Showing papers on "Alkaline phosphatase published in 2020"

RRM2 protects against ferroptosis and is a tumor biomarker for liver cancer.

Abstract: Ferroptosis is the process of cell death triggered by lipid peroxides, and inhibition of glutathione (GSH) synthesis leads to ferroptosis. Liver cancer progression is closely linked to ferroptosis suppression. However, the mechanism by which inhibition of GSH synthesis suppresses potential ferroptosis of liver cancer cells and whether ferroptosis-related liver cancer biomarkers have a promising diagnostic value remain unknown. Ribonucleotide reductase regulatory subunit M2 (RRM2) levels were measured using an enzyme linked immunosorbent assay (ELISA), quantitative RT-PCR (qPCR), immunoblotting (IB) and immunochemistry (IHC). Cell viability and cell death were measured by a CellTiter-Glo luminescent cell viability assay and staining with SYTOX Green followed by flow cytometry, respectively. Metabolites were measured using the indicated kits. The Interaction between glutathione synthetase (GSS) and RRM2 was measured using immunofluorescence (IF), co-immunoprecipitation (co-IP) and the proximal ligation assay (PLA). The diagnostic value was analyzed using the area under the receiver operating characteristic curve (AUC-ROC). Bioinformatics analysis was performed using the indicated database. RRM2 showed specifically elevated levels in liver cancer and inhibited ferroptosis by stimulating GSH synthesis via GSS. Mechanistically, phosphorylation of RRM2 at the Threonine 33 residue (T33) was maintained at normal levels to block the RRM2–GSS interaction and therefore protected RRM2 and GSS from further proteasome degradation. However, under ferroptotic stress, RRM2 was dephosphorylated at T33, thus the RRM2–GSS interaction was promoted. This resulted in the translocation of RRM2 and GSS to the proteasome for simultaneous degradation. Clinically, serum RRM2 was significantly associated with serum alpha-fetoprotein (AFP), carcinoembryonic antigen (CEA), alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), gamma glutamyl transpeptidase (γ-GT), albumin (ALB) and total bilirubin. The AUC-ROC for the combination of RRM2 with AFP was 0.947, with a sensitivity of 88.7% and a specificity of 97.0%, which indicates better diagnostic performance compared to either RRM2 or AFP alone. RRM2 exerts an anti-ferroptotic role in liver cancer cells by sustaining GSH synthesis. Serum RRM2 will be useful as a biomarker to evaluate the degree to which ferroptosis is suppressed and improve diagnostic efficiency for liver cancer.

Osteocalcin promotes bone mineralization but is not a hormone.

Abstract: During the last 24 years the bone and mineral metabolism research community (and the National Institutes of Health and numerous other national and international funding agencies that support it) have devoted a great deal of intellectual energy and resources to some provocative, sometimes controversial, ideas about osteocalcin (OCN). OCN is a 46 amino-acid protein that is produced and secreted almost exclusively by osteoblasts, terminally differentiated cells responsible for the synthesis and mineralization of bone matrix during development of the skeleton and its periodic regeneration throughout life. Osteoblasts originate from mesenchymal progenitors and are short-lived cells that are constantly replaced, depending on the demand for bone formation in a particular location and time [1]. OCN secreted by osteoblasts contains three γ-carboxyglutamic acid residues that confer high affinity to the bone hydroxyapatite matrix. However, when bone is resorbed by osteoclasts, a macrophage-derived cell type, the acidic pH in the resorption compartment causes the carboxyl groups on OCN to be removed, and decarboxylated OCN is released into circulation. The circulating levels of decarboxylated OCN are, therefore, dependent on the rate of bone turnover, also known as remodeling. Originally thought to function exclusively in bone, a more expansive view of decarboxylated OCN as an endocrine hormone has evolved, largely through work of Gerard Karsenty and colleagues and beginning with the description of an OCN knockout mouse 24 years ago [2]. As a hormone, OCN has been proposed to act on multiple end organs and tissues including the pancreas, liver, fat cells, muscle, male gonads, and brain to regulate functions ranging from bone mass accumulation to body weight, adiposity, glucose and energy metabolism, male fertility, brain development, and cognition [2–6]. This idea—that OCN is an endocrine hormone with pleiotropic effects—is widely cited in textbooks and review articles [7,8] and has provided the rationale for numerous human studies on the relationship between OCN and diabetes or obesity. There have been, however, several apparent shortcomings in the “OCN is an endocrine hormone” idea. The number of osteoblasts (and therefore the circulating levels of decarboxylated OCN) inexorably change throughout life as a result of physiologic, adaptive, or pathologic changes of bone itself that can be acute or chronic, systemic or localized, and reversible or irreversible, without changes in the putative extraskeletal targets of decarboxylated OCN. Examples are skeletal development, growth, adaptation of the skeleton to mechanical forces, fracture healing, changing calcium needs, stress, menstrual cycle, pregnancy, lactation, menopause, aging, hyperparathyroidism or hypoparathyroidism, hyperthyroidism, hypercortisolemia, Paget’s disease, bone tumors, etc. Similarly, medications—approved after extensive trials with PLOS GENETICS

Recent advances with alkaline phosphatase isoenzymes and their inhibitors.

Abstract: Alkaline phosphatases are found in different living species and play crucial roles in various significant functions, such as hydrolyzing a variable spectrum of phosphate-containing physiological compounds, contributing to DNA synthesis, bone calcification, and attenuation of inflammation. They are homodimeric enzymes; each subunit contains one magnesium ion and two zinc ions crucial for the catalytic activity of the enzyme. Alkaline phosphatases exist in four distinct isoenzymes (placental, intestinal, germ cell, and tissue nonspecific alkaline phosphatases), which are expressed by four different genes; each one of them has distinguished functions. Any disturbance in the gene expression of alkaline phosphatase eventually induces serious disease conditions. Thus, the need to explore new lead inhibitors has increased recently. In this literature review, we aim to investigate the role of alkaline phosphatase in different diseases and physiological conditions and to study the structure-activity relationships of recently reported inhibitors. We focused on the lead compounds reported in the last 5 years (between 2015 and 2019).

Inhibition of tissue-nonspecific alkaline phosphatase protects against medial arterial calcification and improves survival probability in the CKD-MBD mouse model.

Abstract: Medial arterial calcification (MAC) is a major complication of chronic kidney disease (CKD) and an indicator of poor prognosis. Aortic overexpression of tissue-nonspecific alkaline phosphatase (TNAP) accelerates MAC formation. The present study aimed to assess whether a TNAP inhibitor, SBI-425, protects against MAC and improves survival probability in a CKD-mineral and bone disorder (MBD) mouse model. CKD-MBD mice were divided in three groups: vehicle, SBI-10, and SBI-30. They were fed a 0.2% adenine and 0.8% phosphorus diet from 14 to 20 weeks of age to induce CKD, followed by a high-phosphorus (0.2% adenine and 1.8% phosphorus) diet for another 6 weeks. At 14-20 weeks of age, mice in the SBI-10 and SBI-30 groups were given 10 and 30 mg/kg SBI-425 by gavage once a day, respectively, while vehicle-group mice were given distilled water as vehicle. Control mice were fed a standard chow (0.8% phosphorus) between the ages of 8 and 20 weeks. Computed tomography imaging, histology, and aortic tissue calcium content revealed that, compared to vehicle animals, SBI-425 nearly halted the formation of MAC. Mice in the control, SBI-10 and SBI-30 groups exhibited 100% survival, which was significantly better than vehicle-treated mice (57.1%). Aortic mRNA expression of Alpl, encoding TNAP, as well as plasma and aortic tissue TNAP activity, were suppressed by SBI-425 administration, whereas plasma pyrophosphate increased. We conclude that a TNAP inhibitor successfully protected the vasculature from MAC and improved survival rate in a mouse CKD-MBD model, without causing any adverse effects on normal skeletal formation and residual renal function. © 2019 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.

Hepatotoxic effects of silver nanoparticles on Clarias gariepinus; Biochemical, histopathological, and histochemical studies

Abstract: The current study investigates the hepatotoxic effects of two acute doses of silver nanoparticles (AgNPs) and silver nitrate (AgNO3) on African catfish (Clarias garepinus) using biochemical, histopathological, and histochemical changes and the determination of silver in liver tissue as biomarkers. AgNPs-induced impacts were recorded in some of these characteristics based on their size (20 and 40 nm) and their concentration (10 and 100 μg/L). Concentrations of liver enzymes (Aspartic aminotransferase; AST, Alanine aminotransferase; ALT), alkaline phosphatase (ALP), total lipids (Tl), Glucose (Glu) and Ag-concentration in liver tissue exhibited a significant increase under stress in all exposed groups compared to the control group. The total proteins (Tp), albumin (Al), and globulin (Gl) concentrations exhibited significantly decrease in all treated groups compared to the control group. At tissue and cell levels, histopathological changes were observed. These changes include proliferation of hepatocytes, infiltrations of inflammatory cells, pyknotic nuclei, cytoplasmic vaculation, melanomacrophages aggregation, dilation in the blood vessel, hepatic necrosis, rupture of the wall of the central vein, and apoptotic cells in the liver of AgNPs-exposed fish. As well as the depletion of glycogen content in the liver (feeble magenta coloration) was observed. The size and number of melanomacrophage centers (MMCs) in liver tissue showed highly significant difference in all exposed groups compared to the control group. Recovery period for 15 days led to improved most alterations in the biochemical, histopathological, and histochemical parameters induced by AgNPs and AgNO3. In conclusion, one can assume liver sensitivity of C. garepinus for AgNPs and the recovery period is a must.

Smart Surface-Enhanced Resonance Raman Scattering Nanoprobe for Monitoring Cellular Alkaline Phosphatase Activity during Osteogenic Differentiation.

Abstract: High-efficiency induction of bone marrow mesenchymal stem cells (BMSCs) to osteogenic differentiation in vitro can help solve a series of bone diseases such as bone injury, fracture repair, and ost...

Alkaline phosphatase delivery system based on calcium carbonate carriers for acceleration of ossification

Abstract: Composite bioceramic and hydrogel-based containers harboring alkaline phosphatase are generated through encapsulation of this enzyme by its immobilization into CaCO3-based bioceramic materials in c...

Lycopene and bone: an in vitro investigation and a pilot prospective clinical study.

Abstract: There are several effective therapies for osteoporosis but these agents might cause serious adverse events. Lycopene intake could prevent bone loss, however studies on its effects on bone are scarce. Our aim was to investigate the effects of lycopene on osteoblast cells as well as bone mineral density and bone turnover markers in postmenopausal women. We investigated the effect of lycopene on the Wnt/β-catenin and ERK 1/2 pathways, RUNX2, alkaline phosphatase, RANKL and COL1A of Saos-2. We also carried out a pilot controlled clinical study to verify the feasibility of an approach for bone loss prevention through the intake of a lycopene-rich tomato sauce in 39 postmenopausal women. Lycopene 10 µM resulted in higher β-catenin and phERK1/2 protein Vs the vehicle (p = 0.04 and p = 0.006). RUNX2 and COL1A mRNA was induced by both 5 and 10 µM doses (p = 0.03; p = 0.03 and p = 0.03; p = 0.05) while RANKL mRNA was reduced (p < 0.05). A significant bone density loss was not detected in women taking the tomato sauce while the control group had bone loss (p = 0.002). Tomato sauce intake resulted in a greater bone alkaline phosphatase reduction than the control (18% vs 8.5%, p = 0.03). Lycopene activates the WNT/β-catenin and ERK1/2 pathways, upregulates RUNX2, alkaline phosphatase, COL1A and downregulates RANKL Saos-2. These processes contributed to prevent bone loss in postmenopausal women.

miR-140-3p aggregates osteoporosis by targeting PTEN and activating PTEN/PI3K/AKT signaling pathway.

Abstract: Osteoporosis (OP) is a systemic bone metabolic disorder, which negatively affects the quality of life in the elders and postmenopausal females. Healthy volunteers and postmenopausal females with OP were enrolled in the present study. Bone densitometry (BMD) was detected by dual-energy X-ray absorptiometry (DXA). CD14+PBMCs and C2C12 cells were cultured to induce osteoclast differentiation and osteoblast differentiation, respectively. The interaction between miR‑140-3p and PTEN was predicted and verified by TargetScan 7.2 and dual luciferase reporter assay, respectively. miRNA/RNA level and protein level were detected by quantitative real-time polymerase chain reaction (qRT-PCR) and western blot, respectively. Cell proliferation and apoptosis were detected by 5-ethynyl-2'-deoxyuridine (EdU) staining and flow cytometry, respectively. Cell differentiation of CD14+PBMCs and C2C12 cells were detected by tartrate-resistant acid phosphatase (TRAP) staining and alizarin red staining, respectively. The activity of alkaline phosphatase (ALP) was detected by ALP assay. Differences were observed in age, body mass index (BMI), and BMD between the OP group and the control group. Higher miR‑140-3p level and lower PTEN level were found in PBMCs of OP group compared to control group; there was a negative correlation between them in the serum of OP group. miR-140-3p targeted and downregulated the expression of PTEN. miR-140-3p inhibitor inhibited cell proliferation, differentiation, and promoted cell apoptosis of CD14+PBMCs; while promoted cell proliferation, differentiation and inhibited cell apoptosis of C2C12 cells, by targeting PTEN and inactivating PTEN/PI3K/AKT signaling pathway. These findings suggested a potential therapeutic role of miR-140-3p in the treatment of patients with OP.

Addition of fructose to the maize hyphosphere increases phosphatase activity by changing bacterial community structure

Abstract: Organic forms of phosphorus (P) tend to accumulate in soils with long-term fertilization, however they are generally not easily available to crops. Many crop plants are obligately arbuscular mycorrhizal (AM) and AM fungi are known for not being able to produce phosphatases, the enzyme involved in the mineralization of organic P. AM fungi are able to recruit bacteria in the hyphosphere which can produce phosphatases and access organic P. It is thought that fructose produced by AM fungi acts as a signal to select these bacteria. We designed a system to test whether fructose could stimulate phosphatase activity in the maize hyphosphere. The concentration of fructose produced by AM fungi and the large concentration of fructose equivalent to the entire concentration of hyphal exudate carbon, were added. Small concentrations of fructose did not stimulate phosphatase activity in soil, cause shifts in bacterial community structure or select phosphatase producing species. However, at large concentrations of fructose, phosphatase activity was increased and this was associated with a larger bacterial population and a shift in bacterial community structure towards species which specialized in using fructose and glucose for energy metabolism, such as Saccharibacteria. Nevertheless, there was no specific shift towards species which harbored alkaline phosphatase genes and it is suggested that the change in phosphatase activity was due to subtle shifts in the properties of the phosphatases being produced. These results suggest that fructose was unable to act as a signal to select phosphatase producing bacteria in soil, but could act as an energy source to stimulate phosphatase activity by shifting soil bacterial community structure.

Characteristics of laboratory indexes in COVID-19 patients with non-severe symptoms in Hefei City, China: diagnostic value in organ injuries.

Abstract: This study compared the laboratory indexes in 40 non-severe COVID-19 patients with those in 57 healthy controls. In the peripheral blood system of non-severe symptom COVID-19 patients, lymphocytes, eosinophils, basophils, total procollagen type 1 amino-terminal propeptide, osteocalcin N-terminal, thyroid-stimulating hormone, growth hormone, and insulin-like growth factor-binding protein 3 significantly decreased, and total protein, albumin, alanine transaminase, alkaline phosphatase, γ-glutamyl transferase, activated partial thromboplastin time, prothrombin time, fibrinogen, D-dimer, fibrinogen degradation products, human epididymal protein 4, serum ferritin, and C-reactive protein were elevated. SARS-CoV-2 infection can affect hematopoiesis, hemostasis, coagulation, fibrinolysis, bone metabolism, thyroid, parathyroid glands, the liver, and the reproductive system.

Ag-Ion-Modified Au Nanoclusters for Fluorometric Analysis of Alkaline Phosphatase

Abstract: Alkaline phosphatase (ALP) has been regarded as one vital biomarker in clinical diagnoses and biomedical studies, and fabrication of sensitive and effective approaches for ALP analysis is of extrao...

Single-Molecule Analysis Determines Isozymes of Human Alkaline Phosphatase in Serum

Abstract: Alkaline phosphatase (ALP) is an important biomarker, as high levels of ALP in blood can indicate liver disease or bone disorders However, current clinical blood tests only measure the total concentration of ALP but are unable to distinguish enzyme isotypes Here, we demonstrate a novel and rapid approach to profile various ALP isozymes in blood via a single-molecule-analysis platform The microarray platform provides enzyme kinetics of hundreds of individual molecules at high throughput Using these single molecule kinetics, we characterize the different activity profiles of ALP isotypes By analyzing both healthy and disease samples, we found the single molecule activity distribution of ALP in serum reflects the health status of patients This result demonstrates the potential utility of the method for improving the conventional ALP test, as well as for analyzing other enzymatic biomarkers, including enzyme isotypes

Prognostic value of alkaline phosphatase in hormone-sensitive prostate cancer: a systematic review and meta-analysis.

Abstract: To assess the prognostic value of alkaline phosphatase in patients with hormone-sensitive prostate cancer. A systematic review and meta-analysis was performed using the PUBMED, Web of Science, Cochrane Library, and Scopus in April 2019 according to the Preferred Reporting Items for Systematic Review and Meta-analysis statement. Studies were deemed eligible if they compared hormone-sensitive prostate cancer patients with high vs. low alkaline phosphatase to determine its predictive value for overall survival, cancer-specific survival, and progression-free survival. We performed a formal meta-analysis of these outcomes. 42 articles with 7938 patients were included in the systematic review and 28 studies with 5849 patients for the qualitative assessment. High alkaline phosphatase was associated with worse overall survival (pooled HR 1.72; 95% CI 1.37−2.14) and progression-free survival (pooled HR 1.30; 95% CI 1.10−1.54). In subgroup analyses of patients with “high-volume” and “low-volume”, alkaline phosphatase was associated with the overall survival (pooled HR 1.41; 95% CI 1.21−1.64 and pooled HR 1.64; 95% CI, 1.06−2.52, respectively). In this meta-analysis, elevated serum levels of alkaline phosphatase were associated with an increased risk of overall mortality and disease progression in patients with hormone-sensitive prostate cancer. In contrast, those were not associated with an increased risk of cancer-specific mortality. Alkaline phosphatase was independently associated with overall survival in both patients with “high-volume” and “low-volume” hormone-sensitive prostate cancer. Alkaline phosphatase may be useful for being integrated into prognostic tools that help guide treatment strategy, thereby facilitating the shared decision making process.

Reduced bone resorption and inflammation in apical periodontitis evoked by dietary supplementation with probiotics in rats.

Abstract: AIM To evaluate the relationship between systemic administration of probiotics and inflammation/resorption processes associated with apical periodontitis (AP) in a rat model. METHODOLOGY Twenty-four male Wistar rats were used. AP was induced in the mandibular left/right first molars. The animals were arranged into three groups: Control, Lactobacillus rhamnosus and L. acidophilus. Probiotics were orally administered via gavage (109 colony-forming units (CFU) diluted in 5 mL of water) for 30 days during the development of AP. On the 30th day, blood was collected to analyse the calcium, phosphorus and alkaline phosphatase concentrations in plasma. Then, the animals were euthanized and the jaws removed for micro-computed tomography and immune-histopathological analysis for receptor activator of NF-κB ligand (RANKL), osteoprotegerin (OPG) and tartrate-resistant acid phosphatase (TRAP). After the Shapiro-Wilk test of normality, the Kruskal-Wallis followed by Dunn's test was performed for nonparametric data, and analysis of variance followed by the Tukey test was performed for parametric data (P 0.05). The level of alkaline phosphatase was significantly higher in the groups that consumed probiotics (P < 0.05). A significantly lower volume of bone resorption was observed in groups that consumed probiotics (P < 0.05). The inflammatory infiltrates and the immunolabelling for RANKL and TRAP were significantly lower in probiotic groups when compared to the control (P < 0.05). Also, the OPG was significantly more immunolabelled in the L. acidophilus group than in the L. rhamnosus and control groups (P < 0.05). CONCLUSION Probiotic supplementation through gavage (L. rhamnosus and L. acidophilus) had a significant effect on the reduction of inflammation and bone resorption in apical periodontitis development in rats.

MicroRNA-145 suppresses osteogenic differentiation of human jaw bone marrow mesenchymal stem cells partially via targeting semaphorin 3A.

Abstract: Purpose: Human jaw bone marrow mesenchymal stem cells (h-JBMMSCs) are multipotent progenitor cells with osteogenic differentiation potential MicroRNAs (miRNAs) have emerged as crucial modulators of osteoblast differentiation In this study, we focus on the role of miR-145 and its target protein in osteoblast differentiation of h-JBMMSCs Materials and Methods: h-JBMMSCs were isolated and cultured in osteogenic medium miR-145 mimics and inhibitors were used to elevate and inhibit miR-145 expression, respectively Osteogenic differentiation was determined by Alkaline phosphatase (ALP) and Alizarin red S (ARS) staining, and osteogenic marker detection using quantitative real-time reverse transcription PCR (qRT-PCR) assay Bioinformatic analysis and luciferase reporter assay were used to identify the target gene of miR-145 Results: MiR-145 was down-regulated during osteogenesis of h-JBMMSCs Inhibition of miR-145 promoted osteogenic differentiation of h-JBMMSCs, revealed by enhanced activity of alkaline phosphatase (ALP), greater mineralisation, and increased expression levels of the osteogenic markers, such as Runt-related transcription factor 2 (RUNX2), Osterix (OSX), ALP and COL1A1 MiR-145 could negatively regulate semaphorin3A (SEMA3A), which acts as a positive regulator of osteogenesis MiR-145 inhibitor induced osteogenesis could be partially attenuated by SEMA3A siRNA treatment in h-JBMMSCs Conclusions: Our data show that miR-145 directly targets SEMA3A, and also suggest miR-145 as a suppressor, plays an important role in the osteogenic differentiation of h-JBMMSCs