Phosphatase Activity Research Articles

Faba bean-wheat intercropping controls the occurrence of faba bean Fusarium wilt by improving the microecological environment of rhizosphere soil

BackgroundFusarium wilt is a severe soil-borne disease that affects faba bean production. Faba bean-wheat intercropping is often used to control the occurrence of Fusarium wilt in faba bean. AimsTo evaluate the effects of faba bean-wheat intercropping on the occurrence of faba bean Fusarium wilt and soil microecology. MethodsWe established two planting patterns, faba bean monocropping (M) and faba bean-wheat intercropping (I), to investigate Fusarium wilt occurrence and plant dry weight and assess changes in soil enzyme activities, microbial diversity, and community composition during different stages of disease onset. ResultsIntercropping effectively controlled faba bean Fusarium wilt at the three disease stages and increased the dry weight of faba bean plants. Intercropping promoted the activities of catalase (CAT), urease, sucrase, and acid phosphatase in the rhizosphere soil of faba bean at three disease stages. Bacterial and fungal diversity decreased with disease progression, and intercropping mitigated this trend. Compared with monocropping, intercropping increased the abundance of beneficial bacteria such as Proteobacteria, Actinobacteriota, Gemmatimonadota, Gemmatimonas, Conexibacter, and Sphingomonas, while reducing the abundance of pathogenic fungi such as Alternaria, Cladosporium, and Fusarium. Intercropping also increased the abundance of arbuscular mycorrhiza, soil saprophytes, and undefined saprophytes while decreasing the abundance of plant pathogens. ConclusionFaba bean-wheat intercropping enhanced soil enzyme activities, effective nutrient content, and alpha diversity indices of bacteria and fungi in the rhizosphere soil of faba bean, while promoting the abundance of beneficial bacteria, arbuscular mycorrhizal fungi, as well as both soil and undefined humus. Simultaneously, intercropping reduced the abundance of plant pathogens, facilitated nutrient cycling in the soil, provided sufficient nutrients for crop uptake, and mitigated the toxic effects of hydrogen peroxide on cells. Ultimately, this resulted in a reduced occurrence of Fusarium wilt.

Development of a Bioactive Titanium Surface via Alkalinization and Naringenin Coating for Peri-Implant Repair: In Vitro Study

This study assessed the effects of titanium (Ti) surface modification with sodium hydroxide (NaOH) associated or not with Naringenin (NA) citrus flavonoid-coating on osteoblastic-like cells (Ob) metabolism. Ti discs were submitted to alkalinization by NaOH solution (5 M, 60 °C) for 24 h; then, the discs were impregnated or not with 100 µg/mL of NA and dried for 1 h at room temperature. The chemical composition, surface topography, and NA release were evaluated. For the biological assays, the discs were placed on 24-well cell culture plates and Ob (Saos-2; ATCC HTB-85) was seeded onto the discs. After different periods, cell adhesion and viability, alkaline phosphatase activity (ALP), and mineralized nodules deposition (MND) were assessed. In addition, cells stimulated with tumor necrosis factor-alpha (TNF-α) were submitted to matrix metalloproteinase (MMP)-2 synthesis and ALP gene expression assessment. Since data presented normal distribution and homogeneity (Shapiro-Wilk; Levene), Student’s t-test or one-way ANOVA/post-hoc tests were selected for data analysis (α = 0.05). Higher roughness was observed on Ti discs submitted to NaOH treatment, while the chemical and NA release evaluations indicated the successful adsorption of NA to alkali-treated Ti surface. Higher cell adhesion, cell viability (after 7 days of culture), ALP activity, and MND were observed on Ti NaOH coated with NA compared to the control group (Ti NaOH) (p < 0.05). Moreover, NA coating also promoted decreased MMP-2 synthesis and increased ALP gene expression in the presence of the inflammatory stimulus TNF-α (p < 0.05). The modification of Ti disks with NaOH associated with NA-coating enhanced bone cell metabolism, suggesting that this type of surface modification has a promising potential to accelerate bone repair and formation around dental implants.

Soy Peptide Ameliorate TGF-β1-Mediated Osteoblast Differentiation through Smad and MAPK Signaling Pathways.

The aim of this study was to determine the osteogenic activity and mechanism of soybean peptide VVELLKAFEEKF (SOP) and the potential relationship between SOP and transforming growth factor-β1 (TGF-β1). The results show that SOP promotes MC3T3-E1 cell proliferation by altering cell progression. SOP induced cell differentiation and mineralization in a dose-dependent manner at 0.7-7 μM. Moreover, SOP stimulates osteoblast differentiation, which may be achieved through the activation of p38-MAPK and Smad2/3 signaling pathways. Furthermore, treatment with a TβRI inhibitor (SB525334) inhibited the phosphorylation levels of p38 and Smad2/3, which indicates the involvement of TβRI in the process of osteoblast differentiation caused by SOP. Besides, in non-FBS-cultured MC3T3-E1 cells, SOP and TGF-β1 promoted the phosphorylation of Smad2/3 and alkaline phosphatase (ALP) activity, but the effect was lost when SOP was incubated separately, indicating that SOP stimulated osteoblast differentiation by promoting TGF-β1 activity. In vivo, SOP significantly restores bone mineral density loss and behavioral deficits in a model of glucocorticoid-induced osteoporosis (GIOP) in zebrafish. These results suggest that SOP may have the function of promoting bone remodeling and may be used as a potential active factor for functional food development to prevent osteoporosis.

Promotion of bone osteogenic and regeneration by resveratrol-derived carbon dots in inflammatory environments via a one-step hydrothermal method

In the context of bone fractures, local inflammation complicates the healing process, adversely affecting the recovery of bone defects. This inflammatory response impedes the regenerative potential of stem cells and induces the differentiation of macrophages into the M1 type, thereby inhibiting bone regeneration. To address this issue, we synthesized carbon dots (CDs) using resveratrol and citric acid. These CDs demonstrated excellent water solubility and retained the anti-inflammatory properties of resveratrol. In an inflammatory environment co-cultured with resveratrol carbon dots (RES-CDs), we observed a reduction in inflammatory factors such as iNOS and an increase in anti-inflammatory factors like CD206. Alkaline phosphatase (ALP) activity, Alizarin Red staining, and immunofluorescence assays indicated that MC3T3 cells produced more calcium nodules after co-culture with RES-CDs, along with enhanced expression of bone formation factors BMP-2 and ALP. Additionally, real-time polymerase chain reaction (qRT-PCR) and immunofluorescence analysis demonstrated that RES-CDs could induce bone tissue proliferation and differentiation by upregulating ALP, BMP-2, OCN, and OPN. In a rat cranial defect model, RES-CDs promoted tissue healing and bone proliferation at weeks 1 and 4 post-surgery. These findings underscore the potential of resveratrol carbon dots in promoting bone tissue healing while mitigating inflammation, thereby offering a promising approach for nanocomposite materials in tissue repair.

Flow cytometry FRET reveals post-translational modifications drive Protein Phosphatase-5 conformational changes in mammalian cells.

The serine/threonine Protein Phosphatase-5 (PP5) plays an essential role in regulating hormone and stress-induced signaling networks as well as extrinsic apoptotic pathways in cells. Unlike other Protein Phosphatases, PP5 possesses both regulatory and catalytic domains, and its function is further modulated through post-translational modifications (PTMs). PP5 contains a tetratricopeptide repeat (TPR) domain, which usually inhibits its phosphatase activity by blocking the active site (closed conformation). Certain activators bind to the PP5-TPR domain, alleviating this inhibition and allowing the catalytic domain to adopt an active (open) conformation. While this mechanism has been proposed based on structural and biophysical studies, PP5 conformational changes and activity have yet to be observed in cells. Here, we designed and developed a flow cytometry-based fluorescence resonance energy transfer (FC-FRET) method, enabling real-time observation of PP5 autoinhibition and activation within live mammalian cells. By quantifying FRET efficiency using sensitized emission, we established a standardized and adaptable data acquisition workflow. Our findings revealed that, in a cellular context, PP5 exists in multiple conformational states, none of which alone fully predicts its activity. Additionally, we have demonstrated that PTMs such as phosphorylation and SUMOylation impact PP5 conformational changes, representing a significant advancement in our understanding of its regulatory mechanisms.

Soil function-microbial diversity relationship is impacted by plant functional groups under climate change

Understanding the interactions between plant and soil microbial diversity is crucial for predicting ecosystem responses to environmental changes. While the individual roles of plant and microbial diversity in driving ecosystem functions have been widely investigated, their interplay especially under stress conditions remains largely underexplored. This study investigated how interactions between plant and microbial diversity affect key soil functions during and after drought. We simultaneously manipulated soil microbial diversity and plant species richness, while also considering the influence of plant functional groups (PFGs), to investigate their interactions and effects on key soil functions. Our results revealed independent and interactive effects of plant and microbial diversity in shaping soil functions. Microbial diversity loss significantly altered microbial community structure and impacted microbially-driven soil N and P pools and processes such as N-mineralization. These effects were modulated by plant species richness and varied across different PFGs. The relative influence of plant and microbial diversity on soil functions was context-dependent. Microbial diversity showed stronger effects on specific functions, such as phosphatase activity, and under the drought condition. Plant diversity, particularly through PFGs (e.g. legumes), played an independent role in shaping the microbial-driven soil functions. These findings advance mechanistic insights and highlight the importance of considering both above- and belowground biodiversity, along with their interactions, in shaping soil functions and ecosystem resilience, particularly under environmental stress. Further, it emphasizes the need to explicitly consider PFGs, along with above- and belowground biodiversity, as a strategy for preserving essential belowground functions in the face of ongoing environmental changes.

Safety assessment of three common food additives: the reproductive, oxidative and enzymatic perspective

This investigation was designed to evaluate the reproductive toxicity induced by potassium bromate (at doses of 50, 40, 30, 20, and 10 mg/kg), sodium benzoate (at doses of 5, 4, 3, 2, and 1 mg/kg), and caramel IV (at a dose of 200 mg/kg for 1, 2, 3, 4, and 5 weeks) in male albino mice through the assessments of sperm abnormality and sperm count. The concentrations of the three additives tested were chosen purposefully based on the acceptable daily intake of each of the additives by standard organizations. The level of Total Testosterone (TT), Luteinizing Hormone (LH), and Follicle Stimulating Hormone (FSH), as well as the activities of catalase (CAT), superoxide dismutase (SOD), alkaline phosphatase (ALP), aspartate aminotransferase (AST), and alanine aminotransferase (ALT), were also measured. The results obtained showed that all three tested food additives led to a statistically significant (p < 0.05) increase in abnormal sperm cells among exposed mice. The sperm abnormalities observed include folded sperm, amorphous heads, distal droplets, absent hooks, improper tail attachments, and looped tails. Furthermore, it was observed that only mice exposed to potassium bromate and caramel exhibited a statistically significant decrease in mean sperm count. Alterations in the levels and activities of TT, ALT, FSH, AST, LH, and ALP were also observed to be concentration-dependent in the exposed mice. The findings suggest that the reproductive toxicity and changes in enzyme activities in the treated mice are dependent on the concentration of potassium bromate and sodium benzoate, as well as the duration of exposure to caramel IV. These indicate that high consumption of foods or beverages containing potassium bromate, sodium benzoate, and caramel IV colourant may induce reproductive abnormalities in the consumers.

Role of in-situ electrical stimulation on early-stage mineralization and in-vitro osteogenesis of electroactive bioactive glass composites

Bioactive glass (BAG) has emerged as an effective bone graft substitute due to its diverse qualities of biocompatibility, bioactivity, osteoblast adhesion and enhanced revascularization. However, inferior osteogenic capacity of BAG compared to autologous bone grafts continues to limiting it's wide-spread clinical applications towards repairing of bone fractures and healing. In this study, we have fabricated BAG composites with 0.5 to 2 wt% bismuth ferrite (BF, a multiferroic material) with an aim to generate in-situ electrical charges pertinent to early-stage bone regeneration thus mimicking nature bone, which is a piezoelectric material. The fabricated BAG composites were characterised in terms of microstructures, phase analysis, remanent polarization, wettability and subsequently evaluated for in vitro cell proliferation and osteogenesis with and without magnetic field exposure (200 mT, 30 min./day). Pre-osteoblast cells from mice (MC3T3-E1) seeded on these composites exhibited excellent cell growth without any cytotoxicity, which is further supported by FITC/DAPI staining and a live/dead assay. The results of Alizarin Red S assay and increased levels of Alkaline Phosphatase (ALP) activity, at 21 days of culture, suggest that the BAG-BF composites promote in vitro osteogenic differentiation of pre-osteoblast cells. The enhanced osteogenesis of BAG-BF composites was also confirmed through qRT-PCR analysis, which showed rapid upregulation of osteoblastogenic specific genes namely RunX-2, Collagen-1, Bone Sialo Protein, and ALP after 21 days. Additionally, the osteogenic differentiation was assessed by the Western Blot technique, which revealed significantly higher band intensity of osteogenic markers in BAG-1.5 BF and BAG-2 BF composites than pure BAG. These findings clearly demonstrate that in-situ electrical stimulation and osteoconductive capacity of BF reinforced BAG composites have positive impact on osteoblast cell development, bone formation, and healing.

Nutritional Innovation Using Green Seaweed (Ulva sp.) and Garlic Powder Extracts for White-Leg Shrimp (Litopenaeus vannamei) Challenged by Vibrio harveyi.

This study aimed to determine the antimicrobial effects of ethanolic extracts of Ulva sp. and garlic (Allium sativum) powder ethanolic extracts against Vibrio harveyi in vitro. The stimulatory effects of Ulva sp. extract (UE) and garlic powder extract (GPE) on the growth performance and innate immune responses of white-leg shrimp, Litopenaeus vannamei, and their challenge against V. harveyi infection were also investigated. A commercial shrimp diet (36.1% protein) was enriched with 0.5, 1.0 and 2.0gUE/kg diet and 2, 4 and 6gGPE/kg diet, whereas the control group was free of any supplement. Health juveniles of L. vannamei (average weight 2-3g) were distributed in 21 fiberglass reinforced plastic (FRP) tanks (500-L capacity) at a stocking density of 300 animals/tank to represent each treatment in triplicate. The animals were fed ad libitum on the experimental diets up to satiety four times daily for 60 days. The phytochemical analysis of ethanolic extracts of Ulva sp. and garlic powder evoked their richness of several bioactive compounds showing significant antibacterial activity against V. harveyi. The GPE exhibited a higher inhibition zone than that of the UE. The supplemented diets did not significantly affect weight gain %, final weight, feed conversion ratio, specific growth rate and survival rates of white shrimp compared to those fed on the control diet. Significant increases were observed in total haemocyte count, phagocytosis and phagocytic index of all treatments compared with the control group. There were significant increases in serum total protein, acid phosphatase activity, alkaline phosphatase, lysosomal enzyme activity, phenoloxidase activity and superoxide dismutase activity with offered diets with increasing the levels of ethanolic extracts of Ulva sp. and garlic powder up to 2.0gUE/kg diet and 6gGPE/kg diet, respectively. The ethanolic extraction of Ulva sp. and garlic powder-supplemented diet groups, particularly at treatments of 2.0 and 6gGPE/kg diet, respectively, significantly reduced the shrimp mortality induced by V. harveyi infection when compared with the control group. The net results evoked that ethanolic extraction of Ulva sp. (2.0gUE/kg) and garlic powder (6gGPE/kg diet) enhanced the immune response and disease resistance of the white-leg shrimp, L. vannamei. It is also noted that the GPE is more efficient than the UE in vitro and in vivo investigations.

Intercropping of tobacco and maize at seedling stage promotes crop growth through manipulating rhizosphere microenvironment.

Changes in the rhizosphere microbiome and metabolites resulting from crop intercropping can significantly enhance crop growth. While there has been an increasing number of studies on various crop combinations, research on the intercropping of tobacco and maize at seedling stage remains limited. This study is the first to explore rhizosphere effects of intercropping between tobacco and maize seedling stages, we analyzed the nitrogen, phosphorus and potassium nutrients in the soil, and revealed the important effects on soil microbial community composition and metabolite profiles, thereby regulating crop growth and improving soil balance. Compared with mono-cropping, intercropping increased the biomass of the two crops and promoted the nutrient absorption of nitrogen, phosphorus and potassium. Under intercropping conditions, the activities of sucrase, catalase and nitrate reductase in tobacco rhizosphere soil and the content of available potassium, the activities of nitrate reductase and acid phosphatase in maize rhizosphere soil were significantly increasing. Rhizosphere soil bacterial and fungal communities such as Sphingomonas, Massilia, Humicola and Penicillium respond differently to crop planting patterns, and soil dominant microbial communities are regulated by environmental factors such as pH, Organic Matter, Available Potassium, Nitrate Reductase, and Urease Enzyme. Network analysis showed that soil microbial communities were more complex after intercropping, and the reciprocal relationship between bacteria and fungi was enhanced. The difference of metabolites in soil between intercropping and monocropping system was mainly concentrated in galactose metabolism, starch and sucrose metabolism pathway, and the content of carbohydrate metabolites was significantly higher than that of monocropping soil. Key metabolites such as D-Sucrose, D-Fructose-6-Phosphate, D-Glucose-1-Phosphatel significantly influence the composition of dominant microbial communities such as Sphingomonas and Penicillium. This study explained the effects of intercropping between flue-cured tobacco and maize on the content of soil metabolites and soil microbial composition in rhizosphere soil, and deepened the understanding that intercropping system can improve the growth of flue-cured crops seedlings through rhizosphere effects.

Effects of Taurine and Enzymatic Cottonseed Protein Concentrate Supplementation in Low-Fishmeal Diet on Growth, Liver Antioxidant Capacity, and Intestinal Health of Golden Pompano (Trachinotus ovatus)

This study was conducted to investigate the impacts of the dietary addition of taurine and enzymatic cottonseed protein concentrate (ECPC) in low-fishmeal diet on the growth performance, plasma biochemical indices, hepatic antioxidant capacity, intestinal anti-inflammatory capacity, intestinal microflora, and muscle quality of golden pompano (Trachinotus ovatus). A total of three isonitrogenous diets were given to 225 golden pompanos (5.6 ± 0.14 g). They were randomly divided into nine cages (1.0 m × 1.0 m × 1.5 m; three cages per treatment) with equal stocking numbers of twenty-five fish per cage. The results indicated that the CSM-TC group significantly increased the growth performance of juvenile T. ovatus (p &lt; 0.05). The results indicated that compared with other groups, the addition of 1% ECPC and 0.25% taurine has been found to enhance the WGR (weight gain rate), SGR (specific growth rate), and CF (condition factor). Compared with other groups, the relative expressions of GH, GHR1, GHR2, IGF1, IGF2, and MyoG were significantly higher in fish fed with CSM-TC. The results showed that CSM-TC significantly increased the activities of alkaline phosphatase, complement 3, and complement 4 enzymes (p &lt; 0.05). The results showed that dietary CSM-TC increased the activities of hepatic superoxide dismutase and total antioxidant capacity enzymes. Compared with other groups, the hepatic relative expressions of Nrf2, HO-1, and GSH-Px were significantly higher in fish fed with CSM-TC. The results showed that dietary CSM-TC increased the activities of intestinal chymotrypsin, lipase, and α-amylase enzymes. A CSM-TC diet significantly increased the relative expressions of IL-10, ZO-1, Occludin, Claudin-3, and Claudin-15 (p &lt; 0.05). The results showed that CSM-C significantly increased the index of Ace and Chao1 (p &lt; 0.05). In conclusion, a high-fermented cottonseed meal diet can have detrimental effects on physiological health in golden pompano, while adding 1% ECPC and 0.25% taurine can improve hepatic and intestinal health via attenuating inflammation and oxidative stress.

Exploring the active components and potential mechanisms of Rhodiola heterodonta for alleviating T2DM based on network pharmacology

Type 2 diabetes mellitus (T2DM), a metabolic disease that poses a serious threat to human health, is characterized by sustained hyperglycemia due to dysregulated insulin secretion. Rhodiola heterodonta (R. heterodonta) is an edible plant, and its root is a popular herbal tea for lowering blood sugar in Tajikistan's folk remedy. However, R. heterodonta has not received much attention outside of the former Soviet Union. Furthermore, the hypoglycemic components of R. heterodonta and their underlying mechanisms are poorly understood. In this study, we extracted R. heterodonta roots and obtained R. heterodonta root extract (RHRE) through preliminary purification. RHRE significantly inhibited the activities of α-glucosidase (α-GAA) and protein tyrosine phosphatase 1B (PTP1B), thus highlighting its powerful hypoglycemic effect. Twenty-six compounds were identified in RHRE by LC-MS/MS analysis, with flavonoids and phenolic acids existing as the predominant components. Network pharmacology further predicted that the PI3K/AKT pathway is crucial for the anti-T2DM effects of RHRE, and molecular docking suggested the significant binding affinities between the key components of RHRE and PTP1B. Importantly, RHRE exerted its hypoglycemic effect by modulating the PTP1B/PI3K/AKT/GSK3β pathway in insulin-resistance L6 and L02 cells. Additionally, RHRE exhibited antioxidant activity and potent DPPH and ABTS radical scavenging abilities. Ferroptosis, an emerging modality of cell death driven by ROS-induced lipid peroxidation, is considerably linked to the progression of T2DM. RHRE alleviated oxidative stress and inhibited ferroptosis through the activation of the Nrf2/HO-1/SLC7A11/GPX4 axis and the downregulation of ACSL4 expression. In conclusion, our study indicates that RHRE is a potential source of antioxidant and hypoglycemic bioactive compounds and can be used for the development of health products and functional foods for the prevention of T2DM.

Proteomics analysis reveals the differential protein expression of female and male adult Toxocara canis using Orbitrap Astral analyzer.

Toxocara canis, the most prevalent helminth in dogs and other canines, is one of the socioeconomically important zoonotic parasites, particularly affecting pediatric and adolescent populations in impoverished communities. However, limited information is available regarding the proteomes of female and male adult T. canis. To address this knowledge gap, we performed a comprehensive proteomic analysis to identify the proteins with differential abundance (PDAs) and gender-specifically expressed proteins between the two sexes adult T. canis. The comparative proteomic analysis was carried out by the Orbitrap mass spectrometry (MS) with asymmetric track lossless (Astral) analyzer. The difference analysis was conducted using t-test and the proteins verification was achieved through parallel reaction monitoring (PRM). The potential biological functions of identified adult T. canis proteins and PDAs were predicted by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) databases. The domain, transcription factor and subcellular localization of the identified proteins and PDAs were analyzed by InterPro, AnimalTFDB 4.0 and Cell-mPLOC 2.0 databases, respectively. A total of 8565 somatic proteins of adult T. canis were identified. Compared to male adult, 682 up-regulated PDAs and 844 down-regulated PDAs were identified in female adult with P-values < 0.05 and |log2FC|> 1, including 139 proteins exclusively expressed in female and 272 proteins exclusively expressed in male. The GO annotation analysis using all PDAs revealed that the main biological processes, cellular components and molecular functions corresponded to aminoglycan metabolic process, extracellular region and protein tyrosine phosphatase activity, respectively. The KEGG analysis using all PDAs showed that the pathways were mainly associated with adipocytokine signaling pathway, proximal tubule bicarbonate reclamation and PPAR signaling pathway. This study reveals the differential protein expression between female and male adult T. canis, providing valuable resource for developing the novel intervention strategies against T. canis infection in humans and animals, especially from the perspective of sexual development and reproduction.

Phosphogypsum with Rice Cultivation Driven Saline-Alkali Soil Remediation Alters the Microbial Community Structure.

The improvement of saline-alkali land plays a key role in ensuring food security and promoting agricultural development. Saline soils modifies the response of the soil microbial community, but research is still limited. The effects of applying phosphogypsum with rice cultivation (PRC) on soil physicochemical properties and bacterial community in soda saline-alkali paddy fields in Songnen Plain, China were studied. The results showed that the PRC significantly improved the physicochemical properties of soil, significantly reduced the salinity, increased the utilization efficiency of carbon, nitrogen, and phosphorus, and significantly increased the activities of urease and phosphatase. The activities of urease and phosphatase were significantly correlated with the contents of total organic carbon and total carbon. A redundancy analysis showed that pH, AP, ESP, HCO3-, and Na+ were dominant factors in determining the bacterial community structure. The results showed that PRC could improve soil quality and enhance the ecosystem functionality of soda saline-alkali paddy fields by increasing nutrient content, stimulating soil enzyme activity, and regulating bacterial community improvement. After many years of PRC, the soda-alkali soil paddy field still develops continuously and healthily, which will provide a new idea for sustainable land use management and agricultural development.

Facilitating effects of plant extracts on soil health and replanted Panax ginseng growth in recession soil.

Plant extracts have been shown to be effective agricultural strategies for improving soil fertility and quality, and promoting plant growth in soil degradation remediation. The application of plant extracts improves the material cycle of soil microecology, such as the decomposition of nitrogen, phosphorus, and potassium, while increasing plant resistance. However, there is currently no experiment to demonstrate whether plant extracts have a promoting effect on the growth of ginseng and the mechanism of action. Pot experiments were carried out to investigate the effects of extracts, namely Rubia cordifolia (RC), Schisandra chinensis (SC), and Euphorbia humifusa (EH) on soil properties, enzyme activities, and plant physiological characteristics were evaluated. Results showed that compared with CK, plant extract-related treatments increased soil Organic carbon (OC), Available nitrogen (AN), Available phosphorus (AP) contents, and Soil urease activity. (S-UE), Soil sucrase activity (Soil sucrase), Soil acid phosphatase activity. (S-ACP). Meanwhile, plant extract-related treatments significantly increased plant physiological properties and TP (Total protein) content, and decreased the content of MDA (malondialdehyde) by 15.70% -36.59% and PRO (proline) by 30.13% -148.44%. Furthermore, plant extract-related treatments also significantly promote plant growth and reduce plant incidence, the fresh weight of ginseng increased by 27.80% -52.08%, ginseng root activity increased by 45.13% -90.07%, and ginseng incidence rate decreased by 20.00% -46.67%. Through correlation analysis between fresh weight of ginseng and root parameters and soil index, fresh weight is significantly positively correlated with root diameter, fiber root number, root activity, total protein (TP), catalytic activity (CAT) and superoxide dismutase activity (SOD), H, soil urea activity (S-UE), soil sucrose activity (S-SC), soil acid phosphate activity (S-ACP), and soil laccase activity (SL); The fresh weight was significantly negatively correlated with incidence rate, disease severity index, and malondialdehyde content (MDA). In summary, plant extract-related treatments improve soil quality and promote ginseng growth, further enhancing soil health and plant disease resistance. These findings provide new insights into ginseng cultivation and soil health management and highlight a new approach that can be applied to a wider range of agricultural practices and environmental sustainability.

7740 Adult-Onset Hypophosphatasia Presenting As Low Alkaline Phosphatase Level Following Denosumab Use For Osteoporosis

Abstract Disclosure: J. Ocampo Mascaro: None. P. Wax: None. M. Al Mukaddam: Grant Recipient; Self; Ipsen, Incyte Research Funding. Background: Hypophosphatasia (HPP) is a genetic disease characterized by low activity of the tissue non-specific alkaline phosphatase. It has a broad range of clinical phenotypes, with adults usually presenting with an incidental low alkaline phosphatase (ALP) finding. Differentiating this from other causes of low ALP is challenging, especially when treating osteoporosis, as antiresorptive therapy such as denosumab and bisphosphonates may cause low ALP and carry the potential of increasing risk of atypical femoral fractures in patients with undiagnosed HPP. Clinical Case: A 64-year-old woman with history of nephrolithiasis, hypercalciuria, and osteoporosis (screening DXA lowest T-score -3.3 in femoral neck (FN)) but no prior fractures or dental issues was seen in clinic. Initial bloodwork showed normal range calcium, 25-OH vitamin D, and intact PTH. Pre-treatment ALP was 42 IU/L (N 39-117). Work up for secondary causes of osteoporosis was negative. Three yearly doses of IV zoledronic acid resulted in +4.2% BMD increase in lumbar spine (LS) but no changes in FN. She had no side effects but was found to have a decrease in ALP to 33 IU/L. This normalized back to 42 IU/L during a three-year drug holiday. Follow up DXA noted significant decline in BMD, hence denosumab was initiated. ALP decreased back to the 30-35 IU/L range. Fasting Vitamin B6 level (off B6 supplements for 1 week) was 23.4 ng/ml (N 2.1-21). Urine phosphoethanolamine was mildly elevated at 5 mmol/mol creatinine (N ≤4). Genetic testing evidenced a pathogenic ALPL variant (Exon 6, c.575T&amp;gt;C (p.Met192Thr), heterozygous) compatible with HPP. She had tolerated three doses of denosumab by then with BMD improvement of 5.0% in LS but no changes in FN and no atypical femoral fracture. Denosumab was stopped. Abaloparatide was started for osteoporosis with close monitoring for worsening hypercalciuria. Conclusion: This case demonstrates that adult HPP patients may present with subtle findings and lower range of normal ALP and may tolerate short courses of antiresorptive agents. There is a need for better screening and diagnostic criteria for adult HPP patients whose ALP levels may not be initially low before initiating antiresorptive drugs for osteoporosis. The risk of adverse effects from therapy will likely vary significantly based on the clinical severity/presentation of HPP as some adults with lower ALP ranges might be more susceptible to developing atypical femur fractures (1). Balancing atypical femur fracture risk with the benefits of antiresorptive treatment in decreasing risk of osteoporotic fractures highlights the need for better risk stratification to prevent undertreatment of osteoporosis. Reference: 1. Warren AM, Ebeling PR, Grill V, Seeman E, Sztal-Mazer S. Bilateral atypical femoral fractures during denosumab therapy in a patient with adult-onset hypophosphatasia. Endocrinol Diabetes Metab Case Rep. 2021 Sep 1;2021:21-0096. Presentation: 6/2/2024

12472 Clinical Features Of Adult Hypophosphatasia And Correlation Between Laboratory Findings And Patient’s Symptomatology: A Retrospective Review At The Penn Bone Center

Abstract Disclosure: J. Ocampo Mascaro: None. R. Tran: None. S. Kallish: Consulting Fee; Self; Sanofi, Amicus. Research Investigator; Self; Ipsen, Sanofi, Incyte, Idorsia. Speaker; Self; Sanofi, Amicus. J. Berman: None. M. Al Mukaddam: Grant Recipient; Self; Ipsen, Incyte Research Funding. Hypophosphatasia (HPP) is an inborn error of metabolism resulting in low activity of the enzyme tissue non-specific alkaline phosphatase (ALP). It primarily involves bone and teeth mineralization. HPP has a wide clinical spectrum. It may present with rickets, fractures, failure to thrive, weakness, respiratory complications, and seizures in its more severe perinatal/infantile forms. Childhood forms tend to be milder but may have substantial musculoskeletal complications and early teeth loss. The clinical picture in adults is extremely variable too. It is usually mild with stress fractures and/or teeth loss in most individuals but can be debilitating in others due to musculoskeletal pain and recurrent fractures. This retrospective study describes the characteristics of 35 adult patients (ages 22-74 years) with HPP based on clinical presentation +/- molecular testing at the Penn Bone Center, located at a US-based large academic hospital. Out of 35 patients, 26 had an ALPL variant in genetic testing (12 pathogenic, 5 likely pathogenic, and 9 of uncertain significance), 3 were negative, and 6 did not undergo testing yet. ALP values ranged from 11 to 39 U/L (N: 38-126). Serum fasting vitamin B6 levels (off supplements) ranged from normal range to 18 times the upper limit of normal. Main symptoms reported were fractures (74%), arthralgias/myalgias (51%), diffuse bone pain (29%), atraumatic teeth loss (20%), muscle weakness (14%), and nephrolithiasis (9%). We found no significant negative correlation between ALP levels and number of symptoms (rs -0.026) or age of first symptom occurrence (rs 0.205). We also found no significant positive correlation between B6 elevation and number of symptoms (rs 0.236) or age of first symptom occurrence (rs -0.175). Six patients within our cohort had previous or current treatment with asfotase alfa. This is an FDA-approved therapy for the treatment of perinatal, infantile, and childhood HPP and has been used in adult patients if one of the presenting symptoms was before age 18 years. Response to therapy in our patient cohort was mixed. Three out of six patients discontinued treatment due to lack of clinical benefit. Continued research of the clinical characteristics of adult HPP is needed to better understand its natural history, markers of severity of disease, and how to determine who may potentially benefit from asfostase alfa therapy. Presentation: 6/1/2024

Effects of silybin on growth performance, antioxidant capacity and immunity in juvenile hybrid grouper (Epinephelus fuscoguttatus ♀ × Epinephelus lanceolatus ♂) fed with high-lipid diets

This research examined the impacts of varying silybin dosages in high-lipid diets on the growth, hepatic histology, immunity, and immune-related gene expression of juvenile hybrid grouper (Epinephelus fuscoguttatus ♀ × Epinephelus lanceolatus ♂). The grouper (with an initial body weight of 8.27 ± 0.08 g) were fed diets containing silybin at levels of 0 g/kg (S1, control group), 0.05 g/kg (S2), 0.10 g/kg (S3), 0.15 g/kg (S4), 0.20 g/kg (S5), 0.25 g/kg (S6), and 0.50 g/kg (S7) for 8 weeks. The study results suggest that the silybin-treated groups displayed an initial increase followed by a decrease in final body weight and specific growth rate, with the highest value observed in the group S5. In serum samples, silybin supplementation of high-lipid diets resulted in increased activities of alkaline phosphatase (AKP), superoxide dismutase (SOD) and catalase (CAT) enzymes, decreased activities of aspartate transaminase (AST) and alanine transaminase (ALT) enzymes and increased total antioxidant capacity (T-AOC) content. In liver, the addition of silybin to high-lipid diets increased SOD, CAT, and lysozyme (LYZ) enzyme activities, increased T-AOC and immunoglobulin M (IgM) contents, and decreased reactive oxygen species (ROS) and malondialdehyde (MDA) contents. Contrasted with the control group, the addition of silybin at 0.05–0.50 g/kg enhanced the hepatic histomorphology, for example, ameliorating the indistinctness of cell outlines and diminishing the hepatocyte vacuolation. Silybin treatment significantly upregulated the expression of liver sod, cat, gpx, nrf2, keap1, hsp70, hsp90, tlr22, myd88, il-1β, tnf-α and il-6(P < 0.05). Silybin treatment significantly upregulated the expression of tlr22, myd88, il-1β, tnf-α, and il-6 of head kidney (P < 0.05). After vibrio harveyi challenge, groups S5-S6 showed higher survival than the control. Incorporating silybin into high-lipid diets boosts grouper's growth, antioxidant, and immune abilities. Regression analysis implies adding 0.23 g/kg silybin to the diets of juvenile hybrid grouper.

8711 The Effect of Testosterone Therapy Upon Bone Remodelling in Testosterone Deficient APOE-/- Mice Fed a High Fat Diet

Abstract Disclosure: A. Williamson: None. L. Bateman: None. C. LeMaitre: None. T.H. Jones: None. N. Aberdein: None. D.M. Kelly: None. Introduction: Low testosterone can lead to bone loss in males, but the mechanisms are not fully characterised. This study investigates the effects of testosterone depletion, and subsequent testosterone treatment on parameters of bone remodeling in a murine model of cardiometabolic disease. Methods: At 8 weeks of age, high-fat diet-fed ApoE-/- mice were randomised into three groups: sham surgery + placebo treatment (control, n=9), orchidectomy + placebo treatment (n =8), and orchidectomy + testosterone treatment (n=10). 25 week old mice were sacrificed and tissues collected for analysis. Left tibiae were decalcified, paraffin embedded and sectioned prior to immunostaining of bone turnover markers including receptor activator of nuclear factor κB ligand (RANKL), Osteoprotegerin (OPG), runx2, Alkaline Phosphatase (ALP), adiponectin and marrow lipid content. Right tibiae underwent mechanical 3-point bend testing at 0.05mm/s using a CellScale Univert. Results: µCT demonstrated a significant decrease in trabecular bone volume, number, and bone mineral density (BMD) and increased trabecular thickness in orchidectomised mice compared to controls. These parameters returned to control levels in testosterone treated orchiectomised mice. No significant differences were observed in cortical bone. Lipid deposition within the bone marrow was significantly increased in testosterone deficient mice compared to testosterone replete mice. No significant differences in ALP immunopositivity were detected between groups, and runx2 was significantly decreased in orchiectomised mice treated with testosterone compared to controls. The ratio of RANKL/OPG immunopositivity was significantly increased in testosterone deficient mice compared to testosterone replete mice. 3-point bending demonstrated no significant differences in maximal force between groups despite a downward trend in orchiectomised groups compared to controls. Osteoclast number quantified from TRAP-stained sections was not different between groups. Discussion: Testosterone depletion accelerates trabecular bone loss, mediated by increased osteoclastogenesis via the RANKL/OPG pathway and increased bone marrow adiposity, an effect reversed by testosterone treatment. While this study highlights the benefits of testosterone upon trabecular bone health in male mice these changes may not alter fracture risk as 3-point-bend maximal force was not affected. Presentation: 6/2/2024

7336 Decades In The Dark: The Long Road To Diagnosing And Treating Hypophosphatasia

Abstract Disclosure: V.A. Mendpara: None. G. Madrigal Loria: None. L.Z. Khan: None. Background: Hypophosphatasia (HPP) is a rare bone disorder that results from deficient activity of the tissue non-specific isoenzyme alkaline phosphatase (ALP).[1] This case highlights a patient whose diagnosis of this treatable genetic condition was significantly delayed for almost two decades, despite receiving care at a prominent academic institution. The patient's persistently low ALP levels in routine lab tests were overlooked by multiple clinicians, leading to a delay in her care and eventual treatment. Case presentation: A 61-year-old Caucasian woman presented at the endocrinology clinic with complaints of severe fatigue, generalized pain, multiple childhood fractures, and ongoing dental issues. The rheumatology clinic to which she was referred for polyarthralgia diagnosed her with Hypophosphatasia (HPP) after noting chronically low alkaline phosphatase (ALP) levels from her previous charts ranging from 15 to 24 U/L (normal range: 34 to 123 U/L) over two decades. Her serum calcium, parathyroid hormone (PTH), and 25-hydroxy vitamin D levels were within normal ranges. A DEXA scan revealed normal bone density, effectively ruling out osteoporosis. Further investigation showed elevated Vitamin B6 levels at 219.6 nmol/L (normal range: 20.0 to 125.0 nmol/L), raising suspicion for HPP. The patient's family history provided additional insight, with her mother having a history of short stature and fractures and her son experiencing premature tooth loss at the age of 2. A genetic evaluation identified a heterozygous pathogenic variant in the ALPL gene (c.881A&amp;gt;C, p.Asp294Ala), confirming the diagnosis of autosomal dominant HPP. Initiating treatment with Asfotase alfa, a medication targeting HPP, resulted in a marked improvement in the patient's symptoms. Discussion: This case shows the diagnostic complexities of HPP, stressing the significance of actively screening for this condition via ALP levels in routine laboratory testing or during osteoporosis evaluations. The diagnostic process is critical to distinguish HPP from primary osteoporosis, as treatment and potential risks differ significantly. Inadvertent use of bisphosphonates can worsen hypomineralization and lead to atypical femoral fractures in HPP patients. Asfotase alfa enzyme replacement therapy, available since 2015, offers a targeted approach by replacing the enzyme defect, thereby mitigating symptoms. Heightened awareness among healthcare providers of HPP in adults holds the potential to enhance diagnosis rates and facilitate improved treatment outcomes for this highly treatable condition.[1] Primer on the Metabolic Bone Diseases and Disorders of Mineral Metabolism, Eighth Edition. ASBMR. Presentation: 6/2/2024