Label-Free Three-Dimensional Morphological Characterization of Cell Death Using Holographic Tomography.

This study presents a novel label-free approach for characterizing cell death states, eliminating the need for complex molecular labeling that may yield artificial or ambiguous results due to technical limitations in microscope resolution. The proposed holographic tomography technique offers a label-free avenue for capturing precise three-dimensional (3D) refractive index morphologies of cells and directly analyzing cellular parameters like area, height, volume, and nucleus/cytoplasm ratio within the 3D cellular model. We showcase holographic tomography results illustrating various cell death types and elucidate distinctive refractive index correlations with specific cell morphologies complemented by biochemical assays to verify cell death states. These findings hold promise for advancing in situ single cell state identification and diagnosis applications.

Impact of aging on copper isotopic composition in the murine brain.

Aging is the main risk factor for Alzheimer's disease (AD). AD is linked to alterations in metal homeostasis and changes in stable metal isotopic composition can occur, possibly allowing the latter to serve as relevant biomarkers for potential AD diagnosis. Copper stable isotopes are used to investigate changes in Cu homeostasis associated with various diseases. Prior work has shown that in AD mouse models, the accumulation of 63Cu in the brain is associated with the disease's progression. However, our understanding of how the normal aging process influences the brain's isotopic composition of copper remains limited. In order to determine the utility and predictive power of Cu isotopes in AD diagnostics, we aim-in this study-to develop a baseline trajectory of Cu isotopic composition in the normally aging mouse brain. We determined the copper concentration and isotopic composition in brains of 30 healthy mice (WT) ranging in age from 6 to 12 mo, and further incorporate prior data obtained for 3-mo-old healthy mice; this range approximately equates to 20-50 yr in human equivalency. A significant 65Cu enrichment has been observed in the 12-mo-old mice compared to the youngest group, concomitant with an increase in Cu concentration with age. Meanwhile, literature data for brains of AD mice display an enrichment in 63Cu isotope compared to WT. It is acutely important that this baseline enrichment in 65Cu is fully constrained and normalized against if any coherent diagnostic observations regarding 63Cu enrichment as a biomarker for AD are to be developed.

Copper Isotope Compositions Measured Using a Sapphire Dual Path MC-ICPMS with a Collision/Reaction Cell.

We present a new approach to Cu isotopic measurements using a state-of-the-art Nu Sapphire multicollector inductively coupled plasma source mass spectrometer equipped with a collision/reaction cell (CRC-MC-ICPMS). We investigate the effects of Na doping and Cu concentration mismatch between bracketing standard and unknown samples and demonstrate the efficacy of introducing a He-H2 gas mix into the CRC to efficiently eliminate the sample matrix-based 40Ar23Na+ isobaric interference on 63Cu+. This capability is crucial when measuring samples with high Na/Cu ratios, such as some biological samples, which have significantly different chemical compositions compared to most geological samples. Moreover, considering the necessity of obtaining large data sets for biological samples to ensure reliable interpretations, the implementation of a CRC for mitigating the 40Ar23Na+ interference offers the advantage of minimizing the requirement for extensive Cu chemical separation procedure prior to Cu isotopic measurements. Our results demonstrate that the accurate determination of the δ65Cu values is achievable for samples with Na/Cu concentration ratios of up to ∼65, even when measuring 100 ppb Cu solutions (equivalent to a signal of ∼3.5-4 V total Cu). Furthermore, our results showcase a good short-term repeatability on δ65Cu for pure Cu standard solutions (NIST SRM 976 and Cu-IPGP), typically of 0.05‰ (2 SD) when measuring >50 ppb Cu solutions. Our long-term external reproducibility stands at approximately 0.07‰ (2 SD). This value accounts for the variable Cu concentrations analyzed across the different analytical sequences (from 10 to 100 ppb Cu solutions). To validate the robustness of our analytical method, we first conduct a comparison between data sets from mice brains processed twice through column chemistry using a Thermo Finnigan Neptune MC-ICPMS and a Nu Sapphire CRC-MC-ICPMS in CRC mode. This comparison serves to verify the reliability of our method for measuring Cu isotopic composition using the CRC on samples with a low Na/Cu ratio after traditional chemical processing. Then, we compare the data sets obtained for biological standards (tuna fish ERM-CE 464 (IRMM) and human serum Seronorm Trace Elements Serum L-1) processed either once, or twice, through column chemistry and demonstrate that the CRC allows accurate Cu isotopic measurements of the samples processed only once and therefore with a higher Na/Cu ratio.

Relay-type sensing mode: A strategy to push the limit on nanomechanical sensor sensitivity based on the magneto lever.

Ultrasensitive molecular detection and quantization are crucial for many applications including clinical diagnostics, functional proteomics, and drug discovery; however, conventional biochemical sensors cannot satisfy the stringent requirements, and this has resulted in a long-standing dilemma regarding sensitivity improvement. To this end, we have developed an ultrasensitive relay-type nanomechanical sensor based on a magneto lever. By establishing the link between very weak molecular interaction and five orders of magnitude larger magnetic force, analytes at ultratrace level can produce a clearly observable mechanical response. Initially, proof-of-concept studies showed an improved detection limit up to five orders of magnitude when employing the magneto lever, as compared with direct detection using probe alone. In this study, we subsequently demonstrated that the relay-type sensing mode was universal in application ranging from micromolecule to macromolecule detection, which can be easily extended to detect enzymes, DNA, proteins, cells, viruses, bacteria, chemicals, etc. Importantly, we found that, sensitivity was no longer subject to probe affinity when the magneto lever was sufficiently high, theoretically, even reaching single-molecule resolution. Electronic Supplementary Material: Supplementary material (experimental section) is available in the online version of this article at 10.1007/s12274-022-5049-0.

Exploring COVID-19 transmission patterns and key factors during epidemics caused by three major strains in Asia.

The COVID-19 epidemic has lasted for more than two years since the outbreak in late 2019. An urgent and challenging question is how to systematically evaluate epidemic developments in different countries, during different periods, and to determine which measures that could be implemented are key for successful epidemic prevention. In this study, SBD distance-based K-shape clustering and hierarchical clustering methods were used to analyse epidemics in Asian countries. For the hierarchical clustering, epidemic time series were divided into three periods (epidemics induced by the Original/Alpha, Delta and Omicron variants separately). Standard deviations, the Hurst index, mortality rates, peak value of confirmed cases per capita, average growth rates, and the control efficiency of each period were used to characterize the epidemics. In addition, the total numbers of cases in the different countries were analysed by correlation and regression in relation to 15 variables that could have impacts on COVID-19. Finally, some suggestions on prevention and control measures for each category of country are given. We found that the total numbers of cases per million of a population, total deaths per million and mortality rates were highly correlated with the proportion of people aged over 65 years, the prevalence of multiple diseases, and the national GDP. We also found significant associations between case numbers and vaccination rates, health expenditures, and stringency of control measures. Vaccinations have played a positive role in COVID-19, with a gradual decline in mortality rates in later periods, and are still playing protective roles against the Delta and Omicron strains. The stringency of control measures taken by a government is not an indicator of the appropriateness of a country's response to the outbreak, and a higher index does not necessarily mean more effective measures; a combination of factors such as national vaccination rates, the country's economic foundation and the availability of medical equipment is also needed. This manuscript was submitted as part of a theme issue on "Modelling COVID-19 and Preparedness for Future Pandemics".

Transcriptome profiling revealed the growth superiority of hybrid pufferfish derived from Takifugu obscurus ♀ × Takifugu rubripes ♂.

Hybridization is an efficient method to breed new strains of aquatic animals. In the present study, we produced a hybrid puffer by crossing female obscure puffer with male tiger puffer. The hybrid puffer could live in fresh water like obscure puffer and exhibited growth superiority. The averaged body weight of 4- and 6-month-old hybrid puffer were respectively 38.06% and 38.93% higher than that of obscure puffer. Then, we analyzed the underlying genetic basis for the growth advantage of hybrid puffer by comparative transcriptome analysis. A total number of 4264 and 1285 differentially expressed genes (DEGs) were respectively identified from pituitary and liver transcriptome profiles between hybrid puffer and obscure puffer. Comprehensive analysis showed that the DEGs related with cell proliferation and differentiation, and protein synthesis and export, specifically showed higher expression levels in hybrid puffer, such as "ECM-receptor interaction", "focal adhesion", "protein export" and "protein processing in endoplasmic reticulum". While the DEGs involved in gametogenesis and carbohydrate and energy metabolism highly expressed in obscure puffer, such as "oxidative phosphorylation", "citrate cycle", "progesterone-mediated oocyte maturation" and "oocyte meiosis". Furthermore, a series of candidate genes related to the growth superiority of hybrid puffer were identified, such as fn1a, ptprc, plcg2, igf1, tgfß1, bmp4, abl1, col1a2, col1a1a, and myl9a. These results will be beneficial to understand the molecular basis of growth superiority and helpful to the hybrid breeding of pufferfish.

Bio-informatics and in Vitro Experiments Reveal the Mechanism of Schisandrin A Against MDA-MB-231 cells.

Schisandrin A (SchA) has been reported to have good anti-cancer effects. However, its anti-cancer mechanism in breast cancer remains unknown. This study aimed to explore the mechanism of SchA in breast cancer treatment using bio-informatics analysis and in vitro experiments. The Cancer Genome Atlas (TCGA), Genotype-Tissue Expression (GTEx), Gene Cards, and PharmMapper databases were used to screen the candidate targets of SchA against MDA-MB-231 cells selected as the tested cell line through MTT analysis. The functions and pathways of the targets were identified using Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis and further analyzed using DAVID 6.8.1 database. Network pharmacology analysis revealed 77 candidate targets, 31 signal pathways, and 208 GO entries (P < 0.05). The targets regulated serine-type endopeptidase and protein tyrosine kinase activities, thereby promoting the migration and inhibiting the apoptosis of MDA-MB-231 cells. Comprehensive analysis of the 'Protein-Protein Interaction' (PPI) and 'Component-Targets-Pathways' (C-T-P) networks constructed using Cytoscape 3.7.1 software revealed four core targets: EGFR, PIK3R1, MMP9 and Caspase 3. Their docking scores with SchA were subsequently investigated through molecular docking. The wound healing, Hoechst 33342/PI, and western blot assays confirmed that SchA significantly down-regulated EGFR, PIK3R1, and MMP9, but up-regulated cleaved-caspase 3, thus inhibiting the migration and promoting the apoptosis of MDA-MB-231 cells. Reckoning the findings of the study, SchA is a potential adjuvant treatment for breast cancer.

Holographic tomography: techniques and biomedical applications [Invited].

Holographic tomography (HT) is an advanced label-free optical microscopic imaging method used for biological studies. HT uses digital holographic microscopy to record the complex amplitudes of a biological sample as digital holograms and then numerically reconstruct the sample's refractive index (RI) distribution in three dimensions. The RI values are a key parameter for label-free bio-examination, which correlate with metabolic activities and spatiotemporal distribution of biophysical parameters of cells and their internal organelles, tissues, and small-scale biological objects. This article provides insight on this rapidly growing HT field of research and its applications in biology. We present a review summary of the HT principle and highlight recent technical advancement in HT and its applications.

Influence of noise-reduction techniques in sparse-data sample rotation tomographic imaging.

Data acquisition and processing is a critical issue for high-speed applications, especially in three-dimensional live cell imaging and analysis. This paper focuses on sparse-data sample rotation tomographic reconstruction and analysis with several noise-reduction techniques. For the sample rotation experiments, a live Candida rugosa sample is used and controlled by holographic optical tweezers, and the transmitted complex wavefronts of the sample are recorded with digital holographic microscopy. Three different cases of sample rotation tomography were reconstructed for dense angle with a step rotation at every 2°, and for sparse angles with step rotation at every 5° and 10°. The three cases of tomographic reconstruction performance are analyzed with consideration for data processing using four noise-reduction techniques. The experimental results demonstrate potential capability in retaining the tomographic image quality, even at the sparse angle reconstructions, with the help of noise-reduction techniques.

MicroRNA-29c Acting on FOS Plays a Significant Role in Nonalcoholic Steatohepatitis Through the Interleukin-17 Signaling Pathway.

Nonalcoholic fatty liver disease is the most common hepatic disease in western countries and is even more ubiquitous in Asian countries. Our study determined that TH17/Treg cells were imbalanced in animal models. Based on our interest in the mechanism underlying TH17/Treg cell imbalance in nonalcoholic fatty liver mice, we conducted a joint bioinformatics analysis to further investigate this process. Common gene sequencing analysis was based on one trial from one sequencing platform, where gene expression analysis and enrichment analysis were the only analyses performed. We compared different sequencing results from different trials performed using different sequencing platforms, and we utilized the intersection of these analytical results to perform joint analysis. We used a bioinformatics analysis method to perform enrichment analysis and map interaction network analysis and predict potential microRNA sites. Animal experiments were also designed to validate the results of the data analysis based on quantitative polymerase chain reaction (qPCR) and western blotting. Our results revealed 8 coexisting differentially expressed genes (DEGs) and 7 hinge genes. The identified DEGs may influence nonalcoholic steatosis hepatitis through the interleukin-17 pathway. We found that microRNA-29c interacts with FOS and IGFBP1. Polymerase chain reaction analyses revealed both FOS and microRNA-29c expression in NASH mice, and western blot analyses indicated the same trend with regard to FOS protein levels. Based on these results, we suggest that microRNA-29c acts on FOS via the interleukin-17 signaling pathway to regulate TH17/Treg cells in NASH patients.

6-Gingerol attenuates ventilator-induced lung injury via anti-inflammation and antioxidative stress by modulating the PPARγ/NF-κBsignalling pathway in rats.

Although mechanical ventilation (MV) is indispensable to life-support therapy in critically ill patients, it may promote or aggravatelunginjury known asventilator-inducedlunginjury(VILI). 6-Gingerol is the principal ingredient of ginger with potential anti-inflammatory and antioxidant properties in various diseases. Nevertheless, the role and mechanism of 6-gingerol in the process of VILI has not been explicitly investigated. In the study, we found that pre-treatment with 6-gingerol significantly improved the histological changes and pulmonary oedema, inhibited neutrophil accumulation and the release of early pro-inflammatory cytokines and MPO, and reduced oxidative stress reactions after high MV. Moreover, 6-gingerol treatment also increased PPARγ expression and decreased NF-κB activation in rats subjected to high MV. Furthermore, GW9662, a specific PPARγ inhibitor, was demonstrated to activatethe NF-κB pathway and cancele the protective role of 6-gingerol in VILI. This indicates that 6-gingerol exerted anti-inflammatory and antioxidative stress effects in VILI by activating PPARγ and inhibiting the NF-κBsignalling pathway.

[Identifying Nitrate Sources in a Typical Karst Underground River Basin].

Underground rivers are an important source of groundwater in karst area. Recently, nitrate pollution of underground rivers has become a serious issue. To identify the sources of nitrate in Guancun typical karst underground river basin, stable isotope techniques (δ15N-NO3-, δ18O-NO3-, and δ18O-H2O) were applied in this study. The contribution rates of different nitrate sources in groundwater were quantitatively identified based on the stable isotope analysis in R (SIAR) model, and the influence of land use type on nitrate distribution and source in watershed water was clarified. The results showed that ① nitrate mainly came from fertilizers, soil organic nitrogen, and manure/sewage based to the isotopic composition of nitrate nitrogen and oxygen isotopes. It was revealed that non-point sources significantly contributed to nitrate in waters of the Guancun underground river basin. ② Nitrification dominated the formation process of nitrate in groundwater, and the initial values of nitrogen and oxygen isotopes were not affected by fractionation. ③ Based on SIAR, the contribution of different sources to nitrate in water in the basin varied seasonally, and the contributions of fertilizer, soil organic nitrogen, and manure/sewage to nitrate were 57.07%, 34.06%, and 8.87% in the wet season and 34.14%, 33.02%, and 32.84% in the dry season, respectively. Overall, the present study quantitatively evaluated the temporal variations of nitrate sources in a typical karst groundwater river basin and provided a theoretical foundation for prevention and control management of non-point source pollution and watershed management in karst areas.

Primordial formation of major silicates in a protoplanetary disc with homogeneous 26Al/27Al.

Understanding the spatial variability of initial 26Al/27Al in the solar system, i.e., (26Al/27Al)0, is of prime importance to meteorite chronology, planetary heat production, and protoplanetary disc mixing dynamics. The (26Al/27Al)0 of calcium-aluminum-rich inclusions (CAIs) in primitive meteorites (~5 × 10-5) is frequently assumed to reflect the (26Al/27Al)0 of the entire protoplanetary disc, and predicts its initial 26Mg/24Mg to be ~35 parts per million (ppm) less radiogenic than modern Earth (i.e., Δ'26Mg0 = -35 ppm). Others argue for spatially heterogeneous (26Al/27Al)0, where the source reservoirs of most primitive meteorite components have lower (26Al/27Al)0 at ~2.7 × 10-5 and Δ'26Mg0 of -16 ppm. We measured the magnesium isotope compositions of primitive meteoritic olivine, which originated outside of the CAI-forming reservoir(s), and report five grains whose Δ'26Mg0 are within uncertainty of -35 ppm. Our data thus affirm a model of a largely homogeneous protoplanetary disc with (26Al/27Al)0 of ~5 × 10-5, supporting the accuracy of the 26Al→26Mg chronometer.

Investigation of Microenvironmental Exposures to Particle-Bound Polycyclic Aromatic Hydrocarbons for Elementary School Children.

Polycyclic aromatic hydrocarbons (PAHs) are formed when organic matters incompletely combust and get distributed into the air in the form of vapor or the particular phase of absorption or condensation on the surface of respirable particles. Certain PAHs are considered as carcinogenic and mutagenic, and are primarily associated with the particulate phase. Therefore, the characterization of exposure to particle-bound PAHs (p-PAHs) is critical to assessing the health risks in our daily life. A panel study was conducted during the years 2004 and 2005 to assess microenvironmental exposures to p-PAHs for elementary school children living in Taipei metropolitan area. During the study, integrated filter samples were collected by a dust monitor (model 1.108, Grimm) for 17 p-PAH species analysis using gas chromatography with mass spectrometry (GC/MS). The sampling durations were five days. Overall, 52 samples for children's microenvironmental exposures were included in the data analysis. Results showed that geometric mean (GM) levels (and geometric standard deviation) of p-PAH exposures were 4.443 (3.395) ng/m3 for children. The top three highest proportions of p-PAH components were indeno[1,2,3-cd]pyrene (IND) (21.7%), benzo[g,h,i]perylene (BghiP) (18.5%), and dibenz[a,h]anthracene (DBA) (9.1%), all of which are 5- or 6-ring p-PAHs. In addition, results from diagnostic ratios and principal component analysis (PCA) found that traffic pollution, incense burning, and cooking emission were the major p-PAH exposure sources for children. The total benzo[a]pyrene equivalent (BaPeq) concentration was 1.07 ± 0.80 ng/m3 (mean ± standard deviation), with a GM of 0.84 ng/m3. The GM value of the inhalation carcinogenic risk was 7.31 × 10-5 with the range of 2.23 × 10-5 to 3.11 × 10-4, which was higher than the U.S. Environmental Protection Administration guideline limit of 10-6. DBA accounted for 45.1% of the excess cancer risk, followed by benzo[a]pyrene (BaP) (33.5%) and IND (10.7%). In conclusion, the current study demonstrated that inhalational cancer risk due to the p-PAH exposures for children is not negligible, and more efficient technical and management policies should be adopted to reduce the PAH pollutant sources.

Adaptive wavefront correction structured illumination holographic tomography.

In this study, a novel adaptive wavefront correction (AWC) technique is implemented on a compactly developed structured illumination holographic tomography (SI-HT) system. We propose a mechanical movement-free compact scanning architecture for SI-HT systems with AWC, implemented by designing and displaying a series of computer-generated holograms (CGH) composed of blazed grating with phase Fresnel lens on a phase-only spatial light modulator (SLM). In the proposed SI-HT, the aberrations of the optical system are sensed by digital holography and are used to design the CGH-based AWC to compensate the phase aberrations of the tomographic imaging system. The proposed method was validated using a standard Siemens star target, its potential application was demonstrated using a live candida rugosa sample, and its label-free three-dimensional refractive index profile was generated at its subcellular level. The experimental results obtained reveal the ability of the proposed method to enhance the imaging performance in both lateral and axial directions.

Geochemical behaviors of antimony in mining-affected water environment (Southwest China).

Antimony (Sb) is a harmful element, and Sb pollution is one of the typical environmental issues in China, meaning that understanding of the geochemical behaviors of Sb is the key to control the fate of environmental Sb pollution. Sb tends to migrate in soluble form in the water-sediment system, but the fate of dissolved Sb is poorly known. Duliujiang river basin, located in southwest China, provided us with a natural aqueous environment to study the transport of Sb because of its unique geological and geographical characteristics. Physicochemical properties (pH, EC, Eh, DO, Flux), trace elements (Sb, As, Sr) and main ions (Ca2+, Mg2+, SO42-) concentrations in mining-impacted waters were measured in order to determine their distribution and migration potential. There are three types of water samples; they are main stream waters (pH of 7.33-8.43), tributary waters (pH of 6.85-9.12) and adit waters with pH values ranging from 7.57 to 9.76, respectively. Results showed that adit waters contained elevated concentrations of Sb reaching up to 13350 µg L-1 from the abandoned Sb mines, and mine wastes contained up to 8792 mg kg-1 Sb from the historical mine dumps are the important sources of Sb pollution in the Duliujiang river basin. Dissolved Sb had strong migration ability in streams, while its attenuation mainly depended on the dilution of tributary water with large flow rate. In the exit section of the Duliujiang river basin, which had only 10 µg L-1 of average Sb concentration. The simple deionized water extraction was designed to investigate the ability of Sb likely to dissolve from the mine wastes. The results indicated that a greater solubility of Sb in alkaline (pH of 7.11-8.16) than in acid (pH of 3.03-4.45) mine wastes, suggesting that mine wastes contained high Sb concentrations, could release Sb into solution in the natural river waters. Furthermore, the fate of Sb pollution depends on the comprehensive treatment of abandoned adit waters and mine wastes in the upper reaches of the drainage basin.

An Unbalanced Weighted Sequential Fusing Multi-Sensor GM-PHD Algorithm.

In this paper, we study the multi-sensor multi-target tracking problem in the formulation of random finite sets. The Gaussian Mixture probability hypothesis density (GM-PHD) method is employed to formulate the sequential fusing multi-sensor GM-PHD (SFMGM-PHD) algorithm. First, the GM-PHD is applied to multiple sensors to get the posterior GM estimations in a parallel way. Second, we propose the SFMGM-PHD algorithm to fuse the multi-sensor GM estimations in a sequential way. Third, the unbalanced weighted fusing and adaptive sequence ordering methods are further proposed for two improved SFMGM-PHD algorithms. At last, we analyze the proposed algorithms in four different multi-sensor multi-target tracking scenes, and the results demonstrate the efficiency.

One-shot synthetic aperture digital holographic microscopy with non-coplanar angular-multiplexing and coherence gating.

This paper proposes one-shot synthetic aperture digital holographic microscopy using a combination of angular-multiplexing and coherence gating. The proposed angular-multiplexing technique uses multiple noncoplanar incident beams into the synthetic aperture to create tight packed passbands so as to extend spatial frequency spectrum. Coherence gating is performed to prevent the self-interference among the multiple beams. Based on the design guideline proposed herein, a phase-only spatial light modulator is employed as an adjustable blazed grating to split multiple noncoplanar beams and perform angular-multiplexing, and then using coherence gating based on low-coherence-light, superresolution imaging is achieved after one-shot acquisition.

Integrated dual-tomography for refractive index analysis of free-floating single living cell with isotropic superresolution.

Digital holographic microtomography is a promising technique for three-dimensional (3D) measurement of the refractive index (RI) profiles of biological specimens. Measurement of the RI distribution of a free-floating single living cell with an isotropic superresolution had not previously been accomplished. To the best of our knowledge, this is the first study focusing on the development of an integrated dual-tomographic (IDT) imaging system for RI measurement of an unlabelled free-floating single living cell with an isotropic superresolution by combining the spatial frequencies of full-angle specimen rotation with those of beam rotation. A novel 'UFO' (unidentified flying object) like shaped coherent transfer function is obtained. The IDT imaging system does not require any complex image-processing algorithm for 3D reconstruction. The working principle was successfully demonstrated and a 3D RI profile of a single living cell, Candida rugosa, was obtained with an isotropic superresolution. This technology is expected to set a benchmark for free-floating single live sample measurements without labeling or any special sample preparations for the experiments.