Solutions to a Radical Problem: Overview of Current and Future Treatment Strategies in Leber’s Hereditary Opic Neuropathy

Leber’s Hereditary Optic Neuropathy (LHON) is the most common primary mitochondrial DNA disorder. It is characterized by bilateral severe central subacute vision loss due to specific loss of Retinal Ganglion Cells and their axons. Historically, treatment options have been quite limited, but ongoing clinical trials show promise, with significant advances being made in the testing of free radical scavengers and gene therapy. In this review, we summarize management strategies and rational of treatment based on current insights from molecular research. This includes preventative recommendations for unaffected genetic carriers, current medical and supportive treatments for those affected, and emerging evidence for future potential therapeutics.     

Propagation of Data Error and Parametric Sensitivity in Computable General Equilibrium Models

While computable general equilibrium (CGE) models are a well-established tool in economic analyses, it is often difficult to disentangle the effects of policies of interest from that of the assumptions made regarding the underlying calibration data and model parameters. To characterize the behavior of a CGE model of carbon output with respect to two of these assumptions, we perform a large-scale Monte Carlo experiment to examine its sensitivity to base year calibration data and elasticity of substitution parameters in the absence of a policy change. By examining a variety of output variables at different levels of economic and geographic aggregation, we assess how these forms of uncertainty impact the conclusions that can be drawn from the model simulations. We find greater sensitivity to uncertainty in the elasticity of substitution parameters than to uncertainty in the base-year data as the projection period increases. While many model simulations were conducted to generate large output samples, we find that few are required to capture the mean model response of the variables tested. However, characterizing standard errors and empirical probability distribution functions is not possible without a large number of simulations.     

Nonparallel training for voice conversion based on a parameter adaptation approach

The objective of voice conversion algorithms is to modify the speech by a particular source speaker so that it sounds as if spoken by a different target speaker. Current conversion algorithms employ a training procedure, during which the same utterances spoken by both the source and target speakers are needed for deriving the desired conversion parameters. Such a (parallel) corpus, is often difficult or impossible to collect. Here, we propose an algorithm that relaxes this constraint, i.e., the training corpus does not necessarily contain the same utterances from both speakers. The proposed algorithm is based on speaker adaptation techniques, adapting the conversion parameters derived for a particular pair of speakers to a different pair, for which only a nonparallel corpus is available. We show that adaptation reduces the error obtained when simply applying the conversion parameters of one pair of speakers to another by a factor that can reach 30%. A speaker identification measure is also employed that more insightfully portrays the importance of adaptation, while listening tests confirm the success of our method. Both the objective and subjective tests employed, demonstrate that the proposed algorithm achieves comparable results with the ideal case when a parallel corpus is available.     

Integration of Emerging Biomedical Technologies in Meat Processing to Improve Meat Safety and Quality

Modern‐day processing of meat products involves a series of complex procedures designed to ensure the quality and safety of the meat for consumers. As the size of abattoirs increases, the logistical problems associated with large‐capacity animal processing can affect the sanitation of the facility and the meat products, potentially increasing transmission of infectious diseases. Additionally, spoilage of food from improper processing and storage increases the global economic and ecological burden of meat production. Advances in biomedical and materials science have allowed for the development of innovative new antibacterial technologies that have broad applications in the medical industry. Additionally, new approaches in tissue engineering and nondestructive cooling of biological specimens could significantly improve organ transplantation and tissue grafting. These same strategies may be even more effective in the preservation and protection of meat as animal carcasses are easier to manipulate and do not have the same stringent requirements of care as living patients. This review presents potential applications of emerging biomedical technologies in the food industry to improve meat safety and quality. Future research directions investigating these new technologies and their usefulness in the meat processing chain along with regulatory, logistical, and consumer perception issues will also be discussed.     

Col4a3-/- Mice on Balb/C Background Have Less Severe Cardiorespiratory Phenotype and SGLT2 Over-Expression Compared to 129x1/SvJ and C57Bl/6 Backgrounds

Alport syndrome (AS) is a hereditary renal disorder with no etiological therapy. In the preclinical Col4a3^-/- model of AS, disease progression and severity vary depending on mouse strain. The sodium-glucose cotransporter 2 (SGLT2) is emerging as an attractive therapeutic target in cardiac/renal pathologies, but its application to AS remains untested. This study investigates cardiorespiratory function and SGLT2 renal expression in Col4a3^-/- mice from three different genetic backgrounds, 129x1/SvJ, C57Bl/6 and Balb/C. male Col4a3^-/- 129x1/SvJ mice displayed alterations consistent with heart failure with preserved ejection fraction (HFpEF). Female, but not male, C57Bl/6 and Balb/C Col4a3^-/- mice exhibited mild changes in systolic and diastolic function of the heart by echocardiography. Male C57Bl/6 Col4a3^-/- mice presented systolic dysfunction by invasive hemodynamic analysis. All strains except Balb/C males demonstrated alterations in respiratory function. SGLT2 expression was significantly increased in AS compared to WT mice from all strains. However, cardiorespiratory abnormalities and SGLT2 over-expression were significantly less in AS Balb/C mice compared to the other two strains. Systolic blood pressure was significantly elevated only in mutant 129x1/SvJ mice. The results provide further evidence for strain-dependent cardiorespiratory and hypertensive phenotype variations in mouse AS models, corroborated by renal SGLT2 expression, and support ongoing initiatives to develop SGLT2 inhibitors for the treatment of AS.     

CX3CL1 Action on Microglia Protects from Diet-Induced Obesity by Restoring POMC Neuronal Excitability and Melanocortin System Activity Impaired by High-Fat Diet Feeding

Both hypothalamic microglial inflammation and melanocortin pathway dysfunction contribute to diet-induced obesity (DIO) pathogenesis. Previous studies involving models of altered microglial signaling demonstrate altered DIO susceptibility with corresponding POMC neuron cytological changes, suggesting a link between microglia and the melanocortin system. We addressed this hypothesis using the specific microglial silencing molecule, CX3CL1 (fractalkine), to determine whether reducing hypothalamic microglial activation can restore POMC/melanocortin signaling to protect against DIO. We performed metabolic analyses in high fat diet (HFD)-fed mice with targeted viral overexpression of CX3CL1 in the hypothalamus. Electrophysiologic recording in hypothalamic slices from POMC-MAPT-GFP mice was used to determine the effects of HFD feeding and microglial silencing via minocycline or CX3CL1 on GFP-labeled POMC neurons. Finally, mice with hypothalamic overexpression of CX3CL1 received central treatment with the melanocortin receptor antagonist SHU9119 to determine whether melanocortin signaling is required for the metabolic benefits of CX3CL1. Hypothalamic overexpression of CX3CL1 increased leptin sensitivity and POMC gene expression, while reducing weight gain in animals fed an HFD. In electrophysiological recordings from hypothalamic slice preparations, HFD feeding was associated with reduced POMC neuron excitability and increased amplitude of inhibitory postsynaptic currents. Microglial silencing using minocycline or CX3CL1 treatment reversed these HFD-induced changes in POMC neuron electrophysiologic properties. Correspondingly, blockade of melanocortin receptor signaling in vivo prevented both the acute and chronic reduction in food intake and body weight mediated by CX3CL1. Our results show that suppressing microglial activation during HFD feeding reduces DIO susceptibility via a mechanism involving increased POMC neuron excitability and melanocortin signaling.     

Generalized Approach towards Secretion-Based Protein Production via Neutralization of Secretion-Preventing Cationic Substrate Residues

Many heterologous proteins can be secreted by bacterial ATP-binding cassette (ABC) transporters, provided that they are fused with the C-terminal signal sequence, but some proteins are not secretable even though they carry the right signal sequence. The invention of a method to secrete these non-secretable proteins would be valuable both for understanding the secretory physiology of ABC transporters and for industrial applications. Herein, we postulate that cationic “supercharged” regions within the target substrate protein block the secretion by ABC transporters. We also suggest that the secretion of such substrate proteins can be rescued by neutralizing those cationic supercharged regions via structure-preserving point mutageneses. Surface-protruding, non-structural cationic amino acids within the cationic supercharged regions were replaced by anionic or neutral hydrophilic amino acids, reducing the cationic charge density. The examples of rescued secretions we provide include the spike protein of SARS-CoV-2, glutathione-S-transferase, streptavidin, lipase, tyrosinase, cutinase, growth factors, etc. In summary, our study provides a method to predict the secretability and a tool to rescue the secretion by correcting the secretion-blocking regions, making a significant step in understanding the physiological properties of ABC transporter-dependent protein secretion and laying the foundation for the development of a secretion-based protein-producing platform.     

Timing HIV infection with a simple and accurate population viral dynamics model

Clinical trials for HIV prevention can require knowledge of infection times to subsequently determine protective drug levels. Yet, infection timing is difficult when study visits are sparse. Using population nonlinear mixed-effects (pNLME) statistical inference and viral loads from 46 RV217 study participants, we developed a relatively simple HIV primary infection model that achieved an excellent fit to all data. We also discovered that Aptima assay values from the study strongly correlated with viral loads, enabling imputation of very early viral loads for 28/46 participants. Estimated times between infecting exposures and first positives were generally longer than prior estimates (average of two weeks) and were robust to missing viral upslope data. On simulated data, we found that tighter sampling before diagnosis improved estimation more than tighter sampling after diagnosis. Sampling weekly before and monthly after diagnosis was a pragmatic design for good timing accuracy. Our pNLME timing approach is widely applicable to other infections with existing mathematical models. The present model could be used to simulate future HIV trials and may help estimate protective thresholds from the recently completed antibody-mediated prevention trials.     

Oxygen Reduction Reaction Performance of [MTBD][beti]‐Encapsulated Nanoporous NiPt Alloy Nanoparticles

Recent advances in oxygen reduction reaction catalysis for proton exchange membrane fuel cells (PEMFCs) include i) the use of electrochemical dealloying to produce high surface area and sometimes nanoporous catalysts with a Pt‐enriched outer surface, and ii) the observation that oxygen reduction in nanoporous materials can be potentially enhanced by confinement effects, particularly if the chemical environment within the pores can bias the reaction toward completion. Here, these advances are combined by incorporating a hydrophobic, protic ionic liquid, [MTBD][beti], into the pores of high surface‐area NiPt alloy nanoporous nanoparticles (np‐NiPt/C + [MTBD][beti]). The high O₂ solubility of the [MTBD][beti], in conjunction with the confined environment within the pores, biases reactant O₂ toward the catalytic surface, consistent with an increased residence time and enhanced attempt frequencies, resulting in improved reaction kinetics. Half‐cell measurements show the np‐NiPt/C+[MTBD][beti] encapsulated catalyst to be nearly an order of magnitude more active than commercial Pt/C, a result that is directly translated into operational PEMFCs.