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Sanchez, A.

Publications and source records attributed to Sanchez, A..

At least 19 recordsLinked to original sources

Statistical learning of bacterial growth in combinatorially constructed environments

Optimizing growth conditions and culture media is a major goal in microbiology. A challenge is that nutrients can have complex, non-additive effects on growth. The fact that resources interact with one another has long been known in ecology, but systematic maps of resource interactions at all orders have been lacking and it is not known whether interactions are primarily low order or fundamentally complex. To tackle this problem, we have followed a full factorial design approach and measured the growth of seven different bacterial species in all possible combinations of 8 carbon sources under carbon-limiting conditions. Our approach allows us to directly estimate interactions at all orders. Even though all C-sources stimulate growth on their own as well as in combination with other nutrients, most of them can also have negative effects on growth when they are added to at least some nutrient mixtures. We show that the switch from positive to negative fitness effects is governed by global epistasis among resources. An analysis of variance shows that additive effects and pairwise interactions explain most of the variation in fitness, allowing us to train simple regression models that accurately predict bacterial growth in novel environments. The generality of these findings across seven different bacterial strains belonging to two different families indicates that interactions between carbon sources under carbon-limiting conditions may be generally learnable from a relatively sparse set of constructed environments, enabling the rational optimization of growth conditions.

microbiology↗

Targeted Analysis of >1500 Plasma Proteoforms via Individual Ion Mass Spectrometry

Plasma proteomics has long sought to accessibly sense human biology, precisely detect disease states, and advance diagnostics through clinical translation. In recent years, companies with defined panels such as Olink and Somascan have entered the field to complement bottom-up mass spectrometry (BUP). This study leverages a novel mass spectrometry platform to capture targeted proteoform information lost by mainline antibody-, aptamer-, and BUP-driven workflows. The Plasma Proteoform Assay (PPA) uses Individual Ion Mass Spectrometry (I{superscript 2}MS) to resolve mixtures of intact proteins presented by direct injection. Two proteoform panels, PPA 526 and PPA 1514, were defined from human plasma samples obtained from 81 individuals. The panels quantify 526 proteoforms derived from 59 genes and 1,514 proteoforms from 155 genes, respectively. Reproducibility for both panels showed coefficients of variation below 20% for most proteoforms (59-80%). PPA was benchmarked in studies including subjects with hepatic cirrhosis (N=30) and resilient agers carrying a SERPINE1 (PAI 1) mutation (N=27). PPA signatures distinguished SERPINE1 mutation carriers from affected individuals and provided sufficient resolution to discriminate among cirrhosis disease stages. In summary, we present PPA 526 and PPA 1514, the first scalable plasma proteoform panels capable of tracking hundreds to thousands of targets in a few minutes per sample.

biochemistry↗

On the limits of detection of epistatic higher-order interactions

The function of microbial communities is often dominated by additive and pairwise interactions, raising the question of whether this reflects intrinsic biological simplicity or fundamental limits of detection. Here, we leverage the theory of fitness landscapes to bridge microbial ecology and genetics, and show that this apparent simplicity is a generic consequence of structural and statistical constraints rather than evidence for intrinsically weak higher-order interactions (HOIs). We separate the detectability of individual epistatic interactions from their contribution to functional variance, and demonstrate that local k-order interactions suffer from exponential noise amplification while their contributions to total variance are intrinsically suppressed by combinatorial geometric dilution. Applying this framework to a fully sampled 210 experimental microbial landscape, we find that only first- and second-order interactions are distinguishable from experimental noise. Furthermore, generalized Lotka-Volterra simulations reveal that experimental noise alone can generate the illusion of higher-order structure in communities where all direct mechanistic interactions are pairwise and indirect interactions are weak. Our findings identify universal, order-dependent limits on the quantification of epistasis that apply to high-dimensional landscapes across ecology and genetics, providing a principled foundation for rational community design.

systems biology↗

A Statistical Review of Virus Reduction in Coagulation, Flocculation, and Sedimentation Treatment Processes

Coagulation, flocculation, and sedimentation (CFS) is widely applied as a combined unit process in the treatment of drinking water, wastewater, and recycled water; however, virus reduction through CFS has not been sufficiently characterized to assign pathogen log reduction value (LRV) credits. This study collected data through a systematic review that yielded over 1000 LRVs from 43 manuscripts covering 46 viruses to characterize virus reduction through CFS. The results demonstrate that CFS is effective at reducing viruses, with 68% of virus LRVs greater than 1. A mixed-effects model was used to identify potential mechanisms of virus reduction with ferric and aluminum coagulants, as well as factors associated with variability in performance. Key insights from the model show that virus reduction is: (1) improved at lower pH, similar to natural organic matter (NOM) reduction, (2) lower in secondary effluent than surface water for drinking water treatment, (3) virus-dependent, and (4) dependent on virus enumeration methods, with lower LRVs observed for molecular techniques. These findings demonstrate the potential for CFS to provide consistent and explainable virus reduction, potentially establishing a foundation for regulatory crediting in potable reuse applications. Future crediting frameworks will need to account for the factors impacting performance to accurately quantify and assign credit for virus reduction.

microbiology↗

Long-Read Sequencing of the MUC1 VNTR: Genomic Variation, Mutational Landscape, and Its Impact on ADTKD Diagnosis and Progression

BackgroundADTKD-MUC1 is caused by frameshift mutations in MUC1 gene that produce a frameshifted protein (MUC1fs) toxic to kidney cells. The genes variable number of tandem repeats (VNTR), with high GC content, makes it largely inaccessible to standard sequencing. As a result, both the reference sequence and natural variation in this region remain poorly defined, complicating mutation detection and data interpretation. Standard methods also fail to pinpoint the exact VNTR unit affected, limiting insight into mutation mechanisms and genotype-phenotype correlations. MethodsWe employed Single Molecule, Real-Time (SMRT) sequencing and characterized the genomic sequence of MUC1 in 300 individuals including 279 individuals from 143 families suspected of having ADTKD-MUC1. We compared these results to those obtained using the CLIA-approved mass spectrometry-based probe extension (PE) assay, which specifically detect the most prevalent 59dupC mutation. We correlated the structural features of the MUC1 VNTR with the rate of kidney function decline in affected individuals. ResultsWe identified MUC1 consensus sequences for 205 unique VNTR alleles, with 9 distinct types of frameshift mutations present on 52 distinct mutated VNTR alleles. MUC1 frameshift mutations were identified in 71 of 143 families (50%) with suspected ADTKD, comprising 135 genetically affected individuals (48%). The SMRT assay exhibited complete concordance and revealed that the PE assay is capable of detecting frameshift mutations in approximately 85% of affected families. The constellation of VNTR structures supports a genotype-progression model, in which fast progressors exhibit a significantly lower number of repeat units on the wild-type allele and a higher number of repeats on the mutation-bearing allele, including an increased number of frameshifted repeat units. ConclusionsSMRT sequencing outperforms current diagnostic methods for ADTKD-MUC1 and reveals the prognostic value of VNTR structures. Although their contribution to disease progression is modest ([~]6% variance explained), it remains biologically and clinically meaningful. Key Points 3O_LISingle Molecule, Real-Time (SMRT) sequencing of MUC1 outperforms existing genetic and immunohistochemical methods for ADTKD-MUC1 diagnosis. C_LIO_LICLIA-approved mass spectrometry-based genotyping assay, complemented by SMRT sequencing, can detect nearly all ADTKD-MUC1 patients. C_LIO_LIDisease progression correlates with VNTR pattern: fast progressors have fewer WT repeats and more mutant/frameshifted repeats C_LI

genetics↗

CRISPR screen identifies BAP1 as a deubiquitinase regulating SPIN4 stability

Protein homeostasis is tightly controlled by the coordinated actions of E3 ubiquitin ligases and deubiquitinases (DUBs). We identify Spindlin-4 (SPIN4), a histone H3K4me3 reader, as a substrate regulated by opposing pathways: degradation mediated by the Cullin-RING E3 ligase DCAF16 and stabilization by the DUB BAP1. Through CRISPR-Cas9 knockout screens and biochemical analyses, we demonstrate that DCAF16 promotes SPIN4 degradation, while BAP1 interacts with and stabilizes SPIN4 through its catalytic activity. Inhibition or loss of BAP1 reduces SPIN4 levels, highlighting its critical role in maintaining SPIN4 homeostasis. Proteomics and interactome analyses further support this regulatory axis. These findings reveal a dynamic balance controlling SPIN4 stability, with potential implications for epigenetic regulation and disease processes.

biochemistry↗

DeepSeq: High-Throughput Single-Cell RNA Sequencing Data Labeling via Web Search-Augmented Agentic Generative AI Foundation Models

Generative AI foundation models offer transformative potential for processing structured biological data, particularly in single-cell RNA sequencing, where datasets are rapidly scaling toward billions of cells. We propose the use of agentic generative AI foundation models with real-time web search to automate the labeling of experimental data, achieving up to 82.5% accuracy. This addresses a key bottleneck in supervised learning for structured omics data by increasing annotation throughput without manual curation and human error. Our approach enables the development of virtual cell foundation models capable of downstream tasks such as cell-typing and perturbation prediction. As data volume grows, these models may surpass human performance in labeling, paving the way for reliable inference in large-scale perturbation screens. This application demonstrates domain-specific innovation in health monitoring and diagnostics, aligned with efforts like the Human Cell Atlas and Human Tumor Atlas Network.

bioinformatics↗

NEUROLINGUA: A Neuroimaging Database Tailored to Unravel the Complexity of Multilingual Comprehension

The neural mechanisms underlying language processing involve a well-defined brain network, including mainly left perisylvian areas. Yet, the extent of individual variability remains largely unexplored, particularly in bilingual and multilingual contexts. Differences in linguistic profiles (e.g., age of acquisition, exposure, proficiency) provide an opportunity to assess how network topology is shaped by sociolinguistic factors. To address this, we developed NEUROLINGUA, a comprehensive database of functional and structural MRI data, enriched with sociodemographic, sociolinguistic, and behavioral information. It includes 725 healthy individuals aged 18-82 immersed in a Basque-Spanish multilingual environment, ranging from near-monolinguals to highly proficient multilinguals. Participants completed a functional MRI language localizer with both auditory and visual comprehension tasks, enabling cross-modal comparisons, as well as sentences involving arithmetic problem-solving. Exploratory analyses confirmed associations between structural MRI, sociodemographic, and cognitive measures. We demonstrate that NEUROLINGUAs functional MRI data localize the language comprehension network and thus capture linguistic profile effects. This integrative dataset offers an unparalleled resource to investigate factors influencing language network adaptability and variability in diverse sociolinguistic contexts.

neuroscience↗

A Platform for Mitochondrial Profiling in Enriched Kidney Segments Under Thermodynamic Control

Mitochondrial function varies widely across kidney nephron segments, yet conventional approaches lack the resolution and control needed to assess cell-type-specific bioenergetics in situ. We present a methodological platform that enables segment-resolved profiling of mitochondrial respiration, conductance, and membrane potential in freshly isolated mouse and human nephron segments. Combining mechanical sieving and adhesion-based enrichment with permeabilized high-resolution respirometry, we adapted the creatine kinase clamp to quantify oxygen flux and mitochondrial membrane potential across defined free energies. Using this approach, we found that proximal tubules exhibit high respiratory conductance and dynamic mitochondrial polarization, while distal tubules and glomeruli maintain static membrane potential and low conductance. In a model of adenine-induced nephropathy, only proximal tubule mitochondria showed marked reductions in respiration and ATP production. This segment-specific dysfunction was not detectable in bulk mitochondrial isolates. We then demonstrate that these methods can be applied in human kidney samples. Our approach provides thermodynamically anchored, segment-resolved insight into mitochondrial adaptation under physiological and pathological conditions. It is broadly applicable to other tissues with metabolic heterogeneity and compatible with disease models, genetic tools, and pharmacological interventions. This platform bridges a critical gap between conventional respirometry and functional mitochondrial phenotyping in native tissue structures.

biochemistry↗

Evidence for cPLA2 activation in Alzheimer's Disease Synaptic Pathology

BackgroundSynapses are essential for learning and memory, and their loss predicts cognitive decline in Alzheimers disease (AD). Synaptic loss is associated with excitotoxicity, neuroinflammation, amyloid-{beta}, and tau pathology, but the molecular mechanisms remain unclear. There is an urgent need to identify new targets to modify the disease and slow synaptic loss and cognitive decline. This study examines if calcium-dependent phospholipase A2 (cPLA2) is implicated in AD synaptic loss. cPLA2 catalyzes membrane phospholipids to release arachidonic acid, which can be metabolized into inflammatory eicosanoids. MethodscPLA2 levels were examined in synaptosomes isolated from the postmortem frontal cortex of individuals with no cognitive impairment (NCI), mild cognitive impairment (MCI), and AD dementia from the Religious Orders Study (ROS). Eicosanoids in synaptosomes were analyzed using lipidomics. Immunofluorescent staining investigated cPLA2 interactions with synaptic markers. Human iPSCs-derived neurons were used to study cPLA2 overactivation after exposure to amyloid-{beta} 42 oligomers (A{beta}42O), its relationships with synaptic markers, and the effects of cPLA2 inhibitors. ResultsWe observed elevated cPLA2 (cPLA2 and cPLA2{beta}) in AD synaptosomes and positive correlations with postsynaptic density protein 95 (PSD-95) and cognitive dysfunction. Eicosanoids were increased in AD synaptosomes and correlated with cPLA2, indicating cPLA2 activity at synapses/synaptosomes. Phosphorylated cPLA2 (p-cPLA2) colocalized with PSD-95 in synaptosomes, and with postsynaptic Ca2+/calmodulin-dependent protein kinase II (CaMKII) and dendritic microtubule-associated protein 2 (MAP2) in NCI and AD brains, where their levels were reduced in AD. P-cPLA2 colocalizes with MAP2 at the neuronal soma associated with neuritic plaques and neurodegeneration in AD. A{beta}42O activates cPLA2 in human iPSCs-derived neurons, leading to p-cPLA2 relocation from the cytosol to synaptic and dendritic sites to colocalize with CaMKII and MAP2, resulting in their reduction. P-cPLA2 also colocalized with PSD-95 in A{beta}42O-exposed neurons, accompanied with increased PSD-95 intensity at soma membrane. These processes were reversed by the cPLA2 inhibitor ASB14780. ConclusionscPLA2 overactivation at synapses, dendrites, and excitatory neuronal somas is associated with synaptic loss, neuritic plaques and neurodegeneration, potentially contributing to cognitive decline in AD. Future research needs to explore the role of cPLA2 as a disease-modifying target for AD.

neuroscience↗

Next-Generation Protein Sequencing and individual ion mass spectrometry enable complementary analysis of interleukin-6

The vast complexity of the proteome currently overwhelms any single analytical technology in capturing the full spectrum of proteoform diversity. In this study, we evaluated the complementarity of two cutting-edge proteomic technologies--single-molecule protein sequencing and individual ion mass spectrometry--for analyzing recombinant human IL-6 (rhIL-6) at the amino acid, peptide, and intact proteoform levels. For single-molecule protein sequencing, we employ the recently released Platinum(R) instrument. Next-Generation Protein Sequencing (NGPS) on Platinum utilizes cycles of N-terminal amino acid recognizer binding and aminopeptidase cleavage to enable parallelized sequencing of single peptide molecules. We found that NGPS produces single amino acid coverage of multiple key regions of IL-6, including two peptides within helices A and C which harbor residues that reportedly impact IL-6 function. For top-down proteoform evaluation, we use individual ion mass spectrometry (I2MS), a highly parallelized orbitrap-based charge detection MS platform. Single ion detection of gas-phase fragmentation products (I2MS2) gives significant sequence coverage in key regions in IL-6, including two regions within helices B and D that are involved in IL-6 signaling. Together, these complementary technologies deliver a combined 52% sequence coverage, offering a more complete view of IL-6 structural and functional diversity than either technology alone. This study highlights the synergy of complementary protein detection methods to more comprehensively cover protein segments relevant to biological interactions. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=114 SRC="FIGDIR/small/637157v1_ufig1.gif" ALT="Figure 1"> View larger version (39K): org.highwire.dtl.DTLVardef@82d72aorg.highwire.dtl.DTLVardef@1fc39f3org.highwire.dtl.DTLVardef@40a677org.highwire.dtl.DTLVardef@70f6c3_HPS_FORMAT_FIGEXP M_FIG C_FIG

biochemistry↗

Cooperative Role of PACT and ADAR1 in Preventing Aberrant PKR Activation by Self-Derived Double-Stranded RNA

Double-stranded RNAs (dsRNAs) produced during viral infections are recognized by the innate immune sensor protein kinase R (PKR), triggering a host translation shutoff that inhibits viral replication and propagation. Given the harmful effects of uncontrolled PKR activation, cells must tightly regulate PKR to ensure that its activation occurs only in response to viral infections, not endogenous dsRNAs. Here, we use CRISPR-Translate, a FACS-based genome-wide CRISPR-Cas9 knockout screening method that exploits translation levels as a readout and identifies PACT as a key inhibitor of PKR during viral infection. We find that cells deficient for PACT hyperactivate PKR in response to several different RNA viruses, raising the question of why cells need to limit PKR activity. Our results demonstrate that PACT cooperates with ADAR1 to suppress PKR activation from self-dsRNAs in uninfected cells. The simultaneous deletion of PACT and ADAR1 results in synthetic lethality, which can be fully rescued in PKR-deficient cells. We propose that both PACT and ADAR1 act as essential barriers against PKR, creating a threshold of tolerable levels to endogenous dsRNA in cells without activating PKR-mediated translation shutdown and cell death.

microbiology↗

ATR safeguards replication forks against APOBEC3B-induced toxic PARP1 trapping

ATR is the master safeguard of genomic integrity during DNA replication. Acute inhibition of ATR with ATR inhibitor (ATRi) triggers a surge in origin firing, leading to increased levels of single-stranded DNA (ssDNA) that rapidly deplete all available RPA. This leaves ssDNA unprotected and susceptible to breakage, a phenomenon known as replication catastrophe. However, the mechanism by which unprotected ssDNA breaks remains unclear. Here, we reveal that APOBEC3B is the key enzyme targeting unprotected ssDNA at replication forks, triggering a reaction cascade that induces fork collapse and PARP1 hyperactivation. Mechanistically, we demonstrate that uracils generated by APOBEC3B at replication forks are removed by UNG2, creating abasic sites that are subsequently cleaved by APE1 endonuclease. Moreover, we demonstrate that APE1-mediated DNA cleavage is the critical enzymatic step for PARP1 trapping and hyperactivation in cells, regardless of how abasic sites are generated on DNA. Finally, we show that APOBEC3B-induced toxic PARP1 trapping in response to ATRi drives cell sensitivity to ATR inhibition, creating to a context of synthetic lethality when combined with PARP inhibitors. Together, these findings reveal the mechanisms that cause replication forks to break during replication catastrophe and explain why ATRi-treated cells are particularly sensitive to PARP inhibitors.

molecular biology↗

RPS19 and RPL5, the most commonly mutated genes in Diamond Blackfan anemia, impact DNA double-strand break repair

Diamond Blackfan anemia (DBA) is caused by germline heterozygous loss-of-function pathogenic variants (PVs) in ribosomal protein (RP) genes, most commonly RPS19 and RPL5. In addition to red cell aplasia, individuals with DBA are at increased risk of various cancers. Importantly, the mechanism(s) underlying cancer predisposition are poorly understood. We found that DBA patient-derived lymphoblastoid cells had persistent {gamma}-H2AX foci following ionizing radiation (IR) treatment, suggesting DNA double-strand break (DSB) repair defects. RPS19- and RPL5-knocked down (KD) CD34+ cells had delayed repair of IR-induced DSBs, further implicating these RPs in DSB repair. Assessing the impact of RPS19- and RPL5-KD on specific DSB repair pathways, we found RPS19-KD decreased the efficiency of pathways requiring extensive end-resection, whereas RPL5-KD increased end-joining pathways. Additionally, RAD51 was reduced in RPS19- and RPL5-KD and RPS19- and RPL5-mutated DBA cells, whereas RPS19-deficient cells also had a reduction in PARP1 and BRCA2 proteins. RPS19-KD cells had an increase in nuclear RPA2 and a decrease in nuclear RAD51 foci post-IR, reflective of alterations in early, critical steps of homologous recombination. Notably, RPS19 and RPL5 interacted with poly(ADP)-ribose chains noncovalently, were recruited to DSBs in a poly(ADP)-ribose polymerase activity-dependent manner, and interacted with Ku70 and histone H2A. RPL5s recruitment, but not RPS19s, also required p53, suggesting that RPS19 and RPL5 directly participate in DSB repair via different pathways. We propose that defective DSB repair arising from haploinsufficiency of these RPs may underline the cancer predisposition in DBA. KEY POINTSO_LIRPS19- and RPL5-deficiency alters DNA double-strand break repair, which may underlie cancer predisposition in Diamond Blackfan anemia. C_LIO_LIRPS19 and RPL5 bind PAR chains and are rapidly recruited to sites of DNA double-strand breaks, suggesting active roles in repair. C_LI

molecular biology↗

Six months of physical inactivity is insufficient to cause chronic kidney disease in C57BL/6J mice

Chronic kidney disease (CKD) is a progressive disorder marked by a decline in kidney function. Obesity and sedentary behavior contribute to the development of CKD, though mechanisms by which this occurs are poorly understood. This knowledge gap is worsened by the lack of a reliable murine CKD model that does not rely on injury, toxin, or gene deletion to induce a reduction in kidney function. High-fat diet (HFD) feeding alone is insufficient to cause reduced kidney function until later in life. Here, we employed a small mouse cage (SMC), a recently developed mouse model of sedentariness, to study its effect on kidney function. Wildtype C57BL/6J male mice were housed in sham or SMC housing for six months with HFD in room (22{degrees}C) or thermoneutral (30{degrees}C) conditions. Despite hyperinsulinemia induced by the SMC+HFD intervention, kidneys from these mice displayed normal glomerular filtration rate (GFR). However, the kidneys showed early signs of kidney injury, including increases in Col1a1 and NGAL transcripts, as well as fibrosis by histology, primarily in the inner medullary/papilla region. High-resolution respirometry and fluorometry experiments showed no statistically significant changes in the capacities for respiration, ATP synthesis, or electron leak. These data confirm the technical challenge in modeling human CKD. They further support the notion that obesity and a sedentary lifestyle make the kidneys more vulnerable, but additional insults are likely required for the pathogenesis of CKD.

physiology↗

MUC2 Expression Modulates Immune Infiltration in Colorectal Cancer

IntroductionColorectal cancer (CRC) is a prevalent malignancy with significant morbidity and mortality worldwide. A deeper understanding of the interaction of cancer cells with other cells in the tumor microenvironment is crucial for devising effective therapeutic strategies. MUC2, a major component of the protective mucus layer in the gastrointestinal tract, has been implicated in CRC progression and immune response regulation. MethodIn this study, we sought to elucidate the relationship between MUC2 expression and immune infiltration within CRC, using in-vitro models involving two well-established cell lines, HT-29 and LS-174T. By employing CRISPR-mediated MUC2 knockout, we investigated the influence of MUC2 on tumor immune infiltration and its interplay with T cells and NK cells enriched peripheral blood mononuclear cells (PBMCs) in 3D spheroid cultures. ResultsWhile MUC2 was more abundant in LS-174T cell lines compared to HT-29, its knockout resulted in increased immune infiltration solely in the HT-29 cell line, but not in LS-174T. We revealed that the removal of MUC2 protein was compensated in LS-174T by the expression of other gel forming mucin proteins (Muc6, Muc5B) commonly expressed in gastrointestinal epithelium, while this was not observed in HT-29 cell line. DiscussionWe propose that the role of MUC2 documented in CRC progression can partially be explained by impairing immune infiltration due to physical barrier established by the gel forming proteins such as MUC2 in mucinous CRC. On the other hand, the removal of MUC2 expression can be compensated by alternative gel forming mucin proteins, thereby impeding any increase in tumor immune infiltration.

cancer biology↗

Sex-Specific Changes in Cardiac Function and Electrophysiology During Progression of Adenine-Induced Chronic Kidney Disease in Mice

BACKGROUNDChronic kidney disease (CKD) and cardiovascular disease (CVD) often co-exist and interact; however, notable sex-dependent differences are observed in how these conditions manifest and progress in parallel, despite men and women sharing similar risk factors. Identifying sex-specific diagnostic markers of cardiac structure and function throughout CKD progression could elucidate why the development and progression of these diseases differ by sex. METHODS AND RESULTSAdult, C57BL/6J male and female mice were subjected to a high-adenine (0.2%) diet throughout 12-weeks to induce CKD. Control mice were fed a normal chow diet. Every three weeks, electrocardiogram (ECG) and echocardiogram-based markers of cardiac physiology were evaluated. Adenine-induced CKD showed markers of left ventricular (LV) hypertrophy in male mice only. CKD males had markers indicative of LV systolic and diastolic dysfunction throughout regimen duration, worsening as disease progressed. Adenine males had a prolonged QTc and STc intervals when compared to Adenine females and Control males. Sex-dependent differences in the duration of the Speak-J marker, measured via ECG, was identified, with Adenine males showing increases in duration earlier than Adenine females compared to their Control counterparts. CONCLUSIONSIn this study, we identified sex-dependent differences in cardiac structure, function, and electrophysiology in a mouse model of CKD-induced CVD throughout disease progression. We found that male mice are more prone to developing LV hypertrophy, systolic dysfunction, and diastolic dysfunction, with significant increases in ECG markers indicative of ventricular dysfunction observed in adenine-treated males at late stages of the disease. Additionally, we identified a new ECG parameter, Speak-J duration, that highlights sex-specific cardiac electrophysiological changes, demonstrating the models utility in studying sex-dependent cardiac differences.

pathology↗

Deep Profiling of Plasma Proteoforms with Engineered Nanoparticles for Top-down Proteomics

The dynamic range challenge for detection of proteins and their proteoforms in human plasma has been well documented. Here, we use the nanoparticle protein corona approach to enrich low-abundant proteins selectively and reproducibly from human plasma and use top-down proteomics to quantify differential enrichment for the 2841 detected proteoforms from 114 proteins. Furthermore, nanoparticle enrichment allowed top-down detection of proteoforms between [~]1 {micro}g/mL and [~]10 pg/mL in absolute abundance, providing up to 105-fold increase in proteome depth over neat plasma in which only proteoforms from abundant proteins (>1 {micro}g/mL) were detected. The ability to monitor medium and some low abundant proteoforms through reproducible enrichment significantly extends the applicability of proteoform research by adding depth beyond albumin, immunoglobins and apolipoproteins to uncover many involved in immunity and cell signaling. As proteoforms carry unique information content relative to peptides, this report opens the door to deeper proteoform sequencing in clinical proteomics of disease or aging cohorts.

biochemistry↗