Elucidating Gender-Specific Distribution of Imipramine, Chloroquine, and Their Metabolites in Mice Kidney Tissues through AP-MALDI-MSI.

Knowledge of gender-specific drug distributions in different organs are of great importance for personalized medicine and reducing toxicity. However, such drug distributions have not been well studied. In this study, we investigated potential differences in the distribution of imipramine and chloroquine, as well as their metabolites, between male and female kidneys. Kidneys were collected from mice treated with imipramine or chloroquine and then subjected to atmospheric pressure matrix-assisted laser desorption ionization-mass spectrometry imaging (AP-MALDI-MSI). We observed differential distributions of the drugs and their metabolites between male and female kidneys. Imipramine showed prominent distributions in the cortex and medulla in male and female kidneys, respectively. Desipramine, one of the metabolites of imipramine, showed significantly higher (*** p < 0.001) distributions in the medulla of the male kidney compared to that of the female kidney. Chloroquine and its metabolites were accumulated in the pelvis of both male and female kidneys. Interestingly, they showed a characteristic distribution in the medulla of the female kidney, while almost no distributions were observed in the same areas of the male kidney. For the first time, our study revealed that the distributions of imipramine, chloroquine, and their metabolites were different in male and female kidneys.

Low-Temperature Plasma Pretreatment Enhanced Cholesterol Detection in Brain by Desorption Electrospray Ionization-Mass Spectrometry Imaging.

Cholesterol is a primary lipid molecule in the brain that contains one-fourth of the total body cholesterol. Abnormal cholesterol homeostasis is associated with neurodegenerative disorders. Mass spectrometry imaging (MSI) technique is a powerful tool for studying lipidomics and metabolomics. Among the MSI techniques, desorption electrospray ionization-MSI (DESI-MSI) has been used advantageously to study brain lipidomics due to its soft and ambient ionization nature. However, brain cholesterol is poorly ionized. To this end, we have developed a new method for detecting brain cholesterol by DESI-MSI using low-temperature plasma (LTP) pretreatment as an ionization enhancement. In this method, the brain sections were treated with LTP for 1 and 2 min prior to DESI-MSI analyses. Interestingly, the MS signal intensity of cholesterol (at m/z 369.35 [M + H - H2O]+) was more than 2-fold higher in the 1 min LTP-treated brain section compared to the untreated section. In addition, we detected cholesterol, more specifically excluding isomers by targeted-DESI-MSI in multiple reaction monitoring (MRM) mode and similar results were observed: the signal intensity of each cholesterol transition (m/z 369.4 → 95.1, 109.1, 135.1, 147.1, and 161.1) was increased by more than 2-fold due to 1 min LTP treatment. Cholesterol showed characteristic distributions in the fiber tract region, including the corpus callosum and anterior commissure, anterior part of the brain where LTP markedly (p < 0.001) enhanced the cholesterol intensity. In addition, the distributions of some unknown analytes were exclusively detected in the LTP-treated section. Our study revealed LTP pretreatment as a potential strategy to ionize molecules that show poor ionization efficiency in the MSI technique.

A stroke prediction framework using explainable ensemble learning.

The death of brain cells occurs when blood flow to a particular area of the brain is abruptly cut off, resulting in a stroke. Early recognition of stroke symptoms is essential to prevent strokes and promote a healthy lifestyle. FAST tests (looking for abnormalities in the face, arms, and speech) have limitations in reliability and accuracy for diagnosing strokes. This research employs machine learning (ML) techniques to develop and assess multiple ML models to establish a robust stroke risk prediction framework. This research uses a stacking-based ensemble method to select the best three machine learning (ML) models and combine their collective intelligence. An empirical evaluation of a publicly available stroke prediction dataset demonstrates the superior performance of the proposed stacking-based ensemble model, with only one misclassification. The experimental results reveal that the proposed stacking model surpasses other state-of-the-art research, achieving accuracy, precision, F1-score of 99.99%, recall of 100%, receiver operating characteristics (ROC), Mathews correlation coefficient (MCC), and Kappa scores 1.0. Furthermore, Shapley's Additive Explanations (SHAP) are employed to analyze the predictions of the black-box machine learning (ML) models. The findings highlight that age, BMI, and glucose level are the most significant risk factors for stroke prediction. These findings contribute to the development of more efficient techniques for stroke prediction, potentially saving many lives.

Quantification, characterization and risk assessment of microplastics from five major estuaries along the northern Bay of Bengal coast.

Microplastics (MPs) as hazardous contaminants has drawn the rapid attention of the general public due to their omnipresence and adverse impacts on ecosystems and human health. Despite this, understanding of MPs contamination levels in the estuarine ecosystems along the Bay of Bengal coast remains very limited. This research focused on the presence, spatial distribution, morpho-chemical characteristics and ecological implications of MPs in water and sediment from five key estuaries (Meghna, Karnaphuli, Matamuhuri, Bakkhali, and Naf rivers) within the Bengal delta. Out of the five estuaries, the Meghna exhibited the least amount of MPs in both surface water (150.00 ± 65.62 items/m3) and sediment (30.56 ± 9.34 items/kg). In contrast, the highest occurrence of MPs was recorded in Karnaphuli river water (350.00 ± 69.22 items/m3) and Matamuhuri river sediment (118.33 ± 26.81 items/kg). ANOVA indicated a statistically significant distinction (p < 0.01) among the examined estuaries. Most identified MPs were fibers and < 0.5 mm in size in both water and sediment samples. Transparent MPs were dominant in both water (42.28%) and sediment (45.22%). Besides violet, red, blue, pink and green colored MPs were also observed. Various polymer types, including PE, PP, PET, PS, Nylon, EVA, and ABS, were detected, with PE being the dominant one. Based on the polymer risk index (PHI), the estuaries were classified as hazard level V, signifying a severe level of MP contamination. However, the potential ecological hazardous index (PHI), potential ecological risk index (RI), and pollution load index (PLI) indicated moderate pollution levels. This study offers initial insights into the pollution caused by MPs in major estuaries of Bengal delta, which policymakers can utilize to implement suitable management strategies.

Lipid Polyunsaturated Fatty Acid Chains in Mouse Kidneys Were Increased within 5 min of a Single High Dose Whole Body Irradiation.

To understand the ultra-early reaction of normal organ lipids during irradiation, we investigated the response of lipids, including polyunsaturated fatty acid (PUFA) chains, which are particularly susceptible to damage by ROS, in mice's kidneys, lungs, brains, and livers within 5 min of single high-dose irradiation. In this study, we set up three groups of C56BL/6 male mice and conducted whole-body irradiation with 0 Gy, 10 Gy, and 20 Gy single doses. Kidney, lung, brain, and liver tissues were collected within 5 min of irradiation. PUFA-targeted and whole lipidomic analyses were conducted using liquid chromatography-tandem mass spectrometry (LC-MS/MS). The results showed that PUFA chains of kidney phosphatidylcholine (PC), phosphatidylethanolamine (PE), and triacylglycerol (TG) significantly increased within 5 min of 10 Gy and 20 Gy irradiation. The main components of increased PUFA chains in PC and PE were C18:2, C20:4, and C22:6, and in TG the main component was C18:2. The kidney lipidomes also showed significant changes from the perspective of lipid species, mainly dominated by an increase in PC, PE, TG, and signal lipids, while lipidomes of the lung, brain, and liver were slightly changed. Our results revealed that acute PUFA chains increase and other lipidomic changes in the kidney upon whole-body irradiation within 5 min of irradiation. The significantly increased lipids also showed a consistent preference for possessing PUFA chains. The lipidomic changes varied from organ to organ, which indicates that the response upon irradiation within a short time is tissue-specific.

Detection of Distinct Distributions of Acetaminophen and Acetaminophen-Cysteine in Kidneys up to 10 µm Resolution and Identification of a Novel Acetaminophen Metabolite Using an AP-MALDI Imaging Mass Microscope.

Drug distribution studies in tissue are crucial for understanding the pharmacokinetics and potential toxicity of drugs. Recently, matrix-assisted laser desorption ionization-mass spectrometry imaging (MALDI-MSI) has gained attention for drug distribution studies due to its high sensitivity, label-free nature, and ability to distinguish between parent drugs, their metabolites, and endogenous molecules. Despite these advantages, achieving high spatial resolution in drug imaging is challenging. Importantly, many drugs and metabolites are rarely detectable by conventional vacuum MALDI-MSI because of their poor ionization efficiency. It has been reported that acetaminophen (APAP) and one of its major metabolites, APAP-Cysteine (APAP-CYS), cannot be detected by vacuum MALDI-MSI without derivatization. In this context, we showed the distribution of both APAP and APAP-CYS in kidneys at high spatial resolution (25 and 10 µm) by employing an atmospheric pressure-MALDI imaging mass microscope without derivatization. APAP was highly accumulated in the renal pelvis 1 h after drug administration, while APAP-CYS exhibited characteristic distributions in the outer medulla and renal pelvis at both 30 min and 1 h after administration. Interestingly, cluster-like distributions of APAP and APAP-CYS were observed in the renal pelvis at 10 µm spatial resolution. Additionally, a novel APAP metabolite, tentatively coined as APAP-butyl sulfate (APAP-BS), was identified in the kidney, brain, and liver by combining MSI and tandem MSI. For the first time, our study revealed differential distributions of APAP, APAP-CYS (in kidneys), and APAP-BS (in kidney, brain, and liver) and is believed to enhance the understanding of the pharmacokinetics and potential nephrotoxicity of this drug.

Distribution, characterization and contamination risk assessment of microplastics in the sediment from the world's top sediment-laden estuary.

Microplastics (MPs) have gained a serious attention as an emerging contaminant throughout the world because of their persistence and possible risks to aquatic ecosystems and human well-being. However, knowledge on MPs contamination from sub-tropical coastal systems is limited, and no study has been conducted on the MPs contamination in sediment from one of the highest sediment-laden estuaries, Meghna River, in the world. This is the first study to examine the quantity, morpho-chemical characteristics and contamination risk level of MPs from this large scale river. MPs were extracted from the sediment samples of 10 stations along the banks of the estuary by density separation, and then characterized using a stereomicroscope and Fourier Transform Infrared (FTIR) spectroscopy. The incidence of MPs varied from 12.5 to 55 item/kg dry sediment with an average of 28.67 ± 10.80 item/kg. The majority (78.5%) of the MPs were under 0.5 mm in size, with fibers being the most (74.1%) prevalent MPs type. Polypropylene (PP) was found to be the predominant polymer (53.4%), followed by polyethylene (PE, 20%), polystyrene (PS, 13.3%), and polyvinyl chloride (PVC, 13.3%). The highest occurrence of PP indicted the MPs in the estuary might be originated from clothing and dying industries, fishing nets, food packages, and pulp industries. The sampling stations were contaminated with MPs as shown by the contamination factor (CF) values and pollutant load index (PLI), both of which were >1. This study exposed new insights on the status of MPs in the sediments of the Meghna River, laying the groundwork for future research. The findings will contribute to estimate the global share of MPs to the marine environment.

Microplastics in surface water from a mighty subtropical estuary: First observations on occurrence, characterization, and contamination assessment.

Estuarine contamination by Microplastics (MPs) is a mater of serious concern since these areas offer the society valuable ecosystem, economic, and recreational services such as breeding and feeding ground for fish, carbon fixation, nutrients recycling and port development. The Meghna estuary, located along the Bengal delta coast, provides livelihoods for thousands of peoples in Bangladesh, and served as breeding ground for national fish, Hilsha shad. Therefore, knowledge and understanding on any kind of pollution including MPs of this estuary is essential. In this study, the abundance, characteristics and contamination assessment of MPs from the surface water of a Meghna estuary were investigated for the first time. The results demonstrated that MPs were present in all samples and the abundance ranged from 33.33 to 316.67 item/m3 with a mean value of 128.89 ± 67.94 item/m3. Morphological analyses resulted in four types of MPs such as fibers (87%), fragments (6%), foam (4%), and films (3%) with the majority of these being colored (62%) and smaller (<0.5 mm) in size (88%). On the other hand, FTIR analysis for chemical characteristics confirmed five types of polymers, including polythene (PE), polystyrene (PS), polythene terephthalate (PET), polypropylene (PP), and polyvinyl chloride (PVC). The area was determined to be moderately to severely contaminated with MPs based on contamination factor (CF) values (6.18 ± 2.08 to 2.50 ± 1.0) and the pollutant load index (PLI) value (1.94 ± 0.33) as these values were > 3-6 for CF, and >1 for PLI. These results can be utilized to develop policy for the protection of this important environment.

Endocannabinoid 2-Arachidonoylglycerol Levels in the Anterior Cingulate Cortex, Caudate Putamen, Nucleus Accumbens, and Piriform Cortex Were Upregulated by Chronic Restraint Stress.

Endocannabinoid 2-arachidonoylglycerol (2-AG) has been implicated in habituation to stress, and its augmentation reduces stress-induced anxiety-like behavior. Chronic restraint stress (CRS) changes the 2-AG levels in some gross brain areas, such as the forebrain. However, the detailed spatial distribution of 2-AG and its changes by CRS in stress processing-related anatomical structures such as the anterior cingulate cortex (ACC), caudate putamen (CP), nucleus accumbens (NAc), and piriform cortex (PIR) are still unclear. In this study, mice were restrained for 30 min in a 50 mL-centrifuge tube for eight consecutive days, followed by imaging of the coronal brain sections of control and stressed mice using desorption electrospray ionization mass spectrometry imaging (DESI-MSI). The results showed that from the forebrain to the cerebellum, 2-AG levels were highest in the hypothalamus and lowest in the hippocampal region. 2-AG levels were significantly (p < 0.05) upregulated and 2-AG precursors levels were significantly (p < 0.05) downregulated in the ACC, CP, NAc, and PIR of stressed mice compared with control mice. This study provided direct evidence of 2-AG expression and changes, suggesting that 2-AG levels are increased in the ACC CP, NAc, and PIR when individuals are under chronic stress.

Influence of Y3+ and La3+ ions on the structural, magnetic, electrical, and optical properties of cobalt ferrite nanoparticles.

In the current study, nanocrystalline CoY0.5xLa0.5xFe2-xO4 (where x = 0.00, 0.02, 0.04, 0.06, 0.08, and 0.10) ferrites have been synthesized via a sol-gel auto combustion process. The synthesized powders were pressed into pellet forms and sintered at 900 °C for 4 h in the air. X-ray diffractometry (XRD) confirmed the single-phase cubic spinel structure of the synthesized samples having the mean crystallite domain sizes ranging from 122 and 54 nm. FTIR spectroscopic analyses revealed two strong bands within the range of 600 to 350 cm-1, further confirming the cubic inverse spinel structure of the prepared materials. The surface morphologies and composition were investigated by Field Emission Scanning Electron Microscopy (FE-SEM) and Energy Dispersive X-ray (EDX) Spectroscopy. The magnetic hysteresis curves recorded at room temperature exhibit ferrimagnetic behavior. The highest coercivity (Hc∼1276 Oe) was found at a high doping (x = 0.10) concentration of Y3+ and La3+ in cobalt ferrite. Dielectric constant increase with increased doping concentration whereas real-impedance and dielectric loss decrease with increased in doping concentration and applied frequency. The band gap energy increased from 1.48 to 1.53 eV with increasing Y3+ and La3+concentrations in the UV-Vis region. The elevated levels of magnetic and dielectric substances in the ferrite nanoparticles suggest that the material could be used for magnetic recording media and high-frequency devices.

Trend and gender-based association of the Bangladeshi student suicide during the COVID-19 pandemic: a GIS-based nationwide distribution.

BACKGROUND: Students are one of the most vulnerable groups to suicide. Before the COVID-19 pandemic, a Bangladeshi study was conducted assessing their suicide patterns regarding gender-based associations. But how has the pandemic changed the Bangladeshi students' suicide patterns is not studied yet, which is investigated herein. Besides, for the first time, this study provides GIS-based distribution of suicide cases across the country's administrative district. METHODS: As Bangladesh has no suicide surveillance system, this study utilized media reporting suicide cases following the prior studies. A total of 127 students' suicide cases from March 2020 to March 2021 were finally analyzed after eliminating the duplicate ones, and data were synthesized following the prior studies. Arc-GIS was also used to distribute the suicide cases across the administrative district. RESULTS: Results revealed that female (72.4%; n = 92/127) was more prone to die by suicide than males. About 42.5% of the cases were aged between 14 and 18 years (mean age 16.44 ± 3.512 years). The most common method of suicide was hanging (79.5%; n = 101), whereas relationship complexities (15.7%), being emotional (12.6%), not getting the desired one (11%), conflict with a family member (9.4%), academic failure (9.4%), mental health problem (8.7%), sexual complexities (6.3%), scolded or forbidden by parents (3.9%) were the prominent suicide causalities. In respect to gender and suicide patterns, only the suicide stressor was significantly distributed, whereas the method of suicide was significantly associated with GIS-based distribution. However, a higher number of suicide cases was documented in the capital (i.e. Dhaka) and the northern region than in its surrounding districts. CONCLUSIONS: The findings reported herein are assumed to be helpful to identify the gender-based suicide patterns and suicide-prone regions in the time of the COVID-19 pandemic to initiate suicide prevention programs of the risky students.

Dimethylthiourea Alleviates Drought Stress by Suppressing Hydrogen Peroxide-Dependent Abscisic Acid-Mediated Oxidative Responses in an Antagonistic Interaction with Salicylic Acid in Brassica napus Leaves.

In plants, prolonged drought induces oxidative stress, leading to a loss of reducing potential in redox components. Abscisic acid (ABA) is a representative hormonal signal regulating stress responses. This study aimed to investigate the physiological significance of dimethylthiourea (DMTU, an H2O2 scavenger) in the hormonal regulation of the antioxidant system and redox control in rapeseed (Brassica napus L.) leaves under drought stress. Drought treatment for 10 days provoked oxidative stress, as evidenced by the increase in O2•- and H2O2 concentrations, and lipid peroxidation levels, and a decrease in leaf water potential. Drought-induced oxidative responses were significantly alleviated by DMTU treatment. The accumulation of O2•- and H2O2 in drought-treated plants coincided with the enhanced expression of the NADPH oxidase and Cu/Zn-SOD genes, leading to an up-regulation in oxidative signal-inducible 1 (OXI1) and mitogen-activated protein kinase 6 (MAPK6), with a concomitant increase in ABA levels and the up-regulation of ABA-related genes. DMTU treatment under drought largely suppressed the drought-responsive up-regulation of these genes by depressing ABA responses through an antagonistic interaction with salicylic acid (SA). DMTU treatment also alleviated the drought-induced loss of reducing potential in GSH- and NADPH-based redox by the enhanced expression of glutathione reductase 1 (GR1) and up-regulation of oxidoreductase genes (TRXh5 and GRXC9). These results indicate that DMTU effectively alleviates drought-induced oxidative responses by suppressing ABA-mediated oxidative burst signaling in an antagonistic regulation of SA.

Application of AP-MALDI Imaging Mass Microscope for the Rapid Mapping of Imipramine, Chloroquine, and Their Metabolites in the Kidney and Brain of Wild-Type Mice.

Mass spectrometry imaging (MSI) is well-known for the non-labeling visualization of analytes, including drugs and their metabolites in biological samples. In this study, we applied three different tools of MSI, desorption electrospray ionization (DESI)-MSI, matrix-assisted laser desorption ionization (MALDI)-MSI, and a newly developed atmospheric pressure (AP)-MALDI-MSI known as iMScopeTM QT for rapid mapping of imipramine, chloroquine, and their metabolites in C57BL/6 male wild-type mice. Among three MSI tools, better detection capability for targeted drugs at higher speed (up to 32 pixels/s) was observed in iMScope QT. It revealed that imipramine and its metabolites were significantly accumulated in the renal cortex of mice, but chloroquine and its metabolites were highly accumulated in the renal pelvis and renal medulla of mice. Additionally, a higher accumulation of imipramine was noted in the thalamus, hypothalamus, septum, and hindbrain of mice brains. However, chloroquine and its metabolites showed notable accumulation in the lateral ventricle, fourth ventricle, and fornix of the mice brains. These findings of our study can be helpful in understanding clinically relevant properties, efficacy, and potential side effects of these drugs. Our study also showed the potentiality of iMScope QT for rapid mapping of small drugs and their metabolites in biological samples.

Coenzyme Q10 in the eye isomerizes by sunlight irradiation.

Photoisomerization of lipids has been well studied. As for the eyes, photoisomerization from 11-cis isomer to all-trans-retinal is well-known as the first step of the visual transduction in the photoreceptors. In addition to that, there would be other ocular lipids that undergo photoisomerization, which may be involved in ocular health and function. To explore any photoisomerizable lipids in the eyes, the nonirradiated and sunlight-irradiated eyeball extracts were subjected to liquid chromatography-mass spectrometry analysis, followed by the identification of the decreased lipid species in the irradiated extracts. Surprisingly, more than nine hundred lipid species were decreased in the irradiated extracts. Three lipid species, coenzyme Q10 (CoQ10), triglyceride(58:4), and coenzyme Q9, were decreased both significantly (p < 0.05) and by more than two-fold, where CoQ10 showed the most significant decrease. Later, photoisomerization was identified as the prominent cause underlying the decrease of CoQ10. Interestingly, CoQ10 in the sunlight-irradiated fresh eyeballs was also isomerized. Both the visible light and ultraviolet radiation were capable of producing CoQ10 isomer, while the latter showed rapid action. This study is believed to enhance our understanding of the biochemistry and photodamage of the eye and can potentially contribute to the advancement of opto-lipidomics.

A convenient online desalination tube coupled with mass spectrometry for the direct detection of iodinated contrast media in untreated human spent hemodialysates.

BACKGROUND: Mass spectrometry (MS) analysis using direct infusion of biological fluids is often problematic due to high salts/buffers. Iodinated contrast media (ICM) are frequently used for diagnostic imaging purposes, sometimes inducing acute kidney injury (AKI) in patients with reduced kidney function. Therefore, detection of ICM in spent hemodialysates is important for AKI patients who require urgent continuous hemodiafiltration (CHDF) because it allows noninvasive assessment of the patient's treatment. In this study, we used a novel desalination tube before MS to inject the sample directly and detect ICM. METHODS: Firstly, spent hemodialysates of one patient were injected directly into the electrospray ionization (ESI) source equipped with a quadrupole time-of-flight mass spectrometer (Q-TOF MS) coupled to an online desalination tube for the detection of ICM and other metabolites. Thereafter, spent hemodialysates of two patients were injected directly into the ESI source equipped with a triple quadrupole mass spectrometer (TQ-MS) connected to that online desalination tube to confirm the detection of ICM. RESULTS: We detected iohexol (an ICM) from untreated spent hemodialysates of the patient-administered iohexol for computed tomography using Q-TOF MS. Using MRM profile analysis, we have confirmed the detection of ICM in the untreated spent hemodialysates of the patients administered for coronary angiography before starting CHDF. Using the desalination tube, we observed approximately 178 times higher signal intensity and 8 times improved signal-to-noise ratio for ioversol (an ICM) compared to data obtained without the desalination tube. This system was capable of tracking the changes of ioversol in spent hemodialysates of AKI patients by measuring spent hemodialysates. CONCLUSION: The online desalination tube coupled with MS showed the capability of detecting iohexol and ioversol in spent hemodialysates without additional sample preparation or chromatographic separation. This approach also demonstrated the capacity to monitor the ioversol changes in patients' spent hemodialysates.

iACP-MultiCNN: Multi-channel CNN based anticancer peptides identification.

Cancer is one of the most dangerous diseases in the world that often leads to misery and death. Current treatments include different kinds of anticancer therapy which exhibit different types of side effects. Because of certain physicochemical properties, anticancer peptides (ACPs) have opened a new path of treatments for this deadly disease. That is why a well-performed methodology for identifying novel anticancer peptides has great importance in the fight against cancer. In addition to the laboratory techniques, various machine learning and deep learning methodologies have developed in recent years for this task. Although these models have shown reasonable predictive ability, there's still room for improvement in terms of performance and exploring new types of algorithms. In this work, we have proposed a novel multi-channel convolutional neural network (CNN) for identifying anticancer peptides from protein sequences. We have collected data from the existing state-of-the-art methodologies and applied binary encoding for data preprocessing. We have also employed k-fold cross-validation to train our models on benchmark datasets and compared our models' performance on the independent datasets. The comparison has indicated our models' superiority on various evaluation metrics. We think our work can be a valuable asset in finding novel anticancer peptides. We have provided a user-friendly web server for academic purposes and it is publicly available at: http://103.99.176.239/iacp-cnn/.

H2O2-Responsive Hormonal Status Involves Oxidative Burst Signaling and Proline Metabolism in Rapeseed Leaves.

Drought alters the level of endogenous reactive oxygen species (ROS) and hormonal status, which are both involved in the regulation of stress responses. To investigate the interplay between ROS and hormones in proline metabolism, rapeseed (Brassica napus L.) plants were exposed to drought or exogenous H2O2 (Exo-H2O2) treatment for 10 days. During the first 5 days, the enhanced H2O2 concentrations in drought treatment were associated with the activation of superoxide dismutase (SOD) and NADPH oxidase, with enhanced ABA and SA levels, while that in Exo-H2O2 treatment was mainly associated with SA-responsive POX. During the latter 5 days, ABA-dependent ROS accumulation was predominant with an upregulated oxidative signal-inducible gene (OXI1) and MAPK6, leading to the activation of ABA synthesis and the signaling genes (NCED3 and MYC2). During the first 5 days, the enhanced levels of P5C and proline were concomitant with SA-dependent NDR1-mediated signaling in both drought and Exo-H2O2 treatments. In the latter 5 days of drought treatment, a distinct enhancement in P5CR and ProDH expression led to higher proline accumulation compared to Exo-H2O2 treatment. These results indicate that SA-mediated P5C synthesis is highly activated under lower endogenous H2O2 levels, and ABA-mediated OXI1-dependent proline accumulation mainly occurs with an increasing ROS level, leading to ProDH activation as a hypersensitive response to ROS and proline overproduction under severe stress.

NAD+ Levels Are Augmented in Aortic Tissue of ApoE-/- Mice by Dietary Omega-3 Fatty Acids.

BACKGROUND: Maintaining bioenergetic homeostasis provides a means to reduce the risk of cardiovascular events during chronological aging. Nicotinamide adenine dinucleotide (NAD+) acts as a signaling molecule, and its levels were used to govern several biological pathways, for example, promoting angiogenesis by SIRT1 (sirtuin 1)-mediated inhibition of Notch signaling to rejuvenate capillary density of old-aged mice. NAD+ modulation shows promise in the vascular remodeling of endothelial cells. However, NAD+ distribution in atherosclerotic regions remains uncharacterized. Omega-3 polyunsaturated fatty acids consumption, such as docosahexaenoic acid and eicosapentaenoic acid, might increase the abundance of cofactors in blood vessels due to omega-3 polyunsaturated fatty acids metabolism. METHODS: Apolipoprotein E-deficient (ApoE-/-) mice were fed a Western diet, and the omega-3 polyunsaturated fatty acids-treated groups were supplemented with docosahexaenoic acid (1%, w/w) or eicosapentaenoic acid (1%, w/w) for 3 weeks. Desorption electrospray ionization mass spectrometry imaging was exploited to detect exogenous and endogenous NAD+ imaging. RESULTS: NAD+, NADH, NADP+, NADPH, FAD+, FADH, and nicotinic acid adenine dinucleotide of the aortic arches were detected higher in the omega-3 polyunsaturated fatty acids-treated mice than the nontreated control. Comparing the distribution in the outer and inner layers of the arterial walls, only NADPH was detected slightly higher in the outer part in eicosapentaenoic acid-treated mice. CONCLUSIONS: Supplementation of adding docosahexaenoic acid or eicosapentaenoic acid to the Western diet led to a higher NAD+, FAD+, and their metabolites in the aortic arch. Considering the pleiotropic roles of NAD+ in biology, this result serves as a beneficial therapeutic strategy in the animal model counter to pathological conditions.

Imaging and Manipulation of Plasma Membrane Fatty Acid Clusters Using TOF-SIMS Combined Optogenetics.

The plasma membrane (PM) serves multiple functions to support cell activities with its heterogeneous molecular distribution. Fatty acids (FAs) are hydrophobic components of the PM whose saturation and length determine the membrane's physical properties. The FA distribution contributes to the PM's lateral heterogeneity. However, the distribution of PM FAs is poorly understood. Here, we proposed the FA cluster hypothesis, which suggested that FAs on the PM exist as clusters. By the optogenetic tool translocating the endoplasmic reticulum (ER), we were able to manipulate the distribution of PM FAs. We used time-of-flight combined secondary ion mass spectrometry (TOF-SIMS) to image PM FAs and discovered that PM FAs were presented and distributed as clusters and are also manipulated as clusters. We also found the existence of multi-FA clusters formed by the colocalization of more than one FA. Our optogenetic tool also decreased the clustering degree of FA clusters and the formation probability of multi-FA clusters. This research opens up new avenues and perspectives to study PM heterogeneity from an FA perspective. This research also suggests a possible treatment for diseases caused by PM lipid aggregation and furnished a convenient tool for therapeutic development.

Mass Spectrometry Imaging for Glycome in the Brain.

Glycans are diverse structured biomolecules that play crucial roles in various biological processes. Glycosylation, an enzymatic system through which various glycans are bound to proteins and lipids, is the most common and functionally crucial post-translational modification process. It is known to be associated with brain development, signal transduction, molecular trafficking, neurodegenerative disorders, psychopathologies, and brain cancers. Glycans in glycoproteins and glycolipids expressed in brain cells are involved in neuronal development, biological processes, and central nervous system maintenance. The composition and expression of glycans are known to change during those physiological processes. Therefore, imaging of glycans and the glycoconjugates in the brain regions has become a "hot" topic nowadays. Imaging techniques using lectins, antibodies, and chemical reporters are traditionally used for glycan detection. However, those techniques offer limited glycome detection. Mass spectrometry imaging (MSI) is an evolving field that combines mass spectrometry with histology allowing spatial and label-free visualization of molecules in the brain. In the last decades, several studies have employed MSI for glycome imaging in brain tissues. The current state of MSI uses on-tissue enzymatic digestion or chemical reaction to facilitate successful glycome imaging. Here, we reviewed the available literature that applied MSI techniques for glycome visualization and characterization in the brain. We also described the general methodologies for glycome MSI and discussed its potential use in the three-dimensional MSI in the brain.