@article{AzamRoesslingGeitheetal.2024, author = {Azam, Hafiz Muhammad Husnain and R{\"o}ßling, Rosa Lise and Geithe, Christiane and Khan, Muhammad Moman and Dinter, Franziska and Hanack, Katja and Pr{\"u}ß, Harald and Husse, Britta and Roggenbruck, Dirk and Schierack, Peter and R{\"o}diger, Stefan}, title = {MicroRNA biomarkers as next-generation diagnostic tools for neurodegenerative diseases : a comprehensive review}, series = {Frontiers in Molecular Neuroscience}, volume = {17}, journal = {Frontiers in Molecular Neuroscience}, publisher = {Frontiers}, address = {Lausanne}, issn = {1662-5099}, doi = {10.3389/fnmol.2024.1386735}, year = {2024}, abstract = {Neurodegenerative diseases (NDs) are characterized by abnormalities within neurons of the brain or spinal cord that gradually lose function, eventually leading to cell death. Upon examination of affected tissue, pathological changes reveal a loss of synapses, misfolded proteins, and activation of immune cells all indicative of disease progression before severe clinical symptoms become apparent. Early detection of NDs is crucial for potentially administering targeted medications that may delay disease advancement. Given their complex pathophysiological features and diverse clinical symptoms, there is a pressing need for sensitive and effective diagnostic methods for NDs. Biomarkers such as microRNAs (miRNAs) have been identified as potential tools for detecting these diseases. We explore the pivotal role of miRNAs in the context of NDs, focusing on Alzheimer's disease, Parkinson's disease, multiple sclerosis, Huntington's disease, and Amyotrophic Lateral Sclerosis. The review delves into the intricate relationship between aging and NDs, highlighting structural and functional alterations in the aging brain and their implications for disease development. It elucidates how miRNAs and RNA-binding proteins are implicated in the pathogenesis of NDs and underscores the importance of investigating their expression and function in aging. Significantly, miRNAs exert substantial influence on post-translational modifications (PTMs), impacting not just the nervous system but a wide array of tissues and cell types as well. Specific miRNAs have been found to target proteins involved in ubiquitination or deubiquitination processes, which play a significant role in regulating protein function and stability. We discuss the link between miRNA, PTM, and NDs. Additionally, the review discusses the significance of miRNAs as biomarkers for early disease detection, offering insights into diagnostic strategies.}, subject = {Neurodegenerative diseases; MicroRNA; Diagnostic tools; Therapeutic tools; Protein post-translational modifications; Nervendegeneration; Biomarker; miRNS; Diagnose; Posttranslationale {\"A}nderung}, language = {en} } @article{KhanMushtaqAbbasetal.2024, author = {Khan, Muhammad Moman and Mushtaq, Muhammad Ahmed and Abbas, Nayyar and Fatima, Fariha and Gibbon, Marjorie J. and Schierack, Peter and Mohsin, Mashkoor}, title = {Occurrence, antimicrobial resistance and genomic features of Klebsiella pneumoniae from broiler chicken in Faisalabad, Pakistan}, series = {Frontiers in Veterinary Science}, volume = {11}, journal = {Frontiers in Veterinary Science}, publisher = {Frontiers}, address = {Lausanne}, issn = {2297-1769}, doi = {10.3389/fvets.2024.1433124}, year = {2024}, abstract = {The dissemination of antimicrobial resistance (AMR) in critical priority pathogens is a significant threat. Non-clinical reservoirs of AMR, such as agriculture and food production facilities, may contribute to the transmission of clinically relevant pathogens such as multidrug-resistant (MDR) Klebsiella pneumoniae. There is currently very limited knowledge regarding the population structure and genomic diversity of K. pneumoniae in poultry production in Pakistan. We explored healthy broilers in a commercial farm from Faisalabad, Pakistan, and identified six K. pneumoniae strains from 100 broiler birds. We characterized the strains, determining clonality, virulence and antimicrobial resistance genes using next generation sequencing. The evaluation of antimicrobial susceptibility revealed that all the strains were MDR. Genomic analysis showed that 3/6 strains belonged to ST152, harbouring acquired resistance aminoglycosides [aadA2, aph(4′)-Ia], β-lactams (blaSHV-187, blaLAP2), fosfomycin (fosA6), tetracycline (tetA), trimethoprim (dfrA12), quinolone (qnrS1), sulphonamides (sul2) and phenicol (floR). All the strains harboured the efflux pump genes oqxA, oqxB, emrR, kpnG, kpnH, kpnF, baeR, mtdB and mtdC. All six strains encoded identical virulence profiles possessing six genes, i.e., ureA, iutA, entB, allS, fimH and mrkD. Phylogenomic analysis of the dominant sequence type (ST152) present in our dataset with publicly available genomes showed that the isolates clustered to strains mainly from human sources and could pose a potential threat to food safety and public health. The combination of these findings with antimicrobial use data would allow a better understanding of the selective pressures that may be driving the spread of AMR. This is the first report of MDR K. pneumoniae isolated from broiler hens in Pakistan, and the finding suggests that routine surveillance of WHO critical priority pathogens in such settings would be beneficial to the development of effective control strategies to reduce AMR.}, subject = {Antimicrobial resistance; Poultry; Genomics; K. pneumoniae; Sequence type}, language = {en} } @article{KhanSidorczukBeckeretal.2024, author = {Khan, Muhammad Moman and Sidorczuk, Katarzyna and Becker, Juliane and Aleksandrowicz, Adrianna and Baraniewicz, Karolina and Ludwig, Christina and Ali, Aamir and Kingsley, Robert A. and Schierack, Peter and Kolenda, Rafał}, title = {Characterization of clumpy adhesion of Escherichia coli to human cells and associated factors influencing antibiotic sensitivity}, series = {Microbiology Spectrum}, volume = {12}, journal = {Microbiology Spectrum}, number = {5}, publisher = {American Society for Microbiology (ASM)}, address = {Washington, DC}, issn = {2165-0497}, doi = {10.1128/spectrum.02606-23}, year = {2024}, abstract = {Escherichia coli intestinal infection pathotypes are characterized by distinct adhesion patterns, including the recently described clumpy adhesion phenotype. Here, we identify and characterize the genetic factors contributing to the clumpy adhesion of E. coli strain 4972. In this strain, the transcriptome and proteome of adhered bacteria were found to be distinct from planktonic bacteria in the supernatant. A total of 622 genes in the transcriptome were differentially expressed in bacteria present in clumps relative to the planktonic bacteria. Seven genes targeted for disruption had variable distribution in different pathotypes and non-pathogenic E. coli, with the pilV and spnT genes being the least frequent or absent from most groups. Deletion (Δ) of five differentially expressed genes, flgH, ffp, pilV, spnT, and yggT, affected motility, adhesion, or antibiotic stress. ΔflgH exhibited 80\% decrease and ΔyggT depicted 184\% increase in adhesion, and upon complementation, adhesion was significantly reduced to 13\%. ΔflgH lost motility and was regenerated when complemented, whereas Δffp had significantly increased motility, and reintroduction of the same gene reduced it to the wild-type level. The clumps produced by Δffp and ΔspnT were more resistant and protected the bacteria, with ΔspnT showing the best clump formation in terms of ampicillin stress protection. ΔyggT had the lowest tolerance to gentamicin, where the antibiotic stress completely eliminated the bacteria. Overall, we were able to investigate the influence of clump formation on cell surface adhesion and antimicrobial tolerance, with the contribution of several factors crucial to clump formation on susceptibility to the selected antibiotics.}, subject = {Escherichia coli; Clumpy adhesion; Motility; Antibiotic tolerance; Stress; Escherichia coli; Zelladh{\"a}sion; Ph{\"a}notyp; Genanalyse}, language = {en} } @article{AliKhanKolendaetal.2023, author = {Ali, Aamir and Khan, Muhammad Moman and Kolenda, Rafal and Olowe, Olugbenga Adekunie and Weinreich, J{\"o}rg and Li, Ganwu and Schierack, Peter}, title = {The role of AJB35136 and fdtA genes in biofilm formation by avian pathogenic Escherichia coli}, doi = {10.1186/s12917-023-03672-7}, year = {2023}, abstract = {Background Infections caused by avian pathogenic Escherichia coli (APEC) result in significant economic losses in poultry industry. APEC strains are known to form biofilms in various conditions allowing them to thrive even under harsh and nutrient-deficient conditions on different surfaces, and this ability enables them to evade chemical and biological eradication methods. Despite knowing the whole genome sequences of various APEC isolates, little has been reported regarding their biofilm-associated genes. A random transposon mutant library of the wild-type APEC IMT 5155 comprising 1,300 mutants was analyzed for biofilm formation under nutrient deprived conditions using Videoscan technology coupled with fluorescence microscopy. Seven transposon mutants were found to have reproducibly and significantly altered biofilm formation and their mutated genes were identified by arbitrary PCR and DNA sequencing. The intact genes were acquired from the wild-type strain, cloned in pACYC177 plasmid and transformed into the respective altered biofilm forming transposon mutants, and the biofilm formation was checked in comparison to the wild type and mutant strains under the same conditions. Results In this study, we report seven genes i.e., nhaA, fdeC, yjhB, lysU, ecpR, AJB35136 and fdtA of APEC with significant contribution to biofilm formation. Reintroduction of AJB35136 and fdtA, reversed the altered phenotype proving that a significant role being played by these two O-antigen related genes in APEC biofilm formation. Presence of these seven genes across nonpathogenic E. coli and APEC genomes was also analyzed showing that they are more prevalent in the latter.}, subject = {APEC; Biofilm; Gene; Complementation; Transposon mutant; VideoScan; Genkomplementierung; Transposon-Mutanten; Gefl{\"u}gelkrankheit; Escherichia coli; Biofilm; Komplementation; Transponson; Mutant}, language = {en} } @phdthesis{Khan2023, author = {Khan, Muhammad Moman}, title = {Escherichia coli colonization}, doi = {10.26127/BTUOpen-6442}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:co1-opus4-64429}, school = {BTU Cottbus - Senftenberg}, year = {2023}, abstract = {To increase our understanding of bacterial intestinal colonization in animal populations lacking anthropogenic influence we devised the first objective and studied the diversity of E. coli in pristine wild animal population. Overall, E. coli in cormorants maintained a high diversity under minimal anthropogenic influences. E. coli were isolated from individual birds of two cormorant colonies located on small islands in lakes at least 100 km away from human settlements. Diversity of the isolates was studied using pulsed field gel electrophoresis (PFGE). 137 isolates of cormorant colony-1 and 75 isolates of cormorant colony-2 resulted in 60 and 33 PFGE types, respectively. Representative strains of each PFGE type were analysed in terms of phylogroups, extraintestinal virulence associated genes (exVAGs), adhesion to the chicken intestinal cell line and antimicrobials. Most isolates belonged to phylogroup B1 (68.3\%) followed by B2 and E with B2 harbouring the highest total number of exVAGs per isolate. Surprisingly, a PFGE type with relatively few exVAGs displayed the highest isolation frequency, also showing a high adhesion rate. Comparative analysis of exVAGs to other E. coli populations of wildlife origin revealed that the secreted autotransporter toxin encoding sat gene was only present in cormorants. The second objective was to characterize and identify factors contributing to the novel adhesion phenotype, known as Clumpy adhesion by the strain 4972 E. coli when incubated on human urinary bladder 5637 cell line. Transcriptome and proteome of adhered bacteria clustered together and diverged from bacteria in the supernatant. The clumpy structure was further explored in terms antibiotic stress where sessile structures could tolerate high antibiotic pressure of ampicillin up to 16 mg/ml and gentamicin up to 2 µg/ml. Data analysis unveiled differential expression of 623 genes between bacteria forming clumps and in supernatant. 148 genes were hypothetical and five hypothetical genes i.e., ffp, espX1, pilV, spnT and yggT along with four known genes i.e., dnaK, spy, flgH and fimH were targeted for deletion. Seven genes deletion mutants, three known (i.e., spy, flgH and fimH) and four hypothetical (ffp, pilV, spnT and yggT) genes were successfully generated and analysed for various parameters. ΔflgH showed loss of clumpy phenotype and adhesion ability up to 80\% was regenerated by complementation. When antibiotic stress by ampicillin was applied after clump formation, Δffp depicted no effect of high concentration of ampicillin. In case of ΔpilV, adhesion significantly increased to 151\% and upon complementation, adhesion was reduced to 53\%. ΔspnT during ampicillin stressed adhesion fared almost 8.5 times better than the wild type strain. ΔyggT exhibited high adhesion of 145.5 \% and upon reintroduction, adhesion decreased to 13\%. Overall, we were able characterize a new phenotype and determined the role of genes contributing to clumpy adhesion phenotype.}, subject = {E. coli; Colonization; Adhesion; Antibiotic resistance; Genes; Kolonisation; Antibiotikaresistenz; Gene; Adh{\"a}sion; Tiere; Darmflora; Escherichia coli; Gen; Antibiotikum; Arzneimittelreistenz}, language = {en} }