Epigenetic modulators link mitochondrial redox homeostasis to cardiac function in a sex-dependent manner.

While excessive production of reactive oxygen species (ROS) is a characteristic hallmark of numerous diseases, clinical approaches that ameliorate oxidative stress have been unsuccessful. Here, utilizing multi-omics, we demonstrate that in cardiomyocytes, mitochondrial isocitrate dehydrogenase (IDH2) constitutes a major antioxidative defense mechanism. Paradoxically reduced expression of IDH2 associated with ventricular eccentric hypertrophy is counterbalanced by an increase in the enzyme activity. We unveil redox-dependent sex dimorphism, and extensive mutual regulation of the antioxidative activities of IDH2 and NRF2 by a feedforward network that involves 2-oxoglutarate and L-2-hydroxyglutarate and mediated in part through unconventional hydroxy-methylation of cytosine residues present in introns. Consequently, conditional targeting of ROS in a murine model of heart failure improves cardiac function in sex- and phenotype-dependent manners. Together, these insights may explain why previous attempts to treat heart failure with antioxidants have been unsuccessful and open new approaches to personalizing and, thereby, improving such treatment.

(St)aging in place: Information and communication technologies for a health-centered agile dwelling unit.

As the number of older adults is growing rapidly in the U.S., the need for personalized, innovative, and sustainable Information and Communication Technologies (ICTs) solutions is critical to support individuals' social, emotional, and physical health. Such technology can significantly help older adults' ability to live independently in their homes despite the challenges the aging process may present, referred to as aging or staging in place. In this study, we explored ways to integrate ICTs into Agile Dwelling Units (AgDUs) through affordable, innovative, technology-enabled tools and practices that can be adapted to respond to individual's needs while supporting independent, secure, and engaged healthy living. The technology-enabled and human-centered AgDUs organically transform in response to users' needs. This approach offers a viable solution for older adults at different stages throughout their lifespan to transition into an intimate, technologically-enhanced living environment while allowing for (1) customization to user's needs; (2) cost optimization and maintenance; and (3) accessibility that minimizes gaps in compliance from a provider and user perspectives. Integrating ICTs in AgDUs to support health monitoring and management could reduce forthcoming pressure on the healthcare system and care providers to accommodate the needs of older adults. This approach is described through a collaborative multidisciplinary lens that highlights a partnership between academia, industry experts, and key stakeholders to advance healthy living and extend lifespan through design-build and technology integration. The main goal of this approach is to increase access to health services and optimize healthcare costs.

Neonicotinoid pesticide and nitrate mixture removal and persistence in floating treatment wetlands.

Mesocosm and microcosm experiments were conducted to explore the applicability of floating treatment wetlands (FTWs), an ecologically based management technology, to remove neonicotinoid insecticides and nitrate from surface water. The mesocosm experiment evaluated three treatments in triplicate over a 21-d period. Floating treatment wetland mesocosms completely removed nitrate-N over the course of the experiment even when neonicotinoid insecticides were present. At the completion of the experiment, 79.6% of imidacloprid and degradation byproducts and 68.3% of thiamethoxam and degradation byproducts were accounted for in the water column. Approximately 3% of imidacloprid and degradation byproducts and 5.0% of thiamethoxam and degradation byproducts were observed in above-surface biomass, while ∼24% of imidacloprid and degradation byproducts, particularly desnitro imidacloprid, and <0.1% of thiamethoxam and degradation byproducts were found in the below surface biomass. Further, 1 yr after the experiments, imidacloprid, thiamethoxam, and degradation byproducts persisted in biomass but at lower concentrations in both the above- and below-surface biomass. Comparing the microbial communities of mature FTWs grown in the presence and absence of neonicotinoids, water column samples had similar low abundances of nitrifying Archaeal and bacterial amoA genes (below detection to 104  ml-1 ) and denitrifying bacterial nirK, nirS, and nosZ genes (below detection to 105  ml-1 ). Follow-up laboratory incubations found the highest denitrification potential activities in FTW plant roots compared with water column samples, and there was no effect of neonicotinoid addition (100 ng L-1 ) on potential denitrification activity. Based on these findings, (a) FTWs remove neonicotinoids from surface water through biomass incorporation, (b) neonicotinoids persist in biomass long-term (>1 yr after exposure), and (c) neonicotinoids do not adversely affect nitrate-N removal via microbial denitrification.

Engineered Cas9 extracellular vesicles as a novel gene editing tool.

Extracellular vesicles (EVs) have shown promise as biological delivery vehicles, but therapeutic applications require efficient cargo loading. Here, we developed new methods for CRISPR/Cas9 loading into EVs through reversible heterodimerization of Cas9-fusions with EV sorting partners. Cas9-loaded EVs were collected from engineered Expi293F cells using standard methodology, characterized using nanoparticle tracking analysis, western blotting, and transmission electron microscopy and analysed for CRISPR/Cas9-mediated functional gene editing in a Cre-reporter cellular assay. Light-induced dimerization using Cryptochrome 2 combined with CD9 or a Myristoylation-Palmitoylation-Palmitoylation lipid modification resulted in efficient loading with approximately 25 Cas9 molecules per EV and high functional delivery with 51% gene editing of the Cre reporter cassette in HEK293 and 25% in HepG2 cells, respectively. This approach was also effective for targeting knock-down of the therapeutically relevant PCSK9 gene with 6% indel efficiency in HEK293. Cas9 transfer was detergent-sensitive and associated with the EV fractions after size exclusion chromatography, indicative of EV-mediated transfer. Considering the advantages of EVs over other delivery vectors we envision that this study will prove useful for a range of therapeutic applications, including CRISPR/Cas9 mediated genome editing.

Modelling human liver fibrosis in the context of non-alcoholic steatohepatitis using a microphysiological system.

Non-alcoholic steatohepatitis (NASH) is a common form of chronic liver disease characterised by lipid accumulation, infiltration of immune cells, hepatocellular ballooning, collagen deposition and liver fibrosis. There is a high unmet need to develop treatments for NASH. We have investigated how liver fibrosis and features of advanced clinical disease can be modelled using an in vitro microphysiological system (MPS). The NASH MPS model comprises a co-culture of primary human liver cells, which were cultured in a variety of conditions including+/- excess sugar, fat, exogenous TGFß or LPS. The transcriptomic, inflammatory and fibrotic phenotype of the model was characterised and compared using a system biology approach to identify conditions that mimic more advanced clinical disease. The transcriptomic profile of the model was shown to closely correlate with the profile of patient samples and the model displayed a quantifiable fibrotic phenotype. The effects of Obeticholic acid and Elafibranor, were evaluated in the model, as wells as the effects of dietary intervention, with all able to significantly reduce inflammatory and fibrosis markers. Overall, we demonstrate how the MPS NASH model can be used to model different aspects of clinical NASH but importantly demonstrate its ability to model advanced disease with a quantifiable fibrosis phenotype.

ß-Mannosidosis caused by a novel homozygous intragenic inverted duplication in MANBA.

ß-Mannosidosis is a lysosomal storage disorder characterized by accumulation of disaccharides due to deficiency of the lysosomal enzyme ß-mannosidase. The disease is caused by mutations in MANBA and is extremely rare in humans. Although the clinical presentation is heterogeneous, common symptoms include various degrees of developmental delay, behavioral disturbances, hearing loss, and frequent infections. We report a 15-yr-old girl presenting with mild intellectual disability, sensorineural hearing loss, severe behavioral disturbances, dysmorphic traits, and evolving angiokeratomas. Copy-number variation analysis of next-generation sequencing (NGS) data indicated increased coverage in exons 8-11 of MANBA Low ß-mannosidase activity (1 µkatal/kg protein, refv 25-40) established the diagnosis of ß-mannosidosis. Whole-genome sequencing (WGS) and cDNA analysis revealed a novel homozygous intragenic inverted duplication in MANBA, where a 13.1-kb region between introns 7 and 11 was duplicated and inserted in an inverted orientation, creating a 67-base nonduplicated gap at the insertion point. Both junctions showed microhomology regions. The inverted duplication resulted in exon skipping of exons 8-9 or 8-10. Our report highlights the importance of copy-number variation analysis of data from NGS and in particular the power of WGS in the identification and characterization of copy-number variants.

γ-glutamyl transpeptidase deficiency caused by a large homozygous intragenic deletion in GGT1.

γ-Glutamyl transpeptidase deficiency (glutathionuria, OMIM 231950) is a rare disease, with only six patients reported in the literature, although this condition has probably been underdiagnosed due the difficulty to routinely analyze glutathione in clinical samples and to the fact that no genetic defect has been coupled to the disease so far. We report two siblings with mild psychomotor developmental delay and mild neurological symptoms, who presented a markedly increased excretion of glutathione in urine and a very low γ-glutamyl transpeptidase activity in serum. Whole-genome sequencing revealed the presence of a 16.9 kb homozygous deletion in GGT1, one of the genes encoding enzymes with γ-glutamyl transpeptidase activity in the human genome. Close analysis revealed the presence of a 13 bp insertion at the deletion junction. This is the first report of a genetic variant as the cause of glutathionuria. In addition, genetic characterization of the patients' parents and a healthy sibling has provided direct genetic evidence regarding the autosomal recessive nature of this disease.

Identification of a novel mutation in PEX10 in a patient with attenuated Zellweger spectrum disorder: a case report.

BACKGROUND: The peroxisome biogenesis disorders, which are caused by mutations in any of 13 different PEX genes, include the Zellweger spectrum disorders. Severe defects in one of these PEX genes result in the absence of functional peroxisomes which is seen in classical Zellweger syndrome. These patients present with hypotonia and seizures shortly after birth. Other typical symptoms are dysmorphic features, liver disease, retinal degeneration, sensorineural deafness, polycystic kidneys, and the patient does not reach any developmental milestones. CASE PRESENTATION: We report a case of a patient with Zellweger spectrum disorder due to a novel mutation in the PEX10 gene, presenting with a mild late-onset neurological phenotype. The patient, an Assyrian girl originating from Iraq, presented with sensorineural hearing impairment at the age of 5 followed by sensorimotor polyneuropathy, cognitive delay, impaired gross and fine motor skills, and tremor and muscle weakness in her teens. Analyses of biochemical markers for peroxisomal disease suggested a mild peroxisomal defect and functional studies in fibroblasts confirmed the existence of a peroxisome biogenesis disorder. Diagnosis was confirmed by next generation sequencing analysis, which showed a novel homozygous mutation (c.530 T > G (p.Leu177Arg) (NM_153818.1)) in the PEX10 gene predicted to be pathogenic. CONCLUSIONS: This case highlights the importance of performing biochemical, functional, and genetic peroxisomal screening in patients with clinical presentations milder than those usually observed in Zellweger spectrum disorders.