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Biology subjects

Gorman, B.

Publications and source records attributed to Gorman, B..

4 recordsLinked to original sources

Microbial Influenced Corrosion: A Novel Initiator of In Vivo Spine Rod Fracture

ABSTRACTSpine rod catastrophic failure/rod fracture after implantation is estimated to occur in [~]10% of all cases worldwide with this rate remaining consistent for more than 20 years. To accommodate the spine mechanical environment, metal alloys such as austenitic stainless steels, cobalt-chromium-molybdenum alloys, and /{beta} type titanium alloys such are used as they have a high resistance to fatigue failure. The work presented herein addresses the gap in understanding about what happens in a laboratory environment versus what is happening in human patients that results in metal leaching into tissues as well as a shortened lifespan of a spine rod in a patient population that is not capable of breaking these spine rods from purely mechanical means. Eighty-five (N=85) patients (51 female - 62.9 {+/-} 13.5 years old, 34 male - 62.7 {+/-} 10.9 years old) who had spine revision surgery due to patient mechanical issues, such as loss of sagittal or coronal balance, pain, pedicle screw loosening, hardware associated infection were included in this study. All explanted rods had optical indications of surface modification that were not the result of mechanical damage, e.g. gouges, scratches, notches. The presence of Ti6Al4V rods increases the presence of local tissue concentrations of elemental Ti (190.98 {+/-} 207.84 g/g (ppm)) in all patients over the amount that would normally be present in patients who never had any titanium based orthopedic device implants (spinous muscle, 13.12 {+/-} 11.43 g/g). PQS, HHQ, and NHQ ToF-SIMS molecular signals were found co-located to corrosion pits as well with AHLs and palmitic acids indicating microbial presence and metabolism in all patients. This suggests that MIC can cause in vivo pitting corrosion resulting on rod failure.

bioengineering↗

CRISPR interference in a Streptococcus agalactiae Multi-locus Sequence Type 17 Strain

Group B Streptococcus (GBS), a common colonizer of the human genital and gastrointestinal tracts, is a leading cause of neonatal bacterial meningitis, which can lead to severe neurological complications. The hypervirulent serotype III, sequence type 17 (ST-17) strain COH1 is strongly associated with late-onset disease due to its unique set of virulence factors. However, genetic manipulation of ST-17 strains is notoriously challenging, limiting the ability to study key pathogenic genes. In this study, we developed a CRISPR interference (CRISPRi) system utilizing an endogenous catalytically inactivated Cas9 (dCas9) in the COH1 strain, enabling targeted and tunable gene expression knockdown. We confirmed the efficacy of this system through hemolysis assays, qPCR transcriptional analysis, and in vitro infection models using human brain endothelial cells. The CRISPRi system successfully produced phenotypic knockdowns of essential virulence genes, including pilA, srr2, and iagA, reducing adhesion, invasion, and inflammatory responses at the blood-brain barrier. This platform enables rapid gene knockdowns for functional genomics in ST-17 GBS, enabling high-throughput screening and pathogenesis research. ImportanceGroup B Streptococcus (GBS) remains the worlds leading cause of neonatal meningitis. GBS-host interactions at the blood-brain barrier (BBB) are dependent on bacterial factors, including surface factors and two-component systems. Multi-locus sequence type 17 (ST-17) GBS strains are highly associated with neonatal meningitis, and these strains harbor many virulence factors for infection at the BBB. Historically, these factors have been studied using traditional knockout mutagenesis, which has proven challenging in the most common ST-17 lab strain, COH1. This study utilizes CRISPR interference (CRISPRi) to generate rapid expression knockdown. This study validates a CRISPRi-enabled COH1 dCas9 strain as a versatile tool for probing GBS pathogenesis at the BBB.

microbiology↗

Single-cell and spatiotemporal transcriptomic profiling of brain immune infiltration following Venezuelan equine encephalitis virus infection

Neurotropic alphaviruses such as Venezuelan equine encephalitis virus (VEEV) are critical human pathogens that continually expand to naive populations and for which there are no licensed vaccines or therapeutics. VEEV is highly infectious via the aerosol route and is a recognized weaponizable biothreat that causes neurological disease in humans. The neuropathology of VEEV has been attributed to an inflammatory immune response in the brain yet the underlying mechanisms and specific immune cell populations involved are not fully elucidated. This study uses single-cell RNA sequencing to produce a comprehensive transcriptional profile of immune cells isolated from the brain over a time course of infection in a mouse model of VEEV. Analyses reveal differentially activated subpopulations of microglia, including a distinct type I interferon-expressing subpopulation. This is followed by the sequential infiltration of myeloid cells and cytotoxic lymphocytes, also comprising subpopulations with unique transcriptional signatures. We identify a subpopulation of myeloid cells that form a distinct localization pattern in the hippocampal region whereas lymphocytes are widely distributed, indicating differential modes of recruitment, including that to specific regions of the brain. Altogether, this study provides a high-resolution analysis of the immune response to VEEV in the brain and highlights potential avenues of investigation for therapeutics that target neuroinflammation in the brain. Author SummaryVenezuelan equine encephalitis virus (VEEV) causes brain inflammation in both animals and humans when transmitted by mosquito bite or infectious aerosols. The mechanisms underlying disease caused by VEEV, including the role of the immune response in brain pathology, are not well understood. Here we performed a comprehensive assessment of the immune response to VEEV in the brain over time using two advanced sequencing techniques. Following infection, immune cells infiltrate the brain in a sequential fashion and display different activation profiles. Different types of immune cells also display strikingly different spatial patterns throughout the brain. This study provides the most comprehensive description of the immune response to VEEV in the brain performed to date and advances our understanding of immune-driven neuropathology and identification of therapeutic targets.

immunology↗

Pharmacogenetic allele variant frequencies: An analysis of the VAs Million Veteran Program (MVP) as a representation of the diversity in US population.

We present allele frequencies of pharmacogenomics relevant variants across multiple ancestry in a sample representative of the US population. We analyzed 658,582 individuals with genotype data and extracted pharmacogenomics relevant single nucleotide variant (SNV) alleles, human leukocyte antigens (HLA) 4-digit alleles and an important copy number variant (CNV), the full deletion/duplication of CYP2D6. We compiled distinct allele frequency tables for European, African American, Hispanic, and Asian ancestry individuals. In addition, we compiled allele frequencies based on local ancestry reconstruction in the African-American (2-way deconvolution) and Hispanic (3-way deconvolution) cohorts.

genomics↗