Targeting Macrophages: Therapeutic Approaches in Diabetic Kidney Disease.

Diabetes is not solely a metabolic disorder but also involves inflammatory processes. The immune response it incites is a primary contributor to damage in target organs. Research indicates that during the initial phases of diabetic nephropathy, macrophages infiltrate the kidneys alongside lymphocytes, initiating a cascade of inflammatory reactions. The interplay between macrophages and other renal cells is pivotal in the advancement of kidney disease within a hyperglycemic milieu. While M1 macrophages react to the inflammatory stimuli induced by elevated glucose levels early in the disease progression, their subsequent transition to M2 macrophages, which possess anti-inflammatory and tissue repair properties, also contributes to fibrosis in the later stages of nephropathy by transforming into myofibroblasts. Comprehending the diverse functions of macrophages in diabetic kidney disease and regulating their activity could offer therapeutic benefits for managing this condition.

Insights into the Molecular Mechanisms of NRF2 in Kidney Injury and Diseases.

Redox is a constant phenomenon in organisms. From the signaling pathway transduction to the oxidative stress during the inflammation and disease process, all are related to reduction-oxidation (redox). Nuclear factor erythroid 2-related factor 2 (NRF2) is a transcription factor targeting many antioxidant genes. In non-stressed conditions, NRF2 maintains the hemostasis of redox with housekeeping work. It expresses constitutively with basal activity, maintained by Kelch-like-ECH-associated protein 1 (KEAP1)-associated ubiquitination and degradation. When encountering stress, it can be up-regulated by several mechanisms to exert its anti-oxidative ability in diseases or inflammatory processes to protect tissues and organs from further damage. From acute kidney injury to chronic kidney diseases, such as diabetic nephropathy or glomerular disease, many results of studies have suggested that, as a master of regulating redox, NRF2 is a therapeutic option. It was not until the early termination of the clinical phase 3 trial of diabetic nephropathy due to heart failure as an unexpected side effect that we renewed our understanding of NRF2. NRF2 is not just a simple antioxidant capacity but has pleiotropic activities, harmful or helpful, depending on the conditions and backgrounds.

Discovery of compounds that reactivate p53 mutants in vitro and in vivo.

The tumor suppressor p53 is the most frequently mutated protein in human cancer. The majority of these mutations are missense mutations in the DNA binding domain of p53. Restoring p53 tumor suppressor function could have a major impact on the therapy for a wide range of cancers. Here we report a virtual screening approach that identified several small molecules with p53 reactivation activities. The UCI-LC0023 compound series was studied in detail and was shown to bind p53, induce a conformational change in mutant p53, restore the ability of p53 hotspot mutants to associate with chromatin, reestablish sequence-specific DNA binding of a p53 mutant in a reconstituted in vitro system, induce p53-dependent transcription programs, and prevent progression of tumors carrying mutant p53, but not p53null or p53WT alleles. Our study demonstrates feasibility of a computation-guided approach to identify small molecule corrector drugs for p53 hotspot mutations.

New Insights into the Treatment of Glomerular Diseases: When Mechanisms Become Vivid.

Treatment for glomerular diseases has been extrapolated from the experience of other autoimmune disorders while the underlying pathogenic mechanisms were still not well understood. As the classification of glomerular diseases was based on patterns of juries instead of mechanisms, treatments were typically the art of try and error. With the advancement of molecular biology, the role of the immune agent in glomerular diseases is becoming more evident. The four-hit theory based on the discovery of gd-IgA1 gives a more transparent outline of the pathogenesis of IgA nephropathy (IgAN), and dysregulation of Treg plays a crucial role in the pathogenesis of minimal change disease (MCD). An epoch-making breakthrough is the discovery of PLA2R antibodies in the primary membranous nephropathy (pMN). This is the first biomarker applied for precision medicine in kidney disease. Understanding the immune system's role in glomerular diseases allows the use of various immunosuppressants or other novel treatments, such as complement inhibitors, to treat glomerular diseases more reasonable. In this era of advocating personalized medicine, it is inevitable to develop precision medicine with mechanism-based novel biomarkers and novel therapies in kidney disease.

Relationship Between Serum Concentration of Adrenomedullin and Myocardial Ischemic T Wave Changes in Patients With Lung Cancer.

Background: Patients with lung cancer are at increased risk for the development of cardiovascular diseases. Molecular markers for early diagnosis of cardiac ischemia are of great significance for the early prevention of cardiovascular events in patients with lung cancer. By evaluating the relationship between adrenomedullin (ADM) and myocardial ischemic T wave changes, the clinical value of circulating ADM as a predictor of myocardial ischemia in patients with lung cancer is confirmed. Methods: We enrolled patients with lung cancer and healthy people from 2019 to 2021 and extracted a detailed ECG parameter. After adjustment for potential confounders, logistic regression was used to assess the association of clinical data. We performed analyses on differences in T wave between patients with lung cancer and healthy people, and the relationship between T wave and ADM among patients with lung cancer. Receiver operator characteristic (ROC) curves were drawn to confirm the diagnostic value of biomarkers. Results: After adjusting for potential confounders, the incidence of T wave inversion or flattening in patients with lung cancer was higher than in healthy people (OR: 3.3228, P = 0.02). Also, further analysis of the data of lung cancer patients revealed that the ADM in lung cancer patients with T wave inversion or flat was higher than those with normal T wave (189.8 ± 51.9 vs. 131.9 ± 38.4, p < 0.001). The area under the ROC curve was 0.8137. Conclusion: Among the patients with lung cancer, serum ADM concentration is associated with the incidence of the abnormal T wave. ADM might be a potentially valuable predictor for heart ischemia in patients with lung cancer.

Metabolic remodelling during early mouse embryo development.

During early mammalian embryogenesis, changes in cell growth and proliferation depend on strict genetic and metabolic instructions. However, our understanding of metabolic reprogramming and its influence on epigenetic regulation in early embryo development remains elusive. Here we show a comprehensive metabolomics profiling of key stages in mouse early development and the two-cell and blastocyst embryos, and we reconstructed the metabolic landscape through the transition from totipotency to pluripotency. Our integrated metabolomics and transcriptomics analysis shows that while two-cell embryos favour methionine, polyamine and glutathione metabolism and stay in a more reductive state, blastocyst embryos have higher metabolites related to the mitochondrial tricarboxylic acid cycle, and present a more oxidative state. Moreover, we identify a reciprocal relationship between α-ketoglutarate (α-KG) and the competitive inhibitor of α-KG-dependent dioxygenases, L-2-hydroxyglutarate (L-2-HG), where two-cell embryos inherited from oocytes and one-cell zygotes display higher L-2-HG, whereas blastocysts show higher α-KG. Lastly, increasing 2-HG availability impedes erasure of global histone methylation markers after fertilization. Together, our data demonstrate dynamic and interconnected metabolic, transcriptional and epigenetic network remodelling during early mouse embryo development.

Next-Generation Genome-Scale Metabolic Modeling through Integration of Regulatory Mechanisms.

Genome-scale metabolic models (GEMs) are powerful tools for understanding metabolism from a systems-level perspective. However, GEMs in their most basic form fail to account for cellular regulation. A diverse set of mechanisms regulate cellular metabolism, enabling organisms to respond to a wide range of conditions. This limitation of GEMs has prompted the development of new methods to integrate regulatory mechanisms, thereby enhancing the predictive capabilities and broadening the scope of GEMs. Here, we cover integrative models encompassing six types of regulatory mechanisms: transcriptional regulatory networks (TRNs), post-translational modifications (PTMs), epigenetics, protein-protein interactions and protein stability (PPIs/PS), allostery, and signaling networks. We discuss 22 integrative GEM modeling methods and how these have been used to simulate metabolic regulation during normal and pathological conditions. While these advances have been remarkable, there remains a need for comprehensive and widespread integration of regulatory constraints into GEMs. We conclude by discussing challenges in constructing GEMs with regulation and highlight areas that need to be addressed for the successful modeling of metabolic regulation. Next-generation integrative GEMs that incorporate multiple regulatory mechanisms and their crosstalk will be invaluable for discovering cell-type and disease-specific metabolic control mechanisms.

Localized Proteolysis for the Construction of Intracellular Asymmetry in Escherichia coli.

Protein-level regulations have gained importance in building synthetic circuits, as they offer a potential advantage in the speed of operation compared to gene regulation circuits. In nature, localized protein degradation is prevalent in polarizing cellular signaling. We, therefore, set out to systematically investigate whether localized proteolysis can be employed to construct intracellular asymmetry in Escherichia coli. We demonstrate that, by inserting a cognate cleavage site between the reporter and C-terminal degron, the unstable reporter can be stabilized in the presence of the tobacco etch virus protease. Furthermore, the split protease can be functionally reconstituted by the PopZ-based polarity system to exert localized proteolysis. Selective stabilization of the unstable reporter at the PopZ pole can lead to intracellular asymmetry in E. coli. Our study provides complementary evidence to support that localized proteolysis may be a strategy for polarization in developmental cell biology. Circuits designed in this study may also help to expand the synthetic biology repository for the engineering of synthetic morphogenesis, particularly for processes that require rapid control of local protein abundance.

Association of furosemide or hydrochlorothiazide use with risk of atrial fibrillation post pacemaker implantation among elderly patients.

BACKGROUND: Atrial fibrillation (AF) induced by artificial pacing is directly related to atrial remodeling. Previous basic research has shown that furosemide aggravates pathologic myocardial remodeling while hydrochlorothiazide alleviates it. However, whether furosemide or hydrochlorothiazide plays a role in developing AF after pacemaker implantation remains unknown. The study aims to investigate the association between oral furosemide or hydrochlorothiazide and the risk of developing AF after pacemaker implantation. METHODS: After a review of electronic medical records, elderly patients with pacemaker implantation and without a known baseline history of AF were included and information on their use of daily oral furosemide or hydrochlorothiazide was extracted. New incident AF cases were confirmed via the records of outpatient visits. A Cox proportional-hazards model was used to evaluate the association between daily oral furosemide or hydrochlorothiazide and risk of developing AF after pacemaker implantation, after adjustment for potential confounders. RESULTS: Among a total of 551 patients aged more than 65 years, 157 AF cases were identified after pacemaker implantation during a maximum follow up of 3.0±1.6 years. Of these, 242 had used furosemide and 97 had used hydrochlorothiazide therapy. Patients taking daily oral furosemide had a relatively higher risk of AF after pacemaker implantation [hazard ratio (HR): 1.507, 95% confidence interval (CI): 1.036-2.192; P=0.032] after being adjusted for related disease and prescribed medications, while oral taking of hydrochlorothiazide was shown to be a non-effective factor (HR: 0.666, 95% CI: 0.413-1.074), which had no statistical significance. CONCLUSIONS: Daily oral furosemide might increase the risk of developing AF after pacemaker implantation in elderly patients, while hydrochlorothiazide has no detrimental effect.

Lipid remodeling in response to methionine stress in MDA-MBA-468 triple-negative breast cancer cells.

Methionine (Met) is an essential amino acid and critical precursor to the cellular methyl donor S-adenosylmethionine. Unlike nontransformed cells, cancer cells have a unique metabolic requirement for Met and are unable to proliferate in growth media where Met is replaced with its metabolic precursor, homocysteine. This metabolic vulnerability is common among cancer cells regardless of tissue origin and is known as "methionine dependence", "methionine stress sensitivity", or the Hoffman effect. The response of lipids to Met stress, however, is not well-understood. Using mass spectroscopy, label-free vibrational microscopy, and next-generation sequencing, we characterize the response of lipids to Met stress in the triple-negative breast cancer cell line MDA-MB-468 and its Met stress insensitive derivative, MDA-MB-468res-R8. Lipidome analysis identified an immediate, global decrease in lipid abundances with the exception of triglycerides and an increase in lipid droplets in response to Met stress specifically in MDA-MB-468 cells. Furthermore, specific gene expression changes were observed as a secondary response to Met stress in MDA-MB-468, resulting in a downregulation of fatty acid metabolic genes and an upregulation of genes in the unfolded protein response pathway. We conclude that the extensive changes in lipid abundance during Met stress is a direct consequence of the modified metabolic profile previously described in Met stress-sensitive cells. The changes in lipid abundance likely results in changes in membrane composition inducing the unfolded protein response we observe.

Construction of intracellular asymmetry and asymmetric division in Escherichia coli.

The design principle of establishing an intracellular protein gradient for asymmetric cell division is a long-standing fundamental question. While the major molecular players and their interactions have been elucidated via genetic approaches, the diversity and redundancy of natural systems complicate the extraction of critical underlying features. Here, we take a synthetic cell biology approach to construct intracellular asymmetry and asymmetric division in Escherichia coli, in which division is normally symmetric. We demonstrate that the oligomeric PopZ from Caulobacter crescentus can serve as a robust polarized scaffold to functionalize RNA polymerase. Furthermore, by using another oligomeric pole-targeting DivIVA from Bacillus subtilis, the newly synthesized protein can be constrained to further establish intracellular asymmetry, leading to asymmetric division and differentiation. Our findings suggest that the coupled oligomerization and restriction in diffusion may be a strategy for generating a spatial gradient for asymmetric cell division.

Eight-year dry deposition of atmospheric mercury to a tropical high mountain background site downwind of the East Asian continent.

Atmospheric deposition, either dry or wet, has been identified as an important pathway of mercury (Hg) input to terrestrial and aquatic systems. Although East Asia is the major atmospheric Hg emission source region, very few studies have been conducted to quantify atmospheric Hg deposition in its downwind region. In this study, 8-year (2009-2016) atmospheric Hg dry deposition was reported at the Lulin Atmospheric Background Station (LABS), a high mountain forest site in central Taiwan. Dry deposition of speciated Hg was estimated using a bi-directional air-surface flux exchange model for gaseous elemental mercury (GEM) and dry deposition models for gaseous oxidized mercury (GOM) and particulate-bound mercury (PBM), making use of the monitored speciated atmospheric Hg concentrations. Annual total Hg dry deposition ranged from 51.9 to 84.9 µg m-2 yr-1, with a multi-year average of 66.1 µg m-2 yr-1. Among the three forms of atmospheric Hg, GEM was the main contributor to the total dry deposition, contributing about 77.8% to the total, due to the high density of forest canopy as well as the much higher concentration of GEM than GOM and PBM at LABS. Mercury dry deposition is higher in winter and spring than in summer and fall, partly due to the elevated Hg concentrations associated with air masses from East and Southeast Asia where with high atmospheric Hg emissions. The mean annual dry/wet deposition ratio of 2.8 at LABS indicated that Hg deposition to forest landscape was governed by dry rather than wet deposition.

Temporal changes in atmospheric mercury concentrations at a background mountain site downwind of the East Asia continent in 2006-2016.

Atmospheric mercury (Hg) has been monitored at the Lulin Atmospheric Background Station (LABS) in Taiwan since April 2006 and is still continuing. Here we reported the trend in gaseous elemental Hg (GEM) concentrations at LABS between April 2006 and December 2016, before the Minamata Convention on Mercury entered into force in 2017. Previous research indicated nighttime (0-8 am) data collected at LABS are better representative of regional influence. Therefore, only nighttime GEM data were used for trend analysis. A significant decreasing trend in GEM at a rate of -1.5% yr-1 (-0.022 ng m-3 yr-1, p < 0.01) was found, comparable to the decreasing trends observed in Europe, North America, South Africa, and over the North Atlantic Ocean. Five major GEM source regions to the LABS were identified, including northern Indochina Peninsula, China, Northeast Asia, the Pacific Ocean, and South China Sea. Significant decreasing trends in GEM were found for air masses coming from northern Indochina Peninsula (-0.042 ng m-3 yr-1, -2.6% yr-1, p < 0.01), China (-0.041 ng m-3 yr-1, -2.4% yr-1, p < 0.01), Northeast Asia (-0.031 ng m-3 yr-1, -2.0% yr-1, p < 0.05), and the Pacific Ocean (-0.022 ng m-3 yr-1, -1.7% yr-1, p < 0.05). Decreasing GEM trend (-0.020 ng m-3 yr-1, -1.5% yr-1), but insignificant (p > 0.05), was also found for air masses coming from South China Sea. The decreasing trends observed with air from the Pacific Ocean and South China Sea indicated declining background GEM concentrations in Northern Hemisphere. Decrease in GEM concentrations at the LABS was in agreement with the reduction in atmospheric Hg export from the East Asia continent caused by changes in Hg emission quantity and speciation, and temporal and spatial distribution in emission sources that have been suggested by recent research. Additionally, changes in the frequency distribution of air mass origins and transport paths may also contribute to the changes in GEM concentrations at LABS.

Microhomology-based CRISPR tagging tools for protein tracking, purification, and depletion.

Work in yeast models has benefitted tremendously from the insertion of epitope or fluorescence tags at the native gene locus to study protein function and behavior under physiological conditions. In contrast, work in mammalian cells largely relies on overexpression of tagged proteins because high-quality antibodies are only available for a fraction of the mammalian proteome. CRISPR/Cas9-mediated genome editing has recently emerged as a powerful genome-modifying tool that can also be exploited to insert various tags and fluorophores at gene loci to study the physiological behavior of proteins in most organisms, including mammals. Here we describe a versatile toolset for rapid tagging of endogenous proteins. The strategy utilizes CRISPR/Cas9 and microhomology-mediated end joining repair for efficient tagging. We provide tools to insert 3×HA, His6FLAG, His6-Biotin-TEV-RGSHis6, mCherry, GFP, and the auxin-inducible degron tag for compound-induced protein depletion. This approach and the developed tools should greatly facilitate functional analysis of proteins in their native environment.

Isolation and Characterization of Methionine-Independent Clones from Methionine-Dependent Cancer Cells.

Unlike normal cells, transformed cells are unable to grow when methionine in the growth media is restricted. Reversion to methionine independence is a rare event in transformed and malignant cells. Methionine-independent revertants provide an excellent system to identify metabolic signatures and molecular characteristics associated with methionine dependency of transformed cells. Revertants maintain the genetic background and general growth behavior of the parental cell line, except that they proliferate under methionine restriction such as in methionine-free media supplemented with homocysteine. Here we describe a general approach to generate methionine-independent revertants using the example of the triple-negative breast cancer cell line MDA-MB-468. To validate and characterize reversion we describe assays to evaluate cell proliferation and anchorage-independent growth in soft agar.

A New Monitoring Effort for Asia: The Asia Pacific Mercury Monitoring Network (APMMN).

The Asia Pacific Mercury Monitoring Network (APMMN) cooperatively measures mercury in precipitation in a network of sites operating in Asia and the Western Pacific region. The network addresses significant data gaps in a region where mercury emission estimates are the highest globally, and available measurement data are limited. The reduction of mercury emissions under the Minamata Convention on Mercury also justifies the need for continent-wide and consistent observations that can help determine the magnitude of the problem and assess the efficacy of reductions over time. The APMMN's primary objectives are to monitor wet deposition and atmospheric concentrations of mercury and assist partners in developing their own monitoring capabilities. Network planning began in 2012 with wet deposition sampling starting in 2014. Currently, eight network sites measure mercury in precipitation following standardized procedures adapted from the National Atmospheric Deposition Program. The network also has a common regional analytical laboratory (Taiwan), and quality assurance and data flagging procedures, which ensure the network makes scientifically valid and consistent measurements. Results from our ongoing analytical and field quality assurance measurements show minimal contamination in the network and accurate analytical analyses. We are continuing to monitor a potential concentration and precipitation volume bias under certain conditions. The average mercury concentration in precipitation was 11.3 (+9.6) ng L-1 for 139 network samples in 2018. Concentrations for individual sites vary widely. Low averages compare to the low concentrations observed on the U.S. West Coast; while other sites have average concentrations similar to the high values reported from many urban areas in China. Future APMMN goals are to (1) foster new network partnerships, (2) continue to collect, quality assure, and distribute results on the APMMN website, (3) provide training and share best monitoring practices, and (4) establish a gaseous concentration network for estimating dry deposition.

High-performance InGaN-based green light-emitting diodes with quaternary InAlGaN/GaN superlattice electron blocking layer.

In this study, high-performance InGaN-based green light-emitting diodes (LEDs) with a quaternary InAlGaN/GaN superlattice electron blocking layer (QSL-EBL) have been demonstrated. The band structural simulation was employed to investigate the electrostatic field and carriers distribution, show that the efficiency and droop behavior can be intensively improved by using a QSL-EBL in LEDs. The QSL-EBL structure can reduce the polarization-related electrostatic fields in the multiple quantum wells (MQWs), leading to a smoother band diagram and a more uniform carriers distribution among the quantum wells under forward bias. In comparison with green LEDs with conventional bulk-EBL structure, the light output power of LEDs with QSL-EBL was greatly enhanced by 53%. The efficiency droop shows only 30% at 100 A/cm2 comparing to its peak value, suggesting that the QSL-EBL LED is promising for future white lighting with high performance.

Bridging the "green gap" of LEDs: giant light output enhancement and directional control of LEDs via embedded nano-void photonic crystals.

Green LEDs do not show the same level of performance as their blue and red cousins, greatly hindering the solid-state lighting development, which is the so-called "green gap". In this work, nano-void photonic crystals (NVPCs) were fabricated to embed within the GaN/InGaN green LEDs by using epitaxial lateral overgrowth (ELO) and nano-sphere lithography techniques. The NVPCs act as an efficient scattering back-reflector to outcouple the guided and downward photons, which not only boost the light extraction efficiency of LEDs with an enhancement of 78% but also collimate the view angle of LEDs from 131.5° to 114.0°. This could be because of the highly scattering nature of NVPCs which reduce the interference giving rise to Fabry-Perot resonance. Moreover, due to the threading dislocation suppression and strain relief by the NVPCs, the internal quantum efficiency was increased by 25% and droop behavior was reduced from 37.4% to 25.9%. The enhancement of light output power can be achieved as high as 151% at a driving current of 350 mA. Giant light output enhancement and directional control via NVPCs point the way towards a promising avenue of solid-state lighting.

[Determination of five pesticides in fishpond by SPE-GC/MS].

OBJECTIVE: To establish the solid phase extraction (SPE) with GC/MS technology for fish poisoning cases to determine five pesticides in fishpond. METHODS: By three solid phase extraction column including Oasis HLB cartridge, Bond Elut C18 and SampliQ C18, the recovery rate was compared to extract and purify five pesticides in fishpond. The effects of different kinds and dosages of eluents on extract rate were also reviewed. RESULTS: Using Bond Elut C18 as solid phase extraction column and 3 mL benzene as eluent, the linear range of mass concentration of five pesticides in fishpond was 1-50 µg/mL, and the correlation coefficient was 0.996 2-0.999 6. The limit of detection was 3.4-26 µg/L and the recovery was 61.49%-102.48%. The relative standard deviations was less than or equal to 3.01%. CONCLU-SION: With high sensitivity, good accuracy and precision, SPE-GC/MS has simple and quick operation and less solvent. It can be applied to determination of five pesticides in fishpond.

Improvement of emission uniformity by using micro-cone patterned PDMS film.

Micro-patterned PDMS film was fabricated and combined with LED chip on board (COB) package to improve the emission uniformity of LED chip. The micro scale patterned sapphire substrate (PSS) was used as a mold to fabricate micro-cone patterned PDMS (MC-PDMS) film. A strong scattering effect from this MC-PDMS film can be verified by the high haze ratio and the Bi-directional Transmission effect. The angle dependent color temperature measurement system was used to measure the ΔCCT of COB with and without MC-PDMS. The measurement results indicate that the ΔCCT was reduced from 1025K to 428K. This improvement can effectively eliminate the yellow ring effect of LED chip. This technology can be thus considered as a cost-effective way for the next generation of light source packages.