Assessing the stability of azopolymer nanotopography during live-cell fluorescence imaging.

Introduction: Photomodifiable azopolymer nanotopographies represent a powerful means of assessing how cells respond to rapid changes in the local microenvironment. However, previous studies have suggested that azopolymers are readily photomodified under typical fluorescence imaging conditions over much of the visible spectrum. Here we assess the stability of azopolymer nanoridges under 1-photon and 2-photon imaging over a broad range of wavelengths. Methods: Azopolymer nanoridges were created via microtransfer molding of master structures that were created using interference lithography. The effects of exposure to a broad range of wavelengths of light polarized parallel to the ridges were assessed on both a spinning-disk confocal microscope and a 2-photon fluorescence microscope. Experiments with live Dictyostelium discoideum cells were also performed using alternating cycles of 514-nm light for photomodification and 561-nm light for fluorescence imaging. Results and Discussion: We find that for both 1-photon and 2-photon imaging, only a limited range of wavelengths of light leads to photomodification of the azopolymer nanotopography. These results indicate that nondestructive 1-photon and 2-photon fluorescence imaging can be performed over a considerably broader range of wavelengths than would be suggested by previous research.

The First Alveolar Bone Graft Simulator.

Alveolar bone graft (ABG) surgery in cleft patients is technically challenging. The procedure requires design, dissection and release of soft tissue flaps to create a seal around the bone graft. In addition, visualization during the procedure is challenging within the confines of the cleft. These features make ABG surgery difficult to learn and teach, and it is, therefore, a suitable procedure for the use of a simulator. A high-fidelity cleft ABG simulator was developed using three-dimensional printing, polymer, and adhesive techniques. Simulated ABG surgery was performed by two expert cleft surgeons for a total of five simulation sessions to test the simulator's features and the ability to perform the critical steps of an ABG. ABG surgery was successfully performed on the simulator. The simulations involved interacting with realistic dissection planes as well as multi-layered synthetic soft (periosteum, mucosa, gingiva, adipose tissue) and hard (teeth, bone) tissue. The simulator allowed performance of cleft marginal incisions, dissection, and elevation of a muco-gingival-periosteal flap, creation of nasal upturned and palatal downturned flaps, nasal and palatal side closure, insertion of simulated bone graft material, and advancement of the muco-gingival-periosteal flap for closure of the anterior wall of the cleft. The ABG simulator allowed performance of the critical steps of ABG surgery. This is the first ABG simulator developed, which incorporates the features necessary to practice the procedure from start to finish.

The Gender Self-Report: A multidimensional gender characterization tool for gender-diverse and cisgender youth and adults.

Gender identity is a core component of human experience, critical to account for in broad health, development, psychosocial research, and clinical practice. Yet, the psychometric characterization of gender has been impeded due to challenges in modeling the myriad gender self-descriptors, statistical power limitations related to multigroup analyses, and equity-related concerns regarding the accessibility of complex gender terminology. Therefore, this initiative employed an iterative multi-community-driven process to develop the Gender Self-Report (GSR), a multidimensional gender characterization tool, accessible to youth and adults, nonautistic and autistic people, and gender-diverse and cisgender individuals. In Study 1, the GSR was administered to 1,654 individuals, sampled through seven diversified recruitments to be representative across age (10-77 years), gender and sexuality diversity (∼33% each gender diverse, cisgender sexual minority, cisgender heterosexual), and autism status (> 33% autistic). A random half-split subsample was subjected to exploratory factor analytics, followed by confirmatory analytics in the full sample. Two stable factors emerged: Nonbinary Gender Diversity and Female-Male Continuum (FMC). FMC was transformed to Binary Gender Diversity based on designated sex at birth to reduce collinearity with designated sex at birth. Differential item functioning by age and autism status was employed to reduce item-response bias. Factors were internally reliable. Study 2 demonstrated the construct, convergent, and ecological validity of GSR factors. Of the 30 hypothesized validation comparisons, 26 were confirmed. The GSR provides a community-developed gender advocacy tool with 30 self-report items that avoid complex gender-related "insider" language and characterize diverse populations across continuous multidimensional binary and nonbinary gender traits. (PsycInfo Database Record (c) 2023 APA, all rights reserved).

Bilateral Paragangliomas in the Setting of Autonomic Dysfunction: A Case Report.

In 2018, a 59-year-old female patient presented with hoarseness in her voice, headache, intermittent pain in her right side, difficulty of right arm movement, left side neck pain, difficulty controlling hypertension of unknown etiology, and a large mass on the upper left side of her neck with a smaller mass on the right side. MRI of the neck revealed masses at each carotid bifurcation. These were determined to be bilateral paragangliomas. Paragangliomas are rare tumors, and bilateral ones tremendously so. The patient underwent radiation over 2 years, resulting in the successful shrinking and stabilization of both masses. Since completing radiation, the patient reported improvement in her memory, and her blood pressure has stabilized with medication.

Extreme Enlargement of the Cerebellum in a Clade of Teleost Fishes that Evolved a Novel Active Sensory System.

Brains, and the distinct regions that make up brains, vary widely in size across vertebrates [1, 2]. Two prominent hypotheses have been proposed to explain brain region scaling evolution. The mosaic hypothesis proposes that changes in the relative sizes of particular brain regions are the result of selection acting independently on those regions [2, 3]. The concerted hypothesis proposes that the brain evolves as a coordinated structure due to developmental constraints [4]. These hypotheses have been widely debated [3-7], and recent studies suggest a combination of the two best describes vertebrate brain region scaling [8-10]. However, no study has addressed how the mosaic and concerted models relate to the evolution of novel behavioral phenotypes. We addressed this question using African mormyroid fishes. The mormyroids have evolved a novel active electrosensory system and are well known for having extreme encephalization [11] and a large cerebellum [2, 12], which is cited as a possible example of mosaic evolution [2]. We found that compared to outgroups without active electrosensing, mormyroids experienced mosaic increases in the sizes of the cerebellum and hindbrain, and mosaic decreases in the sizes of the telencephalon, optic tectum, and olfactory bulb. However, the evolution of extreme encephalization within mormyroids was associated with concerted changes in the sizes of all brain regions. This suggests that mosaic evolutionary change in the regional composition of the brain is most likely to occur alongside the evolution of novel behavioral functions, but not with the evolution of extreme encephalization.

Chronic N-methyl-D-aspartate receptor activation induces cardiac electrical remodeling and increases susceptibility to ventricular arrhythmias.

BACKGROUND: N-Methyl-d-aspartate receptors, also known as NMDA Receptors or NMDAR, are glutamate receptors that control calcium ion channels and regulate synaptic plasticity. Acute NMDAR activation can induce ventricular arrhythmias (VAs). However, the influence of chronic NMDAR activation on cardiac electrophysiology remains unknown. METHODS AND RESULTS: Wistar rats were randomly administered 0.9% saline (CTL group), NMDA (N group), or NMDA plus MK801 (N+M group) for 14 days. Compared with the CTL group, the N group displayed elevated heart rate and prolonged QT, QTc, and TpTe intervals in the electrocardiogram (P < 0.05 for all). Then, the S1 S2 , S1 S1 , and Burst pacing were performed to assess the characteristics of repolarization; threshold of action potential duration (APD) alternans; beat-to-beat variability of repolarization (BVR); and VAs susceptibility in the left ventricular. The prolonged APD at 90% repolarization (APD90 ); decreased ERP/APD90 ; increased dispersion of APD90 , ERP, and ERP/APD90 ; decreased threshold of APD alternans; increased BVR; and incidence of VAs were showed in the N group compared with those of the CTL group (P < 0.01 for all). Moreover, chronic NMDA administration reduced the expression of Kv4.2, Kv4.3, KChIP2, and Kv11.1 proteins, and induced mild myocardial interstitial fibrosis (P < 0.01 for all). Importantly, these alterations induced by NMDA were normalized by co-treatment with MK801. CONCLUSION: Chronic NMDAR activation prolonged repolarization, induced electrical instability, and facilitated VAs, which may be associated with reduced Ito and IKr and myocardial fibrosis.

An evolving new paradigm: endothelial cells--conditional innate immune cells.

Endothelial cells (ECs) are a heterogeneous population that fulfills many physiological processes. ECs also actively participate in both innate and adaptive immune responses. ECs are one of the first cell types to detect foreign pathogens and endogenous metabolite-related danger signals in the bloodstream, in which ECs function as danger signal sensors. Treatment with lipopolysaccharide activates ECs, causing the production of pro-inflammatory cytokines and chemokines, which amplify the immune response by recruiting immune cells. Thus, ECs function as immune/inflammation effectors and immune cell mobilizers. ECs also induce cytokine production by immune cells, in which ECs function as immune regulators either by activating or suppressing immune cell function. In addition, under certain conditions, ECs can serve as antigen presenting cells (antigen presenters) by expressing both MHC I and II molecules and presenting endothelial antigens to T cells. These facts along with the new concept of endothelial plasticity suggest that ECs are dynamic cells that respond to extracellular environmental changes and play a meaningful role in immune system function. Based on these novel EC functions, we propose a new paradigm that ECs are conditional innate immune cells. This paradigm provides a novel insight into the functions of ECs in inflammatory/immune pathologies.

A watershed-scale goals approach to assessing and funding wastewater infrastructure.

Capital needs during the next twenty years for public wastewater treatment, piping, combined sewer overflow correction, and storm-water management are estimated to be approximately $300 billion for the USA. Financing these needs is a significant challenge, as Federal funding for the Clean Water Act has been reduced by 70% during the last twenty years. There is an urgent need for new approaches to assist states and other decision makers to prioritize wastewater maintenance and improvements. We present a methodology for performing an integrated quantitative watershed-scale goals assessment for sustaining wastewater infrastructure. We applied this methodology to ten watersheds of the Hudson-Mohawk basin in New York State, USA that together are home to more than 2.7 million people, cover 3.5 million hectares, and contain more than 36,000 km of streams. We assembled data on 183 POTWs treating approximately 1.5 million m(3) of wastewater per day. For each watershed, we analyzed eight metrics: Growth Capacity, Capacity Density, Soil Suitability, Violations, Tributary Length Impacted, Tributary Capital Cost, Volume Capital Cost, and Population Capital Cost. These metrics were integrated into three goals for watershed-scale management: Tributary Protection, Urban Development, and Urban-Rural Integration. Our results demonstrate that the methodology can be implemented using widely available data, although some verification of data is required. Furthermore, we demonstrate substantial differences in character, need, and the appropriateness of different management strategies among the ten watersheds. These results suggest that it is feasible to perform watershed-scale goals assessment to augment existing approaches to wastewater infrastructure analysis and planning.

Caspase-1 recognizes extended cleavage sites in its natural substrates.

OBJECTIVE: The preferred amino acids in the proteolytic sites have been considered to be similar between caspase-1 and caspase-9, which do not support their differential functions in inflammatory pyroptosis and apoptosis. We attempted to solve this problem. METHODS: We analyzed the flanking 20 amino acid residues in the cleavage sites in 34 caspase-1 and 11 capase-9 experimentally identified substrates. RESULTS: This study has made the following findings: first, we verified that caspase-1 and caspase-9 shared 100% aspartic acid in the P1 position. However, the structures in the cleavage sites of most caspase-1 substrates are different from that of caspase-9 substrates in the following three aspects, (a) the amino acid residues with the statistically high frequencies; (b) the hydrophobic amino acid occurrence frequencies; and (c) the charged amino acid occurrence frequencies; second, the amino acid pairs P1-P1' are different; third, our identified cleavage site patterns are useful in the prediction for the 91.4% cleavage sites of 35 new caspase-1 substrates. CONCLUSION: Since most caspase-1 substrates are involved in vascular function, inflammation and atherogenesis, our novel structural patterns for the caspases' substrates are significant in developing new diagnostics and therapeutics.

Adenoviral proteins mimic nutrient/growth signals to activate the mTOR pathway for viral replication.

Like tumor cells, DNA viruses have had to evolve mechanisms that uncouple cellular replication from the many intra- and extracellular factors that normally control it. Here we show that adenovirus encodes two proteins that activate the mammalian target of rapamycin (mTOR) for viral replication, even under nutrient/growth factor-limiting conditions. E4-ORF1 mimics growth factor signaling by activating PI3-kinase, resulting in increased Rheb.GTP loading and mTOR activation. E4-ORF4 is redundant with glucose in stimulating mTOR, does not affect Rheb.GTP levels and is the major mechanism whereby adenovirus activates mTOR in quiescent primary cells. We demonstrate that mTOR is activated through a mechanism that is dependent on the E4-ORF4 protein phosphatase 2A-binding domain. We also show that mTOR activation is required for efficient S-phase entry, independently of E2F activation, in adenovirus-infected quiescent primary cells. These data reveal that adenovirus has evolved proteins that activate the mTOR pathway, irrespective of the cellular microenvironment, and which play a requisite role in viral replication.