Biology subjects

Pongchaikul, P.

Publications and source records attributed to Pongchaikul, P..

4 recordsLinked to original sources

bioRxiv2025

Mechanism of neuroinvasion and interferon response in human choroid plexus organoids during Japanese encephalitis virus infection

Orthoflaviviruses are medically important arthropod-borne pathogens that cause a significant global disease burden. Some of these viruses can cause neurological infections, requiring a clear understanding of how they enter the central nervous system (CNS). The choroid plexus (ChP) is a cerebrospinal fluid (CSF)-producing tissue residing in the brain ventricles. The ChP forms a blood-CSF barrier, which acts as a crucial peripheral-CNS interface and a physical defense against neuroinvasive pathogens. While this tissue is increasingly recognized as a potential entry site for hematogenous pathogens, how orthoflaviviruses overcome the blood-CSF barrier and the corresponding intrinsic immune response in a human model remains critically undefined. This study aimed to investigate the potential of the ChP to serve as a neuroinvasive site for orthoflaviviruses. ChP organoids (ChPOs) derived from human inducible pluripotent stem cells were established and determined for susceptibility to a panel of orthoflaviviruses, namely Japanese encephalitis virus (JEV), Zika virus (ZIKV), and Dengue virus (DENV). These viruses exhibited differential susceptibility to ChPOs, with the pathogenic JEV strain being the most infective. Transcriptomic analysis of ChPOs confirmed a robust interferon (IFN) response to JEV infection. Functional studies further demonstrated that antiviral defenses are crucial for controlling virus infection, and that type I IFN responses are superior to the epithelial-restricted type III IFN responses. Critically, despite the productive release of JEV into the CSF-like fluid within the ChP lumen, selective barrier integrity was maintained, confirming a nondestructive transcellular neuroinvasion mechanism across the blood-CSF barrier. This study highlights a valuable in vitro platform and its potential for studying virus neuroinvasion at the blood-CSF barrier.

microbiology↗

bioRxiv2019

A megaplasmid family responsible for dissemination of multidrug resistance in Pseudomonas

Multidrug resistance (MDR) represents a global threat to health. Although plasmids can play an important role in the dissemination of MDR, they have not been commonly linked to the emergence of antimicrobial resistance in the pathogen Pseudomonas aeruginosa. We used whole genome sequencing to characterize a collection of P. aeruginosa clinical isolates from a hospital in Thailand. Using long-read sequence data we obtained complete sequences of two closely related megaplasmids (>420 kb) carrying large arrays of antibiotic resistance genes located in discrete, complex and dynamic resistance regions, and revealing evidence of extensive duplication and recombination events. A comprehensive pangenomic and phylogenomic analysis indicated that 1) these large plasmids comprise a family present in different members of the Pseudomonas genus and associated with multiple sources (geographical, clinical or environmental); 2) the megaplasmids encode diverse niche-adaptive accessory traits, including multidrug resistance; 3) the pangenome of the megaplasmid family is highly flexible and diverse, comprising a substantial core genome (average of 48% of plasmid genes), but with individual members carrying large numbers of unique genes. The history of the megaplasmid family, inferred from our analysis of the available database, suggests that members carrying multiple resistance genes date back to at least the 1970s.\n\nFundingThis work was supported by the International Pseudomonas Genomics Consortium, funded by Cystic Fibrosis Canada [RCL]; and the Secretaria de Educacion, Ciencia, Tecnologia e Innovacion (SECTEI), Mexico [AC].

microbiology↗

bioRxiv2018

Comparative Genomics of Staphylococcus Reveals Determinants of Speciation and Diversification of Antimicrobial Defense.

The bacterial genus Staphylococcus comprises diverse species with most being described as colonizers of human and animal skin. A relational analysis of features that discriminate its species and contribute to niche adaptation and survival remains to be fully described. In this study, an interspecies, whole-genome comparative analysis of 21 Staphylococcus species was performed based on their orthologues. Three well-defined multi-species groups were identified: group A (including aureus/epidermidis); group B (including saprophyticus/xylosus) and group C (including pseudintermedius/delphini). The machine learning algorithm Random Forest was applied to identify variable orthologues that drive formation of the Staphylococcus species groups A-C. Orthologues driving staphylococcal infrageneric diversity comprised regulatory, metabolic and antimicrobial resistance proteins. Notably, the BraSR (NsaRS) two-component system (TCS) and its associated BraDE transporters that regulate antimicrobial resistance distinguish group A Staphylococcus species from others in the genus that lack the BraSR TCS. Divergence of BraSR and GraSR antimicrobial peptide survival TCS and their associated transporters was observed across the staphylococci, likely reflecting niche specific evolution of these TCS/transporters and their specificities for AMPs. Experimental evolution, with selection for resistance to the lantibiotic nisin, revealed multiple routes to resistance and differences in the selection outcomes of the BraSR- positive species S. hominis and S. aureus. Selection supported a role for GraSR in nisin survival responses of the BraSR-negative group B species S. saprophyticus. Our study reveals diversification of antimicrobial-sensing TCS across the staphylococci and hints at differential relationships between GraSR and BraSR in those species positive for both TCS.\n\nImportanceThe genus Staphylococcus includes species that are commensals and opportunist pathogens of humans and animals. Identifying the features that discriminate species of staphylococci is relevant to understanding niche selection and the structure of their microbiomes. Moreover, the determinants that structure the community are relevant for strategies to modify the frequency of individual species associated with dysbiosis and disease. In this study, we identify orthologous proteins that discriminate genomes of staphylococci. In particular, species restriction of a major antimicrobial survival system, BraSR (NsaRS), to a group of staphylococci dominated by those that can colonize human skin. The diversity of antimicrobial sensing loci was revealed by comparative analysis and experimental evolution with selection for nisin resistance identified the potential for variation in antimicrobial sensing in BraRS-encoding staphylococci. This study provides insights into staphylococcal species diversity.

microbiology↗

bioRxiv2017

Large scale and significant expression from pseudogenes in Sodalis glossinidius - a facultative bacterial endosymbiont

The majority of bacterial genomes have high coding efficiencies, but there are some genomes of intracellular bacteria that have low gene density. The genome of the endosymbiont Sodalis glossinidius contains almost 50% pseudogenes containing mutations that putatively silence them at the genomic level. We have applied multiple omic strategies, combining: Illumina and Pacific Biosciences Single-Molecule Real Time DNA-sequencing and annotation; stranded RNA-sequencing; and proteome analysis to better understand the transcriptional and translational landscape of Sodalis pseudogenes, and potential mechanisms for their control. Between 53% and 74% of the Sodalis transcriptome remains active in cell-free culture. Mean sense transcription from Coding Domain Sequences (CDS) is four-times greater than that from pseudogenes. Comparative genomic analysis of six Illumina-sequenced Sodalis isolates from different host Glossina species shows pseudogenes make up ~40% of the 2,729 genes in the core genome, suggesting that they are stable and/or Sodalis is a recent introduction across the Glossina genus as a facultative symbiont. These data further shed light on the importance of transcriptional and translational control in deciphering host-microbe interactions, and demonstrate that pseudogenes are more complex than a simple degrading DNA sequence. The combination of genomics, transcriptomics and proteomics give a multidimensional perspective for studying prokaryotic genomes with a view to elucidating evolutionary adaptation to novel environmental niches.\n\nImportanceBacterial genomes are generally 1Kb in length, organized efficiently (i.e. with few gaps between genes or operons), and few open reading frames (ORFs) lack any predicted function. Intracellular bacteria have been removed from extracellular selection pressures acting on pathways of declining importance to fitness and thus, these bacteria tend to delete redundant genes in favour of smaller functional repertoires - maintaining genome efficiency. In the genomes of endosymbionts with a recent evolutionary relationship with their host, however, this process of genome reduction is not complete; Genes and pathways may be at an intermediate stage, undergoing mutation linked to reduced selection and small population numbers being vertically transmitted from mother to offspring in their hosts, resulting in an increase in abundance of pseudogenes and reduced coding capacities. A greater knowledge of the genomic architecture of persistent pseudogenes, with respect to their DNA structure, mRNA transcription and even putative translation to protein products, will lead to a better understanding of the evolutionary trajectory of endosymbiont genomes, many of which have important roles in arthropod ecology.

genomics↗