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Coral reefs thrive and provide maximal ecosystem services when they support a multi-level trophic structure and grow in favorable water quality conditions that include high light levels, rapid water flow, and low nutrient levels. Poor water quality and other anthropogenic stressors have caused coral mortality in recent decades, leading to trophic downgrading and the loss of biological complexity on many reefs. Solutions to reverse the causes of trophic downgrading remain elusive, in part because efforts to restore reefs are often attempted in the same diminished conditions that caused coral mortality in the first place.Coral Arks, positively buoyant, midwater structures, are designed to provide improved water quality conditions and supportive cryptic biodiversity for translocated and naturally recruited corals to assemble healthy reef mesocosms for use as long-term research platforms. Autonomous Reef Monitoring Structures (ARMS), passive settlement devices, are used to translocate the cryptic reef biodiversity to the Coral Arks, thereby providing a "boost" to natural recruitment and contributing ecological support to the coral health. We modeled and experimentally tested two designs of Arks to evaluate the drag characteristics of the structures and assess their long-term stability in the midwater based on their response to hydrodynamic forces.We then installed two designs of Arks structures at two Caribbean reef sites and measured several water quality metrics associated with the Arks environment over time. At deployment and 6 months after, the Coral Arks displayed enhanced metrics of reef function, including higher flow, light, and dissolved oxygen, higher survival of translocated corals, and reduced sedimentation and microbialization relative to nearby seafloor sites at the same depth. This method provides researchers with an adaptable, long-term platform for building reef communities where local water quality conditions can be adjusted by altering deployment parameters such as the depth and site.

The utility of single fluorescent protein biosensors (SFPBs) in biological research is offset by the difficulty in engineering these tools. SFPBs generally consist of three basic components: a circularly permuted fluorescent protein, a ligand-binding domain, and a pair of linkers connecting the two domains. In the absence of predictive methods for biosensor engineering, most designs combining these three components will fail to produce allosteric coupling between ligand binding and fluorescence emission. Methods to construct libraries of biosensor designs with variations in the site of GFP insertion and linker sequences have been developed, however, our ability to construct new variants has exceeded our ability to test them for function. Here, we address this challenge by applying a massively parallel assay termed “sort-seq” to the characterization of biosensor libraries. Sort-seq combines binned fluorescence-activated cell sorting, next-generation sequencing, and maximum likelihood estimation to quantify the dynamic range of many biosensor variants in parallel. We applied this method to two common biosensor optimization tasks: choice of insertion site and optimization of linker sequences. The sort-seq assay applied to a maltose-binding protein domain-insertion library not only identified previously described high-dynamic-range variants but also discovered new functional insertion-sites with diverse properties. A sort-seq assay performed on a pyruvate biosensor linker library expressed in mammalian cell culture identified linker variants with substantially improved dynamic range. Machine learning models trained on the resulting data can predict dynamic range from linker sequence. This high-throughput approach will accelerate the design and optimization of SFPBs, expanding the biosensor toolbox.

Cellular metabolism is regulated over space and time to ensure that energy production is efficiently matched with consumption. Fluorescent biosensors are useful tools for studying metabolism as they enable real-time detection of metabolite abundance with single-cell resolution. For monitoring glycolysis, the intermediate fructose 1,6-bisphosphate (FBP) is a particularly informative signal as its concentration is strongly correlated with flux through the whole pathway. Using GFP insertion into the ligand-binding domain of theBacillus subtilistranscriptional regulator CggR, we developed a fluorescent biosensor for FBP termed HYlight. We demonstrate that HYlight can reliably report the real-time dynamics of glycolysis in living cells and tissues, driven by various metabolic or pharmacological perturbations, alone or in combination with other physiologically relevant signals. Using this sensor, we uncovered previously unknown aspects of β-cell glycolytic heterogeneity and dynamics.

Most invasive Brain Computer Interfaces (iBCIs) use spike and Local Field Potentials (LFPs) from the motor or parietal cortices to decode movement intentions. It has been debated whether harvesting signals from other brain areas that encode global cognitive variables, such as the allocation of attention and eye movement goals in a variety of spatial reference frames, may improve the outcome of iBCIs. Here, we explore the ability of LFP signals, sampled from the lateral prefrontal cortex (LPFC) of macaque monkeys, to encode eye-movement intention during the pre-movement fixation period of a delayed saccade task. We use spectral dimensionality reduction to examine the spatiotemporal properties of the extracted non-rhythmic broadband activity and explore its usefulness in decoding saccade goals. The dynamics of the broadband signal in low spatial dimensions across the pre-movement fixation period uncovered saccade target separation; its discriminative potential was confirmed using support vector machine classifications. These findings reveal that broadband LFP from the LPFC can be used to decode intended saccade target location during pre-movement periods. We further provide a general workflow that can be implemented in iBCIs and it is relatively robust to the loss of spikes in individual electrodes.

ContextDeficits in both balance and oculomotor function, including impairments in saccadic eye movements, are observed in approximately 30% of patients postconcussion. Whereas balance and saccadic eye movements are routinely assessed separately, growing evidence suggests that they should be assessed concurrently.ObjectiveTo compare balance measures and saccades between adolescents 1 to 3 months postconcussion and healthy uninjured adolescents.DesignCase-control study.SettingConcussion clinic and 2 private schools.Patients or Other ParticipantsTwenty-five adolescents (10 boys, 15 girls; median [interquartile range (IQR)] age = 14 years [11.5–16 years]) between 1 and 3 months postconcussion (median [IQR] time since injury = 39.5 days [30–56.75 days]) and 33 uninjured adolescents (18 boys, 15 girls; median [IQR] age = 13 years [11.5–14 years]).Main Outcome Measure(s)The center-of-pressure 95% ellipse area and medial-lateral and anterior-posterior velocity and the number of saccades in the dual-task balance conditions including a high cognitive load (cognitive condition), a low cognitive load and a gaze-shifting component (visual condition) or both a high cognitive load and a gaze-shifting component (combined condition).ResultsConcussion-group participants swayed over larger center-of-pressure ellipse areas in the visual (P = .02; effect size = 0.73) and combined (P = .005; effect size = 0.86) conditions but not in the cognitive condition (P = .07; effect size = 0.50). No group differences were identified for anterior-posterior (F1,56 = 2.57, P = .12) or medial-lateral (F1,56 = 0.157, P = .69) velocity. Concussion-group participants also did not perform more saccades than the control-group participants (F1,56 = 2.04, P = .16).ConclusionsPerforming dual-task balance conditions for which the secondary task involved a gaze-shifting component or both a gaze-shifting component and a high cognitive load resulted in greater sway amplitude in adolescents with concussion. However, these larger amounts of postural sway were not associated with increased saccadic eye movements.

Axon degeneration, a hallmark of chemotherapy-induced peripheral neuropathy (CIPN), is thought to be caused by a loss of the essential metabolite nicotinamide adenine dinucleotide (NAD+) via the prodegenerative protein SARM1. Some studies challenge this notion, however, and suggest that an aberrant increase in a direct precursor of NAD+, nicotinamide mononucleotide (NMN), rather than loss of NAD+, is responsible. In support of this idea, blocking NMN accumulation in neurons by expressing a bacterial NMN deamidase protected axons from degeneration. We hypothesized that protection could similarly be achieved by reducing NMN production pharmacologically. To achieve this, we took advantage of an alternative pathway for NAD+ generation that goes through the intermediate nicotinic acid mononucleotide (NAMN), rather than NMN. We discovered that nicotinic acid riboside (NAR), a precursor of NAMN, administered in combination with FK866, an inhibitor of the enzyme nicotinamide phosphoribosyltransferase that produces NMN, protected dorsal root ganglion (DRG) axons against vincristine-induced degeneration as well as NMN deamidase. Introducing a different bacterial enzyme that converts NAMN to NMN reversed this protection. Collectively, our data indicate that maintaining NAD+ is not sufficient to protect DRG neurons from vincristine-induced axon degeneration, and elevating NMN, by itself, is not sufficient to cause degeneration. Nonetheless, the combination of FK866 and NAR, which bypasses NMN formation, may provide a therapeutic strategy for neuroprotection.

Aseptic loosening remains one of the leading causes for failure of total knee arthroplasty (TKA). We sought to identify early radiographic measures that may associate with aseptic tibial component loosening, emphasizing systematic evaluation of the cement mantle.All TKA revisions from 2007 to 2015 with the primary indication of tibial aseptic loosening were identified using in an institutional implant retrieval database. After exclusion criteria, 61 TKAs comprised the study group. A matched control group of 59 TKAs that had not failed at a minimum of 3 years was identified for comparison. Radiographic analysis on all 6-week postoperative radiographs included angulation of components, cement penetration depth, and presence of radiolucency at the implant-cement and bone-cement interfaces. Groups were compared with Student's t-test, chi-squared test, and Mann-Whitney U-test. A final multivariable logistic regression model was formed for the outcome of aseptic loosening.On multivariable analysis, failure was associated with a greater number of zones with cement penetration2 mm (5.6 vs 3.4 zones, odds ratio [OR] 1.89, P.001), increasing percent involvement of radiolucency at the implant-cement interface (8.7% vs 3.1%, OR = 1.15, P = .001), and increased varus alignment of the tibial component (1.5° vs 0°, OR = 1.35, P = .014). A greater number of zones with a radiolucent line at the bone-cement interface did not significantly associate (1.1 vs 0.3, P = .091).Our results suggest that radiographic indicators of poor cement mantle quality associate with later aseptic loosening. This emphasizes the need for surgeons to perform careful cement technique in order to reduce the risk of TKA failure.III (Case-control).