Antibiotic Heteroresistance in Klebsiella pneumoniae

Klebsiella pneumoniae is one of the most common pathogens responsible for infections, including pneumonia, urinary tract infections, and bacteremias. The increasing prevalence of multidrug-resistant K. pneumoniae was recognized in 2017 by the World Health Organization as a critical public health threat. Heteroresistance, defined as the presence of a subpopulation of cells with a higher MIC than the dominant population, is a frequent phenotype in many pathogens. Numerous reports on heteroresistant K. pneumoniae isolates have been published in the last few years. Heteroresistance is difficult to detect and study due to its phenotypic and genetic instability. Recent findings provide strong evidence that heteroresistance may be associated with an increased risk of recurrent infections and antibiotic treatment failure. This review focuses on antibiotic heteroresistance mechanisms in K. pneumoniae and potential therapeutic strategies against antibiotic heteroresistant isolates.

Genetic instability of the Omicron strain of the SARS-CoV-2 virus.

Among the mutations found in the Omicron strain, the results of cytosine deamination dominate. There is a mutation in the nsp14 gene. These two facts suggest that the omicron strain has an impaired repair system. The instability of the genome of the Omicron strain to the action of APOBEC deaminases will most likely lead to the degradation of this strain. However, the same mutations have led to several dangerous properties of the Omicron strain. It is proposed to use the instability of the Omicron strain to deamination of cytosine for the prevention of a severe course of the disease.

Cytomic analysis: a modern universal tool for biomedical and ecological and hygienic research (literature review). Part 1

The understanding of the connection between malignant cell transformation and genetic instability has existed for a long time. Such markers of genetic instability as micronuclei (MN) and nuclear abnormalities - nucleoplasmic bridges (NPM) and nuclear buds are signs of malignant growth. However, they were seen only as a by-product of genetic instability, a convenient tool for its study for a long time. Only the studies of recent decades that used the latest methods of molecular genetic analysis (genome sequencing of an individual cell, long-term intravital microscopy and individual chromosomes labelling, hybridization in situ, etc.) have made it possible to establish that the rearrangements of the genetic material in cancer cells are much deeper and more massive than it thought to be. In addition, MN turned out to play an active role in maintaining the state of chromosomal instability in the cell population. This review outlines the current understanding of the processes leading to the emergence of unstable genomes - the phenomenon of «genomic chaos» and its particular case, chromothripsis. The molecular biological features of MN and their role in cellular life and the life of the whole organism are also considered. The significance of MN as diagnostic and prognostic indicators in oncological, neurodegenerative and many other diseases has been analyzed. Much attention is paid to the use of cytome analysis of peripheral blood lymphocytes and human epithelial cells in medical research. It has been suggested that, when used in medical research, cytome analysis can serve as a tool to identify individuals with higher cancer risk. We used the PubMed, Web of Science, ResearchGate, Scopus, eLibrary databases as the sources of literature.

Benign uterine leiomioma with lung lesions

Clinical cases of uterine leiomyoma with secondary lung involvement are described. The results of X-ray, computerized tomography, and histological examination of lung specimens, as well as those of heterozygosity and microsatellite instability are presented. Our own experience and the few descriptions available to date in the literature confirm the pseudo tumorous nature of benign uterine leiomyoma with lung involvement which should be considered as nodular dyshormonal hyperplasia. The signs of genetic instability ՛s identification do not allow to differentiate clearly between the benign or malignant nature of the disease. The key role in the differential diagnosis of uterine leiomyoma with lung involvement and leiomyosarcoma belongs to unprejudiced clinical observation.

Combinatorial assembly platform enabling engineering of genetically stable metabolic pathways in cyanobacteria

Cyanobacteria are simple, efficient, genetically-tractable photosynthetic microorganisms which in principle represent ideal biocatalysts for CO2 capture and conversion. However, in practice, genetic instability and low productivity are key, linked problems in engineered cyanobacteria. We took a massively parallel approach, generating and characterising libraries of synthetic promoters and RBSs for the cyanobacterium Synechocystis sp. PCC 6803, and assembling a sparse combinatorial library of millions of metabolic pathway-encoding construct variants. Genetic instability was observed for some variants, which is expected when variants cause metabolic burden. Surprisingly however, in a single combinatorial round without iterative optimisation, 80% of variants chosen at random and cultured photoautotrophically over many generations accumulated the target terpenoid lycopene from atmospheric CO2, apparently overcoming genetic instability. This large-scale parallel metabolic engineering of cyanobacteria provides a new platform for development of genetically stable cyanobacterial biocatalysts for sustainable light-driven production of valuable products directly from CO2, avoiding fossil carbon or competition with food production.

Prognostic Immunity and Therapeutic Sensitivity Analyses Based on Differential Genomic Instability-Associated LncRNAs in Left- and Right-Sided Colon Adenocarcinoma

Background: The purpose of our study was to develop a prognostic risk model based on differential genomic instability-associated (DGIA) long non-coding RNAs (lncRNAs) of left-sided and right-sided colon cancers (LCCs and RCCs); therefore, the prognostic key lncRNAs could be identified.Methods: We adopted two independent gene datasets, corresponding somatic mutation and clinical information from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) databases. Identification of differential DGIA lncRNAs from LCCs and RCCs was conducted with the appliance of “Limma” analysis. Then, we screened out key lncRNAs based on univariate and multivariate Cox proportional hazard regression analysis. Meanwhile, DGIA lncRNAs related prognostic model (DRPM) was established. We employed the DRPM in the model group and internal verification group from TCGA for the purpose of risk grouping and accuracy verification of DRPM. We also verified the accuracy of key lncRNAs with GEO data. Finally, the differences of immune infiltration, functional pathways, and therapeutic sensitivities were analyzed within different risk groups.Results: A total of 123 DGIA lncRNAs were screened out by differential expression analysis. We obtained six DGIA lncRNAs by the construction of DRPM, including AC004009.1, AP003555.2, BOLA3-AS1, NKILA, LINC00543, and UCA1. After the risk grouping by these DGIA lncRNAs, we found the prognosis of the high-risk group (HRG) was significantly worse than that in the low-risk group (LRG) (all p < 0.05). In all TCGA samples and model group, the expression of CD8+ T cells in HRG was lower than that in LRG (all p < 0.05). The functional analysis indicated that there was significant upregulation with regard to pathways related to both genetic instability and immunity in LRG, including cytosolic DNA sensing pathway, response to double-strand RNA, RIG-Ⅰ like receptor signaling pathway, and Toll-like receptor signaling pathway. Finally, we analyzed the difference and significance of key DGIA lncRNAs and risk groups in multiple therapeutic sensitivities.Conclusion: Through the analysis of the DGIA lncRNAs between LCCs and RCCs, we identified six key DGIA lncRNAs. They can not only predict the prognostic risk of patients but also serve as biomarkers for evaluating the differences of genetic instability, immune infiltration, and therapeutic sensitivity.

Genetic Instability Due to Spindle Anomalies Visualized in Mutants of Dictyostelium

Aberrant centrosome activities in mutants of Dictyostelium discoideum result in anomalies of mitotic spindles that affect the reliability of chromosome segregation. Genetic instabilities caused by these deficiencies are tolerated in multinucleate cells, which can be produced by electric-pulse induced cell fusion as a source for aberrations in the mitotic apparatus of the mutant cells. Dual-color fluorescence labeling of the microtubule system and the chromosomes in live cells revealed the variability of spindle arrangements, of centrosome-nuclear interactions, and of chromosome segregation in the atypical mitoses observed.

Genetic Stability in Melon Transformation System: Assessment by Flow Cytometry and ISSR Markers

Genetic instability in melon species sometimes occurs as a result of in vitro tissue culture and transformation systems. This study describes a new regeneration technique for agrobacterium-mediated co-culture of muskmelon explants (Cucumis melo L. c.v. ‘Khatooni’). Here, no genetic instability was observed in positive PCR regenerants. 4-day-old cotyledonary explants had been infected with LBA4404 Agrobacterium suspensions. The co-cultivation occurred in the presence of 100mg/l rifampicin and 50mg/l kanamycin. The bacteria contained a binary vector pBI121 carrying nopaline synthase by the promoter-neomycin phosphotransferase gene. The regeneration succeeded 65% in selective MS medium containing N6-benzylaminopurine (600 µg/l), β-naphthoxyacetic acid (25 µg/l) and 50mg/l kanamycin in inoculated 4-day-old cotyledonary explants. According to the polymerase chain reaction analysis of neomycin phosphotransferase II gene, transformation was merely successful (8.4%), indicating a substantial miss on a large number of regenerated plants in the selective medium, as a consequence of PGR and antibiotic imbalances. Inter Single Sequence Repeat markers and flow cytometry analyses were used for evaluating the genetic stability and ploidy level of transplants, respectively. The integrated approach underlined that Agrobacterium inoculation and plant growth regulators were successfully combined in vitro to enable muskmelon transformation.

Mechanisms of genetic instability in a single S-phase following whole genome doubling

Doubling of the full chromosome content -whole genome duplications (WGDs)- is frequently found in human cancers and is responsible for the rapid evolution of genetically unstable karyotypes. It has previously been established that WGDs fuel chromosome instability due to abnormal mitosis owing to the presence of extra centrosomes and extra chromosomes. Tolerance to ploidy changes has been identified in different model organisms and cell types, revealing long term cellular adaptations that accommodate ploidy increase. Importantly, however, the immediate consequences of WGDs as cells become tetraploid are not known. It also remains unknown whether WGD triggers other events leading to genetic instability (GIN), independently of mitosis. In this study, we induced tetraploidy in diploid genetically stable RPE-1 cells and monitored the first interphase. We found that newly born tetraploids undergo high rates of DNA damage during DNA replication. Using DNA combing and single cell sequencing, we show that replication forks are unstable, perturbing DNA replication dynamics and generating under- and over-replicated regions at the end of S-phase. Mechanistically, we found that these defects result from lack of protein mass scaling up at the G1/S transition, which impairs the fidelity of DNA replication. This work shows that within a single interphase, unscheduled tetraploid cells can accumulate highly abnormal karyotypes. These findings provide an explanation for the GIN landscape that favors tumorigenesis after tetraploidization.

Immunological and Biochemical Interplay between Cytokines, Oxidative Stress and Schistosomiasis

The host–parasite schistosome relationship relies heavily on the interplay between the strategies imposed by the schistosome worm and the defense mechanisms the host uses to counter the line of attack of the parasite. The ultimate goal of the schistosome parasite entails five important steps: evade elimination tactics, survive within the human host, develop into adult forms, propagate in large numbers, and transmit from one host to the next. The aim of the parasitized host on the other hand is either to cure or limit infection. Therefore, it is a battle between two conflicting aspirations. From the host’s standpoint, infection accompanies a plethora of immunological consequences; some are set in place to defend the host, while most end up promoting chronic disease, which ultimately crosses paths with oxidative stress and cancer. Understanding these networks provides attractive opportunities for anti-schistosome therapeutic development. Hence, this review discusses the mechanisms by which schistosomes modulate the human immune response with ultimate links to oxidative stress and genetic instability.