Biology subjects

Bailoo, J. D.

Publications and source records attributed to Bailoo, J. D..

4 recordsLinked to original sources

bioRxiv2025

Molecular mechanisms of 10-Butyl Ether Minocycline (BEM), a novel non-antibiotic tetracycline, as a potential treatment for inflammatory and neuroimmune-related disorders.

The pleiotropy of minocycline (MINO), including anti-inflammatory, antioxidant, anti-migratory, anti-MMP, and neuroprotective effects, has been extensively reported. A novel non-antibiotic minocycline derivative, 10-butyl ether minocycline (BEM), was synthesized to retain the pleiotropy of minocycline while minimizing side effects such as antibiotic resistance and gut dysbiosis. Previously, we showed that BEM reduced alcohol consumption in dependent murine and porcine models of Alcohol Use Disorder (AUD). In this study, we investigated the molecular mechanisms of BEM to determine its potential as a therapeutic agent for neuroimmune and inflammatory conditions such as AUD. Here, we report that BEM showed a nearly complete loss of antimicrobial activity against E. coli, S. typhi, and C. albicans. BEM showed a dose-dependent reduction in cell viability as measured by the MTT assay, similarly to MINO. BEM also suppressed LPS-induced microglial activation as shown by reduced Iba1 expression in immunohistochemistry and western blot analyses. Inhibition of MMP-9 by BEM (IC50 = 42.2 {micro}M) was improved compared to MINO (IC50 = 60.3 {micro}M) while MMP-8 inhibition was moderate (IC50: BEM = 69.4 {micro}M; MINO = 45.4 {micro}M). BEM was found to be effective in inhibiting VEGF-induced endothelial cell migration and L-glutamine-induced ROS levels. Limited inhibition of 15-LOX activity was observed (IC50: BEM = 92.6 {micro}M; MINO = 65.6 {micro}M). BEM was not toxic to mitochondria, even at high concentrations (200 {micro}M). By eliminating antimicrobial properties while preserving therapeutic pleiotropy, BEM presents an advancement in the development of a promising candidate with multimodal mechanisms to treat neuroimmune-inflammatory pathologies. ImpactOur current approach to correlate non-antibacterial actions of BEM to MINOs established mechanistic effects will enable the informed use of BEM for several medical indications including for inflammation and neuroimmune conditions. The focus on BEMs multimodal actions and long-term safety during drug discovery represents a paradigm shift toward complex therapeutic drug development and repositioning, improving upon traditional singular high-affinity target-based approaches. Such new drug discovery attempts could potentially enhance treatment relevance in complex disorders with multiple targets and theoretically guide the creation of second-generation analogs. Significance statementWe report mechanisms of action for BEM, a minocycline analog under evaluation for the treatment of Alcohol Use Disorder, which may also show efficacy for other complex disease processes that involve inflammatory or neuroimmune components. We show that BEM had a nearly complete loss of antimicrobial action, yet retained the pleiotropy of MINO, likely making it a better multimodal therapeutic for long-term treatment of complex diseases with neuroimmune-related components. Visual abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=120 SRC="FIGDIR/small/661183v3_ufig1.gif" ALT="Figure 1"> View larger version (54K): org.highwire.dtl.DTLVardef@1ade11corg.highwire.dtl.DTLVardef@961d1borg.highwire.dtl.DTLVardef@15101f8org.highwire.dtl.DTLVardef@158810e_HPS_FORMAT_FIGEXP M_FIG C_FIG

pharmacology and toxicology↗

bioRxiv2024

Suspect screening-data independent analysis workflow for the identification of arsenolipids in marine standard reference materials

There has been limited research into arsenolipid toxicological risks and health-related outcomes due to challenges with their separation, identification, and quantification within complex biological matrices (e.g., fish, seaweed). Analytical approaches for arsenolipid identification such as suspect screening have not been well documented and there are no certified standard reference materials, leading to issues with reproducibility and uncertainty regarding the accuracy of results. In this study, a detailed workflow for the identification of arsenolipids utilizing suspect screening coupled with data independent analysis is presented and applied to three commercially available standard reference materials (Hijiki seaweed, dogfish liver, and tuna). Hexane and dichloromethane/methanol extraction, followed by reversed-phase high-performance liquid chromatography-inductively coupled plasma mass spectrometry and liquid chromatography-electrospray ionization-quadrupole time-of-flight mass spectrometry. Using the workflow developed, mass fragmentation matching, mass error calculations, and retention time matching were performed to identify suspect arsenolipids. Arseno-fatty acids (AsFAs), arsenohydrocarbons (AsHCs), and arsenosugar phospholipids (AsSugPLs) were identified with high confidence; AsHC332, AsHC360, and AsSugPL720 in seaweed, AsHC332 in tuna, and AsFA474 and AsFA502 in the dogfish liver. AsHC332, AsHC360, and AsFA502 were identified as promising candidates for further work on synthesis, quantification using MS/MS, and toxicity testing.

pharmacology and toxicology↗

bioRxiv2023

The Horizontal Ladder Test (HLT) protocol: A novel, optimized, and reliable means of assessing motor coordination in Sus scrofa domesticus

Pigs can be an important model for preclinical biological research, including neurological diseases such as Alcohol Use Disorder. Such research often involves longitudinal assessment of changes in motor coordination as the disease or disorder progresses. Current motor coordination tests in pigs are derived from behavioral assessments in rodents and lack critical aspects of face and construct validity. While such tests may permit for the comparison of experimental results to rodents, a lack of validation studies of such tests in the pig itself may preclude the drawing of meaningful conclusions. Here, we present a novel, optimized, and reliable horizontal ladder test (HLT) test protocol for evaluating motor coordination in pigs and an initial validation of its construct validity using voluntary alcohol consumption as an experimental manipulation. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=157 SRC="FIGDIR/small/571517v2_ufig1.gif" ALT="Figure 1"> View larger version (101K): org.highwire.dtl.DTLVardef@167924eorg.highwire.dtl.DTLVardef@b674f2org.highwire.dtl.DTLVardef@b7c369org.highwire.dtl.DTLVardef@7e5a42_HPS_FORMAT_FIGEXP M_FIG C_FIG

animal behavior and cognition↗

bioRxiv2023

ATP1A1-linked diseases require a malfunctioning protein product from one allele

Heterozygous germline variants in ATP1A1, the gene encoding the 1 subunit of the Na+/K+-ATPase (NKA), have been linked to diseases including primary hyperaldosteronism and the peripheral neuropathy Charcot-Marie-Tooth disease (CMT). ATP1A1 variants that cause CMT induce loss-of-function of NKA. This heterodimeric ({beta}) enzyme hydrolyzes ATP to establish transmembrane electrochemical gradients of Na+ and K+ that are essential for electrical signaling and cell survival. Of the 4 catalytic subunit isoforms, 1 is ubiquitously expressed and is the predominant paralog in peripheral axons. Human population sequencing datasets indicate strong negative selection against both missense and protein-null ATP1A1 variants. To test whether haploinsufficiency generated by heterozygous protein-null alleles are sufficient to cause disease, we tested the neuromuscular characteristics of heterozygous Atp1a1+/- knockout mice and their wildtype littermates, while also evaluating if exercise increased CMT penetrance. We found that Atp1a1+/- mice were phenotypically normal up to 18 months of age. Consistent with the observations in mice, we report clinical phenotyping of a healthy adult human who lacks any clinical features of known ATP1A1-related diseases despite carrying a protein-null early truncation variant, p.Y148*. Taken together, these results suggest that a malfunctioning gene product is required for disease induction by ATP1A1 variants and that if any pathology is associated with protein-null variants, they may display low penetrance or high age of onset.

physiology↗