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Hernandez, S. I.

Publications and source records attributed to Hernandez, S. I..

3 recordsLinked to original sources

Astroglial Dysfunction, Demyelination and Nodular inflammation in Necrotizing Meningoencephalitis

Necrotizing Meningoencephalitis (NME), a form of Meningoencephalitis of Unknown Origin (MUO), is a progressive neuroinflammatory disease that primarily affects young, small-breed dogs. Due to limited understanding of its pathophysiology, early detection and the development of targeted therapies remain challenging. Definitive ante-mortem diagnosis is often unfeasible, and dogs with NME are frequently grouped under the broader MUO category. Our long-term objective is to identify distinct disease mechanisms within each MUO subtype to improve diagnostic accuracy, therapeutic approaches, and prognostic outcomes. To establish unique inflammatory patterns as they relate to neuropathologic changes in NME, we studied we studied the degree of immune cell infiltration, astrogliosis, demyelination, and microglial activation, comparing these factors with granulomatous meningoencephalomyelitis (GME), a closely related MUO subtype. We found that in the leptomeninges, NME is characterized by mild immune cell infiltration, in contrast to the prominent, B cell-rich aggregates seen in GME. In the neuroparenchyma, both diseases exhibit a comparable degree of lymphocyte infiltration; however, demyelination is more pronounced in NME, particularly within the subcortical white matter. Notably, areas of the brain affected by NME display a reduction in astrogliosis, which is associated with a marked decrease in the expression of the water channel protein aquaporin-4 (AQP4), a reduction not observed in GME. Additionally, we found that AQP4 expression levels correlate with the extent of microglial and macrophage activation. These findings suggest that astrocyte dysfunction in regions of microglial inflammation is a driver of NME and with adaptive immune responses likely playing a supportive role.

neuroscience↗

Self-Documenting Plasmids

SO_SCPLOWUMMARYC_SCPLOWPlasmids are the workhorse of biotechnology. These small DNA molecules are used to produce recombinant proteins and to engineer living organisms. They can be regarded as the blueprints of many biotechnology products. It is, therefore, critical to ensure that the sequences of these DNA molecules match their intended designs. Yet, plasmid verification remains challenging. To secure the exchange of plasmids in research and development workflows, we have developed self-documenting plasmids that encode information about themselves in their own DNA molecules. Users of self-documenting plasmids can retrieve critical information about the plasmid without prior knowledge of the plasmid identity. The insertion of documentation in the plasmid sequence does not adversely affect their propagation in bacteria and does not compromise protein expression in mammalian cells. This technology simplifies plasmid verification, hardens supply chains, and has the potential to transform the protection of intellectual property in the life sciences.

synthetic biology↗

Hybrid Sequencing Facilitates Robust De Novo Plasmid Assembly

Despite the wide use of plasmids in research and clinical production, the need to verify plasmid sequences is a bottleneck that is too often underestimated in the manufacturing process. Although sequencing platforms continue to improve, the method and assembly pipeline chosen still influence the final plasmid assembly sequence. Furthermore, few dedicated tools exist for plasmid assembly, especially for de novo assembly. Here, we evaluated short-read, long-read, and hybrid (both short and long reads) de novo assembly pipelines across three replicates of a 24-plasmid library. Consistent with previous characterizations of each sequencing technology, short-read assemblies had issues resolving GC-rich regions, and long-read assemblies commonly had small insertions and deletions, especially in repetitive regions. The hybrid approach facilitated the most accurate, consistent assembly generation and identified mutations relative to the reference sequence. Although Sanger sequencing can be used to verify specific regions, some GC-rich and repetitive regions were difficult to resolve using any method, suggesting that easily sequenced genetic parts should be prioritized in the design of new genetic constructs. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=126 SRC="FIGDIR/small/586694v2_ufig1.gif" ALT="Figure 1"> View larger version (16K): org.highwire.dtl.DTLVardef@13945c1org.highwire.dtl.DTLVardef@11287a8org.highwire.dtl.DTLVardef@1885654org.highwire.dtl.DTLVardef@1dbdbf3_HPS_FORMAT_FIGEXP M_FIG GRAPHICAL ABSTRACT C_FIG

synthetic biology↗