Comprehensive insights into fluorescent probes for the determination nitric oxide for diseases diagnosis.

Nitric oxide (NO) plays an important role in multiple physiological processes of the body involved in regulation, such as cardiovascular relaxation, neural homeostasis, and immune regulation, etc. The real-time monitoring of NO is of great significance in the investigation of related disease mechanisms and the evaluation of pharmacodynamics. Fluorescent probes are considered as a highly promising approach for pharmaceutical analysis and bioimaging due to their non-invasive character, real-time detection, and high sensitivity. However, there are still some challenges in the determination of biological nitric oxide with fluorescent probes, such as low anti-interference ability, poor function modifiability, and low organ specificity. Therefore, it would be beneficial to develop a new generation of fluorescent probes for real-time bioimaging of NO in vivo after this systematic summary.

Practical NIR Assay Derived from Cyanine to Evaluate Intracellular H2S in Living Cell Imaging.

To monitor the biological function of H2S in real time, this investigation demonstrated the design and synthesis of a novel fluorescent probe integrated with cyanine and 2,4-dinitrophenol for the qualitative and quantitative detection of H2S. An NIR sensitive sensor (FS-HS-1) was provided with a straightforward process. Spectroscopy experiments elucidated that FS-HS-1 could selectively detect H2S in a PBS solution (containing 40% acetonitrile) with a 111-fold fluorescence enhancement at 715 nm (ex. 605 nm). The response towards NaHS occurred in less than 2 min, and the detection limit was confirmed to be as low as 4.47 ± 0.11 nmol/L. Furthermore, the probe is capable of monitoring changes in exogenous H2S concentrations within living cells with confocal and 2P imaging.

Recent Progress on Fluorescent Probes in Heavy Metal Determinations for Food Safety: A Review.

One of the main challenges faced in food safety is the accumulation of toxic heavy metals from environmental sources, which can sequentially endanger human health when they are consumed. It is invaluable to establish a practical assay for the determination of heavy metals for food safety. Among the current detection methods, technology based on fluorescent probes, with the advantages of sensitivity, convenience, accuracy, cost, and reliability, has recently shown pluralistic applications in the food industry, which is significant to ensure food safety. Hence, this review systematically presents the recent progress on novel fluorescent probes in determining heavy metals for food safety over the past five years, according to fluorophores and newly emerging sensing cores, which could contribute to broadening the prospects of fluorescent materials and establishing more practical assays for heavy metal determinations.

Molecular Characterization of a New Tetrodotoxin-Binding Protein, Peroxiredoxin-1, from Takifugu bimaculatus.

Pufferfish are considered a culinary delicacy but require careful preparation to avoid ingestion of the highly toxic tetrodotoxin (TTX), which accumulates in certain tissues. In this study, the tissue distribution of peroxiredoxin-1 from Takifugu bimaculatus was investigated. The peroxiredoxin-1 protein was obtained by in vitro recombinant expression and purification. The recombinant protein had a strong ability to scavenge hydroxyl radicals, protect superhelical DNA plasmids from oxidative damage, and protect L929 cells from H2O2 toxicity through in vitro antioxidant activity. In addition, we verified its ability to bind to tetrodotoxin using surface plasmon resonance techniques. Further, recombinant proteins were found to facilitate the entry of tetrodotoxin into cells. Through these analyses, we identified, for the first time, peroxiredoxin-1 protein from Takifugu bimaculatus as a potential novel tetrodotoxin-binding protein. Our findings provide a basis for further exploration of the application of peroxiredoxin-1 protein and the molecular mechanisms of tetrodotoxin enrichment in pufferfish.

Molecular Mechanism of Anti-Inflammatory Activities of a Novel Sulfated Galactofucan from Saccharina japonica.

Seaweed of Saccharina japonica is the most abundantly cultured brown seaweed in the world, and has been consumed in the food industry due to its nutrition and the unique properties of its polysaccharides. In this study, fucoidan (LJNF3), purified from S. japonica, was found to be a novel sulfated galactofucan, with the monosaccharide of only fucose and galactose in a ratio of 79.22:20.78, and with an 11.36% content of sulfate groups. NMR spectroscopy showed that LJNF3 consists of (1→3)-α-l-fucopyranosyl-4-SO3 residues and (1→6)-ß-d-galactopyranose units. The molecular mechanism of the anti-inflammatory effect in RAW264.7 demonstrated that LJNF3 reduced the production of nitric oxide (NO), and down-regulated the expression of MAPK (including p38, ENK and JNK) and NF-κB (including p65 and IKKα/IKKß) signaling pathways. In a zebrafish experiment assay, LJNF3 showed a significantly protective effect, by reducing the cell death rate, inhibiting NO to 59.43%, and decreasing about 40% of reactive oxygen species. This study indicated that LJNF3, which only consisted of fucose and galactose, had the potential to be developed in the biomedical, food and cosmetic industries.

Insights into salvianolic acid B biosynthesis from chromosome-scale assembly of the Salvia bowleyana genome.

Salvia bowleyana is a traditional Chinese medicinal plant that is a source of nutritional supplements rich in salvianolic acid B and a potential experimental system for the exploration of salvianolic acid B biosynthesis in the Labiatae. Here, we report a high-quality chromosome-scale genome assembly of S. bowleyana covering 462.44 Mb, with a scaffold N50 value of 57.96 Mb and 44,044 annotated protein-coding genes. Evolutionary analysis revealed an estimated divergence time between S. bowleyana and its close relative S. miltiorrhiza of ~3.94 million years. We also observed evidence of a whole-genome duplication in the S. bowleyana genome. Transcriptome analysis showed that SbPAL1 (PHENYLALANINE AMMONIA-LYASE1) is highly expressed in roots relative to stem and leaves, paralleling the location of salvianolic acid B accumulation. The laccase gene family in S. bowleyana outnumbered their counterparts in both S. miltiorrhiza and Arabidopsis thaliana, suggesting that the gene family has undergone expansion in S. bowleyana. Several laccase genes were also highly expressed in roots, where their encoded proteins may catalyze the oxidative reaction from rosmarinic acid to salvianolic acid B. These findings provide an invaluable genomic resource for understanding salvianolic acid B biosynthesis and its regulation, and will be useful for exploring the evolution of the Labiatae.

Silver nanocube coupling with a nanoporous silver film for dual-molecule recognition based ultrasensitive SERS detection of dopamine.

Dopamine (DA) is one of the catecholamine neurotransmitters used for the treatment of neural disorders. In this study, a novel sensor based on surface-enhanced Raman scattering (SERS) with dual molecule-recognition for ultrasensitive detection of DA was presented, with a limit of detection (LOD) of 40 fM, without any pretreatment of clinical samples. To realize the sensitive and selective detection of DA in complex samples, the nanoporous silver film (AgNF) surfaces were functionalized with mercaptopropionic acid (MPA) to accurately capture DA, while silver nanocubes (AgNCs) were modified with 4-mercaptobenzene boronic acid (4-MPBA) as a Raman reporter for the quantitative detection of DA. The nanogaps between AgNCs and the AgNF led to the generation of an abundance of hot spots for the SERS signal and thus effectively improved the sensitivity of DA detection. Measurements of DA concentrations in clinical body fluids such as human serum and urine samples are also demonstrated, showing excellent performance for DA detection in a complex environment. Our results demonstrate the promising potential for the ultrasensitive detection of DA for the potential diagnosis of DA-related diseases.

Overexpression of acetyl-CoA carboxylase increases fatty acid production in the green alga Chlamydomonas reinhardtii.

Chlamydomonas reinhardtii is a photosynthetic unicellular model algae with multiple biotechnological advantages, and its fatty acids can be used to produce biofuels. Numerous studies suggest that acetyl-coA carboxylase (ACCa) catalyzes the first committed and rate-limiting step of fatty acid biosynthesis, thereby playing a central role in oil accumulation. Here, we cloned and overexpressed ACCa in C. reinhardtii to directly evaluate its effect on fatty acid synthesis. GC-MS analysis found that the unsaturated FAs contents of the CW15-24 and CW15-85 strains were 55.45% and 56.15%, which were significantly enriched compared to the wild type CW15 (48.39%). Under the optimized conditions, the content of lipid by overexpressed the ACCa gene in the mutant CW15-85 (0.46 g/l) was 1.16-fold greater than control through optimization of N and P sources. Altogether, our data clearly demonstrate that ACCa overexpression in C. reinhardtii can directly increase the synthesis of fatty acids.

Improved bioethanol production using CRISPR/Cas9 to disrupt the ADH2 gene in Saccharomyces cerevisiae.

Bioethanol, as a form of renewable and clean energy, has become increasingly important to the energy supply. One major obstacle in ethanol production is developing a high-capacity system. Existing approaches for regulating the ethanol production pathway are relatively insufficient, with nonspecific genetic manipulation. Here, we used CRISPR/Cas9 technology to disrupt the alcohol dehydrogenase (ADH) 2 gene via complete deletion of the gene and introduction of a frameshift mutation in the ADH2 locus. Sequencing demonstrated the accurate knockout of the target gene with 91.4% and near 100% targeting efficiency. We also utilized genome resequencing to validate the mutations in the ADH2 mutants targeted by various single-guide RNAs. This extensive analysis indicated the mutations in the CRISPR/Cas9-engineered strains were homozygous. We applied the engineered Saccharomyces cerevisiae strains for bioethanol production. Results showed that the ethanol yield improved by up to 74.7% compared with the yield obtained using the native strain. This work illustrates the applicability of this highly efficient and specific genome engineering approach to promote the improvement of bioethanol production in S. cerevisiae via metabolic engineering. Importantly, this study is the first report of the disruption of a target gene, ADH2, in S. cerevisiae using CRISPR/Cas9 technology to improve bioethanol yield.

Two new furaldehyde compounds from the rhizomes of Gastrodia elata.

Two new compounds 5-[4'-(4″-hydroxybenzyl)-3'-hydroxybenzyloxymethyl]-furan-2-carbaldehyde (1) and 5-[4'-(4″-hydroxybenzyl)-3'-hydroxybenzyl]-furan-2-carbal-dehyde (2), together with two known 5-(4-hydroxbenzyloxymethyl)-furan-2-carbaldehyde] (3) and 5-(hydroxymethyl)-2-furaldehyde (4), were isolated from the rhizome of Gastrodia elata. Their structures were elucidated by spectroscopic analysis and comparison of their spectral data with those reported previously. All compounds exhibited weak or no cytotoxicity against human colon carcinoma cell line (HT-29) and human chronic myelogenous leukemia cell line (K-562).

Accurate and rapid mercury susceptibility detection in aquatic samples using fluorescent probe integrated rhodamine with pyridyl isothiocyanate.

Mercury, one of the various harmful metals, is particularly significant in affecting aquatic organisms, currently gaining more attentions and sparking discussions. In response to the limitations of traditional detections, fluorescent probes have emerged as a promising solution with some advantages, such as weaker background interference, shorter processing time, higher accuracy. Thus, a novel fluorescent probe, FS-Hg-1, has been developed for assessing mercury ion (Hg2+) concentrations in aquatic products. This probe displays specific recognition of mercury ions in fluorescence spectra. Notably, FS-Hg-1 exhibits a distinct color change to pink when combined with Hg2+ (with a 948-fold increase in absorption at 568 nm) and a substantial fluorescence change towards Hg2+ (361-fold increase, excitation at 562 nm, emission at 594 nm) in N, N-dimethylformamide. The probe boasts a detection limit of 0.14 µM and rapid reaction with Hg2+ within 10 s, showing an excellent linear correlation with [Hg2+] in the range of 0 to 10 µM. Through thorough analysis using FS-Hg-1, the results align with those from the standard method (P > 0.05), with spiked recovery rates ranging from 108.4% to 113.2%. With its precise recognition, low detection limit, and remarkable sensitivity, this fluorescent assay proves effective in mercury concentration determination in aquatic samples without interference. The potential of FS-Hg-1 is promising for speedy detection of residual Hg2+ and holds significance in ensuring food safety.

Amifostine acts upon mitochondria to stimulate growth of bone marrow and regulate cytokines.

Amifostine is a first-line cytoprotective drug used to prevent radiotherapy-induced or chemotherapy-induced injuries. However, its mechanism of action is not well understood. In this study, freshly harvested bone marrow cells were treated with amifostine and analyzed with a series of mitochondrial indices. In vitro results showed that bone marrow cells treated with amifostine 0.5 h before irradiation (0.5 Gy) experienced several benefits, as compared to vehicle controls, including (1) reduced reactive oxygen species levels, which reduced the production of free radicals; (2) better preservation of mitochondria, as indicated by MitoTracker-positive staining and the increased intensity of staining; (3) reduced apoptosis, as demonstrated by Annexin V staining; and (4) a better proliferation rate, as illustrated by MTT assay. Our in vitro studies showed that amifostine-treated mice exhibited (1) higher ATP production; (2) reduced plasma IL-2 levels, suppressing the immune response triggered by radiotoxicity; and (3) enhanced radiation-induced production of granulocyte colony-stimulating factor. All of these processes benefit recovery from radiation-induced damage.

Interleukin 11 protects bone marrow mitochondria from radiation damage.

Interleukin 11 (IL-11) is a multifunctional cytokine isolated from bone marrow (BM)-derived stromal cells that promotes hematopoiesis and prolongs the life span of lethally irradiated animals. However, the underlying mechanism for the protective effect of IL-11 on BM is unclear. In this study, we explored the effect of IL-11 on irradiated BM cells. Freshly harvested BM cells were pretreated with 20 ng/ml of recombinant IL-11 for 30 min, irradiated with a dose of 0.5 Gy, cultured for 24 h, and then subjected to several assays. In vitro data showed that, as compared to the vehicle controls, IL-11: (1) reduced the production of reactive oxygen species; (2) reduced the alteration of mitochondrial membrane potential; (3) increased MitoTracker staining, suggesting that the number of mitochondria and their functions were better maintained; and (4) reduced apoptosis of BM cells and enhanced BM cell proliferation. In vivo studies of mice pretreated with saline or 100 µg/kg of IL-11 at 12 and 2 h before 10-Gy total body irradiation (TBI) demonstrated that G-CSF and IL-6 were significantly upregulated, whereas IL-2 and IL-4 were reduced. We found that IL-11 protects mitochondrial functions, acts with G-CSF and IL-6 to stimulate the growth of radiation-damaged BM, and reduces the immune response to radiation injury.

Radiation affects the responsiveness of bone marrow to G-CSF.

In this study, we investigated the response of irradiated bone marrow cells to granulocyte colony-stimulating factor (G-CSF). Freshly harvested bone marrow cells were treated with either saline (vehicle control) or 20 ng/ml of G-CSF. Thereafter, cells were separated into nonirradiated (no-IR) and irradiated (IR, 0.5 Gy) groups. IR cells exhibited a higher proliferation rate in response to G-CSF, as compared to the no-IR cells. Reduced levels of reactive oxygen species indicated that G-CSF-treated IR cells produced fewer free radicals, as compared to the no-IR cells. The G-CSF-treated IR cells also had a lower apoptotic rate than their no-IR counterparts. Furthermore, G-CSF-treated IR cells exhibited less alteration of mitochondrial membrane potential, as compared to the no-IR cells. Finally, the mitochondrial number increased in the G-CSF-treated IR cells. The radiation-induced increase in plasma IL-6 in vivo could be enhanced by the administration of G-CSF. The data suggest that radiation potentiates the response of bone marrow cells to G-CSF treatment.

Comparative study of three cultivars of jaboticaba berry: nutrient, antioxidant and volatile compounds.

Jaboticaba is a tropical plant and its fruit rich in nutrients, volatile compounds, and biological activities, which considered to be an edible health benefits plant. Despite its popularity for fresh consumption, jaboticaba is rarely used in intensive processing in China. The content of nutrients and antioxidant in jaboticaba greatly impacts how it is processed healthy food. In this study, we evaluated the nutrients, antioxidant capacity, and volatile compounds of three jaboticaba cultivars including Sabara, Argentina, and Fukuoka, respectively. Our results revealed each variety has its merits. Sabara had an abundance of volatile compounds, a suitable acid-sugar ratio, and a slightly lower antioxidant capacity, making it suitable for fresh consumption. Argentina is the richest in volatile compounds in ripe fruit, but slightly lighter in taste and acid-sugar ratio, making it suitable for dry products. The large size, juicy flesh, low acid-sugar ratio, and less volatile compounds content of Fukuoka also make it suitable for juice processing. Three cultivars of jaboticaba berry exhibited different characteristics, providing reference evidence for the manufacturing and processing of jaboticaba health food.

FGF23 alleviates neuronal apoptosis and inflammation, and promotes locomotion recovery via activation of PI3K/AKT signalling in spinal cord injury.

Fibroblast growth factor 23 (FGF23) regulates neuronal morphology, synaptic growth and inflammation; however, its involvement in spinal cord injury (SCI) remains unclear. Therefore, the present study aimed to investigate the effect of FGF23 on neuronal apoptosis, inflammation and locomotion recovery, as well as its underlying mechanism in experimental SCI models. Primary rat neurons were stimulated with H2O2 to establish an in vitro model of SCI and were then transfected with an FGF23 overexpression (oeFGF23) or short hairpin RNA (shFGF23) adenovirus-associated virus and treated with or without LY294002 (a PI3K/AKT inhibitor). Subsequently, an SCI rat model was constructed, followed by treatment with oeFGF23, LY294002 or a combination of the two. FGF23 overexpression (oeFGF23 vs. oeNC) decreased the cell apoptotic rate and cleaved-caspase3 expression, but increased Bcl-2 expression in H2O2-stimulated neurons, whereas shFGF23 transfection (shFGF23 vs. shNC) exhibited the opposite effect (all P<0.05). Furthermore, FGF23 overexpression (oeFGF23 vs. oeNC) could activate the PI3K/AKT signalling pathway, whereas treatment with the PI3K/AKT inhibitor (LY294002) (oeFGF23 + LY294002 vs. LY294002) attenuated these effects in H2O2-stimulated neurons (all P<0.05). In SCI model rats, FGF23 overexpression (oeFGF23 vs. oeNC) reduced the laceration and inflammatory cell infiltration in injured tissue, decreased TNF-α and IL-1ß levels, and improved locomotion recovery (all P<0.05); these effects were attenuated by additional administration of LY294002 (oeFGF23 + LY294002 vs. LY294002) (all P<0.05). In conclusion, FGF23 alleviated neuronal apoptosis and inflammation, and promoted locomotion recovery via activation of the PI3K/AKT signalling pathway in SCI, indicating its potential as a treatment option for SCI; however, further studies are warranted for validation.

Antarctic krill (Euphausia superba) oil modulatory effects on ethanol-induced acute injury of the gastric mucosa in rats.

Antarctic krill oil (KO) prepared using supercritical carbon dioxide extraction and characterized using gas chromatography-mass spectrometry was used to investigate its preventive effect on ethanol-induced gastric tissue damage in a rat model in vivo. KO characterization showed that 74.96% of the unsaturated fatty acids consist of oleic acid, eicosapentaenoic acid (EPA), and docosahexaenoic acid (DHA). Rats pre-treated with KO (100, 200, and 500 mg/kg) showed mitigated oxidative stress through enhanced antioxidant enzyme superoxide dismutase (SOD) and reducing enzymes malondialdehyde (MDA) and myeloperoxidase (MPO) in gastric mucosal injury induced by ethanol. Additionally, the secretion of pro-inflammatory cytokines (TNF-α, IL-6, and IL-1ß), the expression of the IκBα/NF-κB signaling pathway, and nitric oxide (NO) production was suppressed. The results also demonstrated a significant decrease in histological injury and hemorrhage scores in a dose-dependent manner in the KO range. Therefore, KO has potential as a food supplement to alleviate ethanol-induced acute gastric mucosal injury.

Agar oligosaccharides: A review of preparation, structures, bioactivities and application.

Agar, a gelatinous polysaccharide which is in the cell wall of many red algae, is widely used as food and gelling agent. Agar oligosaccharides (AOs), the hydrolysate of agar, show much more kinds of bio-activities because of its lower molecular weight, better water solubility and higher absorption efficiency. It is indicated that AOs with different structure and degree of polymerization, i.e. series of agaro-oligosaccharides and neoagaro-oligosaccharides, can be obtained under different preparation conditions. In addition, the biological activities of AOs are diversely and closely correlated to the composition and structure. This review aims to comprehensively summarize the preparation, structural characteristics and bio-activities of AOs, so as to provide a reference for applications of AOs as marine natural products in pharmacological, cosmetics and nutraceutical fields.

Lipid and carotenoid production by the Rhodosporidium toruloides mutant in cane molasses.

Cane molasses is beneficial for lipid and carotenoid production in microalgae. We made a survey for the lipid and carotenoid production profile of R. toruloides M18 (MT) with various concentrations of molasses under nitrogen-deficited conditions. The production of α-linolenate and torularhodin from MT were 1.22- and 14.68-fold higher than those of the wild-type strain. We observed that molasses at concentrations of 35 g/L and 70 g/L represented a cheap and environmentally friendly strategy for producing lipids and carotenoids. Transcriptome and WGCNA analysis demonstrated that the genes relevant to the lipid and carotenoid production, including MYB, bHLH, Δ-4 desaturase, Δ-12 desaturase and FA2H, were significantly highly expressed. The results indicated that molasses could represent an inexpensive means for achieving high lipids and carotenoids production in R. toruloides.

Fabrication of Fe3O4/Au@ATP@Ag Nanorod sandwich structure for sensitive SERS quantitative detection of histamine.

We have successfully prepared a highly sensitive sandwich nanosensor combined Fe3O4 and Au@ATP@Ag nanorods for histamine detection based on surface-enhanced Raman spectroscopy (SERS). The Fe3O4 beads with -COOH served as a capture part to enrich histamine. The Au@ATP@Ag core-shell nanorods functionalized with Nalpha,Nalpha-Bis(carboxymethyl)-l-lysine (AB-NTA) were then used to connect with the imidazolyl group of histamine, simultaneously the internal standard 4-aminothiophenol (4-ATP) in the core-shell structure was used as the SERS signal. PLS regression model based on concentration range 10-3-10-8mol/L showed a linear trend with R2 = 0.9907. Our new approach can quickly and reliably determine histamine in fish sample and RAW264.7 cell lysates. This protocol for histamine extraction and SERS analysis enables the development of ultra-sensitive method for histamine detection.