Alkaline Phosphatase Research Articles

Unleashing the Potential of Blackgram: Exploring the Impact of Diverse Nutrient Prescription Techniques on Alfisols

ABSTRACT A study was conducted in Thondamuthur block, Coimbatore, during 2022–2023 and 2023–2024 to evaluate the impact of combining chemical fertilizers with farmyard manure (FYM) and biocompost on soil properties and blackgram yield. Three field experiments were carried out under varying soil organic carbon levels, with soil samples collected after 45 days and analyzed for biological properties. The combined application significantly enhanced soil health, improving bacterial, fungal, and actinomycetes populations, soil respiration, microbial quotient, microbial biomass carbon, and enzymatic activities, including urease, alkaline phosphatase, beta-glucosidase, and dehydrogenase. The STCR-IPNS-1.4 t ha− 1 resulted in the highest blackgram grain yields, ranging from 1314 to 1384 kg ha− 1 in experiment I and 1342–1368 kg ha− 1 in experiment II. During the year 2022–2023, the mean percent increase in grain yield due to STCR-IPNS-1.4 t ha− 1 (with FYM) was 48.2, 38.7, and 58.2 over blanket, blanket + FYM, and farmer’s practice, respectively. Similarly, during the year 2023–2024, the mean percent increase in grain yield due to STCR-IPNS-1.4 t ha− 1 (with biocompost) was 51.3, 43.8, and 63.8 over blanket, blanket + biocompost, and farmer’s practice. These findings highlight the potential of integrating organic and inorganic fertilizers to improve soil health and enhance crop yields in water-scarce regions. By boosting microbial activity and soil fertility, this approach offers a sustainable and cost-effective solution for addressing the challenges of soil degradation, water scarcity, and climate change in arid and semi-arid areas. This method can be adapted to similar regions globally, contributing to long-term agricultural sustainability and resilience.

Cardamom extract alleviates tamoxifen-induced liver damage by suppressing inflammation and pyroptosis pathway

Tamoxifen (TAM) is extensively used to manage estrogen receptor-positive breast cancer. Despite its effectiveness, its administration can negatively impact various organs, including the liver. This research focused on the effects of TAM on the pyroptotic pathway in the liver and evaluated the potential of cardamom extract (CRDE) to lessen hepatic damage of TAM in female rats. Rats received 45 mg/kg of TAM injections for 10 days, while the groups treated with CRDE received 12 ml/kg of CRDE for 20 days, commencing 10 days before TAM administration. TAM exposure resulted in apparent degenerations in hepatic tissue with inflammatory cell infiltration and loss of architectures. Serum levels of liver enzymes including alanine aminotransferase, aspartate aminotransferase and alkaline phosphatase were elevated, along with hepatic oxidative stress, as shown by increased lipid peroxidation with lower levels of reduced glutathione. TAM caused inflammation in the liver tissue as indicated by higher levels of tumor necrosis factor-α and interleukin-6 as well as increased expression of CD68; a phagocytic Kupffer’s cells marker. Additionally, the protein expression analysis revealed a high expression of pyroptotic markers including NLRP3-inflammasome, caspase-1, and gasdermin D. Conversely, CRDE treatment effectively neutralized the biochemical, histological, and protein expression alterations induced by TAM. In conclusion, CRDE demonstrated the potential to protect the liver from TAM-induced damage by regulating mechanisms involving oxidative damage, inflammation, and pyroptosis.

Effect of Luteolin on cadmium-inhibited bone growth via suppressing osteoclastogenesis in laying chickens.

Luteolin (Lut) is a flavonoid derived from several plant sources. Cadmium (Cd) is a widespread environmental contaminant and potential toxin with detrimental effects on animal health. However, the effect of Lut on Cd-induced inhibition of bone growth in laying chickens remains unclear. This study investigates the effects of Lut on Cd-induced inhibition of bone growth in the femur and tibia of laying chickens. A total of sixty 1-day-old green-eggshell yellow feather laying chickens were randomly assigned to four groups after a 5-day of acclimation period: basal diet (Con), cadmium chloride (CdCl2, Cd), Lut, and Lut + Cd. Bone microstructure, serum biomarkers of bone remodeling, the levels of Cd, calcium (Ca), phosphorus (P), and trace metal elements were assessed using the micro-computed tomography (Micro-CT), enzyme-linked immunosorbent assay (ELISA), and microwave digestion, respectively. Bone remodeling biomarkers, late endosomal/lysosomal adaptor and MAPK and mTOR activator 1 (LAMTOR1), as well as the phosphorylation of AMP-activated protein kinase α (AMPKα) and protein kinase B (Akt), were quantified using the qRT-PCR and western blot. The results indicated that Lut effectively mitigated Cd-induced bone mass loss compared to the Cd group, resulting in increased bone volume (BV), bone surface/bone volume (BS/BV), connectivity density (Conn.Dn), and the length and weight of the femur and tibia in laying chickens. Mechanistically, compared to Cd group, Lut restored the ratio of osteoprotegerin (OPG)/receptor activator of NF-κB ligand (RANKL) in serum and bone tissue, enhanced the expression of bone morphogenetic protein-2 (BMP-2), runt-related transcription factor 2 (Runx2), osteocalcin (OCN), and Osterix (OSX), while reducing the levels of Ca, Cd, and alkaline phosphatase (ALP) activity, as well as the expression of osteopontin (OPN), c-Fos, osteoclast stimulatory-transmembrane protein (OC-STAMP), tartrate-resistant acid phosphatase (TRAP), cathepsin K (CTSK), matrix metalloprotein-9 (MMP-9), LAMTOR1, and the phosphorylation of AMPKα and Akt. Therefore, Lut alleviates Cd-induced damage to the femur and tibia of chickens by promoting osteogenesis and inhibiting osteoclastogenesis, positioning Lut as a potential therapeutic plant extract for enhancing bone growth in laying chickens.

Paracrine Influence of Masquelet's Induced Membrane on Marrow Derived Mesenchymal Stem Cells.

Masquelet's induced membrane technique (MIMT) is a staged surgical procedure that leverages the foreign body induced membrane (IM) that forms around a cement spacer placed into a segmental bone defect to support subsequent bone grafting. The mechanisms by which the IM supports bone consolidation are not fully understood. We present an indirect coculture system for studying IM-MSC interactions using a rat model of MIMT. Compared to control cells, MSC Tnap (alkaline phosphatase) was induced by 4- but not 8-week IM. MSC Spp1 (Osteopontin) was attenuated by both 4- and 8-week IM. Although Tnfrsf11b (osteoprotegrin) in MSC exposed to IM was not different from control cells, it was induced by 8-week IM compared to 4-week IM. MSC Tnfsf11 (RANKL) was reduced by 4-week and 8-week IM. MSC Thbs2 (Tsp2) was induced by 8-week but not 4-week IM. Ablation of macrophages in IM blocked the induction of Thbs2 by 8-week IM. MSC Col1a1 expression was not affected under any condition tested. TMT proteomics analysis of IM-conditioned medium revealed 150 unique secreted proteins, 7 of which were differentially abundant (fold change ≥ 2 and FDR corrected p ≤ 0.05) in 8-week versus 4-week IM secretomes. All differentially abundant proteins were elevated in medium conditioned by 8-week IM. Our data suggest that factor(s) secreted by IM resident cells affect MSC gene expression, and that duration of IM development influences the potency and nature of this paracrine effect. Patient-specific factors including age and interval between MIMT surgeries may affect IM biological potency and graft to bone consolidation.

Dose response effects of dietary clove and peppermint oils on the growth performance, physio-metabolic response, feed utilization, immunity, and organ histology in African catfish (clarias gariepinus).

Phytochemicals and essential oils have been widely used as growth promoters in aquaculture. However, the optimal dose of a blend of essential oils for promoting the growth and health of African catfish (Clarias gariepinus) remains largely unknown. Therefore, the present study aimed to assess the dose-response effects of dietary supplementation with a blend of clove and peppermint oils (CPO) on growth indices, feed utilization, physio-metabolic responses, immunity, and organ histology in African catfish. Catfish (105.1 ± 0.5g) were fed diets containing 0.0 (control; CPO0), 1.0 (CPO1), 2.0 (CPO2), or 3.0 mL CPO/kg diet for two months. The results displayed that fish performance and feed intake significantly improved with increasing CPO levels in a linear manner (P < 0.01). Additionally, the specific growth rate and hemoglobin (R2 = 84.6%) levels showed quadratic improvements with the optimal doses at 1.12 mL and 1.6 mL/kg diet, respectively. The inclusion of dietary CPO linearly affected aspartate aminotransferase (AST; R2 = 93.1%), alanine aminotransferase (ALT; R2 = 97.7%), alkaline phosphatase (ALP; R2 = 97.0%), and glucose (R2 = 85.9%) levels in the plasma of catfish (P < 0.001). Increasing the concentrations of dietary CPO led to a quadratic decrease in creatinine (R2 = 98.7%), uric acid (R2 = 94.8%), and cortisol (R2 = 87.1%; P < 0.001), with optimal doses confirmed at 1.75, 2.7, and 1.3-mL CPO per kg of diet, respectively. Dietary CPO has a significant modulatory impact on immune-antioxidant variables in African catfish in a dose-dependent manner. Increasing CPO in the fish diets resulted in a quadratic increase in the levels of immunoglobulin G (IgG; R2 = 98.8%), lysozyme activity (R2 = 93.9%), and total antioxidant capacity (R2 = 84.9%). The corresponding dose-response curves displayed that the optimal doses were at 1.85, 2.1, and 1.8 mL/kg of diet. Furthermore, superoxide dismutase (SOD), and reduced glutathione (GSH) levels were significantly improved in all CPO-treated groups in a quadratic manner, with optimal doses at 1.25 mL (R2 = 90.6%) and 1.55 mL (R2 = 89.7%) per kg of diet. Catfish fed diets supplemented with CPO showed gradual improvements in hepatocytes (HP), and blood vessels (BV) with an increased number of binucleated cells. Moreover, catfish fed diets containing CPO had improvement in the tubular epithelium, Bowman's capsule, and a few melanomacrophage areas. This study revealed that CPO supplementation at an optimal dose of 1-2 mL/kg of diet significantly improved growth performance, feed utilization, metabolic processes, and immune and antioxidant functions in African catfish.

The protective role of SGLT2 inhibitors on aortic aneurysm mediated by oxidative stress and inflammation in type 2 diabetes mellitus

BackgroundSodium-glucose transport protein 2 inhibitors (SGLT2i) have been widely used to treat patients with type 2 diabetes mellitus (T2DM) and have demonstrated protective effects against certain cardiovascular diseases. However, no clinical research has been conducted to explore the relationship between SGLT2i and the risk of aortic aneurysm (AA).MethodsWe extracted and analyzed the data of 4964 patients with T2DM from the First Affiliated Hospital of Zhengzhou University during July 2017 to January 2023. Multivariate Cox models, interaction analysis and Kaplan–Meier curves were performed to approximate the associations of SGLT2i therapy on the risk of AA. A sensitivity analysis was performed to test the robustness of results. Mediation analyses explored the roles of inflammatory (neutrophils, lymphocytes, C-reactive protein and alkaline phosphatase) and oxidative stress (gamma glutamyl transferase, total bilirubin, and uric acid) markers in the associations between SGLT2i and AA.ResultsA total of 1942 SGLT2 inhibitor (SGLT2i) users (39.12%) and 3022 non-SGLT2 inhibitor (NonSGLT2i) users were included in final analysis. After full adjustment for potential risk factors, SGLT2i patients were associated with a lower risk of aortic aneurysm (HR, 95% CI 0.91, 0.89–0.98, p = 0.001). Dapagliflozin showed the greatest difference for reduction of aortic aneurysm incidence (HR, 95% CI 0.84, 0.80–0.95, p = 0.011). Subgroup analysis indicated that use of SGLT2i lower the risk of aortic aneurysm in some subgroups of T2DM patients. The sensitivity analysis demonstrated the robustness of the results. CRP, lymphocytes, neutrophils, and uric acid were significantly associated with both SGLT2i and AA, with mediation proportions of 13.35%, 8.83%, 9.67% and 31.17%, respectively.ConclusionsOur study suggested that patients using SGLT2i may have a lower risk of aortic aneurysm, and this effect could potentially be mediated by inflammation and oxidative stress. Further mechanistic and prospective studies are required to verify this association.Graphical abstract

Priapism-related biomarkers in sickle cell disease: a systematic review.

Priapism is a major clinical complication of sickle cell disease (SCD), with severe sexual, reproductive, and mental health impact. There are currently no consensus diagnostic biomarkers for identifying individuals with SCD at risk of priapism before its occurrence. To systematically review the biochemical, hematological, imaging, genetic, and rheological parameters associated with priapism occurrence among individuals with SCD. A systematic literature search for studies investigating the association of biochemical, hematological, rheological, imaging, rheological, and genetic parameters with the occurrence of priapism in individuals with SCD was performed in the MEDLINE, Embase, and Cochrane databases, using the following terms: "priapism," "sickle cell," "biomarker," "marker," "laboratory," "radiographic," "diagnostic," and "predictive." A systematic review of the identified studies was conducted to describe the landscape of priapism-related biomarkers in individuals with SCD. A total of 358 studies were identified, of which 14 studies were eventually selected for evidence synthesis. The selected studies were published between 2005 and 2023, with authorship spanning five continents. We identified multiple clinical parameters investigated as potential biomarker candidates for their association with priapism occurrence in patients with SCD. We classified these into biochemical (lactate dehydrogenase, bilirubin, aspartate transaminase, alanine transaminase, alkaline phosphatase, nitric oxide metabolites, interleukin 6), hematological (hemoglobin concentration, mean corpuscular volume, mean corpuscular hemoglobin, reticulocyte count, leukocyte count), genetic (Klotho, TGFBR3, QAP1, ITGAV, LNC02537, NAALADL2), rheological (red blood cell deformability, aggregation index, augmentation index), and imaging parameters. However, the results were often contradictory and do not support the clinical application of any of the investigated parameters. Several clinical and laboratory parameters have been associated with priapism occurrence in SCD; however, contradictory findings across geographical locations paint an unreliable picture of their clinical utility. Additional studies are needed to generate enough level 1 evidence in support of any of the current candidates.

Variation in soil enzyme activity with amendments of biochar and polyacrylamide in coal gangue soils

Coal gangue is a prevalent solid waste material, and its utilization presents an urgent challenge. This study investigates the impact of incorporating biochar (BC) and polyacrylamide (PAM) into a composite matrix of coal gangue soil (CGS). We conducted incubation experiments to evaluate the physicochemical properties and enzyme activity of CGS with different ratios of BC (1%, 2%, and 5% designated as B1, B2, and B5) and PAM (0.02%, 0.05%, and 0.10% designated as P2, P5, and P10), compared to a control (CK) with no amendments. The results indicate that (1) the lowest bulk density was observed in the B5P10 treatment. The organic carbon content in B5P10 increased by 57.98% compared to the CK. (2) The activities of α-glucosidase, N-acetyl-glucosidase, and alkaline phosphatase in the B5P10 treatment increased by 112.34%, 110.77%, and 52.40%, respectively. The geometric mean values of enzyme activities showed no significant differences among the treatments B2P5, B2P10, B5P2, B5P5, and B5P10, but these were significantly higher than those in the CK. (3) The parameters of pH, active carbon, field capacity, and available phosphorus were identified as the main factors affecting enzyme activity. CGS incorporating 2% BC and 0.05% PAM is recommended for soil reconstruction in mining regions.

Multifaceted skeletal effects of sevelamer carbonate in a secondary hyperparathyroidism model.

Hyperphosphatemia leads to abnormal mineralization of bones and soft tissues in patients with chronic kidney disease-induced secondary hyperparathyroidism (CKD-SHPT). Sevelamer lowers phosphate levels by binding to dietary phosphate in the gastrointestinal system, forming new bone and reducing the risk of renal osteodystrophy and fracture. However, the influence of sevelamer carbonate (SevC) on bone microarchitecture, material qualities, and mechanical behavior is unknown in CKD-SHPT conditions. We utilized a rat model of CKD-induced hyperphosphatemia by feeding a 1.8% high-phosphate diet to 5/6 nephrectomized rats to test the effects of SevC on skeletal quality and strength, employing microCT, Fourier transform infrared spectroscopy (FTIR), 3-point bending, nanoindentation, and compression tests. SevC preserved mineral homeostasis and reduced PTH, and FGF-23 levels in CKD-SHPT rats. SevC mitigated the serum renal parameters, pyrophosphate levels, and indole acetic acid. In CKD-SHPT rats, SevC reduced hyperphosphatemia, improved the mineralization defect, and upregulated mineralization-promoting genes like ankyrin-1, ectonucleotide-pyrophosphatase/phosphodiesterase-1, tissue non-specific alkaline phosphatase, phosphate-regulating endopeptidase X-linked, dentin matrix protein-1, and matrix extracellular phosphoglycoprotein. In the cortical bones of CKD-SHPT rats, SevC increased cortical mass and thickness, decreased porosity by likely decreasing cortical bone remodeling induced by high PTH, and increased osteocyte preservation. SevC mitigated all of the alterations in the mineral and matrix composition of CKD-SHPT rats, including decreased collagen-maturity, mineral-to-matrix ratio, and increased carbonate substitution of hydroxyapatite crystals. SevC enhanced bone strength and mechanical behavior in CKD-SHPT rats at a macro (three-point bending) and nano (nanoindentation) scales. These findings in CKD-SHPT rats suggest that SevC improves bone mechanical properties at various levels by decreasing serum pyrophosphate, empty lacunae, and enhancing renal clearance of indole acetic acid, organized mineral-matrix deposition, and osteocyte number by suppressing cortical remodeling.

Effects of short-term dietary oxytetracycline treatment in the farmed fish Piaractus mesopotamicus.

Oxytetracycline (OTC), a key antibiotic used in global aquaculture, has still unclear ecotoxicological effects. In this study, freshwater fish Piaractus mesopotamicus were fed with diets containing 750 mg kg-1 either pure OTC (ATB1) or commercial OTC (ATB2) for 10 days (treatment period-TP), followed by a 21-day withdrawal period (depuration period-DP). Fish fed with ATB2 showed decreased hematocrit (at DP) and increased glucose levels (TP and DP). In general, catalase activity increased in the liver, gills, and muscle of OTC-treated individuals at both TP and DP, particularly with ATB2. Similarly, glutathione S-transferase activity rose in the brain, gills, and muscle (TP and DP). Conversely, alkaline phosphatase activity in the liver decreased in both treated groups (TP and DP). Additionally, only ATB2 induced lactate dehydrogenase in fish muscle after 1-day depuration. Principal components analysis (PCA) identified most antioxidant enzymes, hematocrit, weight, length, and mean corpuscular hemoglobin concentration as key biomarkers, distinguishing ATB2 from control fish. These results indicate that the dietary therapeutic dose of OTC caused adverse effects in P. mesopotamicus. Differences in biomarker responses between ATB1 and ATB2 might be linked to unknown compounds in the commercial formulation, potentially influencing biological responses or altering OTC bioavailability. Further research on the toxicity of antimicrobial impurities and degradation compounds should accompany enhanced quality control measures in aquaculture to guarantee sustainable and safe products.

Hypophosphatemic rickets in an Italian multicentric cohort of 24 subjects: a clinical and molecular characterisation.

Rickets is a rare bone disorder due to altered calcium, vitamin D, and phosphorus metabolism, caused by nutritional deficiencies or, in 13% of cases, genetic origin. Few data are available on an Italian cohort of rickets. Twenty-four patients with confirmed low serum phosphorus levels and reduced renal tubular phosphate reabsorption were recruited from different tertiary care centres over the last 5 years. Biochemical, clinical, and anamnestic data were also collected. DNA was extracted and subjected to targeted next-generation sequencing. Twenty-four single-nucleotide variants were identified in the PHEX (eight pathogenic, five likely pathogenic, three variants of uncertain significance), CYP27B1 (two pathogenic, four likely pathogenic), and SLC34A3 (one pathogenic, one likely pathogenic) genes. Five large genomic deletions involving one or more PHEX exons were detected. Eight of 20 PHEX and both SLC34A3 variants were novel, and segregation analysis identified 11 familial and three de novo cases. Biochemical data confirmed high serum alkaline phosphatase and low 25-hydroxyvitamin D3 levels, whereas the main clinical manifestations were short stature (76.1%), bone deformities (85.7%), musculoskeletal pain (71.4%), and muscle weakness (55.5%). Our study provides clinical and genetic descriptions of rickets in a cohort of Italian patients. Moreover, we expanded the spectrum of mutations associated with the genetic forms of this disorder and suggested a high-throughput sequencing approach to provide a molecular diagnosis for adequate follow-up of patients.

Adsorption of Serum Fetuin onto Octacalcium Phosphate and Its Relation to Osteogenic Property

This study aimed to investigate how the chemical elements in relation to octacalcium phosphate (OCP) hydrolysis affect the osteoblastic differentiation in the presence of serum fetuin. The adsorption of fetuin onto OCP was examined in buffers having different degrees of supersaturation (DS) with respect to OCP and hydroxyapatite (HA) at pH 7.4 and 37 °C. The osteoblastic differentiation of mesenchymal stem cells (MSCs) was evaluated in cultures with OCP and 0 to 0.8 mg/mL of fetuin. The amount of fetuin adsorbed increased with increasing DS in the buffer. In the MSC culture, the coexistence of OCP and 0.2–0.4 mg/mL of fetuin close to serum level increased alkaline phosphatase activity; however, the activity was suppressed by 0.2–0.8 mg/mL of fetuin. Transmission electron microscopy revealed de novo crystal formation on OCP in supersaturated buffer and culture media with respect to OCP and HA at lower fetuin concentrations. Infrared spectroscopy and DS estimation indicate that the hydrolysis of OCP with de novo apatite formation was promoted in the culture media at 0.2–0.4 mg/mL of fetuin. These results suggest that OCP may promote osteoblastic differentiation if the suitable conditions are attained regarding the chemical elements and fetuin adsorption around OCP.

Circadian rhythm and the influence of light on parameters related to calcium metabolism in stroke patients admitted for rehabilitation

Hospitalized stroke patients are at high risk of developing circadian disruption due to lack of natural sunlight. This may affect the circadian rhythm of the calcium metabolism. This study is a secondary explorative analysis from a Randomized Controlled Trial. Acute stroke patients requiring a minimum of two weeks of rehabilitation were randomized to an Intervention unit (IU) equipped with naturalistic light or a Control unit (CU) with standard indoor lighting. Blood was drawn across 24 h at inclusion and discharge in 45 patients, 25 from the IU and 20 from the CU. Calcium showed significant rhythmicity at inclusion and discharge in both groups. Alkaline phosphatase, parathyroid hormone (PTH), and Vitamin D exhibited no significant rhythmicity at inclusion or discharge in either group while phosphate exhibited rhythmicity at discharge in the CU. PTH levels were elevated in the CU group compared to the IU group at time of discharge. Of the measured parameters, only calcium exhibited circadian rhythmicity after stroke. Naturalistic light did not have any influence on the rhythmicity, indicating that light may not be the main circadian regulator of the circadian oscillations that regulate calcium metabolism. PTH seems to be decreased by naturalistic light.

Ultra-Sensitive Nanoplatform for Detection of Brain-Derived Neurotrophic Factor Using Silica-Coated Gold Nanoparticles with Enzyme-Formed Quantum Dots

A fluorescence-based detection platform was developed for brain-derived neurotrophic factor (BDNF), a key biomarker of Alzheimer’s disease (AD). This platform utilizes localized surface plasmon resonance effects resulting from the interactions between silica-coated gold nanoparticles (Au@SiO2) and enzymatically synthesized quantum dots (QDs). The gold nanoparticles were silica coated via the hydrolysis of tetraethyl orthosilicate, which allowed for precise control over the distance between the nanoparticles and QDs and refined the dynamics of fluorescence quenching and enhancement. Antibody conjugation was performed via sequential amination and carboxylation, followed by EDC/NHS coupling. BDNF was detected across a range of concentrations, from 1 ng/mL to 1 ng/mL, using an alkaline phosphatase (ALP)-conjugated polyclonal antibody targeting a secondary epitope of BDNF. The enzymatic hydrolysis of p-nitrophenyl phosphate by immobilized ALP led to the formation of cadmium sulfide QDs, with the fluorescence intensity correlating directly with the BDNF concentration. This platform offers a refined and precise method for detecting BDNF and is a reliable tool for the early diagnosis of AD.

Soyasaponin Bb has poor absorption and bioavailability in Sprague–Dawley rats and Caco-2 intestinal epithelial cell model

Soyasaponin Bb has various health-promoting bioactivities. However, the bioavailability of soyasaponin Bb is not fully understood. This study aimed to explore the absorption and metabolism of soyasaponin Bb by using both in vivo and in vitro methods. Soyasaponin Bb (100 mg/kg) was orally administrated in Sprague–Dawley (SD) rats, and the content of soyasaponin Bb and soyasapogenol B in plasma, urine and feces were determined by HPLC–MS/MS. The Caco-2 intestinal epithelial cell model was established by culturing on Transwell plates and assessing through cell morphology, transepithelial electrical resistance, alkaline phosphatase activity, and phenol red flux. Then, the apical (AP) to basolateral (BL) transport or uptake of soyasaponin Bb in the model were determined. In SD rats, soyasaponin Bb reached a maximum of 19.8 ng/mL in plasma and showed two material peaks. The cumulative excretion of soyasaponin Bb at 168 h was (0.0022 ± 0.0006) % in urine and (0.36 ± 0.21) % in feces. Soyasapogenol B was not detected in plasma and urine, but had a cumulative excretion of (0.45 ± 0.29) % in feces at 168 h. Culturing Caco-2 cells on Transwell plates for 21 days formed good intestinal epithelial monolayers. The apparent permeability (Papp) of both AP-BL and BL-AP transport of soyasaponin Bb in the Caco-2 cell models was less than 1.0 × 10–6 cm/s, and the efflux ratio (ER) was 0.5 ± 0.2. The uptake of soyasaponin Bb in the AP-BL cells was significantly higher (p < 0.05) than that in the BL-AP cells. In conclusion, soyasaponin Bb has poor absorption and bioavailability in SD rats and Caco-2 intestinal epithelial cell model.

Taxifolin attenuates gentamicin-induced hepatotoxicity via modulation of oxidative stress, inflammation, and apoptosis in mice: potential involvement of Nrf2/HO-1 signaling

ABSTRACT The natural flavonoid taxifolin (TX) is known for its anti-inflammatory and antioxidant potential. Gentamicin (GEN) is a popular antibiotic agent that is associated with hepatotoxicity. The study assessed the hepatoprotective impacts of TX against GEN-linked liver injury in mice. Mice were orally administered with TX (25 and 50 mg/kg/day) for 14 days followed by GEN administration (100 mg/kg/day, i.p.) from the 8th to 14th day. GEN treatment significantly increased lactate dehydrogenase, transaminases (AST and ALT), and alkaline phosphatase levels in mouse serum, in addition to various histopathological liver changes. Higher levels of protein carbonyl and malondialdehyde and lower levels of superoxide dismutase, catalase, and glutathione were noted in the livers of GEN-administered mice. GEN administration also showed enhanced interleukins (IL-6 and IL-1β) and TNF-α levels and NF-κB p65 expression in mouse liver. TX treatment effectively ameliorated tissue injury, attenuated levels of MDA, protein carbonyl, and pro-inflammatory mediators, and improved antioxidants in GEN-treated mice. Increased caspase-3 and Bax protein expressions with decreased Bcl-2 levels were noted in the GEN-administrated mice, which were largely alleviated by TX pretreatment. Moreover, TX restored hepatic Nrf2/HO-1 signaling in GEN-administrated mice to counter GEN toxicity. These findings suggest TX as a promising adjuvant to prevent drug-associated tissue injury.

Metformin alleviates doxorubicin-induced hepatic damage by modulating oxidative stress: a molecular, biochemical, and histopathological approach in a rat model.

Doxorubicin (DOX) is one of the most commonly prescribed anti-cancer drugs. However, DOX-induced hepatotoxicity is a dose-limiting side effect. This study aimed to clarify the potential protective effects of metformin on DOX-induced hepatotoxicity in rats. The animals were divided into six groups (n = 6 each): Control Group, DOX group, metformin 200mg/kg group, DOX + metformin 50mg/kg group, DOX + metformin 100mg/kg group, and DOX + metformin 200mg/kg group. Hepatic injury was induced by a single intraperitoneal injection of DOX (20mg/kg). The activities of aspartate aminotransferase (AST), alanine aminotransferase (ALT), and alkaline phosphatase (ALP) in serum were determined. Furthermore, the hepatic histopathological changes were evaluated. To identify the markers of oxidative stress, the level of malondialdehyde (MDA) and the activities of antioxidant enzymes such as superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) in liver tissue were measured. Results showed that DOX provoked a marked elevation in ALT, AST, and ALP serum levels. In addition, oxidative stress was significantly boosted in DOX-treated rats compared to control rats. All these were abolished with the metformin administration. Histological examination also showed that metformin could substantially reduce DOX-induced alterations. The most prominent effect was observed by high-dose metformin.

Qi-Gui-Jian-Gu Decoction Accelerates Osteogenesis and Fracture Healing by Activating the Wnt/β-Catenin Signaling Pathway.

Qi-Gui-Jian-Gu decoction (QGJG), as a clinical empirical formula, has clinical benefits in promoting bone formation, but the underlying mechanism for its application in treating fractures has not been investigated. The potential therapeutic target and signaling pathway of QGJG for treating fractures were analyzed by network pharmacology. In vitro, we used bone marrow mesenchymal stem cells (MSCs) to evaluate osteogenic differentiation and mineralization by alizarin red staining, quantitative real-time polymerase chain reaction (qRT-PCR), western blot (WB), and immunofluorescence staining. In vivo, the 8w male SPF C57BL/6J mouse femoral fracture model was constructed, and the therapeutic effects of QGJG were evaluated. By network pharmacology analysis, we found that glycogen synthase kinase 3 beta (GSK3β) was a potential therapeutic target of QGJG for treating fractures. The canonical Wnt signaling pathway was selected as the potential molecular mechanism. QGJG was confirmed to upregulate the mRNA levels of alkaline phosphatase (ALP) and bone morphogenetic protein 2 (BMP2), thereby promoting osteogenic differentiation and mineralization. Mechanistically, QGJG inhibited GSK3β while increasing p-Ser9-GSK3β to increase β-catenin protein expression and its nuclear translocation, implying the activation of the canonical Wnt signaling pathway. In vivo, QGJG administration promoted fracture healing, as demonstrated by the up-regulation of OPN and Osx, and accelerated the progression of ossification at 2 and 3 weeks after surgery. QGJG promotes osteogenic differentiation and fracture healing by activating the canonical Wnt pathway.

Sericin’s Potential in Osteoporosis Management: The Roles of L-Serine and D-Serine in Bone Metabolism Regulation

Background: Osteoporosis is a bone remodeling disease characterized by an imbalance between bone formation and resorption, leading to bone fragility. Current treatments focus on bone resorption suppression but often have adverse effects. This study aimed to explore the potential of sericin, a silkworm-derived protein, as a dual-action therapeutic agent that enhances bone formation through its component L-serine and inhibits bone resorption via D-serine, which is derived from L-serine by the action of serine racemase. Methods: Cellular experiments were conducted to evaluate the effects of L-serine on osteoblast differentiation and D-serine on osteoclast inhibition. Serum levels of D-serine were measured following sericin administration in an osteoporosis animal model. μ-CT analysis assessed trabecular and cortical bone quality, and bone-related protein expression was analyzed using immunoprecipitation-based high-performance liquid chromatography (IP-HPLC). Results: L-serine significantly upregulated osteogenic markers, including alkaline phosphatase (ALP), Runx2, osterix, and Col1a1, in osteoblasts (p &lt; 0.05). D-serine inhibited osteoclast activation by suppressing cathepsin K expression (p &lt; 0.001). Sericin feeding elevated serum D-serine levels (p &lt; 0.001) and upregulated bone-related proteins such as BMP-2, osterix, and Runx2. Micro-computed tomography (μ-CT) analysis revealed significant improvements in trabecular bone parameters in the OVX-sericin group, including increased trabecular bone volume (Tb.BV/TV; p &lt; 0.05) and reduced trabecular separation (Tb.Sp; p &lt; 0.05), compared to the OVX and OVX-amino acid groups. Cortical bone parameters, including cortical bone volume (Ct.BV/TV) and cortical area (Ct.Ar), did not significantly differ among OVX groups, but all were lower than in the sham group (p &lt; 0.05). Conclusions: This study demonstrates that sericin modulates bone metabolism by enhancing osteoblast activity through L-serine and inhibiting osteoclastogenesis via D-serine. Sericin supplementation improved trabecular bone mass in an osteoporosis model, highlighting its potential for bone health.

Transcriptomic and metabolomic analysis of recalcitrant phosphorus solubilization mechanisms in Trametes gibbosa

IntroductionPhosphorus (P) is a crucial growth-limiting nutrient in soil, much of which remains challenging for plants to absorb and use. Unlike chemical phosphate fertilizers, phosphate-solubilizing microorganisms (PSMs) offer a means to address available phosphorus deficiency without causing environmental harm. PSMs possess multiple mechanisms for phosphorus solubilization. Although the phosphorus-solubilizing mechanisms of phosphate-solubilizing bacteria (PSB) have been well characterized, the mechanisms utilized by phosphate-solubilizing fungi (PSF) remain largely unexplored.MethodsThis study isolated a PSF strain, Trametes gibbosa T-41, from soil and evaluated its phosphorus solubilizing capacity with organic (calcium phytin; Phytin-P) and inorganic (tricalcium phosphate; Ca-P) phosphorus sources. The phosphorus solubilization, enzyme activity, and organic acid production of T-41 were measured. And the P-solubilizing mechanism conducted by transcriptomic and metabolomic analyses.Results and discussionT-41 exhibited varying phosphorus solubilizing capacity when grown with organic (calcium phytin; Phytin-P) and inorganic (tricalcium phosphate; Ca-P) phosphorus sources (109.80 ± 8.9 mg/L vs. 57.5 ± 7.9 mg/L, p &amp;lt; 0.05). Compared with the Ca-P treatment, T-41 demonstrated a stronger alkaline phosphatase (ALP) production capacity under Phytin-P treatment (34.5 ± 1.2 μmol/L/h vs. 19.8 ± 0.8 μmol/L/h, p &amp;lt; 0.05). Meanwhile, the production of oxalic acid, maleic acid, and succinic acid was higher under Phytin-P treatment (p &amp;lt; 0.05). Transcriptomic and metabolomic analysis revealed that different phosphorus sources altered metabolic pathways such as galactose metabolism, glyoxylate and dicarboxylic acid metabolism, and ascorbate and aldolate metabolism. Key metabolites like myo-inositol, 2-oxoglutarate, and pyruvate were found to impact the performance of T. gibbosa T-41 differently under the two P sources. Notably, synthesis in Ca-P vs. Pytin-P, T-41 upregulated genes involved in myo-inositol synthesis, potentially enhancing its P-solubilizing ability. These results provide new insights into the molecular mechanisms of PSF at the transcriptomic and metabolomic levels, laying a theoretical foundation for the broader application of PSF as bio-phosphorus fertilizers in the future.