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Searcy, J.

Publications and source records attributed to Searcy, J..

4 recordsLinked to original sources

Fungal:bacterial biomass balance links environmental gradients to soil respiration across a forest-to-marsh transition

Soil microbes regulate whether carbon is retained in soils or returned to the atmosphere through respiration, but the extent to which microbial community characteristics improve the prediction of heterotrophic soil CO2 production beyond predictions by environmental controls remains unclear. We tested this across a topographically structured forest-to-marsh gradient in coastal Oregon by measuring heterotrophic soil respiration, soil physicochemical properties, PLFA-based microbial biomass, metagenomic taxonomic composition, and functional gene-based trait indicators. Across the gradient, soil moisture increased from forest to marsh, while mineral soil and organic matter C:N decreased. These environmental shifts were accompanied by strong but uneven microbial responses: total microbial, fungal, and bacterial biomass declined from forest to marsh, taxonomic composition showed the strongest structuring by environmental conditions, and functional gene-based indicators showed mixed relationships with the gradient. Environmental model comparisons identified soil moisture and organic layer C as the strongest baseline predictors of respiration. Among microbial descriptors, only a small subset improved respiration prediction beyond this environmental baseline. The fungal:bacterial (F:B) biomass ratio produced the largest increase in model fit and the greatest reduction in AICc, whereas the best taxonomic and functional-gene predictors yielded more minor gains. Our results showed that the microbial descriptors most responsive to environmental gradients were not the ones most useful for predicting soil respiration; instead, a relatively simple biomass-partitioning metric captured respiration-relevant microbial variation more effectively than finer taxonomic and genomic descriptors. This suggests that F:B ratio may be especially useful for representing respiration-relevant microbial variation in local landscape-scale studies and for future carbon cycle models applied across heterogeneous ecosystem transition zones.

ecology↗

A semi-automated pipeline for quantitation of Pax7+, myonuclei, and cross-sectional area by fiber type

Manual analysis of skeletal muscle cross-sections is time-consuming and subject to error and user bias. To overcome these limitations, we developed and validated a semi-automated, quantitative, and reproducible image-analysis pipeline specifically tailored to quantify Pax7+ satellite cells, myonuclei, and cross-sectional area by fiber type. The workflow combines FIJI/ImageJ-based image preprocessing with CellProfiler, Cellpose, and a custom Python script to process and analyze immunohistological images of muscle tissue cross-sections. Outcomes include Pax7+ satellite cells and myonuclei quantified per fiber by fiber type, along with cross-sectional area, perimeter, and fiber type classification. This semi-automated approach provides a robust and efficient platform for high-throughput analysis of muscle tissue cross-sections from large datasets. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=141 SRC="FIGDIR/small/729866v1_ufig1.gif" ALT="Figure 1"> View larger version (44K): org.highwire.dtl.DTLVardef@e85bfdorg.highwire.dtl.DTLVardef@ef75e0org.highwire.dtl.DTLVardef@123e461org.highwire.dtl.DTLVardef@166a304_HPS_FORMAT_FIGEXP M_FIG C_FIG

cell biology↗

A semi-automated pipeline for morphological analysis of myonuclei along single muscle fibers

Manual quantitation of skeletal muscle myonuclear number, spatial orientation, and morphology is time-consuming and subject to error and bias. To overcome these limitations, we developed and validated a semi-automated, quantitative, and reproducible image-analysis pipeline. The workflow combines FIJI-based preprocessing with custom Python scripts to process immunohistological images of individual muscle fibers, enabling high-resolution and scalable quantification of nuclei. Analyses incorporate morphometric parameters including nuclear position, shape, and three-dimensional orientation, as well as centroid-to-skeleton distance and nearest-neighbor relationships to capture spatial patterns of myonuclear organization along the fiber. Outputs include per-fiber and biopsy-level summaries integrated with Imaris metrics. This semi-automated approach provides a robust and efficient platform for high-throughput analysis of myonuclear number and structural features across large single fiber datasets.

cell biology↗

Transient Acute Neuronal Activation Response Caused by High Concentrations of Oligonucleotides in the Cerebral Spinal Fluid

Oligonucleotide (ON) therapeutics are promising as a disease-modifying therapy for central nervous system disorders. Intrathecal ON administration into the cerebral spinal fluid is a safe and effective delivery mode to the CNS. However, preclinical studies have shown acute toxicities following high-dose central ON delivery. Here we characterize a transient neurobehavioral change peaking 15 minutes after ON dosing and resolving after 120 minutes. Symptoms include shaking, muscle twitching, cramping, hyperactivity, stereotypic movements, hyperreactivity, vocalizations, tremors, convulsions, and seizures. These are collectively referred here as the acute neuronal activation response. Acute neuronal activation is observed in rats, mice, and non-human primates and is quantifiable using a simple scoring system. It is distinct from acute sedation seen with some phosphorothioate-modified antisense oligonucleotides, characterized by loss of spinal reflexes, ataxia, and sedation. The acute neuronal activation response is largely sequence-independent and is driven by ON chelation of divalent cations, particularly influenced by the divalent cations-to-ON ratio in the dosing solution. Acute neuronal activation can be safely mitigated by adjusting this ratio through magnesium supplementation in the ON formulation. We provide a comprehensive framework for quantifying and mitigating the acute neuronal activation response caused by high concentrations of centrally delivered ON therapeutics in preclinical species. GRAPHICAL ABSTRACT O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=79 SRC="FIGDIR/small/638138v1_ufig1.gif" ALT="Figure 1"> View larger version (32K): org.highwire.dtl.DTLVardef@b18796org.highwire.dtl.DTLVardef@13c9898org.highwire.dtl.DTLVardef@1463019org.highwire.dtl.DTLVardef@ffe719_HPS_FORMAT_FIGEXP M_FIG C_FIG

neuroscience↗