Oak Crest Institute of Science

This research examines the relative influence of the internal combustion engine (ICE) on automobile performance compared to turbochargers and transmissions. The study draws from historical developments, case studies, and modern technological trends to demonstrate that ICE remains the primary determinant of power, efficiency, and driving dynamics. Data collection relied on scholarly databases, historical archives, and peer-reviewed journals, focusing on advances in engine design, turbocharging methods, and transmission systems. Analysis was conducted thematically to identify patterns in technological synergy and performance outcomes. The findings reveal that while turbochargers and transmissions enhance performance, they operate within the boundaries set by the ICEs capabilities. Historical evidence, from muscle cars to modern hybrid supercars, supports the engines central role. Even in the age of electrification, ICE innovation continues through synthetic fuels, hydrogen combustion, and efficiency optimization, ensuring relevance in performance, heavy-duty, and enthusiast markets. This paper concludes that the ICE not only contributes to vehicle performance but defines it, making it the benchmark against which all future automotive technologies will be measured.

The SARS-CoV-2 infection kinetics in a real-world, clinical setting represent a knowledge gap in understanding the underlying COVID-19 pathogenesis. There are scant reports on the dynamics describing the two principal components of the viral life cycle, namely the rapid proliferation and slower clearance phases. Here, we present results from an ongoing workplace clinical surveillance study where two vaccinated participants became infected with SARS-CoV-2 Omicron variant (BA.1. lineage). The subjects were followed longitudinally at high temporal resolution allowing the kinetics of both viral phases to be characterized. The viral doubling times in the proliferation phase (3.3-3.5 h) and maximum measured viral loads were similar to those observed for unvaccinated individuals infected with an earlier SARS-CoV-2 strain. However, the clearance phase was much shorter in the current study and unexpectedly displayed a multimodal profile. Longitudinal whole genome SARS-CoV-2 sequencing identified a stable mutation that arose in one of the participants over the 2-week period of positivity. Our small study provides a rare insight into the clinical SARS-CoV-2 dynamics holding significance to public health measures and the biology underlying COVID-19.

SummaryLeber’s hereditary optic neuropathy (LHON), a disease associated with a mitochondrial DNA mutation, is characterized by blindness due to degeneration of retinal ganglion cells (RGCs) and their axons, which form the optic nerve. We show that a sustained pathological autophagy and compartment-specific mitophagy activity affects LHON patient-derived cells and cybrids, as well as induced pluripotent-stem-cell-derived neurons. This is variably counterbalanced by compensatory mitobiogenesis. The aberrant quality control disrupts mitochondrial homeostasis as reflected by defective bioenergetics and excessive reactive oxygen species production, a stress phenotype that ultimately challenges cell viability by increasing the rate of apoptosis. We counteract this pathological mechanism by using autophagy regulators (clozapine and chloroquine) and redox modulators (idebenone), as well as genetically activating mitochondrial biogenesis (PGC1-α overexpression). This study substantially advances our understanding of LHON pathophysiology, providing an integrated paradigm for pathogenesis of mitochondrial diseases and druggable targets for therapy.

Abstract A comprehensive understanding of the SARS-CoV-2 infection dynamics and the ensuing host immune responses is needed to explain the pathogenesis as it relates to viral transmission. Here, we report results from an ongoing, longitudinal, workplace clinical surveillance study that address knowledge gaps surrounding SARS-CoV-2 in vivo kinetics, particularly in the earliest stages after exposure. Nine study participants who developed COVID-19 between November, 2020 and March, 2021 were monitored at high temporal resolution for three months in terms of viral loads as well as associated inflammatory biomarker and antibody responses. Analysis of the resulting datasets, supported by Bayesian modeling, allowed the underlying kinetic processes to be described, yielding a number of unexpected findings. Early viral replication is more rapid (median doubling time, 3.1 hours) than previously thought, shrinking the window between exposure and viral shedding. Results from our small study give a rare insight into the life-cycle of COVID-19 infection and hold a number of important biological, clinical, and public health implications.

ABSTRACTIn developing and mature nervous systems, diverse neuronal subtypes innervate common targets to establish and maintain functional neural circuits. A major challenge towards understanding the structural and functional architecture of neural circuits is to separate these inputs and determine their intrinsic and heterosynaptic relationships. The Drosophila larval neuromuscular junction is a powerful model system to study these questions, where two glutamatergic motor neurons, the strong phasic-like Is and weak tonic-like Ib, co-innervate individual muscle targets to coordinate locomotor behavior. However, complete neurotransmission from each input has never been electrophysiologically separated. We have developed a botulinum neurotoxin, BoNT-C, that eliminates both spontaneous and evoked neurotransmission without perturbing synaptic growth or structure, enabling the first approach that accurately isolates input-specific neurotransmission. Selective expression of BoNT-C in Is or Ib motor inputs disambiguates functional properties of each input. Importantly, the composite values of Is and Ib neurotransmission can be fully recapitulated by isolated physiology from each input. Finally, selective silencing by BoNT-C does not induce heterosynaptic structural or functional plasticity at the convergent input. Thus, BoNT-C establishes the first approach to cleanly separate neurotransmission between tonic vs phasic neurons and defines heterosynaptic plasticity rules in a powerful model glutamatergic circuit.

Abstract Geological conditions play a significant role in prebiotic/abiotic organic chemistry, especially when reactive minerals are present. Previous studies of the prebiotic synthesis of amino acids and other products in mineral‐containing systems have shown that a diverse array of compounds can be produced, depending on the experimental conditions. However, these previous experiments have not simulated the effects of varying geochemical conditions, in which factors such as pH, iron redox state, or chemical concentrations may vary over time and space in a natural environment. In geochemical systems that contain overlapping gradients, many permutations of individual conditions could exist and affect the outcome of an organic reaction network. We investigated reactions of pyruvate and glyoxylate, two compounds that are central to the emergence of metabolism, in simulated geological gradients of redox, pH, and ammonia concentration. Our results show that the positioning of pyruvate/glyoxylate reactions in this environmental parameter space determines the organic product distribution that results. Therefore, the distribution pattern of amino acids and alpha‐hydroxy acids produced prebiotically in a system reflects the specific reaction conditions, and would be distinct at various locations in an environment depending on local geochemistry. This is significant for origin of life chemistry in which the composition and function of oligomers could be affected by the environmentally driven distribution of monomers available. Also, for astrobiology and planetary science where organic distribution patterns are sometimes considered as a possible biosignature, it is important to consider environmentally driven abiotic organic reactions that might produce similar effects.

Transportersare specialized integral membrane proteins, whichmediate the passage of virtually all molecules through cell membranes.They are expressed in a broad range of human and animal tissues andplay important roles in both normal and disease states. For thesereasons, they are evaluated when developing and testing drugs. Twomajor families of drug transporters, the adenosine 5′-triphosphate-bindingcassette and solute carrier transporters (SLC), have critical rolesin the absorption, distribution, metabolism, and elimination of drugs.The SLC family contains known nucleoside transporters and thereforeare important when nucleoside analogs are used as drugs to preventor treat viral infections. In this study, we wanted to determine ifit was possible to locate one member of the SLC family, the humanconcentrative nucleoside transporter 3 (CNT3) in human vaginal epithelialcells. The CNT3 protein has important roles in drug delivery, subsequentdrug tissue distribution, and, hence, efficacy. Vaginal epithelialcells, taken from two human volunteers (one Caucasian and one AfricanAmerican), were labeled for light and electron microscopy, with acommercial antibody to a cytoplasmic domain of CNT3, the protein productof the SLC28A3 gene. Fluorescent secondary antibodies or protein A-goldwere used to detect antibody binding. By electron microscopy, goldparticle binding was quantified to determine labeling specificity.By light microscopy, positive labeling with anti-CNT3 antibodies wasdetected on human vaginal epithelial cells, but specificity to anyintracellular structure was not easily determined, most likely a resultof specimen preparation. Electron microscopy revealed that the CNT3transporter protein was present predominantly on microvilli locatedon one side of some human vaginal epithelial cells. Quantificationconfirmed specific anti-CNT3 labeling over human vaginal epithelialcell microvilli. The CNT3 protein, present in the microvilli of humanvaginal epithelial cells, may have a role in redistributing nucleosidehomologues delivered to the vaginal tract. Transporter proteins suchas CNT3 could shuttle nucleosides and their analogs through the vaginalepithelium to immune cells located in lower cell layers. Outer layersof cells, which are eventually shed from the epithelium, may removeaccumulated nucleoside drug analogs from the vaginal tract.

Research on neuropeptide function has advanced rapidly, yet there is still no spatio-temporally resolved method to measure the release of neuropeptides in vivo. Here we introduce Neuropeptide Release Reporters (NPRRs): novel genetically-encoded sensors with high temporal resolution and genetic specificity. Using the Drosophila larval neuromuscular junction (NMJ) as a model, we provide evidence that NPRRs recapitulate the trafficking and packaging of native neuropeptides, and report stimulation-evoked neuropeptide release events as real-time changes in fluorescence intensity, with sub-second temporal resolution.

Research on neuropeptide function has advanced rapidly, yet there is still no spatio-temporally resolved method to measure the release of neuropeptides in vivo. Here we introduce Neuropeptide Release Reporters (NPRRs): novel genetically-encoded sensors with high temporal resolution and genetic specificity. Using the Drosophila larval neuromuscular junction (NMJ) as a model, we provide evidence that NPRRs recapitulate the trafficking and packaging of native neuropeptides, and report stimulation-evoked neuropeptide release events as real-time changes in fluorescence intensity, with sub-second temporal resolution.

The ubiquitin ligase Highwire restrains synaptic growth and promotes evoked neurotransmission at NMJ synapses in Drosophila Highwire regulates synaptic morphology by downregulating the MAP3K Wallenda, but excess Wallenda signaling does not account for the decreased presynaptic release observed in highwire mutants. Hence, Highwire likely has a second substrate that inhibits neurotransmission. Highwire targets the NAD+ biosynthetic and axoprotective enzyme dNmnat to regulate axonal injury responses. dNmnat localizes to synapses and interacts with the active zone protein Bruchpilot, leading us to hypothesize that Highwire promotes evoked release by downregulating dNmnat. Here, we show that excess dNmnat is necessary in highwire mutants and sufficient in wild-type larvae to reduce quantal content, likely via disruption of active zone ultrastructure. Catalytically active dNmnat is required to drive defects in evoked release, and depletion of a second NAD+ synthesizing enzyme is sufficient to suppress these defects in highwire mutants, suggesting that excess NAD+ biosynthesis is the mechanism inhibiting neurotransmission. Thus, Highwire downregulates dNmnat to promote evoked synaptic release, suggesting that Highwire balances the axoprotective and synapse-inhibitory functions of dNmnat.