Autofluorescence is a biomarker of neural stem cell activation state.

Neural stem cells (NSCs) must exit quiescence to produce neurons; however, our understanding of this process remains constrained by the technical limitations of current technologies. Fluorescence lifetime imaging (FLIM) of autofluorescent metabolic cofactors has been used in other cell types to study shifts in cell states driven by metabolic remodeling that change the optical properties of these endogenous fluorophores. Using this non-destructive, live-cell, and label-free strategy, we found that quiescent NSCs (qNSCs) and activated NSCs (aNSCs) have unique autofluorescence profiles. Specifically, qNSCs display an enrichment of autofluorescence localizing to a subset of lysosomes, which can be used as a graded marker of NSC quiescence to predict cell behavior at single-cell resolution. Coupling autofluorescence imaging with single-cell RNA sequencing, we provide resources revealing transcriptional features linked to deep quiescence and rapid NSC activation. Together, we describe an approach for tracking mouse NSC activation state and expand our understanding of adult neurogenesis.

Occupational Therapy: An Essential Component of Support for Young Children With Cancer.

We established a multidisciplinary early childhood clinic to support infants and toddlers receiving cancer treatment. The access to occupational therapy (OT) in this population is unknown. To describe the access to OT for infants and toddlers with cancer. We retrospectively reviewed medical records to determine the frequency and characteristics of children with cancer who were referred to OT. Demographic data, medical information, and frequency of referrals were extracted from September 2015 to September 2018. Of the 134 patients, 112 (83.6%) received an OT evaluation: 88.4% were referred for hospital-based OT services; 42.0% were recommended for services upon returning home. Between-group comparisons revealed significant differences in referrals for home- or community-based OT services based on age and disease. OT is crucial to treatment plans that address the developmental needs of young pediatric oncology patients during hospitalization and thereafter. Recommendations for monitoring this vulnerable population are provided.

Interneuronal dynamics facilitate the initiation of spike block in cortical microcircuits.

Pyramidal cell spike block is a common occurrence in migraine with aura and epileptic seizures. In both cases, pyramidal cells experience hyperexcitation with rapidly increasing firing rates, major changes in electrochemistry, and ultimately spike block that temporarily terminates neuronal activity. In cortical spreading depression (CSD), spike block propagates as a slowly traveling wave of inactivity through cortical pyramidal cells, which is thought to precede migraine attacks with aura. In seizures, highly synchronized cortical activity can be interspersed with, or terminated by, spike block. While the identifying characteristic of CSD and seizures is the pyramidal cell hyperexcitation, it is currently unknown how the dynamics of the cortical microcircuits and inhibitory interneurons affect the initiation of hyperexcitation and subsequent spike block.We tested the contribution of cortical inhibitory interneurons to the initiation of spike block using a cortical microcircuit model that takes into account changes in ion concentrations that result from neuronal firing. Our results show that interneuronal inhibition provides a wider dynamic range to the circuit and generally improves stability against spike block. Despite these beneficial effects, strong interneuronal firing contributed to rapidly changing extracellular ion concentrations, which facilitated hyperexcitation and led to spike block first in the interneuron and then in the pyramidal cell. In all cases, a loss of interneuronal firing triggered pyramidal cell spike block. However, preventing interneuronal spike block was insufficient to rescue the pyramidal cell from spike block. Our data thus demonstrate that while the role of interneurons in cortical microcircuits is complex, they are critical to the initiation of pyramidal cell spike block. We discuss the implications that localized effects on cortical interneurons have beyond the isolated microcircuit and their contribution to CSD and epileptic seizures.

The dynamic range of voltage-dependent gap junction signaling is maintained by Ih-induced membrane potential depolarization.

Like their chemical counterparts, electrical synapses show complex dynamics such as rectification and voltage dependence that interact with other electrical processes in neurons. The consequences arising from these interactions for the electrical behavior of the synapse, and the dynamics they create, remain largely unexplored. Using a voltage-dependent electrical synapse between a descending modulatory projection neuron (MCN1) and a motor neuron (LG) in the crustacean stomatogastric ganglion, we find that the influence of the hyperpolarization-activated inward current (Ih) is critical to the function of the electrical synapse. When we blocked Ih with CsCl, the apparent voltage dependence of the electrical synapse shifted by 18.7 mV to more hyperpolarized voltages, placing the dynamic range of the electrical synapse outside of the range of voltages used by the LG motor neuron (-60.2 mV to -44.9 mV). With dual electrode current- and voltage-clamp recordings, we demonstrate that this voltage shift is not due to a change in the properties of the gap junction itself, but is a result of a sustained effect of Ih on the presynaptic MCN1 axon terminal membrane potential. Ih-induced depolarization of the axon terminal membrane potential increased the electrical postsynaptic potentials and currents. With Ih present, the axon terminal resting membrane potential is depolarized, shifting the dynamic range of the electrical synapse toward the functional range of the motor neuron. We thus demonstrate that the function of an electrical synapse is critically influenced by a voltage-dependent ionic current (Ih).NEW & NOTEWORTHY Electrical synapses and voltage-gated ionic currents are often studied independently from one another, despite mounting evidence that their interactions can alter synaptic behavior. We show that the hyperpolarization-activated inward ionic current shifts the voltage dependence of electrical synaptic transmission through its depolarizing effect on the membrane potential, enabling it to lie within the functional membrane potential range of a motor neuron. Thus, the electrical synapse's function critically depends on the voltage-gated ionic current.

Characterization of Focal Liver Masses: A Multicenter Comparison of Contrast-Enhanced Ultrasound, Computed Tomography, and Magnetic Resonance Imaging.

OBJECTIVES: To demonstrate the usefulness of contrast-enhanced ultrasound (CEUS) for the evaluation of focal liver masses via a direct comparison to standard ultrasound and computed tomography/magnetic resonance imaging (CT/MRI). METHODS: A cohort of 214 patients with previously undiagnosed focal liver masses were included from 5 different centers. Each patient was imaged using CEUS and CT and/or MRI. Anonymized and randomized images were interpreted by 4 separate blind readers from 3 of the participating centers (2 readers for CEUS and 2 readers for CT/MRI). Readers were blinded to patient demographics and past medical history. Readers were asked to decide if the lesion was benign or malignant, provide a final diagnosis for the lesion, and provide a confidence interval. Results were compared to truth standard from pathology or expert consensus. RESULTS: In determination of malignancy, CEUS had a sensitivity of 95%, specificity of 82%, PPV of 82%, NPV of 95%, statistically better than standard ultrasound (sensitivity 82%, specificity 56%, PPV 60%, NPV 78%) with P < .01 and not statistically different from CT (sensitivity 90%, specificity 73% PPV 81%, NPV 86%) or MRI (sensitivity 85%, specificity 79%, PPV 68%, NPV 91%) with P ≥ .01. In assigning a final diagnosis, CEUS had an accuracy of 78% statistically better than standard ultrasound (46%) with P < .01 and not statistically different from CT (68%) or MRI (71%) with P > .01. CONCLUSIONS: In the evaluation of focal liver lesions, both for determination of malignancy and in accuracy of final diagnosis, CEUS performs better than standard ultrasound and at least equivalent to both CT and MRI.

GDNF drives rapid tubule morphogenesis in a novel 3D in vitro model for ADPKD.

Cystogenesis is a morphological consequence of numerous genetic diseases of the epithelium. In the kidney, the pathogenic mechanisms underlying the program of altered cell and tubule morphology are obscured by secondary effects of cyst expansion. Here, we developed a new 3D tubuloid system to isolate the rapid changes in protein localization and gene expression that correlate with altered cell and tubule morphology during cyst initiation. Mouse renal tubule fragments were pulsed with a cell differentiation cocktail including glial-derived neurotrophic factor (GDNF) to yield collecting duct-like tubuloid structures with appropriate polarity, primary cilia, and gene expression. Using the 3D tubuloid model with an inducible Pkd2 knockout system allowed the tracking of morphological, protein, and genetic changes during cyst formation. Within hours of inactivation of Pkd2 and loss of polycystin-2, we observed significant progression in tubuloid to cyst morphology that correlated with 35 differentially expressed genes, many related to cell junctions, matrix interactions, and cell morphology previously implicated in cystogenesis.This article has an associated First Person interview with the first author of the paper.

Conformer-Specific Photolysis of Pyruvic Acid and the Effect of Water.

The conformer-specific reactivity of gas-phase pyruvic acid following the S1(nπ*) ← S0 excitation at λmax = 350 nm (290-380 nm) and the effect of water are investigated for the two lowest energy conformers. Conformer-specific gas-phase pyruvic acid photolysis rate constants and their respective populations are measured by monitoring their distinct vibrational OH-stretching frequencies. The geometry, relative energies, fundamental vibrational frequencies, and electronic transitions of the pyruvic acid conformers and their monohydrated complexes are calculated with density functional theory and ab initio methods. Results from experiment and theory show that the more stable conformer with an intramolecular hydrogen bond dominates the gas-phase photolysis of pyruvic acid. Water greatly affects the gas-phase pyruvic acid conformer population and photochemistry through hydrogen bonding interactions. The addition of water decreases the gas-phase relative population of the more stable conformer and decreases the molecule's gas-phase photolysis rate constants. The theoretical results show that even a single water molecule interrupts the intramolecular hydrogen bond, which is essential for the efficient photodissociation of gas-phase pyruvic acid. Results of this study suggest that the aqueous-phase photochemistry of pyruvic acid proceeds through hydrogen-bonded conformers lacking an intramolecular hydrogen bond.

Atmospheric Simulation Chamber Studies of the Gas-Phase Photolysis of Pyruvic Acid.

Pyruvic acid is an atmospherically abundant α-keto-acid that degrades efficiently from the troposphere via gas-phase photolysis. To explore conditions relevant to the environment, 2-12 ppm pyruvic acid is irradiated by a solar simulator in the environmental simulation chamber, CESAM. The combination of the long path length available in the chamber and its low surface area to volume ratio allows us to quantitatively examine the quantum yield and photochemical products of pyruvic acid. Such details are new to the literature for the low initial concentrations of pyruvic acid employed here. We determined photolysis quantum yields of ϕobsN2 = 0.84 ± 0.1 in nitrogen and ϕobsAir = 3.2 ± 0.5 in air, which are higher than those reported by previous studies that used higher partial pressures of pyruvic acid. The quantum yield greater than unity in air is due to secondary chemistry, driven by O2, that emerges under the conditions in these experiments. The low concentration of pyruvic acid and the resulting oxygen effect also alter the product distribution such that acetic acid, rather than acetaldehyde, is the primary product in air. These results indicate that tropospheric pyruvic acid may degrade in part via photoinduced mechanisms that are different than previously expected.

Contrast-enhanced US Approach to the Diagnosis of Focal Liver Masses.

Focal liver lesions are commonly encountered and often demonstrate nonspecific findings at initial imaging. Although most incidentally discovered liver lesions are benign, their noninvasive diagnosis is necessary, especially if they are large or atypical. Imaging characterization of focal liver lesions and exclusion of malignancy are of prime importance, particularly in high-risk populations. Contrast agent-enhanced ultrasonography of liver lesions is both accurate and reproducible for evaluation of benign and malignant liver tumors. Use of an imaging algorithm and a controlled sonographic technique, including dedicated arterial phase cine imaging and imaging every 30 seconds in the portal venous phase and the delayed (or late) phase, is essential for accurate characterization. This algorithmic analysis of focal liver lesions focuses first on the determination of malignancy by imaging the portal venous phase and the late phase; washout in these phases correlates with a malignant tumor, and sustained enhancement in these phases is suggestive that a lesion is benign. In addition, the timing and the intensity of washout differentiate hepatocellular malignancies from nonhepatocellular malignancies. Nonhepatocellular tumors demonstrate early and strong washout, whereas hepatocellular malignancies show delayed and weak washout. Subsequent analysis of dynamic real-time enhancement patterns in the arterial phase demonstrates specific enhancement patterns of common benign and malignant focal liver lesions. Hemangiomas show classic peripheral nodular enhancement, and spoke-wheel centrifugal enhancement is suggestive of focal nodular hyperplasia. Hepatic adenomas may show centripetal filling. However, arterial phase enhancement in malignancy has less specificity. Online supplemental material is available for this article. ©RSNA, 2017 •.

Multiphase Photochemistry of Pyruvic Acid under Atmospheric Conditions.

Aerosol and molecular processing in the atmosphere occurs in a complex and variable environment consisting of multiple phases and interfacial regions. To explore the effects of such conditions on the reactivity of chemical systems, we employ an environmental simulation chamber to investigate the multiphase photolysis of pyruvic acid, which photoreacts in the troposphere in aqueous particles and in the gas phase. Upon irradiation of nebulized pyruvic acid, acetic acid and carbon dioxide are rapidly generated, which is consistent with previous literature on the bulk phase photolysis reactions. Additionally, we identify a new C6 product, zymonic acid, a species that has not previously been reported from pyruvic acid photolysis under any conditions. Its observation here, and corresponding spectroscopic signatures, indicates it could be formed by heterogeneous reactions at the droplet surface. Prior studies of the aqueous photolysis of pyruvic acid have shown that high-molecular-weight compounds are formed via radical reactions; however, they are inhibited by the presence of oxygen, leading to doubt as to whether the chemistry would occur in the atmosphere. Identification of dimethyltartaric acid from the photolysis of multiphase pyruvic acid in air confirms radical polymerization chemistry can compete with oxygen reactions to some extent under aerobic conditions. Evidence of additional polymerization within the particles during irradiation is suggested by the increasing viscosity and organic content of the particles. The implications of multiphase specific processes are then discussed within the broader scope of atmospheric science.

Gas-Phase Photolysis of Pyruvic Acid: The Effect of Pressure on Reaction Rates and Products.

In this work, we investigate the impact of pressure and oxygen on the kinetics of and products from the gas-phase photolysis of pyruvic acid. The results reveal a decrease in the photolysis quantum yield as pressure of air or nitrogen is increased, a trend not yet documented in the literature. A Stern-Volmer analysis demonstrates this effect is due to deactivation of the singlet state of pyruvic acid when the photolysis is performed in nitrogen, and from quenching of both the singlet and triplet state in air. Consistent with previous studies, acetaldehyde and CO2 are observed as the major products; however, other products, most notably acetic acid, are also identified in this work. The yield of acetic acid increases with increasing pressure of buffer gas, an effect that is amplified by the presence of oxygen. At least two mechanisms are necessary to explain the acetic acid, including one that requires reaction of photolysis intermediates with O2. These findings extend the fundamental understanding of the gas-phase photochemistry of pyruvic acid, highlighting the importance of pressure on the photolysis quantum yields and products.

Bifurcation transitions in gap-junction-coupled neurons.

Here we investigate transitions occurring in the dynamical states of pairs of distinct neurons electrically coupled, with one neuron tonic and the other bursting. Depending on the dynamics of the individual neurons, and for strong enough coupling, they synchronize either in a tonic or a bursting regime, or initially tonic transitioning to bursting via a period doubling cascade. Certain intrinsic properties of the individual neurons such as minimum firing rates are carried over into the dynamics of the coupled neurons affecting their ultimate synchronous state.

Work-related musculoskeletal disorders in ultrasound: Can you reduce risk?

Work-related musculoskeletal disorders are a common cause of pain and sickness absence for ultrasound practitioners. This article aims to provide background information about factors increasing the chance of developing work-related musculoskeletal disorders and potential ways to reduce risk. Factors influencing ultrasound professionals' likelihood of developing work-related musculoskeletal disorders include poor posture, repetitive movements, transducer pressure and poor grip, stress, workload, limited support or sense of control and other psychosocial factors. The impact of these risk factors on the health and well being of ultrasound practitioners can be reduced by following recommendations published by professional bodies and the Health and Safety Executive. Ultrasound practitioners should remember that optimising the examination should not be at the detriment of their health. Some hints and tips to reduce the chance of developing work-related musculoskeletal disorders are provided.

Photochemical kinetics of pyruvic acid in aqueous solution.

Pyruvic acid in the atmosphere is found in both the gas and aqueous phases, and its behavior gives insight into that of other α-keto acids. Photolysis is a significant degradation pathway for this molecule in the environment, and in aqueous solution the major photoproducts are higher-molecular-weight compounds that may contribute to secondary organic aerosol mass. The kinetics of the aqueous-phase photolysis of pyruvic acid under aerobic and anaerobic conditions was investigated in order to calculate the first-order rate constant, Jaq, in solution. Analysis of the exponential decay of pyruvic acid was performed by monitoring both pyruvic acid and its photolytic products over the course of the reaction by (1)H NMR spectroscopy. Detection of major and minor products in the 0.1, 0.05, and 0.02 M pyruvic acid photolyses clearly demonstrates that the primary reaction pathways are highly dependent on the initial pyruvic acid concentration and the presence of dissolved oxygen. The Jaq values were calculated with approximations based on the dominant pathways for limiting cases of the mechanism. Finally, a model study using the calculated rate constants demonstrates the importance of aqueous-phase photolysis as a sink for pyruvic acid in the atmosphere, compared with gas-phase photolysis and OH oxidation.

Adenosine antagonists reverse the cataleptic effects of haloperidol: implications for the treatment of Parkinson's disease.

The effects of adenosine antagonists were compared in two rodent models of Parkinsonian symptoms. In the first experiment the dopamine D2 antagonist, haloperidol, was used to induce catalepsy. It was found that treatment with the non-selective adenosine antagonist caffeine significantly reduced catalepsy at each dose. Treatment with the selective A1 antagonist CPT also produced a significant reduction in catalepsy, as did treatment with the selective A2A antagonist SCH58261. In the second experiment haloperidol was used to suppress locomotor activity in an open field test. Treatment with caffeine significantly increased locomotion reduced by haloperidol, but not at all doses tested. Treatment with CPT also increased haloperidol-suppressed locomotor activity in dose-dependent manner. Surprisingly, treatment with SCH58261 did not significantly increase locomotor activity in animals treated with haloperidol at any dose tested. While some of these results were unexpected, the overall pattern suggests that adenosine antagonists would be useful as therapies for Parkinsonian patients as they appear to increase movement. The results also suggest that in acute timelines A1 antagonists may be more beneficial than previously supposed.

Stationary sources of airborne lead: a comparison of emissions data for southern California.

Estimates for the air releases of lead from stationary point sources are considered for the South Coast Air Basin of California. We have examined four databases published by U.S. Environmental Protection Agency, the California Air Resources Board, and the South Coast Air Quality Management District. Our analysis indicates that none of the databases includes every emitting facility in the South Coast Air Basin of California and that other discrepancies among the databases exist. Additionally, the data have been analyzed for temporal variation, and some of the California Air Resources Board data are not current. The South Coast Air Quality Management District inventory covers 12 times more facilities in 2001 than in 1996. From this analysis, we conclude that all four of the databases would benefit by sharing data, increasing transparency, analyzing uncertainty, and standardizing emission estimation methods.

The role of resuspended soil in lead flows in the California South Coast Air Basin.

The inputs and outputs of airborne lead in the South Coast Air Basin of California (SOCAB) are quantified according to standard mass balance calculations. Results for 2001 show that approximately 49,000 kg of lead exitthe Basin each year, but traditional sources contribute only about 6500 kg of lead each year. We resolve this discrepancy through a simple computer model that quantifies the resuspension of lead-containing particles. Our results suggest that these lead particles were deposited during the years of leaded gasoline use and that resuspension is responsible for generating an additional 54,000 kg of airborne lead each year. This agrees roughly with estimated outputs. Thus, we conclude that resuspension, although an insignificant source of airborne lead during the era of leaded fuel, became a principal source in the SOCAB as lead emissions from vehicles declined. The results of the resuspension model further suggest that soil lead levels will remain elevated for many decades, in which case resuspension will remain a major source well into the future.