Search

Sheng-Te Hung,1 Yi-Chen Cheng,2 Chieh-Chen Wu,2 Chun-Hsien Su1,2 1Graduate Institute of Sports Coaching Science, College of Kinesiology and Health, Chinese Culture University, Taipei, 111396, Taiwan; 2Department of Exercise and Health Promotion, College of Kinesiology and Health, Chinese Culture University, Taipei, 111396, TaiwanCorrespondence: Chun-Hsien Su, Department of Exercise and Health Promotion, College of Kinesiology and Health, Chinese Culture University, 55, Hwa-Kang Road, Shilin Distract, Taipei City, 111396, Taiwan, Tel +886-2-28610511 ext.45001, Fax +886-2-28617084, Email chsu@ulive.pccu.edu.twAbstract: This review examines the impact of physical activity, nutrition, and sleep evaluations on the physical wellness (PW) and overall well-being of older individuals. A comprehensive search was conducted in databases like PubMed, Google Scholar, and EBSCO Information Services. The search spanned from January 2000 to December 2022, resulting in 19,400 articles, out of which 98 review articles met the inclusion criteria. Through the analysis of these articles, key characteristics of the literature were summarized, and opportunities to enhance the practical application of physical activity (PA), nutrition, and sleep evaluations in the daily lives of older persons were identified. Regular physical activity is crucial for older persons to maintain their physical, mental, and emotional well-being and prevent age-related health issues. Older persons have specific nutritional needs, including increased protein, vitamin D, calcium, and vitamin B12 intake. Poor sleep quality in older persons is associated with negative health outcomes such as cognitive decline, physical disability, and mortality. This review emphasizes the significance of considering physical wellness as a fundamental element for achieving holistic well-being in older persons and highlights the importance of physical activity, nutrition, and sleep evaluations in improving their overall health and well-being. By understanding and implementing these findings, we can enhance the quality of life and promote healthy aging in older persons.Keywords: wellness, healthy aging, physical activity, nutrition, sleep

Ding-Ping Sun,1,2 Chih-Chan Lin,3 Shih-Ting Hung,3 Yi-Yu Kuang,3 You-Cheng Hseu,4– 6 Chia-Lang Fang,7,8 Kai-Yuan Lin3,9 1Department of Surgery, Chi Mei Medical Center, Tainan, Taiwan; 2Department of Food Science and Technology, Chia Nan University of Pharmacy and Science, Tainan, Taiwan; 3Department of Medical Research, Chi Mei Medical Center, Tainan, Taiwan; 4Department of Cosmeceutics, China Medical University, Taichung, Taiwan; 5Department of Health and Nutrition Biotechnology, Asia University, Taichung, Taiwan; 6Chinese Medicine Research Center, China Medical University, Taichung, Taiwan; 7Department of Pathology, School of Medicine, College of Medicine, Taipei Medical University, Taipei, Taiwan; 8Department of Pathology, Taipei Medical University Hospital, Taipei Medical University, Taipei, Taiwan; 9Department of Biotechnology, Chia Nan University of Pharmacy and Science, Tainan, TaiwanCorrespondence: Kai-Yuan Lin; Chia-Lang Fang Tel +886-6-281-2811; +886-2-27361661Fax +886-6-2832639; +886-2-23770054Email d84306@yahoo.com.tw; ccllfang@tmu.edu.twIntroduction: Gastric cancer (GC), one of the most prevalent malignancies, is the third-leading cause of cancer-related deaths globally. The aim of this study is to investigate the involvement of non-structural maintenance of chromosomes condensin I complex subunit G (NCAPG) in the prognosis of GC.Methods: Western blotting and immunostaining were employed to measure the NCAPG level in gastric tissues and cells. Kaplan–Meier analysis was applied to analyze the prognostic value of NCAPG in GC. RNA interference was applied to investigate the influence of the NCAPG silencing on GC cell growth and spread.Results: NCAPG overexpression was associated with several clinicopathologic characteristics, including nodal status (P = 0.0378), distant metastasis (P = 0.0088), staging (P = 0.0230), vascular invasion (P = 0.0012), and disease-free survival (P = 0.004). Kaplan–Meier analysis revealed that NCAPG overexpression was positively correlated to poor GC patients disease-free and overall survival (P = 0.004 and P < 0.001, respectively). Univariate Cox regression analysis showed that the overexpression of NCAPG was a prognostic biomarker of GC (P = 0.005). In cultured GC cells, the knockdown of NCAPG suppressed cell proliferation, migration and invasion. Meanwhile, further studies revealed that the NCAPG silencing induces the G0/G1 cell cycle arrest and accordingly represses cell division. Finally, Western blotting showed that NCPAG knockdown dysregulated cell cycle- and epithelial–mesenchymal transition-related molecules.Conclusion: Overall, the results reveal that NCAPG overexpression is a candidate prognostic biomarker and potential therapeutic target in GC.Keywords: NCAPG, gastric cancer, prognosis, proliferation, invasion

This study introduces an innovative method for reconstructing metamers, the aim is to identify metamerism when utilising white LEDs with high and low colour rendering. Two types of white LEDs were utilised as experimental light sources with two colour rendering indexes: high colour rendering (International Commission on Illumination (CIE) Ra of 95, TM-30 Rf of 92) and low colour rendering (CIE Ra of 78, TM-30 Rf of 78). Employ 741 training colours, principal component analysis was applied to extract basis function. Subsequently, 18 chromatic samples on ColorChecker colour chart were employed as target colours for spectrum reconstruction. Five sets of constraint were established to effectively filter potential metamers which are capable of inducing metamerism. Ultimately, three sets metamers were successfully reconstruct to differentiate white LEDs with high colour rendering, while an additional three sets of metamers were developed to distinguish white LEDs with low colour rendering. This outcome demonstrates the efficient method in reconstructing metamers to identify metamerism under high and low colour rendering white LEDs. [ABSTRACT FROM AUTHOR]

Despite extensive research efforts to explain the Mandarin Chinese particle le, confusion persists in the absence of a unitary theory and sufficient empirical evidence. This study provides a unitary account of le by adopting a usage-based constructionist approach, one that liberates grammatical aspect from, and is able to accommodate, lexical aspect. We argue that le participates in two distinct family resemblance constructions of aspect construal associated with two distinct sentential positions. The clause-internal le construction construes the closing or final boundary of an event and the clause-final le construction construes the opening or initial boundary of an event. Corpus analysis showed that the two aspect constructions have distinct patterns in natural language uses that are consistent with the proposed construals. Results from elicited response data showed that native speakers paid attention to construction-level formal and semantic cues in making family resemblance judgments about tokens of the two constructions. This study has both theoretical and methodological implications for crosslinguistic research on grammatical aspect in relation to lexical aspect and for usage-based constructionist approaches to grammatical categories beyond aspect.

Water behavior in various natural and manufactured settings is influenced by confinement in organic or inorganic frameworks and the presence of solutes. Here, the effects on dynamics from both confinement and the addition of solutes are examined. Specifically, water and ion dynamics in concentrated (2.5-4.2 m) aqueous magnesium chloride solutions confined in mesoporous silica (2.8 nm pore diameter) were investigated using polarization selective pump-probe and 2D infrared spectroscopies. Fitting the rotational and spectral diffusion dynamics measured by the vibrational probe, selenocyanate, with a previously developed two-state model revealed distinct behaviors at the interior of the silica pores (core state) and near the wall of the confining framework (shell state). The shell dynamics are noticeably slower than the bulk, or core, dynamics. The concentration-dependent slowing of the dynamics aligns with behavior in the bulk solutions, but the spectrally separated water-associated and Mg 2+ -associated forms of the selenocyanate probe exhibit different responses to confinement. The disparity in the complete reorientation times is larger upon confinement, but the spectral diffusion dynamics become more similar near the silica surface. The length scales that characterize the transition from surface-influenced to bulk-like behavior for the salt solutions in the pores are discussed and compared to those of pure water and an organic solvent confined in the same pores. These comparisons offer insights into how confinement modulates the properties of different liquids.

Modulation enhanced single-molecule localization microscopy (meSMLM), where emitters are sparsely activated with sequentially applied patterned illumination, increases the localization precision over single-molecule localization microscopy (SMLM). The precision improvement of modulation enhanced SMLM is derived from retrieving the position of an emitter relative to individual illumination patterns, which adds to existing point spread function information from SMLM. Here, we introduce SpinFlux: modulation enhanced localization for spinning disk confocal microscopy. SpinFlux uses a spinning disk with pinholes in its illumination and emission paths, to sequentially illuminate regions in the sample during each measurement. The resulting intensity-modulated emission signal is analyzed for each individual pattern to localize emitters with improved precision. We derive a statistical image formation model for SpinFlux and we quantify the theoretical minimum localization uncertainty in terms of the Cramér-Rao lower bound. Using the theoretical minimum uncertainty, we compare SpinFlux to localization on Fourier reweighted image scanning microscopy reconstructions. We find that localization on image scanning microscopy reconstructions with Fourier reweighting ideally results in a global precision improvement of 2.1 over SMLM. When SpinFlux is used for sequential illumination with three patterns around the emitter position, the localization precision improvement over SMLM is twofold when patterns are focused around the emitter position. If four donut-shaped illumination patterns are used for SpinFlux, the maximum local precision improvement over SMLM is increased to 3.5. Localization of image scanning microscopy reconstructions thus has the largest potential for global improvements of the localization precision, where SpinFlux is the method of choice for local refinements., Competing Interests: The authors have no conflicts to disclose., (© 2024 The Author(s).)

Image scanning microscopy (ISM) achieves resolution beyond the diffraction limit by a factor of 2. However, prior ISM research predominantly employs scalar diffraction theory, neglecting critical physical effects such as polarization, aberrations, and Stokes shift. This paper presents a comprehensive vectorial ISM point spread function (PSF) model that accounts for these phenomena. By considering the effect of polarization in emission and excitation paths, as well as aberrations and Stokes shift, our model provides a more accurate representation of ISM. We analyze the differences between scalar and vectorial theories in ISM and investigate the impact of pinhole size and aberration strength on resolution. At a numerical aperture of 1.2, the full width half maximum (FWHM) discrepancy between scalar and vectorial ISM PSFs can reach 45 nm, representing a 30 % deviation from the vectorial model. Additionally, we explore multiphoton excitation in ISM and observe increased FWHM for 2-photon and 3-photon excitation compared to 1-photon excitation. The FWHM of the 2-photon excitation ISM PSF increases by 20 % and the FWHM of the 3-photon excitation ISM PSF increases by 28 % compared to the 1-photon excitation ISM. In addition, we found that the optimal sweep factor for 2-photon ISM is 1.22, and the optimal sweep factor of 3-photon ISM is 1.12 instead of the 2 predicted by the one-photon scalar ISM theory. Our work improves the understanding of ISM and contributes to its advancement as a high-resolution imaging technique.

Three dimensional modulation-enhanced single-molecule localization techniques, such as ModLoc, offer advancements in axial localization precision across the entire field of view and axial capture range, by applying phase shifting to the illumination pattern. However, this improvement is limited by the pitch of the illumination pattern that can be used and requires registration between separate regions of the camera. To overcome these limitations, we present ZIMFLUX, a method that combines astigmatic point-spread-function (PSF) engineering with a structured illumination pattern in all three spatial dimensions. In order to achieve this we address challenges such as optical aberrations, refractive index mismatch, supercritical angle fluorescence (SAF), and imaging at varying depths within a sample, by implementing a vectorial PSF model. In scenarios involving refractive index mismatch between the sample and immersion medium, the astigmatic PSF loses its ellipticity at greater imaging depths, leading to a deterioration in axial localization precision. In contrast, our simulations demonstrate that ZIMFLUX maintains high axial localization precision even when imaging deeper into the sample. Experimental results show unbiased localization of 3D 80 nm DNA-origami nanostructures in SAF conditions with a 1.5-fold improvement in axial localization precision when comparing ZIMFLUX to conventional SMLM methods that rely solely on astigmatic PSF engineering.

Genetic mutations that cause adult-onset neurodegenerative diseases are often expressed during embryonic stages, but it is unclear whether they alter neurodevelopment and how this might influence disease onset. Here, we show that the most common cause of frontotemporal dementia (FTD) and amyotrophic lateral sclerosis (ALS), a repeat expansion in C9ORF72, restricts neural stem cell proliferation and reduces cortical and thalamic size in utero. Surprisingly, a repeat expansion-derived dipeptide repeat protein (DPR) not known to reduce neuronal viability plays a key role in impairing neurodevelopment. Pharmacologically mimicking the effects of the repeat expansion on neurodevelopment increases susceptibility of C9ORF72 mice to motor defects. Thus, the C9ORF72 repeat expansion stunts development of the brain regions prominently affected in C9ORF72 FTD/ALS patients., Competing Interests: Declaration of interests J.K.I. is a co-founder of AcuraStem, Inc. and Modulo Bio. J.K.I. is on the scientific advisory board of AcuraStem, Inc.; Spinogenix; and Vesalius Therapeutics. J.K.I. is an employee of BioMarin Pharmaceutical. J.K.I. is a co-founder of Modulo Bio and serves on the scientific advisory board of Spinogenix. Named companies were not involved in this project., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)

While motor and cortical neurons are affected in C9orf72 amyotrophic lateral sclerosis and frontotemporal dementia (ALS/FTD), it remains largely unknown if and how non-neuronal cells induce or exacerbate neuronal damage. We differentiated C9orf72 ALS/FTD patient-derived induced pluripotent stem cells into microglia (iPSC-MG) and examined their intrinsic phenotypes. Similar to iPSC motor neurons, C9orf72 ALS/FTD iPSC-MG mono-cultures form G 4 C 2 repeat RNA foci, exhibit reduced C9orf72 protein levels, and generate dipeptide repeat proteins. Healthy control and C9orf72 ALS/FTD iPSC-MG equally express microglial specific genes and perform microglial functions, including inflammatory cytokine release and phagocytosis of extracellular cargos, such as synthetic amyloid beta peptides and healthy human brain synaptoneurosomes. RNA sequencing analysis revealed select transcriptional changes of genes associated with neuroinflammation or neurodegeneration in diseased microglia yet no significant differentially expressed microglial-enriched genes. Moderate molecular and functional differences were observed in C9orf72 iPSC-MG mono-cultures despite the presence of C9orf72 pathological features suggesting that a diseased microenvironment may be required to induce phenotypic changes in microglial cells and the associated neuronal dysfunction seen in C9orf72 ALS/FTD neurodegeneration., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Lorenzini, Alsop, Levy, Gittings, Lall, Rabichow, Moore, Pevey, Bustos, Burciu, Bhatia, Singer, Saul, McQuade, Tzioras, Mota, Logemann, Rose, Almeida, Gao, Marks, Donnelly, Hutchins, Hung, Ichida, Bowser, Spires-Jones, Blurton-Jones, Gendron, Baloh, Van Keuren-Jensen and Sattler.)

The dynamics of aqueous magnesium chloride solutions, from relatively dilute (0.5 m) to near saturated (4.2 m) concentrations, were investigated using ultrafast two dimensional infrared and polarization selective pump-probe spectroscopies. The experiments were performed on two spectrally distinct nitrile stretch frequencies of the selenocyanate vibrational probe, corresponding to the CN nitrogen lone pair being associated with water and with Mg 2+ . No chemical exchange of the two species was observed over the experimental time scale (∼100 ps), enabling straightforward analysis of their dynamics. The dynamics reported by the Mg 2+ -associated peak are slower than those of the water-associated peak, suggesting that the immediate environment of the hydrated Mg 2+ is different from the rest of the solution. Notably, the Mg 2+ -associated peak displays three spectral diffusion time scales, the slowest being ∼30 ps, while the water-associated peak decays as a faster biexponential. From the complete orientational relaxation time and hydrodynamic theory, a magnesium hydration number of six was obtained, which is in good agreement with NMR and X-ray diffraction studies. This hydration number holds for all concentrations until near saturation, when the linewidths and the dynamics deviate from linear trends, indicative of Mg 2+ solvation structure changes resulting from a shortage of water molecules needed for full solvation.

Optofluidic devices have revolutionized the manipulation and transportation of fluid at smaller length scales ranging from micrometers to millimeters. We describe a dedicated optical setup for studying laser-induced cavitation inside a microchannel. In a typical experiment, we use a tightly focused laser beam to locally evaporate the solution laced with a dye resulting in the formation of a microbubble. The evolving bubble interface is tracked using high-speed microscopy and digital image analysis. Furthermore, we extend this system to analyze fluid flow through fluorescence-Particle Image Velocimetry (PIV) technique with minimal adaptations. In addition, we demonstrate the protocols for the in-house fabrication of a microchannel tailored to function as a sample holder in this optical setup. In essence, we present a complete guide for constructing a fluorescence microscope from scratch using standard optical components with flexibility in the design and at a lower cost compared to its commercial analogues., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2023 The Author(s).)

Pectus excavatum (PE), a chest-wall deformity that can compromise cardiopulmonary function, cannot be detected by a radiologist through frontal chest radiography without a lateral view or chest computed tomography. This study aims to train a convolutional neural network (CNN), a deep learning architecture with powerful image processing ability, for PE screening through frontal chest radiography, which is the most common imaging test in current hospital practice. Posteroanterior-view chest images of PE and normal patients were collected from our hospital to build the database. Among them, 80% were used as the training set used to train the established CNN algorithm, Xception, whereas the remaining 20% were a test set for model performance evaluation. The performance of our diagnostic artificial intelligence model ranged between 0.976-1 under the receiver operating characteristic curve. The test accuracy of the model reached 0.989, and the sensitivity and specificity were 96.66 and 96.64, respectively. Our study is the first to prove that a CNN can be trained as a diagnostic tool for PE using frontal chest X-rays, which is not possible by the human eye. It offers a convenient way to screen potential candidates for the surgical repair of PE, primarily using available image examinations.

Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease that results from many diverse genetic causes. Although therapeutics specifically targeting known causal mutations may rescue individual types of ALS, these approaches cannot treat most cases since they have unknown genetic etiology. Thus, there is a pressing need for therapeutic strategies that rescue multiple forms of ALS. Here, we show that pharmacological inhibition of PIKFYVE kinase activates an unconventional protein clearance mechanism involving exocytosis of aggregation-prone proteins. Reducing PIKFYVE activity ameliorates ALS pathology and extends survival of animal models and patient-derived motor neurons representing diverse forms of ALS including C9ORF72, TARDBP, FUS, and sporadic. These findings highlight a potential approach for mitigating ALS pathogenesis that does not require stimulating macroautophagy or the ubiquitin-proteosome system., Competing Interests: Declaration of interests J.K.I. and S.-T.A. are co-founders of AcuraStem, Inc. S.-T.A., W.-H.C., S.M., and S.H. are employees of AcuraStem, Inc. J.K.I. is a co-founder of Modulo Bio, serves on the scientific advisory boards of AcuraStem, Spinogenix, Synapticure, and Vesalius Therapeutics, and is employed at BioMarin Pharmaceutical. B.V.Z. is a co-founder of ZZ Biotech and chairman of its scientific advisory board. J.A.P. is a co-founder of Modelis. F.-B.G. receives research funding from Stealth BioTherapeutics., (Copyright © 2023 Elsevier Inc. All rights reserved.)

Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease caused by many diverse genetic etiologies. Although therapeutics that specifically target causal mutations may rescue individual types of ALS, such approaches cannot treat most patients since they have unknown genetic etiology. Thus, there is a critical need for therapeutic strategies that rescue multiple forms of ALS. Here, we combine phenotypic chemical screening on a diverse cohort of ALS patient-derived neurons with bioinformatic analysis of large chemical and genetic perturbational datasets to identify broadly effective genetic targets for ALS. We show that suppressing the gene-encoding, spliceosome-associated factor SYF2 alleviates TDP-43 aggregation and mislocalization, improves TDP-43 activity, and rescues C9ORF72 and causes sporadic ALS neuron survival. Moreover, Syf2 suppression ameliorates neurodegeneration, neuromuscular junction loss, and motor dysfunction in TDP-43 mice. Thus, suppression of spliceosome-associated factors such as SYF2 may be a broadly effective therapeutic approach for ALS., Competing Interests: Declaration of interests J.K.I. and S.A. are co-founders of AcuraStem, Inc. S.A., W.-H.C., S.M., and S.H. are employees of AcuraStem, Inc. J.K.I. is a co-founder of Modulo Bio, serves on the scientific advisory boards of AcuraStem, Spinogenix, Synapticure, and Vesalius Therapeutics, and is employed by BioMarin Pharmaceutical. H.G. is a co-founder of Exai Bio and Vevo Therapeutics and serves on the scientific advisory boards of Exai Bio and Verge Genomics. J.K.I. and G.R.L. are inventors on a patent application (PCT/US2021/014541) that has been filed related to this work., (Copyright © 2023 Elsevier Inc. All rights reserved.)

Colorectal cancer (CRC) is one of the types of cancers with a high incidence and is ranked the 3rd among men and 2nd among women worldwide. The purpose of this study was to investigate the correlation between non-SMC condensin I complex subunit G (NCAPG) and the prognosis of CRC and its function in CRC cells. The expression of NCAPG in colorectal tissues and cells was detected by immunoblotting and immunohistochemistry. Kaplan-Meier analysis was used to analyze the correlation between NCAPG and CRC prognosis. RNAi technology was used to investigate how NCAPG inhibition affected the proliferation and migration of CRC cells. Overexpression of NCAPG was positively correlated with several clinicopathologic characteristics, including T stage (P = 0.0198), M stage (P = 0.0005), and TNM stage (P < 0.0001). Kaplan-Meier analysis showed that the overexpression of NCAPG was also negatively correlated with disease-free survival and overall survival. In the culture of CRC cells, the knockdown of NCAPG inhibited the proliferation, migration, and invasion of the cells. Meanwhile, it was also found that NCAPG knockdown could interfere with G2/M-G1 transition in the cell cycle, resulting in the inhibition of cell proliferation. The overexpression of NCAPG may serve as a candidate biomarker for CRC prognosis. NCAPG is also a potential therapeutic target for CRC., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 The Authors. Published by Elsevier GmbH.. All rights reserved.)

Apolipoprotein E4 (APOE4), the main susceptibility gene for Alzheimer's disease, leads to blood-brain barrier (BBB) breakdown in humans and mice. Remarkably, BBB dysfunction predicts cognitive decline and precedes synaptic deficits in APOE4 human carriers. How APOE4 affects BBB and synaptic function at a molecular level, however, remains elusive. Using single-nucleus RNA-sequencing and phosphoproteome and proteome analysis, we show that APOE4 compared with APOE3 leads to an early disruption of the BBB transcriptome in 2-3-mo-old APOE4 knock-in mice, followed by dysregulation in protein signaling networks controlling cell junctions, cytoskeleton, clathrin-mediated transport, and translation in brain endothelium, as well as transcription and RNA splicing suggestive of DNA damage in pericytes. Changes in BBB signaling mechanisms paralleled an early, progressive BBB breakdown and loss of pericytes, which preceded postsynaptic interactome disruption and behavioral deficits that developed 2-5 mo later. Thus, dysregulated signaling mechanisms in endothelium and pericytes in APOE4 mice reflect a molecular signature of a progressive BBB failure preceding changes in synaptic function and behavior., (© 2022 Barisano et al.)

Single-molecule localization microscopy (SMLM) enables the high-resolution visualization of organelle structures and the precise localization of individual proteins. However, the expected resolution is not achieved in tissue as the imaging conditions deteriorate. Sample-induced aberrations distort the point spread function (PSF), and high background fluorescence decreases the localization precision. Here, we synergistically combine sensorless adaptive optics (AO), in-situ 3D-PSF calibration, and a single-objective lens inclined light sheet microscope (SOLEIL), termed (AO-SOLEIL), to mitigate deep tissue-induced deteriorations. We apply AO-SOLEIL on several dSTORM samples including brains of adult Drosophila . We observed a 2x improvement in the estimated axial localization precision with respect to widefield without aberration correction while we used synergistic solution. AO-SOLEIL enhances the overall imaging resolution and further facilitates the visualization of sub-cellular structures in tissue., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Hung, Llobet Rosell, Jurriens, Siemons, Soloviev, Kapitein, Grußmayer, Neukomm, Verhaegen and Smith.)

In this review, ruthenium nanoparticles (Ru NPs)-based functional nanomaterials have attractive electrocatalytic characteristics and they offer considerable potential in a number of fields. Ru-based binary or multimetallic NPs are widely utilized for electrode modification because of their unique electrocatalytic properties, enhanced surface-area-to-volume ratio, and synergistic effect between two metals provides as an effective improved electrode sensor. This perspective review suggests the current research and development of Ru-based nanomaterials as a platform for electrochemical (EC) sensing of harmful substances, biomolecules, insecticides, pharmaceuticals, and environmental pollutants. The advantages and limitations of mono-, bi-, and multimetallic Ru-based nanocomposites for EC sensors are discussed. Besides, the relevant EC properties and analyte sensing approaches are also presented. On the basis of these insights, we highlighted recent results for synthesizing techniques and EC environmental pollutant sensors from the perspectives of diverse supports, including graphene, carbon nanotubes, silica, semiconductors, metal sulfides, and polymers. Finally, this work overviews the modern improvements in the utilization of Ru-based nanocomposites on the basis for electroanalytical sensors as well as suggestions for the field's future development.

The approximately linear scaling of fluorescence quantum yield (ϕ) with fluorescence lifetime (τ) in fluorescent proteins (FPs) has inspired engineering of brighter fluorophores based on screening for increased lifetimes. Several recently developed FPs such as mTurquoise2, mScarlet, and FusionRed-MQV which have become useful for live cell imaging are products of lifetime selection strategies. However, the underlying photophysical basis of the improved brightness has not been scrutinized. In this study, we focused on understanding the outcome of lifetime-based directed evolution of mCherry, which is a popular red-FP (RFP). We identified four positions (W143, I161, Q163, and I197) near the FP chromophore that can be mutated to create mCherry-XL (eXtended Lifetime: ϕ = 0.70; τ = 3.9 ns). The 3-fold higher quantum yield of mCherry-XL is on par with that of the brightest RFP to date, mScarlet. We examined selected variants within the evolution trajectory and found a near-linear scaling of lifetime with quantum yield and consistent blue-shifts of the absorption and emission spectra. We find that the improvement in brightness is primarily due to a decrease in the nonradiative decay of the excited state. In addition, our analysis revealed the decrease in nonradiative rate is not limited to the blue-shift of the energy gap and changes in the excited state reorganization energy. Our findings suggest that nonradiative mechanisms beyond the scope of energy-gap models such the Englman-Jortner model are suppressed in this lifetime evolution trajectory.

Modulation enhanced single-molecule localization microscopy (meSMLM) methods improve the localization precision by using patterned illumination to encode additional position information. Iterative meSMLM (imeSMLM) methods iteratively generate prior information on emitter positions, used to locally improve the localization precision during subsequent iterations. The Cramér-Rao lower bound cannot incorporate prior information to bound the best achievable localization precision because it requires estimators to be unbiased. By treating estimands as random variables with a known prior distribution, the Van Trees inequality (VTI) can be used to bound the best possible localization precision of imeSMLM methods. An imeSMLM method is considered, where the positions of in-plane standing-wave illumination patterns are controlled over the course of multiple iterations. Using the VTI, we analytically approximate a lower bound on the maximum localization precision of imeSMLM methods that make use of standing-wave illumination patterns. In addition, we evaluate the maximally achievable localization precision for different illumination pattern placement strategies using Monte Carlo simulations. We show that in the absence of background and under perfect modulation, the information content of signal photons increases exponentially as a function of the iteration count. However, the information increase is no longer exponential as a function of the iteration count under non-zero background, imperfect modulation, or limited mechanical resolution of the illumination positioning system. As a result, imeSMLM with two iterations reaches at most a fivefold improvement over SMLM at 8 expected background photons per pixel and 95% modulation contrast. Moreover, the information increase from imeSMLM is balanced by a reduced signal photon rate. Therefore, SMLM outperforms imeSMLM when considering an equal measurement time and illumination power per iteration. Finally, the VTI is an excellent tool for the assessment of the performance of illumination control and is therefore the method of choice for optimal design and control of imeSMLM methods., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2022 Biophysical Society. Published by Elsevier Inc. All rights reserved.)

The presence of dark states causes fluorescence intermittency of single molecules due to transitions between "on" and "off" states. Genetically encodable markers such as fluorescent proteins (FPs) exhibit dark states that make several super-resolved single-molecule localization microscopy (SMLM) methods possible. However, studies quantifying the timescales and nature of dark state behavior for commonly used FPs under conditions typical of widefield and total internal reflection fluorescence (TIRF) microscopy remain scarce and pre-date many new SMLM techniques. FusionRed is a relatively bright red FP exhibiting fluorescence intermittency and has thus been identified as a potential candidate for SMLM. We herein characterize the rates for dark-state conversion and the subsequent ground-state recovery of FusionRed and its 2.5-fold brighter descendent FusionRed L175M M42Q (FusionRed-MQ) at low irradiances (1-10 W cm -2 ), which were previously unexplored experimental conditions. We characterized the kinetics of dark state transitions in these two FPs by using single molecule blinking and ensemble photobleaching experiments bridged with a dark state kinetic model. We find that at low irradiances, the recovery process to the ground state is minimally light-driven and FusionRed-MQ has a 1.3-fold longer ground state recovery time indicating a conformationally restricted dark-state chromophore in comparison to FusionRed. Our studies indicate that the brighter FusionRed-MQ variant exhibits higher dark state conversion rates with longer ground state recovery lifetimes, thus it is potentially a better candidate for SMLM applications than its progenitor FusionRed.

High-NA light sheet illumination can improve the resolution of single-molecule localization microscopy (SMLM) by reducing the background fluorescence. These approaches currently require custom-made sample holders or additional specialized objectives, which makes the sample mounting or the optical system complex and therefore reduces the usability of these approaches. Here, we developed a single-objective lens-inclined light sheet microscope (SOLEIL) that is capable of 2D and 3D SMLM in thick samples. SOLEIL combines oblique illumination with point spread function PSF engineering to enable dSTORM imaging in a wide variety of samples. SOLEIL is compatible with standard sample holders and off-the-shelve optics and standard high NA objectives. To accomplish optimal optical sectioning we show that there is an ideal oblique angle and sheet thickness. Furthermore, to show what optical sectioning delivers for SMLM we benchmark SOLEIL against widefield and HILO microscopy with several biological samples. SOLEIL delivers in 15 μ m thick Caco2-BBE cells a 374% higher intensity to background ratio and a 54% improvement in the estimated CRLB compared to widefield illumination, and a 184% higher intensity to background ratio and a 20% improvement in the estimated CRLB compared to HILO illumination., Competing Interests: The authors declare no conflicts of interest., (© 2022 Optica Publishing Group under the terms of the Optica Open Access Publishing Agreement.)

The noncoding genome is substantially larger than the protein-coding genome but has been largely unexplored by genetic association studies. Here, we performed region-based rare variant association analysis of >25,000 variants in untranslated regions of 6,139 amyotrophic lateral sclerosis (ALS) whole genomes and the whole genomes of 70,403 non-ALS controls. We identified interleukin-18 receptor accessory protein (IL18RAP) 3' untranslated region (3'UTR) variants as significantly enriched in non-ALS genomes and associated with a fivefold reduced risk of developing ALS, and this was replicated in an independent cohort. These variants in the IL18RAP 3'UTR reduce mRNA stability and the binding of double-stranded RNA (dsRNA)-binding proteins. Finally, the variants of the IL18RAP 3'UTR confer a survival advantage for motor neurons because they dampen neurotoxicity of human induced pluripotent stem cell (iPSC)-derived microglia bearing an ALS-associated expansion in C9orf72, and this depends on NF-κB signaling. This study reveals genetic variants that protect against ALS by reducing neuroinflammation and emphasizes the importance of noncoding genetic association studies., (© 2022. The Author(s), under exclusive licence to Springer Nature America, Inc.)

Background: Globally, gastric cancer is ranked 4th and 3rd in terms of incidence and mortality rate among all cancer types. This study aimed to examine the relationship between G protein-coupled receptor kinase 3 (GRK3) and gastric cancer prognosis and investigate the role of GRK3 in gastric cancer carcinogenesis. Methods: GRK3 level in gastric tissues and cells were determined using immunohistochemistry and immunoblotting. Kaplan-Meier analysis with the log-rank test was employed to evaluate the relationship between GRK3 expression and gastric cancer prognosis. RNAi technology was applied to examine the effects of GRK3 inhibition on gastric cancer proliferation and spread. Results: GRK3 overexpression was correlated significantly with lymphatic metastasis (P = 0.0011), distant metastasis (P < 0.0001), TNM stage (P = 0.0035), and vascular invasion (P = 0.0025). Kaplan-Meier survival analysis showed that the disease-free survival and overall survival of patients with high GRK3 expression were significantly shorter than those of patients with low GRK3 expression. Multivariate Cox regression analysis also showed that the overexpression of GRK3 was an independent prognostic biomarker of gastric cancer (P = 0.029). In cultured gastric cancer cells, GRK3 knockdown inhibited cell proliferation, migration, and invasion. Further analysis revealed that more GRK3-knockdown cells were in G0/G1 phase and few cells were in S phase, thereby inhibiting cell proliferation. Conclusions: GRK3 overexpression can be a candidate biomarker for gastric cancer prognosis. GRK3 is also a potential therapeutic target for gastric cancer., Competing Interests: Competing Interests: The authors have declared that no competing interest exists., (© The author(s).)

Single-photon avalanche diode (SPAD) arrays can be used for single-molecule localization microscopy (SMLM) because of their high frame rate and lack of readout noise. SPAD arrays have a binary frame output, which means photon arrivals should be described as a binomial process rather than a Poissonian process. Consequentially, the theoretical minimum uncertainty of the localizations is not accurately predicted by the Poissonian Cramér-Rao lower bound (CRLB). Here, we derive a binomial CRLB and benchmark it using simulated and experimental data. We show that if the expected photon count is larger than one for all pixels within one standard deviation of a Gaussian point spread function, the binomial CRLB gives a 46% higher theoretical uncertainty than the Poissonian CRLB. For typical SMLM photon fluxes, where no saturation occurs, the binomial CRLB predicts the same uncertainty as the Poissonian CRLB. Therefore, the binomial CRLB can be used to predict and benchmark localization uncertainty for SMLM with SPAD arrays for all practical emitter intensities.

Enhancement of processes ranging from gas sorption to ion conduction in a liquid can be substantial upon nanoconfinement. Here, the dynamics of a polar aprotic solvent, 1-methylimidazole (MeIm), in mesoporous silica (2.8, 5.4, and 8.3 nm pore diameters) were examined using femtosecond infrared vibrational spectroscopy and molecular dynamics simulations of a dilute probe, the selenocyanate (SeCN - ) anion. The long vibrational lifetime and sensitivity of the CN stretch enabled a comprehensive investigation of the relatively slow time scales and subnanometer distance dependences of the confined dynamics. Because MeIm does not readily donate hydrogen bonds, its interactions in the hydrophilic silanol pores differ more from the bulk than those of water confined in the same mesopores, resulting in greater structural order and more dramatic slowing of dynamics. The extent of surface effects was quantified by modified two-state models used to fit three spatially averaged experimental observables: vibrational lifetime, orientational relaxation, and spectral diffusion. The length scales and the models (smoothed step, exponential decay, and simple step) describing the transitions between the distinctive shell behavior at the surface and the bulk-like behavior at the pore interior were compared to those of water. The highly nonuniform distributions of the SeCN - probe and antiparallel layering of MeIm revealed by the simulations guided the interpretation of the results and development of the analytical models. The results illustrate the importance of electrostatic effects and H-bonding interactions in the behavior of confined liquids.

Gastric cancer (GC) is one of the most common malignant tumors worldwide and has high rates of morbidity and mortality. This study investigated the role of Krüppel-like factor 16 (KLF16) in GC. Real-time polymerase chain reaction, Western blotting, and immunohistochemistry were used to examine the expression of KLF16 in gastric cells and tissues. Gene overexpression and silencing were applied to study the involvement of KLF16 in GC cell growth and metastasis along with its underlying mechanism. The results indicate that KLF16 overexpression is significantly associated with nodal status, distant metastasis, staging, degree of differentiation, vascular invasion, and patient survival. Multivariate Cox proportional hazards regression model analysis revealed that the overexpression of KLF16 is an independent prognostic biomarker of GC. The in vitro study revealed that up-regulated KLF16 accelerates cell growth and metastasis, whereas the inhibition of KLF16 suppresses these cellular activities. The results of an animal study also indicated that the overexpression and silencing of KLF16 accelerate and repress xenograft proliferation and metastasis. Further studies of affected cell growth and metastasis revealed that KLF16 modulates the cell cycle and epithelial-mesenchymal transition through transcriptional regulation of microfibrillar-associated protein 5. Collectively, these results reveal that KLF16 overexpression is a potential prognostic biomarker and therapeutic target for the treatment of GC., Competing Interests: None., (AJCR Copyright © 2021.)

Benzene complex formation and dissociation dynamics with silanols on the amorphous silica surfaces of nanoporous SiO 2 , from a benzene/carbon tetrachloride solution, were measured by the growth of off-diagonal peaks in the two-dimensional infrared (2D IR) chemical exchange spectrum of the isolated Si-OD stretch. The presence of two types of isolated silanols, termed type I and II, was revealed, with dissociation time constants of 82 and 4.0 ps, respectively. The type I silanols are associated with the main IR absorption feature in the Si-OD stretching region, while the type II silanols give rise to a broader shoulder to lower frequency. Polarization selective pump-probe (PSPP) measurements provided the vibrational lifetimes and orientational relaxation rates of the two silanols in the CCl 4 (free) and benzene (complex) environments. The type II silanols constitute roughly 30% of the isolated silanol population and exhibit a substantially faster rate of vibrational relaxation, making the type I dynamics the dominant contribution to the PSPP and 2D IR signals. From the measured dissociation times, the enthalpies of formation for the two surface complexes were obtained, with the formation of the type I complex being significantly more exothermic. As the type II site is preferentially removed from the amorphous silica surface with increasing activation temperature, the results provide a reasonable explanation for the increased exothermicity of benzene adsorption on silica with increasing activation temperature in previous calorimetry experiments.

Obtaining a tiny focal spot is desired for super resolution. We do a vectorial numerical analysis of the linearly, circularly, and radidally polarized electromagnetic fields being focused through a dielectric micro/nanoparticle of size comparable to the wavelength. We find tiny focal spots (up to ∼0.05 λ 2 ) can be obtained behind micro/nanoparticles of various shapes, e.g. spherical, disk-shaped, and cuboid micro/nanoparticles. Furthermore, we also investigate the influence of the misalignment of a real lens system on the tiny focal spots. We find that tiny focal spots can still be generated even though they are distorted due to the misalignment.

The development of fluorescent proteins (FPs) has revolutionized biological imaging. FusionRed, a monomeric red FP (RFP), is known for its low cytotoxicity and correct localization of target fusion proteins in mammalian cells but is limited in application by low fluorescence brightness. We report a brighter variant of FusionRed, "FR-MQV," which exhibits an extended fluorescence lifetime (2.8 ns), enhanced quantum yield (0.53), higher extinction coefficient (∼140 000 M -1 cm -1 ), increased radiative rate constant, and reduced nonradiative rate constant with respect to its precursor. The properties of FR-MQV derive from three mutations-M42Q, C159V, and the previously identified L175M. A structure-guided approach was used to identify and mutate candidate residues around the para-hydroxyphenyl and the acylimine sites of the chromophore. The C159V mutation was identified via lifetime-based flow cytometry screening of a library in which multiple residues adjacent to the para-hydroxyphenyl site of the chromophore were mutated. The M42Q mutation is located near the acylimine moiety of the chromophore and was discovered using site-directed mutagenesis guided by X-ray crystal structures. FR-MQV exhibits a 3.4-fold higher molecular brightness and a 5-fold increase in the cellular brightness in HeLa cells [based on fluorescence-activated cell sorting (FACS)] compared to FusionRed. It also retains the low cytotoxicity and high-fidelity localization of FusionRed, as demonstrated through assays in mammalian cells. These properties make FR-MQV a promising template for further engineering into a new family of RFPs.