Fraction F1 Research Articles

Assessments of anti-breast cancer activity and profiling of active compounds using LC-MS/MS from the Indonesian Agelas nakamurai

Breast cancer is now one of the leading causes of death in women worldwide. Finding a cure is, therefore, critical for women’s health. Marine sponge-derived drugs have gained interest in breast cancer treatment. The development of marine natural products into cancer therapy includes several stages. It starts with the screening using the cytotoxic test method, a preclinical trial using the animal mode, and clinical trial phases I and II. In this study, we focus on the basic cytotoxic test to find the new source of anti-breast cancer compounds derived from Agelas nakamurai to know the potential for further investigation to become lead compounds. However, the anti-breast cancer potential of the Indonesian marine sponge A. nakamurai has never been studied. This study aimed to evaluate the anti-breast cancer potential of extract and fraction of A. nakamurai against epithelial human breast cancer cells (MDA-MB-231 and MCF-7 cells line) and to preliminary profile the known active compounds. The maceration using methanol was used for extraction and followed by partitioning with varied solvents [ethyl acetate (EtOAc), butanol, and water]. Fractionation was done using normal phase open column chromatography, and liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS) was used for analyzing compounds. MDA-MB-231, MCF-7, and HEK-293 cell lines were used for biological activities. The chromatographic separation of the EtOAc fraction led to the F7 subfraction which strongly inhibits proliferation against the MDA-MD-231 (IC50: 10.677 μg/mL), MCF-7 (IC50: 15.154 μg/mL), and HEK-293 cells line. The LC-MS/MS data of active fraction F7 contained gelatin-D and ageloxime-D. This study reported that fraction 7 of A. nakamurai showed strong activity against MDA-MB-231, MCF-7, and HEK-293 cell lines. Further analysis will be carried out to know the mechanisms of action of active compounds.

Abelmoschus esculentus Improves Hippocampal Function Associated with Dipeptidyl Peptidase‐4 in High Fat Diet‐Fed db/db Mice

Hippocampal function can be impaired by diabetes mellitus (DM) and obesity. Abelmoschus esculentus (AE) fractions reportedly mitigate the symptoms of Alzheimer’s disease (AD) by downregulating dipeptidyl peptidase‐4 (DPP‐4)‐mediated insulin resistance. AE extracted by alcohol (fraction 1, F1) and water (fraction 2, F2) contained quercetin glycosides and polysaccharides, respectively. In this study, we investigated whether AE affects hippocampal function in in vitro and in vivo systems. Our results indicate that F1 or F2 enhanced neurogenesis and synapse formation in palmitate‐treated hippocampal neural cells, presumably by downregulating DPP‐4. In db/db mice fed with high fat diet, the hippocampal insulin resistance correlated spatial recognition, with fraction F2 improving hippocampal function. Of note, the alteration of neurogenesis seems interconnecting with changes in gut microbiota. In summary, AE can improve hippocampal function, attenuate insulin resistance, and promote neurogenesis by regulating DPP‐4. AE, particularly F2, has the potential to serve as an adjuvant in preventing DM‐associated AD.

Cholesterol-lowering Effect of Protein Hydrolysates from Lemongrass (Cymbopogon citratus Stapf.)

Lemongrass (Cymbopogon citratus Stapf.) has been used in the Philippines for cooking and as an herb to treat a variety of ailments including hypertension and related cardiovascular diseases (CVDs). This study determined the potential of peptides obtained from the hydrolysis of lemongrass proteins to lower cholesterol in vitro and in an animal model. Proteins were extracted and digested in vitro simulating gastrointestinal conditions. Protein hydrolysates were collected and fractionated using RP-SPE column, and assayed for HMG-CoA reductase inhibitory activity against pravastatin as a control drug. Tannin-free protein extract (TFPE) and total protein hydrolysates (TPC) were administered to Sprague-Dawley rats maintained on a high-fat high-cholesterol diet for two weeks. TFPE yield was 0.03%. Simulated gastrointestinal digestion of the TFPE resulted in 74% yield of protein hydrolysates. Three fractions were obtained from RP-SPE elution of the protein hydrolysates, each with potent HMG-CoA inhibitory activity. The F3 fraction had the highest inhibition of HMG-CoA reductase (IC50 = 0.76 ppm, comparable to pravastatin (IC50 = 0.25 ppm). The inhibitory activity of the fractions was further confirmed through significant serum cholesterol reduction (p < 0.05) in Sprague-Dawley rats. Thus, protein hydrolysates from lemongrass have potential cholesterol-lowering effects in vitro through HMG-CoA reductase inhibition and in vivo through significant reduction of cholesterol levels in an animal model. Protein hydrolysates from lemongrass dietary proteins may serve as promising functional foods for the prevention of CVD risk.

A hybrid artificial intelligence algorithm for fault diagnosis of hot rolled strip crown imbalance

In the production process of hot continuous rolling, due to the imbalance between the number of normal cases and fault cases, the traditional supervised learning methods are often unable to deal with them efficiently. In order to address this problem, a new Hybrid Artificial Intelligence Algorithm is proposed, which is based on Light Gradient Boosting Machine (LightGBM) and combines resampling technique with Support Vector Machine (SVM) to deal with multi-class imbalance problems. Specifically, Support Vector Machines-Synthetic Minority Oversampling Technique (SVM-SMOTE) is used to increase the training data, so as to change the distribution of training data and improve the accuracy of the model, and the Particle Swarm Optimization (PSO) algorithm is used to find the hyper parameters of the model to determine the optimal hyper parameters combination. At the same time, the method of SHapley Additive exPlanation (SHAP) based on game theory is used to reveal the factors affecting the convexity of strip steel plate in hot rolling process and its interpretability. In check to see the validity of the model, experiments were carried out on UCI data set and hot rolling production data set, and multi-model comparison was carried out. The values of Geometric mean (G-mean), macroscopic F1 fraction (F1-Macro) and Matthews correlation coefficient (MCC) tested on the hot rolling data set are 0.992, 0.992 and 0.987 respectively. The model has a good effect. The results show that the model is significantly better than traditional models in solving the problem of strip convexity diagnosis during hot rolling process.

CLINICAL PROTOCOL FOR THE PREPARATION AND ELECTRON MICROSCOPIC ANALYSIS OF THE OBTAINED PRODUCTS OF AUTOLOGOUS MESOCONCENTRATE – PLASMA RICH IN GROWTH FACTORS (PRGF)

Introduction. Ensuring control over regenerative processes by using the patient's blood is a unique concept of an autogenous transplant product, a universal and safe method of application that contains growth factors and cytokines stored in the fibrin matrix and provides stimulating properties of tissue regeneration through the processes of angiogenesis, proliferation, cell migration, and extracellular matrix synthesis.
 The aim. To substantiate the effectiveness of the protocol for collecting, preparing, and forming autocellular mesoconcentrate products in the analysis of the results of their electron microscopic examination.
 Materials and methods. Based on our own clinical experience of using the Endoret-PRGF technique (Human Technology, BTI, Spain) to collect, prepare, and form autocellular transplant products rationally, we used the method of morphological study of objects using a stream of electrons that passed through thin films under high voltage, allowing us to study the structure of these objects at the macromolecular and subcellular levels – transmission electron microscopy, which contributed to the study of the density (number of occurrences in 10 μm2 ), diameter (Ø) of the formed fibrin fibers in the PRGF – F1 and F2 fractions of mesoconcentrate products, namely, isolating membranes (M) and obturating blocks (B). An equally important step in PRGF technology is to follow the step-by-step instructions for preparing and using autocellular transplants.
 Results. Protocol for the collection, preparation, and formation of autocellular transplants using the KMU15 kit (Human Technology, BTI) for the application of the Endoret – PRGF technique, platelet-rich plasma, as a unique and first scientifically proven technique recognized worldwide and patented by the Institute of Human Biotechnology, Spain. The results of the clinical application of mesoconcentrate products, namely isolating membranes widely used in targeted tissue regeneration and obturating blocks, confirm their biological effectiveness. The results of the median (Me) and interquartile range (IQR) during the statistical analysis were obtained, where F1-M = 0.196 (0.176; 0.286) compared to F1-B = 0.344 (0.325; 0.394); F2-M = 0.180 (0.168; 0.214) - F2-B = 0.254 (0.202; 0.338), which gives us the right to assert the reliability of the data set and confirms the quantitative characterization of interfraction differences, and, accordingly, the quality in the sequence of protocol stages.
 Conclusions. The protocol for the efficiency of autocellular graft collection and formation is synchronized with the analysis of electron microscopic images characterizing the density and diameter of fibrin fibers, which differ in the fractional products of the mesoconcentrate and are higher in the obturating blocks than in the isolating membranes, indicating its completeness and compliance.

Antibacterial Activity and Untargeted Metabolomics Profiling of Acalypha arvensis Poepp.

The search for potent antimicrobial compounds is critical in the face of growing antibiotic resistance. This study explores Acalypha arvensis Poepp. (A. arvensis), a Caribbean plant traditionally used for disease treatment. The dried plant powder was subjected to successive extractions using different solvents: hexane (F1), dichloromethane (F2), methanol (F3), a 50:50 mixture of methanol and water (F4), and water (F5). Additionally, a parallel extraction was conducted using a 50:50 mixture of methanol and chloroform (F6). All the fractions were evaluated for their antimicrobial activity, and the F6 fraction was characterized using untargeted metabolomics using SPME-GC×GC-TOFMS. The extracts of A. arvensis F3, F4, and F5 showed antibacterial activity against Staphylococcus aureus ATCC 25923 (5 mg/mL), MRSA BA22038 (5 mg/mL), and Pseudomonas aeruginosa ATCC 27853 (10 mg/mL), and fraction F6 showed antibacterial activity against Staphylococcus aureus ATCC 29213 (2 mg/mL), Escherichia coli ATCC 25922 (20 mg/mL), Pseudomonas aeruginosa ATCC 27853 (10 mg/mL), Enterococcus faecalis ATCC 29212 (10 mg/mL), Staphylococcus aureus 024 (2 mg/mL), and Staphylococcus aureus 003 (2 mg/mL). Metabolomic analysis of F6 revealed 2861 peaks with 58 identified compounds through SPME and 3654 peaks with 29 identified compounds through derivatization. The compounds included methyl ester fatty acids, ethyl ester fatty acids, terpenes, ketones, sugars, amino acids, and fatty acids. This study represents the first exploration of A. arvensis metabolomics and its antimicrobial potential, providing valuable insights for plant classification, phytochemical research, and drug discovery.

Discovery and Characterization of a Dual-Function Peptide Derived from Bitter Gourd Seed Protein Using Two Orthogonal Bioassay-Guided Fractionations Coupled with In Silico Analysis.

The hydrolysate of bitter gourd seed protein, digested by the combined gastrointestinal proteases (BGSP-GPs), exhibited the most potent inhibition on angiotensin-I-converting enzyme (ACE) with an IC50 value of 48.1 ± 2.0 µg/mL. Using two independent bioassay-guided fractionations, fraction F5 from reversed-phase chromatography and fraction S1 from strong cation exchange chromatography exhibited the highest ACE inhibitory (ACEI) activity. Three identical peptides were simultaneously detected from both fractions and, based on the in silico appraisal, APLVSW (AW6) was predicted as a promising ACEI peptide. Their dipeptidyl peptidase-IV (DPP4) inhibitory (DPP4I) activity was also explored. The IC50 values of AW6 against ACE and DPP4 were calculated to be 9.6 ± 0.3 and 145.4 ± 4.4 µM, respectively. The inhibitory kinetics and intermolecular interaction studies suggested that AW6 is an ACE competitive inhibitor and a DPP4 non-competitive inhibitor. The quantities of AW6 in BGSP-GP hydrolysate, fractions F5 and S1, were also analyzed using liquid chromatography-tandem mass spectrometry. Notably, AW6 could resist hydrolysis in the human gastrointestinal tract according to the result of the simulated gastrointestinal digestion. To the best of our knowledge, this is the first discovery and characterization of a dual-function (ACEI and DPP4I activities) peptide derived from bitter gourd seed protein.

Ion-exchange purification, nano-HPLC–MS/MS identification and molecular dynamics simulation of novel umami peptides from fermented grain wine (Huangjiu)

Huangjiu is a traditional Chinese fermented rice wine with abundant peptides, but the umami peptides are little known due to the wine’s complex matrix. Herein, the target umami fraction was enriched with sensory-guided separation coupled with ultrafiltration, ion-exchange, and gel chromatography purification. A total of 1901 peptides were identified from gel F1 fraction with nano-HPLC–MS/MS. Eight novel umami peptides DWDPHGL, VPEPEPH, LTPEPEH, GANQLDPR, FREE, LDFR, QEGW, and RDH were filtered by virtual screening and sensory evaluation. They exhibited thermal stability at 373.15 K and the taste thresholds in water were between 0.29 and 0.65 mmol/L. Among them, RDH was crucial, due to a relatively high dose over threshold (DoT) factor (1.12). Molecular docking results showed that residues ASP147, SER148, ARG151, TYR220, SER276, and ALA302 contributed most to the interaction of peptide and T1R1. Moreover, molecular dynamics simulation and weak interaction analysis suggested the important role of intramolecular forces in peptide thermal stability.

Phenolic Compounds in Fractionated Blackcurrant Leaf Extracts in Relation to the Biological Activity of the Extracts.

The aim of this study was to determine the relationship between antioxidant and anticancer properties of extracts from blackcurrant (Ribes nigrum L.) leaves and their fractions and chemical contents. Dried ethanolic extract was divided into three fractions using solid phase extraction: aqueous (F1), 40% MeOH (F2), and 70% MeOH (F3). Both the extract and the fractions were analyzed in terms of antiradical activity (DPPH• and ABTS+•), total phenolic compounds, and total flavonoids. The antitumor potential of the fractions was evaluated in vitro on human colorectal (HCT 116) and prostate (PC-3) cancer cells. Phenolics were identified using HPLC-QTOF-MS, and twelve compounds were quantified by HPLC-DAD. Finally, principal component analysis was carried out to assess the relationship between the tested factors. The results confirmed that blackcurrant leaves are a rich source of phenolics with high antioxidant activity and anticancer properties. It was demonstrated that the F2 fraction had the highest content of phenolics and the highest antiradical activity. Additionally, only this fraction showed cytotoxic activity against HCT 116 cells. It was confirmed that both the blackcurrant leaf extract and its fractions are a promising source of condensed active compounds and can be used as natural functional food additives.

Reconsidering mercury sources and exposure pathways to bivalves: Insights from mercury stable isotopes

Identifying mercury (Hg) sources and exposure pathways to bivalves, particularly in relation to sediment, is important for expanding the utility of bivalves as a monitoring organism for sediment quality. Here we use Hg isotope ratios to decipher Hg sources accumulated into bivalves by conducting field studies and in situ experiments. In the first part of this study, we characterized Hg isotope ratios in individual geochemical fractions of riverine sediment, contaminated by liquid Hg in South Korea (Hyeongsan River; HS). Asian clams (Corbicula fluminea) were then deployed at the contaminated sites to evaluate the isotopic turnover. Over the two-month period, the isotope ratios of the clams shifted toward the labile/exchangeable Hg pools (F1, F2 fractions) of the sediment. Conversely, in the control site where sediment Hg is low, we observed similar Hg isotope ratios between Asian clams and the samples of precipitation and dissolved phase of water column. In East Fork Poplar Creek, (Oak Ridge) U.S., Asian clams also displayed similar Hg isotope ratios with the dissolved phase of water column, which have undergone substantial in-stream processing or input from Hg-contaminated groundwater from the hyporheic zones and riparian tributary during high hydrologic flow seasons. Our study demonstrates that the dissolved Hg phases within the water column, whether originating via sediment diffusion or derived externally, act as the primary source and exposure pathways to bivalves. The results of our study also shed new light to the prior Hg isotope measurement in bivalves collected from estuarine, lake, and coastal systems, which showed significant isotopic deviation from bulk sediment. The fact that bivalves are sensitive to in situ and external dissolved Hg phases provides additional insight into the existing biomonitoring program, which uses bivalves as a bioindicator for sediment quality.

Spatial and temporal dynamics and fluxes estimation of manganese fractions in sediments from the Pearl River Estuary, southern China

Sequential extraction was used to study the historical dynamics and fluxes of the chemical fractions of manganese (Mn) in sediments collected from the Pearl River Estuary (PRE), southern China. Results revealed that the proportion of Mn associated with different fractions decreased in the order of acid-soluble fraction (F1) > reducible fraction (F2) > residual fraction (F4) > oxidizable fraction (F3). F1 (47%) was the dominant Mn fraction, indicating the strong bioavailability of Mn to aquatic organisms in the PRE. In addition, the Mn fraction F2 was present at an average rate of 27.93 % in surface sediments, indicating that it could be mobilized when environmental conditions become increasingly reducing or oxidizing. The decline in Mn fraction fluxes after 2006 indicated that the region has partially decreased due to the removal of heavily polluting industries and effective control of sewage discharge.

Endothelium-dependent vasorelaxation effects of F5 fraction of Crinum amabile chloroform extract

BackgroundVascular dysfunction can lead to many health problems including hypertension and heart disease. The complexities of vascular dysfunction and vascular disorder-related diseases have prompted the search for many new biologically active compounds in the efforts of resolving the problems. Previous studies have shown that Amaryllidaceae alkaloids exert multiple biological activities, including the vasorelaxation effect. Crinum amabile, which is a family member of Amaryllidaceae, is believed to possess a promising pharmacological activity as a vasorelaxant.MethodThe vasorelaxation activities of Crinum amabile extracts and fractions were determined using in vitro models of phenylephrine pre-contracted intact and denuded rat aortic rings. The mechanistic pathways of vasorelaxation were investigated by pre-treatment of endothelium-intact rat aortic rings with L-NG Nitro Arginine Methyl Ester (L-NAME), methylene blue (MB), indomethacin, atropine and propranolol, respectively.ResultsThe results showed that chloroform extract (CE) of Crinum amabile exhibited the highest vasorelaxation activity, and further fractionation of CE found that its F5 fraction exerted the strongest activity. An in-depth study on the mechanistic pathway revealed that the endothelium-dependent vasorelaxation induced by F5 fraction was primarily achieved through stimulation of prostaglandin (PGI2) production and partially associated with NO/cGMP activation pathway.ConclusionFindings from this study suggest that Crinum amabile can serve as a promising candidate for the discovery and development of the new vasorelaxant drug.

The Effects of the Fraction Isolated from Iranian Buthotus shach Scorpion Venom on Synaptic Plasticity, Learning, Memory, and Seizure Susceptibility.

Epilepsy, as a neurological disease, can be defined as frequent seizure attacks. Further, it affects many other aspects of patients' mental activities, such as learning and memory. Scorpion venoms have gained notice as compounds with potential antiepileptic properties. Among them, Buthotus schach (BS) is one of the Iranian scorpions studied by Aboutorabi et al., who fractionated, characterized, and tested this compound using electrophysiological techniques in brain slices (patch-clamp recording). In the present study, the fraction obtained from gel electrophoresis was investigated through behavioral and electrophysiological assays. At first, ventricular cannulation was performed in rats, and then the active fraction (i.e., F3), carbamazepine, and the vehicle were microinjected into the brain before seizure induction by the subcutaneous (SC) injection of pentylenetetrazol (PTZ). Seizure behaviors were scaled according to Racine stages. Memory and learning were evaluated using the Y-maze and passive avoidance tests. Other groups entered evoked field potential recording after microinjection and seizure induction. Population spike (PS) and field excitatory postsynaptic potential (fEPSP) were measured. The F3 fraction could prevent the fifth stage and postpone the third stage of seizure compared to the control (carbamazepine) group. There was no significant improvement in memory and learning in the group treated with the F3 fraction. Also, PS amplitude and fEPSP slope increased significantly, and long-term potentiation was successfully formed after the high-frequency stimulation of the performant pathway. Our results support the antiepileptic effects of the F3 fraction of BS venom, evidenced by behavioral and electrophysiological studies. However, the effects of this fraction on memory and learning were not in the same direction, suggesting the involvement of two different pathways.

Isolation, identification, and preparation of tyrosinase inhibitory peptides from Pinctada martensii meat.

Recently, natural tyrosinase inhibitors have gained attention in clinical cosmetology research. In this study, the enzymatic hydrolysis of Pinctada martensii meat by protease from Bacillus licheniformis, 401 peptides with tyrosinase inhibitory were identified after isolated by ultrafiltration and Sephadex G-15 from the fraction F4. The peptide effects on the tyrosinase activity and structure were evaluated using molecular docking. Three synthetic peptides classified as W1 (WDRPKDDGGSPIK), W2 (DRGYPPVMF), and W3 (SGGGGGGGLGSGGSIRSSY), which had the lowest binding energies were selected for in vitro synthesis and biological activity investigation. The W3 peptide (5mg/mL) had the highest tyrosinase activity, SPF, DPPH, and ABTS clearance values, and total antioxidant capacity. W3 did not affect the survival rate of mouse melanoma B16-F10 cells (1.0-5.0mg/mL) but decreased the melanin content. Hence, W3 could be suitable for multifunctional tyrosinase inhibition and provides a novel method to use marine organisms as natural tyrosinase inhibitor sources.

Structure and emulsifying properties of unprecedent glucomannan oligo- and polysaccharides from Amazonia Acrocomia aculeata palm fruit

Acrocomia aculeata fruit pulp contains oil (4.1–82.8 % fresh matter) and carbohydrates (6.6–98.0 % fresh matter). To date, only the oil fraction is valorized because very little is known about the nature of carbohydrates. This study explores new ways of adding value to this pulp by developing simple and efficient extraction processes for its carbohydrate components and characterizing their structure and physicochemical properties over two harvest periods. A water-soluble monosaccharide fraction F1 (solubility limit (SL): 98.5–99.3 g/L) (yield: 21 % dry pulp (DP)), a water-soluble polysaccharide fraction F2 (SL: 93.3–95.3 g/L) (yield: 26 % DP) and two additional water-insoluble polysaccharide fractions F3 and F4 (SL: <8 g/L) (yields: 10 and 19 % DP, respectively) were isolated. NMR structural characterizations of fraction F2 revealed it to be a linear glucomannan with β-(1 → 4) osidic linkages between d-Manp and d-Glcp residues. F2 is unique for its d-Manp/d-Glcp ratio of 3:1 and the position of its acetyl group (13–14 %, C-2 d-Manp). Finally, the polysaccharide showed a molecular weight (Mw) variation ranging from 8.2 × 104 to 1.1 × 103 Da over the two harvest periods, with remarkable emulsifying properties associated with a low Mw of F2 (stability >6 months, 1 % w/v in a water-in-oil emulsion).

Screening and mechanism of six angiotensin I-converting eneyme inhibitory peptides from protein hydrolysate of Coregonus peled visceral: A combined in computer and in vitro study

Coregonus peled visceral can be regarded as a rich source of bioactive peptides. The purpose of this study is to screen peptides with high angiotensin I-converting eneyme (ACE) inhibitory activity from protein hydrolysate of Coregonus peled visceral prepared by trypsin, and analyze the inhibitory mechanisms of prepared peptides. The results showed that the UF-4 (MW < 3 kDa) had the highest ACE inhibitory activity of 73.55% (2 mg/ml). UF-4 was separated by Sephadex G-25 column and collected three fractions (F1, F2 and F3), and 397 peptides were detected from F1 fraction, and 6 peptides were screened by silico methods as potential antihypertensive peptides. FPGAKP formed 9 hydrogen bond interactions with Ala354, Tyr523, Glu384 in S1 pocket; His353 in S2 pocket; 5 hydrogen bond interactions with Thr282, Ala356, Arg522, Arg522, and His387 near the active sites. FPGAKP has the highest ACE inhibitory activity (IC50 = 0.039 mg/ml) and exhibits competitive inhibition. The total content of β-sheet and α-helix in the secondary structure of FPGAKP was the highest (80.22%). Studies have shown that the ACE inhibitory activity of the peptide mainly depends on the number of hydrogen bonds formed with the key active site of ACE and the content of β-sheet and α-helix. The results indicated that FPGAKP has the value of further research as a substitute for antihypertensive drugs.

Antiproliferative Activity of Mycelium vs. Fruiting Body: Ganoderma subincrustatum and G. weberianum from Sonora, Mexico

The genus Ganoderma has been little studied in arid areas worldwide. Ganoderma subincrustatum and Ganoderma weberianum strains were obtained from the Sonoran Desert, Sonora, Mexico. Ganoderma spp. synthesize triterpenoids such as ganoderic acids with antiproliferative activity because they inhibit specific targets, induce apoptosis, and increase the activity of killer cells. Mycelium and fruiting body chloroform extracts from G. subincrustatum and G. weberianum were tested on HeLa, A549, L-929, and RAW 264.7 cell lines. Extracts from the fruiting body present higher antiproliferative activity than mycelium. All extracts induced vesicle and cellular debris formation in all cell lines, being non-selective for cancerous cells. Chloroform extract from G. subincrustatum fruiting bodies presented higher activity against all cell lines. Fractions F7 and F15 from this extract exhibited an IC50 of 37.9 and 41.9 µg/mL on the A549 cell line, respectively; however, chloroform crude extract showed higher activity (IC50 of &lt;25 µg/mL) in all cell lines. Flow cytometry assays of F7 revealed cell death by apoptosis in A549 cells. NMR suggested the presence of ganoderic acids in F7. In future research, it will be interesting to characterize these fractions (metabolites, their bioactivities, and mechanism of action).

Co-pyrolysis of textile dyeing sludge/litchi shell and CaO: Immobilization of heavy metals and the analysis of the mechanism

To relieve the secondary contamination of heavy metals (HMs), the synergistic effect of co-pyrolysis of textile dyeing sludge (DS)/litchi shell (LS) and CaO on the migration of HMs was demonstrated in this study. The proportions of Cu, Zn, Cr, Mn, and Ni in the F4 fraction increased to 75%, 55%, 100%, 50%, and 62% at the suitable CaO dosages. When 10% CaO was added, the RI value of DLC-10% was reduced to 7.89, indicating low environmental risk. The characterizations of the physicochemical properties of biochar provided support for the HMs immobilization mechanism. HMs combined with inorganic minerals or functional groups to form new stable HMs crystalline minerals and complexes to achieve immobilization of HMs. The pH value and pore structure also play an important role in improving the immobilization performance of HMs. In conclusion, the results provided a new direction for the subsequent harmless treatment of HMs-enriched waste.

Mining Xanthine Oxidase Inhibitors from an Edible Seaweed Pterocladiella capillacea by Using In Vitro Bioassays, Affinity Ultrafiltration LC-MS/MS, Metabolomics Tools, and In Silico Prediction.

The prevalence of gout and the adverse effects of current synthetic anti-gout drugs call for new natural and effective xanthine oxidase (XOD) inhibitors to target this disease. Based on our previous finding that an edible seaweed Pterocladiella capillacea extract inhibits XOD, XOD-inhibitory and anti-inflammatory activities were used to evaluate the anti-gout potential of different P. capillacea extract fractions. Through affinity ultrafiltration coupled with liquid chromatography tandem mass spectrometry (LC-MS/MS), feature-based molecular networking (FBMN), and database mining of multiple natural products, the extract's bioactive components were traced and annotated. Through molecular docking and ADMET analysis, the possibility and drug-likeness of the annotated XOD inhibitors were predicted. The results showed that fractions F4, F6, F4-2, and F4-3 exhibited strong XOD inhibition activity, among which F4-3 reached an inhibition ratio of 77.96% ± 4.91% to XOD at a concentration of 0.14 mg/mL. In addition, the P. capillacea extract and fractions also displayed anti-inflammatory activity. Affinity ultrafiltration LC-MS/MS analysis and molecular networking showed that out of the 20 annotated compounds, 8 compounds have been previously directly or indirectly reported from seaweeds, and 4 compounds have been reported to exhibit anti-gout activity. Molecular docking and ADMET showed that six seaweed-derived compounds can dock with the XOD activity pocket and follow the Lipinski drug-like rule. These results support the value of further investigating P. capillacea as part of the development of anti-gout drugs or related functional foods.

Impact of inclusion of post-spermatic ejaculate fraction in boar seminal doses on sperm metabolism, quality, and interaction with uterine fluid

Boar ejaculate is composed of sperm cells and seminal plasma (SP) and is emitted in different fractions (pre-sperm fraction; spermatic-rich fraction; intermediate fraction; post-spermatic fraction), with different composition of SP and volume, which could influence the sperm quality during seminal doses preparation, conservation, and interaction with the female reproductive tract. In artificial insemination (AI) centers, seminal doses are usually prepared with the spermatic-rich and intermediate fractions, but the inclusion of other ejaculate fractions, although controversial, is beginning to be applied. The objective was to evaluate the synergic effect of accumulative ejaculated fractions on sperm functionality during seminal doses preparation, throughout storage and after incubation with uterine fluid (UF). For this purpose, a total of 57 ejaculates were collected, and the following experimental groups were prepared (n = 19 per group): (F1) spermatic-rich fraction; (F2) F1 plus intermediate fraction; (F3) F2 plus post-spermatic fraction. Each group was stored for 5 days at ∼16 °C, and the following parameters were evaluated: sperm metabolism of pure and diluted semen (day 1), sperm quality parameters (days 1, 3, 5), thermal-resistance test (TRT) and incubation with uterine fluid (UF) (day 5). Sperm metabolic rates between accumulative ejaculate fractions from pure and diluted semen did not show differences. Also, sperm quality parameters were not affected by the ejaculate fraction during storage. However, sperm subjected to TRT showed similar results except for progressive motility, which was better in F2 and F3 than F1. When sperm were incubated with UF, the quality decreased in each group, but sperm from F2 and F3 were less affected than those from F1. In conclusion, the post-spermatic fraction can be included in seminal doses for their use in AI-centers, with functionality of sperm of different SP origins not being impaired throughout the storage, and responding better to thermal and UF stress. However, further research in AI-centers is necessary to test the sperm behaviour under presented conditions.