Mitochondrial Transfer Improves Cardiomyocyte Bioenergetics and Viability in Male Rats Exposed to Pregestational Diabetes

Offspring born to diabetic or obese mothers have a higher lifetime risk of heart disease. Previously, we found that rat offspring exposed to late-gestational diabetes mellitus (LGDM) and maternal high-fat (HF) diet develop mitochondrial dysfunction, impaired cardiomyocyte bioenergetics, and cardiac dysfunction at birth and again during aging. Here, we compared echocardiography, cardiomyocyte bioenergetics, oxidative damage, and mitochondria-mediated cell death among control, pregestational diabetes mellitus (PGDM)-exposed, HF-diet-exposed, and combination-exposed newborn offspring. We hypothesized that PGDM exposure, similar to LGDM, causes mitochondrial dysfunction to play a central, pathogenic role in neonatal cardiomyopathy. We found that PGDM-exposed offspring, similar to LGDM-exposed offspring, have cardiac dysfunction at birth, but their isolated cardiomyocytes have seemingly less bioenergetics impairment. This finding was due to confounding by impaired viability related to poorer ATP generation, more lipid peroxidation, and faster apoptosis under metabolic stress. To mechanistically isolate and test the role of mitochondria, we transferred mitochondria from normal rat myocardium to control and exposed neonatal rat cardiomyocytes. As expected, transfer provides a respiratory boost to cardiomyocytes from all groups. They also reduce apoptosis in PGDM-exposed males, but not in females. Findings highlight sex-specific differences in mitochondria-mediated mechanisms of developmentally programmed heart disease and underscore potential caveats of therapeutic mitochondrial transfer.     

A Review of Murine Cytomegalovirus as a Model for Human Cytomegalovirus Disease—Do Mice Lie?

Since murine cytomegalovirus (MCMV) was first described in 1954, it has been used to model human cytomegalovirus (HCMV) diseases. MCMV is a natural pathogen of mice that is present in wild mice populations and has been associated with diseases such as myocarditis. The species-specific nature of HCMV restricts most research to cell culture-based studies or to the investigation of non-invasive clinical samples, which may not be ideal for the study of disseminated disease. Initial MCMV research used a salivary gland-propagated virus administered via different routes of inoculation into a variety of mouse strains. This revealed that the genetic background of the laboratory mice affected the severity of disease and altered the extent of subsequent pathology. The advent of genetically modified mice and viruses has allowed new aspects of disease to be modeled and the opportunistic nature of HCMV infection to be confirmed. This review describes the different ways that MCMV has been used to model HCMV diseases and explores the continuing difficulty faced by researchers attempting to model HCMV congenital cytomegalovirus disease using the mouse model.     

Plasma Amyloid-Beta Levels in a Pre-Symptomatic Dutch-Type Hereditary Cerebral Amyloid Angiopathy Pedigree: A Cross-Sectional and Longitudinal Investigation

Plasma amyloid-beta (Aβ) has long been investigated as a blood biomarker candidate for Cerebral Amyloid Angiopathy (CAA), however previous findings have been inconsistent which could be attributed to the use of less sensitive assays. This study investigates plasma Aβ alterations between pre-symptomatic Dutch-type hereditary CAA (D-CAA) mutation-carriers (MC) and non-carriers (NC) using two Aβ measurement platforms. Seventeen pre-symptomatic members of a D-CAA pedigree were assembled and followed up 3–4 years later (NC = 8; MC = 9). Plasma Aβ1-40 and Aβ1-42 were cross-sectionally and longitudinally analysed at baseline (T1) and follow-up (T2) and were found to be lower in MCs compared to NCs, cross-sectionally after adjusting for covariates, at both T1(Aβ1-40: p = 0.001; Aβ1-42: p = 0.0004) and T2 (Aβ1-40: p = 0.001; Aβ1-42: p = 0.016) employing the Single Molecule Array (Simoa) platform, however no significant differences were observed using the xMAP platform. Further, pairwise longitudinal analyses of plasma Aβ1-40 revealed decreased levels in MCs using data from the Simoa platform (p = 0.041) and pairwise longitudinal analyses of plasma Aβ1-42 revealed decreased levels in MCs using data from the xMAP platform (p = 0.041). Findings from the Simoa platform suggest that plasma Aβ may add value to a panel of biomarkers for the diagnosis of pre-symptomatic CAA, however, further validation studies in larger sample sets are required.     

Exploiting a Single-Crystal Environment to Minimize the Charge Noise on Qubits in Silicon

Electron spins in silicon offer a competitive, scalable quantum-computing platform with excellent single-qubit properties. However, the two-qubit gate fidelities achieved so far have fallen short of the 99% threshold required for large-scale error-corrected quantum computing architectures. In the past few years, there has been a growing realization that the critical obstacle in meeting this threshold in semiconductor qubits is charge noise arising from the qubit environment. In this work, a notably low level of charge noise of S₀ = 0.0088 ± 0.0004 μeV² Hz⁻¹ is demonstrated using atom qubits in crystalline silicon, achieved by separating the qubits from surfaces and interface states. The charge noise is measured using both a single electron transistor and an exchange-coupled qubit pair that collectively provide a consistent charge noise spectrum over four frequency decades, with the noise level S₀ being an order of magnitude lower than previously reported. Low-frequency detuning noise, set by the total measurement time, is shown to be the dominant dephasing source of two-qubit exchange oscillations. With recent advances in fast (≈μs) single-shot readout, it is shown that by reducing the total measurement time to ≈1 s, 99.99% two-qubit $\text{◂√▸}\sqrt{SWAP}$ gate fidelities can be achieved in single-crystal atom qubits in silicon.     

Knock-Down of HDAC2 in Human Induced Pluripotent Stem Cell Derived Neurons Improves Neuronal Mitochondrial Dynamics,Neuronal Maturation and Reduces Amyloid Beta Peptides

Histone deacetylase 2 (HDAC2) is a major HDAC protein in the adult brain and has been shown to regulate many neuronal genes. The aberrant expression of HDAC2 and subsequent dysregulation of neuronal gene expression is implicated in neurodegeneration and brain aging. Human induced pluripotent stem cell-derived neurons (hiPSC-Ns) are widely used models for studying neurodegenerative disease mechanisms, but the role of HDAC2 in hiPSC-N differentiation and maturation has not been explored. In this study, we show that levels of HDAC2 progressively decrease as hiPSCs are differentiated towards neurons. This suppression of HDAC2 inversely corresponds to an increase in neuron-specific isoforms of Endophilin-B1, a multifunctional protein involved in mitochondrial dynamics. Expression of neuron-specific isoforms of Endophilin-B1 is accompanied by concomitant expression of a neuron-specific alternative splicing factor, SRRM4. Manipulation of HDAC2 and Endophilin-B1 using lentiviral approaches shows that the knock-down of HDAC2 or the overexpression of a neuron-specific Endophilin-B1 isoform promotes mitochondrial elongation and protects against cytotoxic stress in hiPSC-Ns, while HDAC2 knock-down specifically influences genes regulating mitochondrial dynamics and synaptogenesis. Furthermore, HDAC2 knock-down promotes enhanced mitochondrial respiration and reduces levels of neurotoxic amyloid beta peptides. Collectively, our study demonstrates a role for HDAC2 in hiPSC-neuronal differentiation, highlights neuron-specific isoforms of Endophilin-B1 as a marker of differentiating hiPSC-Ns and demonstrates that HDAC2 regulates key neuronal and mitochondrial pathways in hiPSC-Ns.     

The impact of zinc neutralization on the structure and dynamics of precise polyethylene acrylic acid ionomers: A solid-state 13C NMR study

Solid-state ¹³C NMR experiments were performed on a series of linear poly(ethylene-co-acrylic acid) (E-AA) copolymers with the carboxylic acid group spaced precisely or randomly along the polyethylene backbone in order to determine the impact that molar percent acid content and acid group spacing have on the structure and dynamics of these materials. The impact of Zn-neutralization is investigated in precise E-AA ionomers. ¹³C cross-polarization (CP) NMR experiments with ¹H dipolar or ¹H spin-lock filters were utilized to identify the amorphous or crystalline components in the materials. Constraints extracted from the filtering experiments were used to fit quantitative NMR data for samples that contained both crystalline and amorphous regions. All random copolymers contained crystalline phases, while only the precise sample with the longest polyethylene run between acid groups (twenty CH₂ groups) contained a crystalline component. The crystalline phase in this precise copolymer decreased with increasing zinc content.     

Sustainable reduction of bioreactor contamination in an industrial fermentation pilot plant

Facility experience primarily in drug-oriented fermentation equipment (producing small molecules such as secondary metabolites, bioconversions, and enzymes) and, to a lesser extent, in biologics-oriented fermentation equipment (producing large molecules such as recombinant proteins and microbial vaccines) in an industrial fermentation pilot plant over the past 15 years is described. Potential approaches for equipment design and maintenance, operational procedures, validation/verification testing, medium selection, culture purity/sterility analysis, and contamination investigation are presented, and those approaches implemented are identified. Failure data collected for pilot plant operation for nearly 15 years are presented and best practices for documentation and tracking are outlined. This analysis does not exhaustively discuss available design, operational and procedural options; rather it selectively presents what has been determined to be beneficial in an industrial pilot plant setting. Literature references have been incorporated to provide background and context where appropriate.     

Food worker hand washing practices: an observation study

Improvement of food worker hand washing practices is critical to the reduction of foodborne illness and is dependent upon a clear understanding of current hand washing practices. To that end, this study collected detailed observational data on food worker hand washing practices. Food workers (n = 321) were observed preparing food, and data were recorded on specific work activities for which hand washing is recommended (e.g., food preparation, handling dirty equipment). Data were also recorded on hand washing behaviors that occurred in conjunction with these work activities. Results indicated that workers engaged in approximately 8.6 work activities per hour for which hand washing is recommended. However, workers made hand washing attempts (i.e., removed gloves, if worn, and placed hands in running water) in only 32% of these activities and washed their hands appropriately (i.e., removed gloves, if worn, placed hands in running water, used soap, and dried hands) in only 27% of these work activities. Attempted and appropriate hand washing rates varied by work activity--they were significantly higher in conjunction with food preparation than other work activities (46 versus < or = 37% for attempted hand washing; 41 versus < or = 30% for appropriate hand washing) and were significantly lower in conjunction with touching the body than other work activities (13 versus > or = 27% for attempted hand washing; 10 versus > or = 23% for appropriate hand washing). Attempted and appropriate hand washing rates were significantly lower when gloves were worn (18 and 16%) than when gloves were not worn (37 and 30%). These findings suggest that the hand washing practices of food workers need to be improved, glove use may reduce hand washing, and restaurants should consider reorganizing their food preparation activities to reduce the frequency with which hand washing is needed.