbapenemase detection assays.
METHODS
Eighty-one clinical P. mirabilis isolates with high-level resistance at least to ampicillin (>32 mg/L) or previous detection of carbapenemases were selected and investigated by three susceptibility testing methods (microdilution, automated susceptibility testing, and disk diffusion), six phenotypic carbapenemase assays (CARBA NP, modified carbapenemase inactivation method [CIM], modified zinc-supplemented CIM, simplified CIM, faropenem, and carbapenem-containing agar), two immunochromatographic assays, and whole-genome sequencing.
RESULTS
Carbapenemases were detected in 43 of 81 isolates (OXA-48-like [n = 13]; OXA-23 [n = 12]; OXA-58 [n = 12]; New Delhi metallo-β-lactamase (NDM) [n = 2]; Verona integron-encoded metallo-β-lactamase (VIM) [n = 2]; Imipenemase (IMP) [n = 1]; Klebsiella pneumoniae carbapenemase (KPC) [n = 1]). Carbapenemase-producing Proteus were frequently susceptible to ertapenem (26/43; 60%), meropenem (28/43; 65%), ceftazidime (33/43; 77%), and some even to piperacillin-tazobactam (9/43; 21%). Sensitivity/specificity of phenotypic tests were 30% (CI: 17-46%)/89% (CI: 75-97%) for CARBA NP, 74% (CI: 60-85%)/82% (CI: 67-91%) for faropenem, 91% (CI: 78-97%)/82% (CI: 66-92%) for simplified CIM, and 93% (CI: 81-99%)/100% (CI: 91-100%) for modified zinc-supplemented CIM. An algorithm for improved detection was developed, which demonstrated sensitivity/specificity of 100% (CI: 92-100%)/100% (CI: 91-100%) on the 81 isolates, and 100% (CI: 29-100%)/100% (CI: 96-100%) in a prospective analysis of additional 91 isolates. Interestingly, several OXA-23-producing isolates belonged to the same clonal lineage reported previously from France.
DISCUSSION
Current susceptibility testing methods and phenotypic tests frequently fail to detect carbapenemases in P. mirabilis, which could result in inadequate antibiotic treatment. In addition, the non-inclusion of bla in many molecular carbapenemase assays further impedes their detection. Therefore, the prevalence of carbapenemases in P. mirabilis is likely underestimated. With the herein proposed algorithm, carbapenemase-producing Proteus can be easily identified.
Topics: Humans; Proteus mirabilis; Bacterial Proteins; beta-Lactamases; Anti-Bacterial Agents; Algorithms; Zinc; Microbial Sensitivity Tests
PubMed: 37271195
DOI: 10.1016/j.cmi.2023.05.032 -
Orphanet Journal of Rare Diseases Feb 2024Proteus syndrome is an ultra-rare mosaic overgrowth disorder. Individuals with Proteus syndrome can develop emphysematous and cystic changes of the lung that may lead to...
BACKGROUND
Proteus syndrome is an ultra-rare mosaic overgrowth disorder. Individuals with Proteus syndrome can develop emphysematous and cystic changes of the lung that may lead to progressive respiratory symptoms and require surgical intervention. This retrospective study seeks to quantify the radiographic features of Proteus syndrome-associated lung disease using computed tomography (CT) of the chest. The first method derives a Cystic Lung Score (CLS) by using a computer-aided diagnostic tool to quantify the fraction of cystic involvement of the lung. The second method yields a Clinician Visual Score (CVS), an observer reported scale of severity based on multiple radiographic features. The aim of this study was to determine if these measurements are associated with clinical symptoms, pulmonary function test (PFT) measurements, and if they may be used to assess progression of pulmonary disease.
RESULTS
One hundred and thirteen imaging studies from 44 individuals with Proteus syndrome were included. Dyspnea and oxygen use were each associated with higher CLS (p = 0.001 and < 0.001, respectively) and higher CVS (p < 0.001 and < 0.001). Decreases in percent predicted FVC, FEV, and DLCO each correlated with increased CLS and CVS. The annual increase of CLS in children, 5.6, was significantly greater than in adults, 1.6. (p = 0.03). The annual increase in CVS in children, 0.4, was similar to adults, 0.2 (p = 0.36).
CONCLUSIONS
Proteus syndrome-associated lung disease is progressive. The rate of cystic progression is increased in children. Increased scores in CLS and CVS were associated with clinical symptoms and decreased pulmonary function. Both methods were able to detect change over time and were associated with clinically meaningful outcomes which may enable their use in interventional studies.
Topics: Adult; Child; Humans; Proteus Syndrome; Retrospective Studies; Lung; Tomography, X-Ray Computed; Lung Diseases
PubMed: 38321508
DOI: 10.1186/s13023-023-03013-9 -
Frontiers in Cellular and Infection... 2022Indwelling urinary catheterization can lead to the development of catheter-associated urinary tract infections (CAUTIs), an important type of nosocomial infection, as... (Review)
Review
Indwelling urinary catheterization can lead to the development of catheter-associated urinary tract infections (CAUTIs), an important type of nosocomial infection, as well as other medical issues among institutionalized adults. Recently, was highlighted as the important cause of CAUTIs. The pathogenicity of is dependent on two multicellular types of surface colonization: the adherence and swarming motility. Adhesion, mostly mediated by fimbrial and nonfimbrial adhesins, is important for the initiation of biofilm formation. Moreover, the production of urease frequently results in biofilm crystallization, which leads to the blockage of catheters. The heterologous polymeric matrix of the biofilm offers protection against antibiotics and the host immune system. displays remarkable motility abilities. After contact with solid surfaces, hyper-flagellated cells are able to rapidly migrate. The importance of swarming motility in CAUTIs development remains controversial; however, it was indicated that swarming cells were able to co-express other virulence factors. Furthermore, flagella are strong immunomodulating proteins. On the other hand, both biofilm formation and swarming motility implicates multiple inter- and intraspecies interactions, which might contribute to the pathogenicity.
Topics: Anti-Bacterial Agents; Humans; Life Style; Proteus mirabilis; Urease; Urinary Tract Infections; Virulence Factors
PubMed: 36118021
DOI: 10.3389/fcimb.2022.864305 -
Human Molecular Genetics Sep 2019Proteus syndrome is a mosaic, progressive overgrowth disorder caused by a somatic activating variant c.49G > A p.(E17K) in AKT1. The presentation in affected...
Proteus syndrome is a mosaic, progressive overgrowth disorder caused by a somatic activating variant c.49G > A p.(E17K) in AKT1. The presentation in affected individuals is variable, with a diversity of tissues demonstrating abnormalities. Common manifestations include skin and bony overgrowth, vascular malformations (VMs), cysts and benign tumors. We used two methods to create mouse models that had endogenously-regulated mosaic expression of the Proteus syndrome variant. Variant allele fractions (VAFs) ranged from 0% to 50% across numerous tissues in 44 Proteus syndrome mice. Mice were phenotypically heterogeneous with lesions rarely observed before 12 months of age. VMs were the most frequent finding with a total of 69 found in 29 of 44 Proteus syndrome mice. Twenty-eight cysts and ectasia, frequently biliary, were seen in 22 of 44 Proteus syndrome mice. Varying levels of mammary hyperplasia were seen in 10 of 16 female Proteus syndrome mice with other localized regions of hyperplasia and stromal expansion noted in several additional animals. Interestingly, 27 of 31 Proteus syndrome animals had non-zero blood VAF that is in contrast to the human disorder where it is rarely seen in peripheral blood. Identification of variant-positive cells by green fluorescent protein (GFP) staining in chimeric Proteus syndrome mice showed that in some lesions, hyperplastic cells were predominantly GFP/Akt1E17K-positive and showed increased pAKT signal compared to GFP-negative cells. However, hyperplastic mammary epithelium was a mixture of GFP/Akt1E17K-positive and negative cells with some GFP/Akt1E17K-negative cells also having increased pAKT signal suggesting that the variant-positive cells can induce lesion formation in a non-cell autonomous manner.
Topics: Alleles; Animals; Biopsy; Disease Models, Animal; Genetic Association Studies; Genetic Loci; Genetic Predisposition to Disease; Genotype; Humans; Mice; Mutation; Phenotype; Proteus Syndrome; Proto-Oncogene Proteins c-akt
PubMed: 31194862
DOI: 10.1093/hmg/ddz116 -
Virus Research Mar 2023The crystalline formation of biofilms by Proteus blocks the urine flow which often complicates the health care of catheterized patients. Bacteriophages has been...
The crystalline formation of biofilms by Proteus blocks the urine flow which often complicates the health care of catheterized patients. Bacteriophages has been highlighted as a promising tool to control biofilm-mediated bacterial infections. Here, we isolated and characterized two newly isolated lytic phages capable of infecting clinical isolates of P. mirabilis and P. vulgaris. Moreover, insights regarding the biological and molecular characterization were analysed. Both RP6 and RP7 phages showed a Proteus-genus-specific profile, administering no lytic activity against other family of Enterobacteriaceae. The optimal MOI value of the RP6 and RP7 phages were determined as 0.1 and 0.01, respectively. The one-step growth curve showed that RP6 and RP7 phages have a short latent period of 20 min and large burst size of 220-371 PFU/ML per infected host cell. Bacteria growth was reduced immediately after the phages were added, which is shown by the optical density (OD) measurement after 24 hr. Proteus phage RP6 and RP7 were found to eradicate both the planktonic and mature biofilms produced by the Proteus isolates tested. Genome sequence of Proteus phage RP6 was found to be 58,619 bp, and a G-C content of 47%. Also, Proteus phage RP7 genome size was 103,593 bp with G-C ratio of 38.45%. A total of 70 and 172 open reading frame (ORF) was encoded in RP6 and RP7 phage genomes, respectively. Interestingly, there were no tRNA encoded by Proteus phage RP6 genome even though there is a significant G-C content difference between the phage and its host. Additionally, the exhibition of highly lytic activity and absence of virulence and antibiotic-resistant genes in both Proteus RP6 and RP7 phages emphasized that this newly isolated phages are promising for potential therapeutic phages.
Topics: Humans; Bacteriophages; Sewage; Proteus; Proteus mirabilis; Genome, Viral; Biofilms; Hospitals
PubMed: 36717023
DOI: 10.1016/j.virusres.2023.199049 -
European Journal of Obstetrics,... Feb 2022Urinary tract infections (UTI) among women form a substantial part of medical practice and both patients and medical professionals have an interest in non-antibiotic...
OBJECTIVE
Urinary tract infections (UTI) among women form a substantial part of medical practice and both patients and medical professionals have an interest in non-antibiotic treatments and preventative measures. This research provides preliminary data on a multi-functional composition, DAPAD, which explored several biologic activities of relevance to UTI.
STUDY DESIGN
This formulation included D-mannose, citric acid, three prebiotic compounds, and extracts of dandelion and astragalus. Studies performed employed 4 bacterial strains that have relevance to UTI including E. coli, Proteus mirabilis, Streptococcus agalactiae and Enterococcus faecalis.
RESULTS
Key findings from in vitro studies included: DAPAD at full- and half-strength inhibited growth of all UTI bacteria. Evidence for D-mannose agglutination of E. coli was demonstrated. D-mannose also showed unexpected effects on bacterial membrane integrity with vital staining and modest growth restriction. We did not demonstrate growth inhibition by dandelion or astragalus extracts but the latter showed diminished cytokine elaboration by bladder epithelial cells.
CONCLUSION
DAPAD is a multifunctional composition that may warrant further development as a UTI treatment or preventive if supported by clinical evaluation.
Topics: Anti-Bacterial Agents; Enterococcus faecalis; Escherichia coli; Female; Humans; Proteus mirabilis; Urinary Tract Infections
PubMed: 34979364
DOI: 10.1016/j.ejogrb.2021.12.031 -
Scientific Reports Feb 2021Muscle is an important structural tissue in aquatic animals and it is susceptible to bacterial and fungal infection, which could affect flesh quality and health. In this...
Investigation of Proteus vulgaris and Elizabethkingia meningoseptica invasion on muscle oxidative stress and autophagy in Chinese soft-shelled turtle (Pelodiscus sinensis).
Muscle is an important structural tissue in aquatic animals and it is susceptible to bacterial and fungal infection, which could affect flesh quality and health. In this study, Chinese soft-shelled turtles were artificially infected with two pathogens, Proteus vulgaris and Elizabethkingia meningoseptica and the effects on muscle nutritional characteristics, oxidative stress and autophagy were assayed. Upon infection, the muscle nutritional composition and muscle fiber structure were notably influenced. Meanwhile, the mRNA expression of Nrf2 was down-regulated and Keap1 up-regulated, thus resulting in a decrease in antioxidant capacity and oxidative stress. However, with N-acetylcysteine treatment, the level of oxidative stress was decreased, accompanied by significant increases in antioxidant enzyme activities and the mRNA levels of SOD, CAT, GSTCD, and GSTO1. Interestingly, there was a significant increase in autophagy in the muscle tissue after the pathogen infection, but this increase could be reduced by N-acetylcysteine treatment. Our findings suggest that muscle nutritional characteristics were dramatically changed after pathogen infection, and oxidative stress and autophagy were induced by pathogen infection. However, N-acetylcysteine treatment could compromise the process perhaps by decreasing the ROS level and regulating Nrf2-antioxidant signaling pathways.
Topics: Acetylcysteine; Animals; Antioxidants; Autophagy; China; Flavobacteriaceae; Flavobacteriaceae Infections; Muscles; Oxidative Stress; Proteus vulgaris; Signal Transduction; Turtles
PubMed: 33574492
DOI: 10.1038/s41598-021-83388-6