Levels Of Proteases Research Articles

N-Acetylneuraminic Acid Inhibits Melanogenesis via Induction of Autophagy

N-acetylneuraminic acid (Neu5Ac) is the predominant form of sialic acid present in the glossy swiftlet (Collocalia esculenta). It is also the only form of sialic acid detected in the human body. In this study, we investigated the mechanism underlying melanogenesis inhibition by Neu5Ac. We discovered that a reduction in tyrosinase protein levels led to an inhibition of melanin production by Neu5Ac. Additionally, the mRNA and protein levels of ubiquitin-specific protease (USP5) and microtubule-associated protein 1 light chain 3 (LC3)-II increased, while those of p62 decreased, indicating enhanced autophagic activity. Lysosomal cathepsin L2 protein levels also increased, and immunostaining revealed colocalization of lysosomal membrane protein (LAMP)-1 and tyrosinase. Additionally, levels of chaperonin containing T-complex polypeptide (CCT), implicated in increased autophagic flux, were elevated. Altogether, these findings suggest that tyrosinase-containing coated vesicles are transported by Neu5Ac into the autophagic degradation pathway, suppressing mature melanosome generation. This process involves increased USP5 levels preventing recognition of polyubiquitin by proteasomes. Furthermore, elevated CCT3 protein levels may enhance autophagic flux, leading to the incorporation of tyrosinase-containing coated vesicles into autophagosomes. These autophagosomes then fuse with lysosomes for cathepsin L2–mediated degradation. Thus, our findings suggest that Neu5Ac reduces tyrosinase activity and inhibits melanosome maturation by promoting selective autophagic degradation of abnormal proteins by p62.

COVID-19 PBMCs are doubly harmful, through LDN-mediated lung epithelial damage and monocytic impaired responsiveness to live Pseudomonas aeruginosa exposure.

Although many studies have underscored the importance of T cells, phenotypically and functionally, fewer have studied the functions of myeloid cells in COVID disease. In particular, the potential role of myeloid cells such as monocytes and low-density neutrophils (LDNs) in innate responses and particular in the defense against secondary bacterial infections has been much less documented. Here, we compared, in a longitudinal study, healthy subjects, idiopathic fibrosis patients, COVID patients who were either hospitalized/moderate (M-) or admitted to ICU (COV-ICU) and patients in ICU hospitalized for other reasons (non-COV-ICU). We show that COVID patients have an increased proportion of low-density neutrophils (LDNs), which produce high levels of proteases (particularly, NE, MMP-8 and MMP-9) (unlike non-COV-ICU patients), which are partly responsible for causing type II alveolar cell damage in co-culture experiments. In addition, we showed that M- and ICU-COVID monocytes had reduced responsiveness towards further live Pseudomonas aeruginosa (PAO1 strain) infection, an important pathogen colonizing COVID patients in ICU, as assessed by an impaired secretion of myeloid cytokines (IL-1, TNF, IL-8,…). By contrast, lymphoid cytokines (in particular type 2/type 3) levels remained high, both basally and post PAO1 infection, as reflected by the unimpaired capacity of T cells to proliferate, when stimulated with anti-CD3/CD28 beads. Overall, our results demonstrate that COVID circulatory T cells have a biased type 2/3 phenotype, unconducive to proper anti-viral responses and that myeloid cells have a dual deleterious phenotype, through their LDN-mediated damaging effect on alveolar cells and their impaired responsiveness (monocyte-mediated) towards bacterial pathogens such as P. aeruginosa.

Evaluation of Himalayan nettle (Urtica ardens) extract meal in common carp (Cyprinus carpio) feed: Impact on growth performance, digestive enzyme activity and antioxidant capacity

This research aims to evaluate the impact of an extract derived from the leaves of Himalayan nettle (Urtica ardens) leaf extract on common carp, Cyprinus carpio, growth, digestive enzymes, fish body carcass composition, antioxidant system, and water quality parameters. For a duration of 90 days, experimental diets containing nettle extract were given to 150 fish (12.38 ± 0.53 g) in 15 tanks (10 fish per tank). The five groups in triplicate were: control or T0 (untreated), T1 (fed 0.3% extract), T2 (0.6% extract), T3 (0.9% extract), and T4 (1.2% extract). Results revealed that nettle extract in the diet increases common carp growth up to 0.9% concentration of extract. Its feed conversion ratio decreases, while weight gain and specific growth rate rise significantly compared to the control diet. No mortality was observed during the 90-day experiment. Dietary nettle extract did not affect whole-body fibre and crude ash percentages (p > 0.05). Nettle-fed and control groups showed significant differences in crude protein, body fat, and total body moisture (p < 0.05). After a 90-day feeding trial, 0.9% nettle extract-fed fish had higher protease, amylase, lipase, and cellulase levels than the control group (p < 0.05). When nettle content reached 1.2%, enzyme activity decreased. Orthophosphate production peaked at 4 and 8 hours after feeding, while ammonia excretion peaked at 2 and 6 hours. The control group excreted the most ammonia and orthophosphate, while the 0.9% group excreted the least. Liver and kidney SOD levels were higher in common carp fed a 0.6% nettle extract diet. Feeding with a 0.9% nettle extract diet improved hepatic lipid peroxidation, gut and liver glutathione. Gut and kidney SOD, catalase, LPO, and GSH increased dramatically on the 1.2% diet. Statistically, the 0.9% nettle extract (T3) group showed the overall best results compared to the control diet. These findings demonstrate the potential of nettle extract as a dietary supplement in aquaculture.

Phytosterol supplementation enhances the growth performance, feed utilization, antioxidant status and glucose metabolism of juvenile largemouth bass (Micropterus salmoides) fed a high-starch diet

IntroductionPrevious studies found that phytosterols could influence growth performance, feed utilization and lipid metabolism as well as improve the antioxidant capacity of animals.MethodsTo investigate the effects of dietary phytosterol supplementation on juvenile largemouth bass (Micropterus salmoides) fed a high-starch diet, a 56- day feedingtrial was conducted with four dietary feeds for juvenile largemouth bass: extruded floating feed isonitrogenous and isoenergetic diets were formulated to contain 10% and 15% α-starch; on the basis of a 15% α-starch diet, two other diets were formulated with supplementation of 0.1% and 0.5% phytosterol, respectively. After the feeding trials, the survival rate, weight gain and specific growth rate, feed conversion ratio, intraperitoneal fat ratio, feed intake, protein efficiency ratio and activities of three digestive enzymes, as well as the concentrations of nine plasma biochemical indices, hepatic enzyme activities and glycogen contents, were measured and calculated, and the data were statistically analyzed.ResultsThe results of the present study showed that the survival rate, weight gain and specificgrowthrates were significantly greater in plants fed high-starch diets supplemented with phytosterols. As the supplemental phytosterol concentration increased, the feed conversion ratio and intraperitoneal fat ratio significantly decreased; the protease and lipase levels in the pyloric zone markedly increased; the plasma cholesterol, triglyceride, glucose, malondialdehyde, aspartate transaminase and alanin transaminase levels significantly decreased; the glucokinase and pyruvate kinase levelsmarkedly increased; and the hepatic glycogen content significantly decreased.DiscussionIn summary, dietary phytosterol supplementation promoted the growth performance, feed utilization and antioxidant status of juvenile largemouth bass fed a highstarch diet; enhanced glucose utilization and metabolism; and alleviated the negative stimulation of glycemia stress in M. salmoide fed a high-starch diet.

Apparent and True Digestibility in Clarias gariepinus, Burchell, 1822 FED Soyabean Meal Based Diets Supplemented with Protease

Soyabean meal (SBM) could be a suitable replacement for the expensive fishmeal but for its low digestibility in fish. However, protease supplementation could improve digestibility of SBM based diets. Therefore, apparent and true digestibility in Clarias gariepinus fed SBM based diets supplemented with protease were investigated. Six diets containing varied inclusion levels of protease (ppm) in solvent extracted soybean-based diets (SESBD) were formulated; Control (without protease), SS100 (100), SS200 (200), SS300 (300), SS400 (400), SS500 (500). The Clarias gariepinus (n=720) weighing 12.00±0.10g were fed to satiation with the diets for 12 weeks. Each treatment was in triplicate. Protease supplementation significantly (P&lt;0.05) influenced the weight gain of C. gariepinus with the higher value in fish fed SS400 (44.63±3.13) and least value in control diet (32.03±0.65). FCR had a significantly (P&lt;0.05) least value in C. gariepinus fed SS400 (1.62±0.18) but similar (P&gt;0.05) to C. gariepinus diet SS300 (1.88±0.11). Supplemental protease in soyabean-based diet improved (P&lt;0.05) apparent protein digestibility of C. gariepinus on diet SS400 (87.38±1.10) but similar (P&gt;0.05) to C. gariepinus on diet SS200 (87.29±0.97). Also, true lysine digestibility was enhanced with protease supplementation in soybean-based diet with the higher value in diet SS200 (95.90±0.07). Optimal FCR occurred SESBD of 350ppm dietary inclusion (R2= 0.8147) of protease. The findings suggest that, protease supplementation in solvent extracted soyabean based diet could improve growth performance and amino acid digestibility in C. gariepinus at 350ppm inclusion level.

Single-Cell Transcriptome Analysis Reveals Dynamic Populations of Vascular Cells in Neointimal Hyperplasia.

Neointimal hyperplasia (NIH) is the pathological basis of vascular injury disease. Vascular cells are the dominant cells in the process of NIH, but the extent of heterogeneity amongst them is still unclear. A mouse model of NIH was constructed by inducing carotid artery ligation. Single-cell sequencing was then used to analyze the transcriptional profile of vascular cells. Cluster features were determined by functional enrichment analysis, gene set scoring, pseudo-time analysis, and cell-cell communication analysis. Additionally, immunofluorescence staining was conducted on vascular tissues from fibroblast lineage-traced (PdgfraDreER-tdTomato) mice to validate the presence of Pecam1+Pdgfra+tdTomato+ cells. The left carotid arteries (ligation) were compared to right carotid arteries (sham) from ligation-induced NIH C57BL/6 mice. Integrative analyses revealed a high level of heterogeneity amongst vascular cells, including fourteen clusters and seven cell types. We focused on three dominant cell types: endothelial cells (ECs), vascular smooth muscle cells (vSMCs), and fibroblasts. The major findings were: (1) four subpopulations of ECs, including ECs4, mesenchymal-like ECs (ECs1 and ECs2), and fibro-like ECs (ECs3); (2) four subpopulations of fibroblasts, including pro-inflammatory Fibs-1, Sca1+ Fibs-2, collagen-producing Fibs-3, and mesenchymal-like Fibs-4; (3) four subpopulations of vSMCs, including vSMCs-1, vSMCs-2, vSMCs-3, and vSMCs-3-derived vSMCs; (4) ECs3 express genes related to extracellular matrix (ECM) remodeling and cell migration, and fibro-like vSMCs showed strong chemokine secretion and relatively high levels of proteases; (5) fibro-like vSMCs that secrete Vegfa interact with ECs mainly through vascular endothelial growth factor receptor 2 (Vegfr2). This study presents the dynamic cellular landscape within NIH arteries and reveals potential relationships between several clusters, with a specific focus on ECs3 and fibro-like vSMCs. These two subpopulations may represent potential target cells for the treatment of NIH.

PSII-17 Effect of protease supplementation on amino acids digestibility of soybean meal fed to growing pigs of different ages

Abstract Protease supplementation can increase amino acids (AA) availability; however, the effectiveness of protease in swine diets varies and has been associated with the type and dose of protease, dietary ingredients, and potentially animal age. This study investigated the interactive effect of protease inclusion level and pig age on standardized ileal digestibility (SID) of protein (CP) and AA of soybean meal (SBM) fed to growing pigs. A corn starch–based diet was formulated with SBM as the sole source of AA and containing titanium as an indigestible marker. Protease was supplemented at 0, 15,000; 30,000 and 45,000 NFP/kg of feed (ProAct 360). An N-free diet was used to estimate basal ileal endogenous AA losses. Ten cannulated pigs (body weight = 21 kg ± 2 kg) were assigned to experimental diets in a duplicate 5 × 5 Latin square design. In Phase I (23 to 30 kg-pigs, 90 ± 17 d of age), ileal digesta was collected in 5 periods of 7 d (5 d adaptation and 2 d ileal digesta collection). In Phase II (50 to 65 kg-pigs. 140 ± 17 d of age), using the same diets, ileal digesta was collected in 5 more periods of 7 d. Pigs were fed at 4% of body weight (BW) adjusted at the end of each period. Diets and ileal digesta were analyzed for CP, AA, and titanium and SID was calculated. Orthogonal polynomial contrasts were used to determine the linear and quadratic effects of protease supplementation. The combined data were analyzed as 4 x 2 factorial, including the fixed effects of protease inclusion (excluding N-free diet), age, and their interactions. In phase I, increasing levels of protease resulted in linear improved (P &amp;lt; 0.1) of SID for the majority of indispensable AA (Except Arg, His, Met + Cys, and Trp) and the average of dispensable AA. In phase II, the SID of Ile, Leu, Met, Met + Cys, Val, the average of indispensable AA, and the majority of dispensable AA was quadratically improved. Overall (Phase I and II), the SID of Ile, Leu, Met + Cys, Val, Cys, Glu, Tyr, and the average of dispensable AA was quadratically improved (P &amp;lt; 0.1). In most cases, the supplementation of 30,000 NFP/kg of feed resulted in the greatest AA digestibility improvements. However, the linear response in the first phase for some AA suggests that diets for younger pigs could be supplemented with a greater level (45,000 NFP/kg of feed). There were no interactions between protease levels and age. Younger pigs had improved (P &amp;lt; 0.1) SID of CP and most AA (Except Cys, Gly, and Ser). In conclusion, dietary protease supplementation can improve SID of AA in soybean mean independent of pig age. Pigs age can influence AA digestibility where younger pigs have greater AA digestibility.

The characteristics of microbiome in the upper respiratory tract of COVID-19 patients

BackgroundCo-infection with other pathogens in coronavirus disease 2019 (COVID-19) patients exacerbates disease severity and impacts patient prognosis. Clarifying the exact pathogens co-infected with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is premise of the precise treatment for COVID-19 patients.MethodsSputum samples were collected from 17 patients in the COVID-19 positive group and 18 patients in the COVID-19 negative group. DNA extraction was performed to obtain the total DNA. Sequencing analysis using 16S and ITS rRNA gene was carried out to analyze the composition of bacterial and fungal communities. Meanwhile, all the samples were inoculated for culture.ResultsWe did not observe significant differences in bacterial composition between the COVID-19 positive and negative groups. However, a significantly higher abundance of Candida albicans was observed in the upper respiratory tract samples from the COVID-19 positive group compared to the COVID-19 negative group. Moreover, the Candida albicans strains isolated from COVID-19 positive group exhibited impaired secretion of aspartyl proteinases.ConclusionCOVID-19 positive patients demonstrate a notable increase in the abundance of Candida albicans, along with a decrease in the levels of aspartyl proteinases, indicating the alteration of microbiota composition of upper respiratory tract.

Approaches to Control and Monitor Protease Levels in Chronic Wounds

AbstractSome wounds do not follow the typical healing trajectory and show very little, if any, progress. They dreadfully threaten the patients' quality of life and may even lead to limb amputation and death. Several interrelated systemic and local factors may cause a delay in wound healing. The microenvironment of these wounds can be different from wounds that heal spontaneously or with standard treatment. As discussed in the present work, proteases’ levels or activity can be elevated in some cases of chronic ulcers; infection can aggravate the situation. A detailed overview of the proteases’ effect on wound healing is presented in this work. Assuming untimely or irregularly excessive proteolytic activity plays a pathological role, some researchers targeted attenuation of protease level as a strategy to promote healing. This can be done via different approaches, including physical removal, inhibiting their activity, and controlling their expression directly through gene silencing or indirectly, for instance, by resolving infection. Apart from targeting protease modulation as a therapeutic strategy, many studies have examined the protease state to gain insights into the underlying mechanisms by which a remedy, dressing, or specific therapy has affected the healing process. This study aims to provide an overview of the studies in these two categories. Of course, various proteases are involved in the healing process, but in each study, usually, only one or some of them are investigated; matrix metalloproteinase‐2 (MMP‐2), MMP‐9, and human neutrophil elastase (HNE) are among them. Furthermore, in some cases, cumulative proteolytic activity is evaluated in terms of gelatinolytic, collagenolytic, or caseinolytic activity, which is also absolutely beneficial. Some research groups have considered the potential of proteases as a diagnostic or prognostic biomarker and have tried to develop some sensors or indicators; this topic is also covered in the present work.

Exploring the Mechanism of Acupuncture's Influence on the Protease Activity and Free Radical Damage in Synovial Fluid of Rheumatoid Arthritis Rats Induced by Type II Collagen from the Release of Active Oxygen.

Acupuncture was studied to investigate the mechanism of its effect on protease vitality and free radical damage in Type I CIA rats induced by type II collagen. The study divided rats into a control group (injected with physiological saline, n = 10), a model group (injected with type II collagen, n = 10), and an intervention group (injected with type II collagen + acupuncture ST36 and GB39, 3 times a week, for a total of 4weeks, n = 10) based on the different injected drugs. Then, various indicators of the mice were experimentally tested using joint index scoring, H&E histological staining, protein blotting, and immunohistochemistry staining methods. Acupuncture ST36 and GB39 can reduce arthritis scores, histological staining scores, and increase MVD in CIA rats. And reduce protease levels, alleviate inflammation, synovial hyperplasia, and angiogenesis. In addition, the intervention group TNF-α, IL-1β and IL-6 mRNA were reduced, and the clearance rates of hydrogen peroxide free radicals and nitric oxide free radicals were increased. The expression levels of ROS and MDA decrease, while the expression levels of SOD increase It has been proved that acupuncture at ST36 and GB39 can inhibit the release of ROS, reduce protease activity, inflammation, synovial hyperplasia, angiogenesis and free radical damage, thus reducing the severity of CIA (Collagen-Induced Arthritis) in rats.

Nano calcium carbonate improves wheat nitrogen accumulation and grain yield by enhancing soil nitrogen supply and flag leaf photosynthetic characteristics

ContextOver-application of chemical fertilizers to attain optimal crop yields has resulted in an environmental issue of global concern. To mitigate this, reducing nitrogen (N) application and enhancing fertilizer use efficiency is crucial, and nano-fertilizers offer a promising solution. However, there is a paucity of studies examining the use of nano-fertilizers in fields. ObjectivesThis study aimed to assess the impact of different dosages of nano calcium carbonate (NCC) mixed with compound fertilizer (CF) on soil N supply, enzyme activity, microbial ecology, as well as photosynthetic characteristics, N accumulation and yield of wheat plants. MethodsThe effects were examined through a soil column leaching experiment and a two-year field experiment conducted from 2020 to 2022. Five treatment groups were established, namely CK (CF), T1 (CF + 0.15% NCC), T2 (CF + 0.30% NCC), T3 (CF + 0.45% NCC), and T4 (CF + 0.60% NCC), to determine the optimal ratio of NCC to CF. ResultsAll NCC treatments resulted in reduced leaching of soluble N. Nevertheless, the 0.45% and 0.60% NCC treatments (T3 and T4) posed a risk of migration through water runoff, leading to nutrient losses. The simultaneous application of NCC and CF increased the content of soil nitrate N (NO3--N), as well as the activity levels of urease and protease during the anthesis stage. Additionally, it elevated the abundance of various microbial communities, including Actinobacteria, Acidobacteria, Chloroflexi, Cyanobacteria, and Nitrospirae. NCC application reduced the content of Na+/K+ and malondialdehyde (MDA), while elevating the activities of flag leaf superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), nitrate reductase (NR), and glutamine synthetase (GS). Additionally, it increased stomatal conductance (Gs) and net photosynthetic rate (Pn). The 0.30% NCC treatment showed significantly higher N accumulation in the stem, leaf, and spike compared to CK at the maturity stage. Moreover, 0.30% NCC treatment exhibited a significant increase in the number of grains per spike and higher yields compared to CK. In particular, 0.30% and 0.45% NCC treatments accomplished yield boosts of 29.6–34.7% and 19.9–26.0%, respectively. ConclusionNCC enhanced wheat plant N accumulation and yield through regulation of N release rate and increased soil N content, enzyme activity, and microbial community diversity and abundance. Additionally, NCC boosted flag leaf photosynthesis rate during the anthesis to maturity stage of wheat. This research highlights the potential of NCC to improve wheat crop production. And 0.30% NCC is the optimal application concentration for NCC.

Oxidative Stress-Mediated Repression of Virulence Gene Transcription and Biofilm Formation as Antibacterial Action of Cinnamomum burmannii Essential Oil on Staphylococcus aureus.

This work aimed to identify the chemical compounds of Cinnamomum burmannii leaf essential oil (CBLEO) and to unravel the antibacterial mechanism of CBLEO at the molecular level for developing antimicrobials. CBLEO had 37 volatile compounds with abundant borneol (28.40%) and showed good potential to control foodborne pathogens, of which Staphylococcus aureus had the greatest inhibition zone diameter (28.72 mm) with the lowest values of minimum inhibitory concentration (1.0 μg/mL) and bactericidal concentration (2.0 μg/mL). To unravel the antibacterial action of CBLEO on S. aureus, a dynamic exploration of antibacterial growth, material leakage, ROS formation, protein oxidation, cell morphology, and interaction with genome DNA was conducted on S. aureus exposed to CBLEO at different doses (1/2-2×MIC) and times (0-24 h), indicating that CBLEO acts as an inducer for ROS production and the oxidative stress of S. aureus. To highlight the antibacterial action of CBLEO on S. aureus at the molecular level, we performed a comparative association of ROS accumulation with some key virulence-related gene (sigB/agrA/sarA/icaA/cidA/rsbU) transcription, protease production, and biofilm formation in S. aureus subjected to CBLEO at different levels and times, revealing that CBLEO-induced oxidative stress caused transcript suppression of virulence regulators (RsbU and SigB) and its targeted genes, causing a protease level increase destined for the biofilm formation and growth inhibition of S. aureus, which may be a key bactericidal action. Our findings provide valuable information for studying the antibacterial mechanism of essential oil against pathogens.

The incorporation of sumac seed powder (Rhus coriaria L.) into the diet of quail breeders as a novel feed additive

A total of 150 adult quails, aged 8 wk, were divided into 5 groups to study the effect of sumac seed powder on reproductive and productive parameters, egg quality, digestive enzymes, and quail breeders' blood profiles. Dietary supplements containing sumac powder were formulated as follows: group 1 (G1) (control, only basal diet); group 2 (G2) (basal diet + 1 g sumac powder/kg diet); group 3 (G3) (basal diet + 2 g sumac powder/kg diet); group 4 (G4) (basal diet + 3 g sumac powder/kg diet); and group 5 (G5) (basal diet + 4 g sumac powder/kg diet). The feed conversion ratio was significantly higher at all levels of sumac powder (P < 0.05) compared to the control group (G1). Overall, during the study (8-16 wk), quail-fed 3 g sumac powder/kg diet (G4) showed no significant increase (P > 0.05) in the feed intake compared to the control group. Sumac powder supplementation significantly (P < 0.05) increased egg number, egg weight, egg mass, fertility, and hatchability. While supplementing with sumac powder did not impact other egg quality parameters, it did significantly (P < 0.05) increase yolk percentage, Haugh unit, and unit surface shell weight. Furthermore, when compared to the control group (G1), birds given 2, 3, or 4 g of sumac powder/kg diet showed a significant improvement (P < 0.05) in hematological parameters such as red blood cells, white blood cells, and hemoglobin, as well as a decrease in glucose levels. Feeding quail with a 3 g sumac powder/kg diet (G4) resulted in significantly (P < 0.05) higher globulin levels and improved albumin/globulin ratio compared to other treatments and control (G1). Sumac powder intake significantly (P < 0.05) reduced plasma lipid profile, liver enzymes (aspartate aminotransferase, and alanine aminotransferase), and kidney functions (creatinine, and urea). Furthermore, the supplementation of sumac powder resulted in a substantial increase (P < 0.05) in the levels of amylase, lipase, and protease. Sumac powder administration also significantly (P < 0.05) improves immunity by boosting IgM, IgG, IgA, and lysozyme levels in quail breeders' plasma. Supplementing with sumac powder, on the other hand, increased levels of reduced glutathione, total antioxidant capacity, catalase, and superoxide dismutase. The results of the current study indicated that the addition of 1, 2, 3, and 4 g of sumac powder to the diet of Japanese quail breeders led to improvements in egg quality, digestive enzymes, reproductive and productive performances, and most blood hematological and biochemical parameters.

Platinum-Loaded Cerium Oxide Capable of Repairing Neuronal Homeostasis for Cerebral Ischemia-Reperfusion Injury Therapy.

Effective neuroprotective agents are required to prevent neurological damage caused by reactive oxygen species (ROS) generated by cerebral ischemia-reperfusion injury (CIRI) following an acute ischemic stroke. Herein, it is aimed to develop the neuroprotective agents of cerium oxide loaded with platinum clusters engineered modifications (Ptn-CeO2). The density functional theory calculations show that Ptn-CeO2 could effectively scavenge ROS, including hydroxyl radicals (·OH) and superoxide anions (·O2 -). In addition, Ptn-CeO2 exhibits the superoxide dismutase- and catalase-like enzyme activities, which is capable of scavenging hydrogen peroxide (H2O2). The in vitro studies show that Ptn-CeO2 could adjust the restoration of the mitochondrial metabolism to ROS homeostasis, rebalance cytokines, and feature high biocompatibility. The studies in mice CIRI demonstrate that Ptn-CeO2 could also restore cytokine levels, reduce cysteine aspartate-specific protease (cleaved Caspase 3) levels, and induce the polarization of microglia to M2-type macrophages, thus inhibiting the inflammatory responses. As a result, Ptn-CeO2 inhibits the reperfusion-induced neuronal apoptosis, relieves the infarct volume, reduces the neurological severity score, and improves cognitive function. Overall, these findings suggest that the prominent neuroprotective effect of the engineered Ptn-CeO2 has a significant neuroprotective effect and provides a potential therapeutic alternative for CIRI.

Galactinol Regulates JA Biosynthesis to Enhance Tomato Cold Tolerance.

Low temperatures can inhibit plant growth and development and reduce fruit yield. This study demonstrated that the expression of AnGolS1 from Ammopiptanthus nanus (A. nanus) encoding a galactinol synthase enhanced tomato cold tolerance. In AnGolS1-overexpressing plants, the jasmonic acid (JA) biosynthesis substrates 13-hydroperoxylinolenicacid and 12,13-epoxylinolenicacid were significantly accumulated, and the expression levels of the ethylene response factor (SlERF4-7) and serine protease inhibitor (SlSPI5) were increased. We speculated that there may be correlations among galactinol, ethylene signaling, the protease inhibitor, protease, and JA levels. The expression levels of SlERF4-7 and SlSPI5 as well as the JA content were significantly increased under exogenous galactinol treatment. Additionally, the expression of SlSPI5 was reduced in SlERF4-7-silenced plants, and SlERF4-7 was confirmed to bind to the dehydration-responsive element (DRE) of the SlSPI5 promoter. These results suggest that SlSPI5 is a target gene of the SlERF4-7 transcription factor. In addition, SlSPI5 interacted with cysteine protease (SlCPase), while SlCPase interacted with lipoxygenase (SlLOX5) and allene oxide synthase (SlAOS2). When SlCPase was silenced, JA levels increased and plant cold tolerance was enhanced. Therefore, galactinol regulates JA biosynthesis to enhance tomato cold tolerance through the SlERF4-7-SlSPI5-SlCPase-SlLOX5/SlAOS2 model. Overall, our study provides new perspectives on the role of galactinol in the JA regulatory network in plant adaptation to low-temperature stress.

Sweet potato (Ipomoea Batatas (L.) Lam.): A study on physiological and biochemical properties

This study investigates the physiological and biochemical properties of different sweet potato (Ipomoea batatas (L.) Lam.) varieties grown in the Samarkand soil-climatic conditions and assesses the impact of these varieties on soil enzyme activities. The red, yellow, and white varieties were evaluated for their carbohydrate content, vitamin levels, antioxidant activities, growth parameters, root system development, and enzyme activities, including protease, amylase, and cellulase. Biochemical analysis revealed that the red variety had the highest carbohydrate content (25%) with significant levels of sucrose, glucose, and fructose. It also exhibited the highest vitamin C (30 mg/100g) and vitamin A (8 mg/100g) concentrations as determined by High-Performance Liquid Chromatography (HPLC). Antioxidant activity, measured using the DPPH assay, was highest in the red variety (85% scavenging activity). Physiological analysis showed that the red variety had superior growth parameters with a height of 35 cm, root length of 20 cm, and biomass of 150 g. It also demonstrated the most developed root system with a root weight of 50 g and root volume of 40 cm³. Enzyme activity assays indicated that the red variety had the highest levels of protease (85 U/mg), amylase (70 U/mg), and cellulase (65 U/mg). The study highlights the significant variations among sweet potato varieties in terms of their nutritional and health-promoting characteristics. The red variety emerged as the most beneficial, exhibiting superior nutritional content, antioxidant activity, and enzyme activities that enhance soil health. These findings underscore the potential of red sweet potatoes in promoting sustainable agriculture and improving food security in regions with challenging environmental conditions.

Comparison of thrombomodulin, vWF, and ADAMTS13 levels between preeclampsia and normal pregnancy

To determine and compare plasma thrombomodulin, von Willebrand factor and von Willebrand factorcleaving protease levels between pre-eclamptic and healthy pregnant females. The cross-sectional, comparative study was conducted at the Department of Haematology, University of Health Sciences, Lahore, Pakistan, from November 2019 to December 2020, and comprised pregnant females who were divided into healthy pregnant group A and pre-eclamptic group B. Plasma thrombomodulin and von Willebrand factor-cleaving protease levels were determined by using commercially available enzyme-linked immunosorbent assay kit, and von Willebrand factor level was determined by using immuno-turbidimetric assay kit. Data was analysed using SPSS 25. Of the 88 participants, there were 44(50%) females with mean age 25.5±6 years in group A and 44(50%) in group B with mean age 26±5 years. Median thrombomodulin level in group B was significantly higher than group A (p=0.003). Median von Willebrand factor-cleaving protease levels were lower in group B compared to group A (p=0.838). A significant difference in von Willebrand factor level was observed between the groups (p=0.038). Females with pre-eclampsia had significantly higher plasma levels of von Willebrand factor and thrombomodulin than healthy pregnant subjects.

Effects of dietary common juniper (Juniperus communis) essential oil on growth, immunity, antioxidant status, and disease resistance in rainbow trout, Oncorhynchus mykiss

The main aim of this study is to evaluate the effects of common juniper (Juniperus communis) essential oil (JCEO) on growth, digestive enzymes, biochemical profile, immune and antioxidant system, and liver enzymes in the rainbow trout, Oncorhynchus mykiss. 240 fish (10.63 ± 0.06 g) were fed for 60 days in 12 thanks (20 fish per tank) with experimental diets containing JCEO including: control or T1 (untreated), T2 (fed 0.5% JCEO), T3 (1% JCEO), T4 (2% JCEO). Results revealed an increase in final weight (FW) and weight gain (WG) in all treatments, with a significant difference with the control group (P < 0.05). Feed conversion ratio (FCR) decreased, after addition of JCEO compared to the control group (P < 0.05). Amylase, protease and lipase activity levels increased in all groups (T2, T3 and T4 groups), which were significantly different from the control (P < 0.05). The T2 group had the highest total protein (TP) level in serum compared to the control, and all experimental treatments exhibited a significant difference with the control (P < 0.05). The activity of antioxidant enzymes in serum, superoxide dismutase (SOD) and catalase (CAT) increased compared to the control group, with the highest levels in the T4 group. Inclusion of JCEO to the diet significantly reduced the levels of alkaline phosphatase (ALP) and alanine aminotransferase (ALT) activity in blood compared to the control (P < 0.05). The highest level of serum lysozyme activity and total immunoglobulin (total Ig) were observed in T3 group, which displayed a significant difference with the control (P < 0.05). Serum alternative complement activity (ACH50) increased in the supplemented fish compared to the control (P < 0.05). The fish survival increased after challenge with Yersinia ruckeri in JCEO-supplemented fish. In conclusion, JCEO at dietary level of 1% could improve the nutrient digestion and absorption and provide better growth, as well as reinforce the immune system in rainbow trout.

A modified protocol for studying filaggrin degradation using a reconstructed human epidermis model under low and high humidity.

Filaggrin (FLG) is an essential protein that plays a vital role in maintaining skin barrier function and moisture levels, allowing the skin to adapt to dry environments. However, the precise temporal dynamics of FLG metabolism in the human epidermis remain poorly understood, and suitable tools to study these time-dependent effects are currently lacking. To investigate the molecular mechanisms and time course of FLG metabolism and skin barrier function under high- and low-humidity conditions, utilizing a reconstructed epidermis model. EpiSkin specimens cultured under humid or dry conditions for varying durations (2-48 h) were compared by assessing FLG degradation and skin barrier formation using immunofluorescence staining and western blotting. Under conditions of low humidity, the proteolysis of FLG in EpiSkin increased between 4 and 12 h and was accompanied by elevated levels of cysteine-aspartic protease (caspase)-14. The expression of peptidyl arginine deiminase 1 and calpain 1 also increased at 4 h. However, after 24 h, the expression of these three FLG-degrading proteins significantly decreased. Conversely, the levels of pyrrolidone-5-carboxylic acid and urocanic acid initially decreased at 2 h and then increased between 12 and 24 h. Additionally, the expression of skin barrier proteins, such as FLG, transglutaminase 5, loricrin and zonula occludens-1, decreased starting from 12 h. Notably, epidermal cell viability and activity were also inhibited. We propose a reliable and ethical model to study the temporal dynamics of FLG metabolism and its role in skin barrier function. Using a commercially reconstructed epidermis to mimic dry skin formation obviates the need for animal and human testing.

Micafungin exerts antitumor effect on breast cancer and osteosarcoma through preventing EMT in tumor cells in an USP7/AKT/GSK-3β pathway-dependent manner.

Breast cancer and osteosarcoma are common cancers in women and children, respectively, but ideal drugs for treating patients with breast cancer or osteosarcoma remain to be found. Micafungin is an antifungal drug with antitumor activity on leukemia. Based on the notion of drug repurposing, this study aims to evaluate the antitumor effects of micafungin on breast cancer and osteosarcoma in vitro and in vivo, and to elucidate the underlying mechanisms. Five breast cancer cell lines (MDA-MB-231, BT-549, SK-BR-3, MCF-7, and 4T1) and one osteosarcoma cell line (143B) were chosen for the in vitro studies. Micafungin exerted an inhibitory effect on the viability of all cell lines, and MCF-7 cells were most sensitive to micafungin among the breast cancer cell lines. In addition, micafungin showed an inhibitory effect on the proliferation, clone formation, and migration in MCF7 and 143B cells. The inhibitory effect of micafungin on the growth of breast cancer and osteosarcoma was further confirmed with xenograft tumor mouse models. To explore the underlying mechanisms, the effect of micafungin on epithelial-mesenchymal transition (EMT) was examined. As expected, the levels of matrix metalloproteinase 9 and vimentin in MCF-7 and 143B cells were notably reduced in the presence of micafungin, concomitant with the decreased levels of ubiquitin-specific protease 7 (USP7), p-AKT, and p-GSK-3β. Based on these observations, we conclude that micafungin exerts antitumor effect on breast cancer and osteosarcoma through preventing EMT in an USP7/AKT/GSK-3β pathway-dependent manner.