Changes In Alkaline Phosphatase Activity Research Articles

In Vitro Proliferation of MG-63 Cells in Additively Manufactured Ti-6Al-4V Biomimetic Lattice Structures with Varying Strut Geometry and Porosity.

Lattice structures have demonstrated the ability to provide secondary stability in orthopedic implants by promoting internal bone growth. In response to the growing prevalence of lattices in orthopedic design, we investigated the effects of porosity and unit cell geometry in additively manufactured Ti-6Al-4V biomimetic lattice structures on the osteogenesis of human MG-63 osteoblastic cell lines in vitro. We analyzed glucose consumption, alkaline phosphatase (ALP) concentration, and end-of-culture cell count as markers for osteogenic growth. Two different strut geometries were utilized (cubic and body-centered cubic), along with four different pore sizes (400, 500, 600, and 900 µm, representing 40-90% porosity in a 10 mm cube), in addition to a solid specimen. Structural characterization was performed using scanning electron microscopy. The results indicated that lattices with a 900 µm pore size exhibited the highest glucose consumption, the greatest change in ALP activity, and the highest cell count when compared to other pore sizes. Cubic 900 µm lattice structures outperformed other specimens in facilitating the maturation of viable MG-63 cells from the formation to the mineralization phase of bone remodeling, offering the most promise for osseointegration in additively manufactured titanium implants in the future. However, irrespective of a particular pore size or unit cell geometry, it was found that all the lattices were capable of promoting osteogenic growth due to surface roughness in the printed parts.

Biochemical Markers of Liver Damage in Cattle Naturally Infected with Fasciola Hepatica

Abstract Fascioliasis is a worldwide spread parasitic disease with a high impact on the economy and public health in many countries. Therefore, it is important to apply prevention, effective treatment, and education campaigns, to prevent the disease. This work aimed to determine the biochemical markers of liver damage caused by Fasciola hepatica during 21 days (D0 – D21) post albendazole (ABZ) therapy in the blood sera. This study included 24 cows naturally infected with Fasciola hepatica, randomly divided into a control group (n = 12) and a group treated per os with ABZ (7.5 mg per kg body weight) (n = 12). The activity of hepatic enzymes – aspartate aminotransferase (AST), alanine aminotransferase (ALT) and cholestatic enzymes – gamma-glutamyl transferase (GGT), and alkaline phosphatase (ALP), were measured in both groups. The activity of AST has significantly increased in the course of the experiment (D0 – D21) in both groups (P < 0.001). ALT activity during D0 – D21 was significantly decreased in both groups (P < 0.001). Activity of GGT was not significantly changed in the control group in the course of the experiment (P > 0.05). Contrarily, the activity of GGT in the treated group was significantly decreased (P < 0.01) during D0 – D21. The activity of GGT in the blood sera of the treated group was significantly lower in comparison with the control group in D7 and D21 (P < 0.01). This decreased GGT activity in the blood sera of the treated group may be caused by the effect of albendazole resulting in fasciola elimination and bile flow improvement. The results of ALP determination show no significant changes in ALP activity between groups. This can lead to the conclusion that the most useful marker of cholestasis caused by F. hepatica is the GGT activity and may help to evaluate the ABZ therapy effect in cattle.

Functional and biochemical characteristics of the muscle system in children with type I diabetes

Background. The purpose of the study was to determine possible markers of skeletal muscle damage in children with type 1 diabetes mellitus (T1DM) and their relationship with the features of disease course. Materials and methods. The observation group consisted of 98 children with type 1 diabetes mellitus: the first group included 22 people without disorders of the muscular system; the second — 42 patients with dynapenia; the third — 34 children with diabetic myopathy. Control group — 30 relatively healthy children. Assessment of the static endurance of skeletal muscles, determination of the level of creatine kinase, aspartate aminotransferase, alkaline phosphatase, lactate dehydrogenase, periostin and cardiotrophin-1 in blood serum were performed in all patients. Results. The conducted studies demonstrate that children with diabetes, regardless of the structural and functional state of their muscular system, have signs of skeletal muscle damage, which were most expressed in diabetic myopathy and progressed with maximal deterioration of glycemic control. It was found that the highest content of alkaline phosphatase was characteristic of children from group 1, while in patients with diabetic myopathy its serum content was not statistically different from that of controls. These disorders occurred against the background of changes in alkaline phosphatase activity, the level of which was highest in children from group 1, while in patients with diabetic myopathy, its serum content was not statistically different from that of controls. At the same time, during the course of diabetic myopathy in children with T1DM, there was an increase in lactate dehydrogenase activity by 1.2 times (p < 0.01) and cardiotrophin-1 by 300 times (p < 0.01) compared to the corresponding indicator of the control group. Serum periostin level was increased in all patients with T1DM. Its maximum values were determined in group 1, whose periostin concentration exceeded control indicators by 103 times (p < 0.01). With deterioration of skeletal muscle state, there was a gradual decrease in periostin serum level, but in patients with dynapenia, it was 35.5 times higher than in the control group (p < 0.05) and 19.2 times higher in those with diabetic myopathy (p < 0.05). Conclusions. The course of type 1 diabetes in children is accompanied by skeletal muscle damage, the first clinical sign of which is a decrease in the static muscle endurance against the background of worsening disease course. Alkaline phosphatase, lactate dehydrogenase, periostin, and cardiotrophin-1 are biochemical markers of skeletal muscle damage in children with type 1 diabetes. A common feature of the changes in the specified indicators is their increase; however, each clinical condition of the skeletal muscles corresponds to its own configuration of changes in the abovementioned markers.

Ratiometric detection and imaging of endogenous alkaline phosphatase activity by fluorescein-coumarin-based fluorescence probe

Alkaline phosphatase (ALP) is a type of enzyme that widely exists in various tissues of the human body; it plays an important role in regulating many cell functions. The development of a sensitive and accurate tool to detect the changes of ALP activity in organisms can contribute to research in the fields of biochemistry, cytology, clinical medicine, etc. In this paper, a small organic molecule-based ratiometric fluorescent probe (FCP) was designed based on the hydroxyl electron-donating group in fluorescein-coumarin protected by the phosphate group. ALP can trigger the fluorescence change through the enzyme-catalyzed cleavage of phosphoryl ester groups, and the ratio of ALP can be measured at wavelengths of 465 nm and 530 nm. The probe had high selectivity and sensitivity to ALP, and the detection limit measured under the optimal conditions in an aqueous medium reached 0.006 mU/mL. The ALP activity of human serum samples was determined using the probe and found to be in good agreement with that measured using commercial ALP kits. Finally, the probe was also successfully applied to image ALP in living hepatocytes with good selectivity and sensitivity.

BIOCHEMICAL PARAMETERS CHANGES OF JAW BONE REMODELING IN EXPERIMENTAL ANIMALS UNDER THE INFLUENCE OF THE SUGGESTED PASTE, INTENDED FOR LOCAL TREATMENT OF CHRONIC GRANULATING PERIODONTITIS OF PERMANENT TEETH IN CHILDREN

The aim of the research was to study in an experiment on rats the biochemical parameters of the jaw bone remodeling under the influence of the developed paste, intended for the local treatment of chronic granulating periodontitis of permanent teeth in children.
 The experimental research was performed on 36 young rats (males) of the Wistar line (130-150g body weight), which were kept in the vivarium. The animals were divided into 2 groups: an experimental and a control one - 18 rats each. They were operated in sterile conditions under ketamine anesthesia. After applying the alcohol to the skin, a scalpel was used to make an incision along the lower edge of the mandible in the area of the canines and a longitudinal incision of the buccal muscle, then access to the bone was bluntly created. Using a sharp dental probe, a bone defect was modeled, in which the animals of the experimental group were injected with a paste based on comfrey tincture and calcium hydroxide. The control group was operated without the application of paste, the bone defect healed under a blood clot. The surgical wound was sutured in layers with polyamide thread and treated with 1% alcohol solution of diamond green.
 Blood sampling in experimental animals of both groups (6 rats each) was performed on 7th, 14th and 28th day after surgery. Animals were removed from the experiment by decapitation under ketamine anesthesia. Experimental animals were operated on and removed from the experiment during one day, under the same conditions.
 Determination of the amount of calcium and inorganic phosphorus in the blood serum of rats was carried out using standard kits of research by "SIMKO Ltd” company, Lviv. The activity of acidic and alkaline phosphatases in the serum was determined using kits of research by BIO-Ld-TEST company, Lachem (Czech Republic) and "Simbj Ltd", Lviv, respectively. In order to determine these parameters in the serum of normal rats, we took blood from 6 healthy young animals that did not participate in the experiment and were on the diet of the vivarium.
 The research found out that in the healing of artificial bone defects in the lower jaws of rats of the experimental group there were significant fluctuations in calcium and phosphorus levels and changes in the activity of acid phosphatase and alkaline phosphatase in the serum. The amount of Ca and P in the blood of rats decreased sufficiently. Alkaline phosphatase activity increased in both groups on the 7th and 14th day of the experiment, but the difference between the indicators in the groups was 38.35% (p <0.05) and 13.16% (p <0, 01) respectively. This was probably due to the rapid remission of inflammation at the site of surgery in the experimental group, as the paste we developed, which was used to fill a bone defect in this group of animals, has anti-inflammatory properties. Subsequent changes in alkaline phosphatase activity occurred similarly in both groups. This testified to the intensification of bone remodeling processes, as well as bone regeneration and mineralization under the influence of the paste we have developed.
 Our biochemical researches of the blood of experimental animals confirmed the previously obtained results of morphological and densitometric studies and showed that the recovery of bone wounds of the lower jaws of rats occurs almost equally in the experimental and control groups, and paste based on comfrey tincture and calcium hydroxide promotes bone regeneration at the site of the defect and stimulates osteosynthesis.

Selected abstracts presented at the annual conference of the Canadian Society of Clinical Chemists (CSCC), 2020

Alkaline phosphatase (ALP) serves as a biomarker for diagnosing several types of diseases in adults; nonetheless, its use is restricted in children because of changes in ALP activity during different physiological phases. The present study aimed to investigate ALP activity and its dynamics in children of different ages to establish the reference values for ALP activity in children.Total 167,625 samples of children aged 0–18 years were enrolled in this study. ALP activity was measured using the 4-nitrophenyl-1-phosphate disodium salt (4-NPP)–2-amino-2-methyl-1-propanol (AMP) method with an automatic biochemical analyzer. Patients with known diagnoses that may affect ALP activity were excluded. A percentile curve was plotted using MATLAB software, and the curve was fitted using the skewness–median–coefficient of variation (LMS) method.ALP activity reached the highest peak at 12–13 years of age and then gradually decreased to the lowest peak at 18–19 years of age in boys, whereas it reached the highest at 10–11 years and then gradually reduced to the lowest at 17–18 years in girls. Furthermore, the highest peak of ALP activity appeared substantially earlier in children of either sex in China than in those in Germany.We showed the dynamics of ALP activity in both boys and girls between the ages of 0 and 18 years in China and compared the difference in ALP activity between children in China and Germany. Our findings provide a reference for clinicians.

An in-vitro medical device material biocompatibility study using primary cell cultures of rat osteoblasts

<p class="abstract"><strong>Background:</strong> We questioned if simple chemical methods could be applied as a possible biocompatibility test for biomaterials.</p><p class="abstract"><strong>Methods:</strong> In a qualitative experiment, osteoblasts cells harvested from newly born (3-5) day old sprague-dawley rats were cultivated in growth medium in a controlled environment in the presence of titanium (Ti) (American Elements®), cobalt-chromium (Co-Cr) (Nobilium®), bio-activated rattan wood, orthopaedic bone cement (CEMEX®), implant fixtures (Astra TDC), and resorbable suture material (Poly-Gly-Lac) (VICRYL®). Sample aliquots were withdrawn periodically over 16 days. One and two-dimensional polyacrylamide gel electrophoresis (PAGE) as well as isoelectric focusing (IEF) produced spots that were subjected to enzyme digestion and molecular weight determination. In a follow-up quantitative experiment, the same samples, except for those containing CEMEX and VICRYL, were prepared. The alkaline phosphatase (ALP) activity was monitored continuously using the colorimetric Stanbio® kits. The ALP activities at days 2, 4, 8, 12, and 16, designated as a longitudinal variable, TIME, were analysed, using SPSS V.22, as a mixed ANOVA model with TIME as a repeated measure and the material type as an independent factor.</p><p class="abstract"><strong>Results:</strong> The IEF and second dimension PAGE produced an additional spot for Ti at pH(I) of 5-6. This was identified as IQUB_RAT IQ, a ubiquitin-like domain, molecular weight 2.6 kDa. This method was able to find finite differences in osteoblast activity after initial exposure to a foreign body. Both the ALP activity changes from one day to the next for all materials and the TIME-material type interaction effects were significant (p=0.000).</p><p class="abstract"><strong>Conclusions:</strong> This technique is suitable for use with human cell lines or clones. Experiments like these reduce the need for animal testing. </p>

All-trans retinoic acid and vascular endothelial growth factor induced the directional osteogenic differentiation of mouse embryonic fibroblasts

To investigate the effect of all-trans retinoic acid (ATRA) and vascular endothelial growth factor (VEGF) on the osteogenic differentiation of mouse embryonic fibroblasts (MEFs). The fetal mice in the uterus of NIH pregnant mice (pregnancy 12-15 days) were collected, and the heads and hearts etc. were removed. Then MEFs were separated from the rest tissues of the fetal mice and cultured by trypsin digestion and adherent culture. HEK-293 cells were used to obtain recombinant adenovirus-red fluorescent protein (Ad-RFP) and Ad-VEGF by repeatedly freezing and thawing. Alkaline phosphatase (ALP) staining and quantitative detection were used to detect the changes of ALP activity in MEFs applied with ATRA or VEGF alone or combined use of ATRA and VEGF on the 3rd and 5th days. The cultured 3rd to 4th generation MEFs were divided into groups A, B, C, and D, and were cultured with DMSO plus Ad-RFP, ATRA, Ad-VEGF, ATRA plus Ad-VEGF, respectively. Real-time fluorescence quantitative PCR (qRT-PCR) was used to detect the mRNA expressions of osteogenic markers including ALP, collagen type Ⅰ, osteopontin (OPN), osteocalcin (OCN), and angiogenic markers including VEGF, angiopoietin 1 (ANGPT1), and endomucin (EMCN) on the 3rd and 7th days. Immunohistochemical staining was used to detect the protein expressions of OPN and VEGF on the 3rd, 5th, and 7th days in each group. Alizarin red staining was used to detect calcium salt deposition levels in each group at 14 and 21 days after osteogenic induction. Fifteen athymic female nude mice aged 4 to 6 weeks were randomly divided into 3 groups and 5 mice in each group. Then MEFs treated with ATRA, Ad-VEGF, and ATRA plus Ad-VEGF were injected subcutaneously into the dorsal and ventral sides, respectively. X-ray observation, gross observation, and histological staining (Masson, HE, and Safranin O-fast green stainings) were performed at 5 weeks after implantation to observe the ectopic bone formation in nude mice in each group. MEFs were successfully isolated and cultured. The acquired Ad-RFP and Ad-VEGF were successfully transfected into MEFs with approximately 50% and 20% transfection rates. ALP activity tests showed that ATRA or Ad-VEGF could enhance ALP activity in MEFs ( P<0.05), and ATRA had a stronger effect than Ad-VEGF; and the combined use of ATRA and Ad-VEGF significantly enhanced the ALP activity in MEFs ( P<0.05). qRT-PCR test showed that the combined use of ATRA and Ad-VEGF also increased the relative mRNA expressions of early-stage osteogenesis-related markers ALP, OPN, and collagen type I ( P<0.05); the relative mRNA expressions of angiogenesis-related markers VEGF, EMCN, and ANGPT1 increased at 7 days ( P<0.05). Immunohistochemical staining showed that ATRA combined with Ad-VEGF not only enhanced OPN protein expression, but also increased VEGF protein expression on 7th day. Alizarin red staining showed that the application of ATRA or Ad-VEGF induced weak calcium salt deposition, and the combined use of ATRA and Ad-VEGF significantly enhanced the effect of calcium salt deposition in MEFs. The results of implantation experiments in nude mice showed that X-ray films observation revealed obvious bone mass in the ATRA plus Ad-VEGF group, and the bone was larger than that in other groups. Histological staining showed a large amount of collagen and mature bone trabeculae, bone matrix formation, and gray-green collagen bone tissue, indicating that the combined use of ATRA and Ad-VEGF significantly enhanced the osteogenic effect of MEFs in vivo. The combined use of ATRA and VEGF can induce the osteogenic differentiation of MEFs.

LncRNA TERC alleviates the progression of osteoporosis by absorbing miRNA-217 to upregulate RUNX2.

To elucidate the role of telomerase RNA elements (TERC) in alleviating osteoporosis (OP) by absorbing microRNA-217 (miRNAs-217) to regulate runt-related transcription factor 2 (RUNX2) level. The serum levels of TERC and miRNA-217 in OP patients and healthy controls were determined. During the osteogenic process, the relative levels of alkaline phosphatase (ALP), RUNX2, and Osterix were determined in hMSCs. The regulatory effects of TERC, miRNA-217, and RUNX2 on ALP and RUNX2 levels in hMSCs were examined by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) and Western blot. In addition, the changes in ALP activity and calcification ability in hMSCs influenced by them were assessed through ALP activity determination and alizarin red staining, respectively. The interaction of TERC/miRNA-217/RUNX2 regulatory loop and its role in influencing hMSCs osteogenesis were assessed by Dual-Luciferase Reporter Gene Assay and a series of rescue experiments, respectively. The downregulated TERC and upregulated miRNA-217 were identified in the serum of the OP patients. Consistently, the downregulated TERC and upregulated miRNA-217 were discovered in in vitro osteogenic process of hMSCs. The silence of TERC, or RUNX2 downregulated ALP and RUNX2 levels, decreased ALP activity and attenuated the calcification ability in hMSCs. The overexpression of miRNA-217 gave similar results. The binding relationship in TERC/miRNA-217/RUNX2 regulatory loop was verified. At last, rescue experiments suggested that TERC accelerated hMSCs osteogenesis by absorbing miRNA-217 to upregulate RUNX2. The serum level of TERC is lowly expressed in OP patients. TERC influences hMSCs osteogenesis by absorbing miRNA-217 to upregulate RUNX2, thus alleviating the progression of OP.

Non-thermal effects of 0.1 THz radiation on intestinal alkaline phosphatase activity and conformation**Project supported by the National Natural Science Foundation of China (Grant No. 61675151).

Alkaline phosphatase (ALP) plays an integral role in the metabolism of liver and development of the skeleton in humans. To date, the interactions between different-duration terahertz (THz) radiation and ALP activities, as well as the influence mechanism are still unclear. In this study, using the para-nitro-phenyl-phosphate (pNPP) method, we detect changes in ALP activities during 40-minute THz radiation (0.1 THz, 13 mW/cm2). It is found that the activity of ALP decreases in the first 25 min, and subsequently increases in the later 15 min. Compared with the activity of ALP being heated, the results suggest that short-term terahertz radiation induces a decrease in enzyme activity through the non-thermal mechanism. In order to explore the non-thermal effects of THz radiation on ALP, we focus on the impacts of 0.1 THz radiation for 20 min on the activity of ALP in different concentrations. The results reveal that the activity of ALP decreases significantly after exposure to THz radiation. In addition, it could be deduced from fluorescence, ultraviolet-visible (UV-vis), and THz spectra results that THz radiation has induced changes in ALP structures. Our study unlocks non-thermal interactions between THz radiation and ALP, as well as suggests that THz spectroscopy is a promising technique to distinguish ALP structures.

Effects of sinusoidal electromagnetic fields at different exposure times on osteoblastic activity and the underlying signal transduction mechanisms

Objective To investigate the signal transduction mechanisms of time-dependent effects of 50 Hz 1.8 mT sinusoidal electromagnetic fields(SEMFs)on osteoblastic activity. Methods Newborn rat calvarial osteoblasts(ROBs)were treated with 50 Hz 1.8mT SEMFs for 30, 60, 90, 120, and 150 min, respectively.Intracellular alkaline phosphatase(ALP)activity was assayed, and protein expression of Runt-related transcription factor 2(Runx2), Smad1/5/8 and mitogen-activated protein kinases(MAPKs)were examined by Western blot.The Dual-Luciferase Reporter Assay System was used to measure the activity of the Wnt pathway.Immunofluorescence staining and laser confocal microscopy were applied to examine the nuclear translocation of Smad1/5/8 and β-catenin.Changes in ALP activity were determined after inhibiting p38 MAPK using a specific inhibitor. Results ALP activity of ROBs increased after 30, 60, 90, 120 and 150 min of SEMFs treatment, compared with the control group(51.41±5.21, 59.47±4.02, 67.56±4.68, 63.69±3.92, and 58.16±3.61 vs.45.53±3.24), and reached the highest value at 90 min and then started to decline.Protein expression of Runx-2, p-Smad1/5/8, p-p38 and β-catenin increased after SEMFs treatment, and reached the highest value at 90 min and then gradually returned to baseline levels.The values for Wnt pathway activity for the control group and with SEMFs treatment at 30, 60, 90, 120 and 150 min were 0.49±0.06, 0.52±0.09, 0.75±0.05, 0.77±0.42, 0.58±0.08 and 0.42±0.09, respectively.Wnt pathway activity, the nuclear translocation of β-catenin and Smad1/5/8 reached the highest level at 90 min.Values of ALP activity in the control group and the SEMFs group were 44.60±3.84 and 71.54±7.34, respectively, before specifically inhibiting p38 MAPK, and were 52.08±0.83 and 52.15±10.77, respectively, after p38 MAPK inhibition, indicating that ALP activity could not be increased with inhibition. Conclusions 50 Hz 1.8 mT SEMFs increase osteoblastic activity by activating the BMP-2/Smad1/5/8, Wnt/β-catenin and p38 MAPK signal pathways.The optimal duration of treatment is 90 min per day. Key words: Electromagnetic fields; Osteoblasts; Alkaline phospohatase; Signal transduction

Promoting Proliferation and Differentiation of Pre-Osteoblasts MC3T3-E1 Cells Under Combined Mechanical and Electrical Stimulation.

Electrical or mechanical stimulation has been applied widely in bone fracture treatment or bone tissue construction. The effect of combined mechanical and electrical stimulation to osteoblasts or bone tissue has not been deeply studied, which would provide a more actual simulation of in vivo microenvironment. In this study, a cell culture device that can simultaneously load mechanical and electrical stimulation on cells was self-designed and fabricated. The combined laminar shear stress and electrical stimulation could be supplied to MC3T3-E1 cells seeded on the nano-crystalline fluorine-doped tin oxide glass plate. Cell counting kit-8 assay was used to study the proliferation of MC3T3-E1 cells. The changes in alkaline phosphatase activities, genes expression of collagen-I (Col-I) and fibronectin and concentration of Col-I were also measured. Compared with single stimulation, the combined stimulation obviously enhanced proliferation, differentiation and synthesis of extracellular matrix of pre-osteoblasts in a greater extent. The combined mechanical and electrical stimulation provide a new way to promote in vitro cultivation of cells and tissues, which will be very significant in the development of tissue engineering.

Метаболічний і функціональний стан печінки за умов комбінованої дії свинцю і фтору на фоні застосування біопротекторів

Introduction. The potential danger of combined effect of lead and fluorine to human health motivates the search and experimental substantiation of physiologically acceptable ways of bioprophylaxis directed to preventing or reducing the negative effect on the organism. Objective: the study of the dynamics of the metabolic and functional state of the liver under conditions of combined effect of lead and fluorine without application of bioprotectors and on the background of bioprotectors application. Material and Methods. 4 series of researches were carried out on white rats, each according to the orthogonal planning scheme 22. Lead and fluorine were administered by intragastric within 30 days: in the first series — without bioprotectors application, in the second series — on the background of pectin application, in the third series — pectin and calcium application, in the fourth series — the complex of pectin, calcium and triovit application. The level of diene conjugates, the active products of thiobarbituric acid, the activities of superoxide dis-mutase and catalase, the index of general antioxidant activity in the liver tissue, the activities of alanine aminotransferases, aspartate aminotransferases and alkaline phosphatase in blood serum, concentration of urea were determined at the 15th and 30th day of the experiments, the integral coefficient of antioxidant defense and the type of combined action were calculated. Results. The combined effect of lead and fluorine leads to an imbalance between the indicators of lipoperox-idation and antioxidant activity of the liver tissue, the increased activities of alanine aminotransferases and aspartate aminotransferases, the phase changes in alkaline phosphatase activity, and urea concentration in serum. The application of pectin did not affect the indicators. Pectin and calcium have improved the mechanisms of antioxidant system and the activity of enzymes. The complex of pectin, calcium and triovit normalizes metabolic processes in the liver tissue and improves its functional state. The type of the combined action depends on the registered effect, the duration of the stress factor and the applied bioprotektor. Conclusion. The optimal prophylactic effect under the combined action of lead and fluorine is achieved with the help of a complex of pectin, calcium and triovit. Key Words: lead, fluorine, combined effect, pectin, calcium, triovit, indicators of lipid peroxidation, antioxidant defense, enzyme activity.

Protective effect of endoplasmic reticulum stress on calcification of VSMCs induced by high glucose and its effect on expression of GRP78, CHOP, and Caspase-12

This article investigates the role of endoplasmic reticulum stress (ERS) in high-glucose-induced vascular smooth muscle cells (VSMCs) calcification and to explore its effects on the expression of GRP78, CHOP, and Caspase-12.VSMCs were treated with high glucose (35 mmoL/L) to induce diabetes to see whether high glucose can induce ERS. Changes in alkaline phosphatase activity, calcium deposition, and runt-related transcription factor 2 (Runx2) were measured. After 5 days’ treatment, the alkaline phosphatase activity, calcium content, and Runx2 expression of the bone differentiation marker protein were all up-regulated in the β-glycerophosphate group, the high-glucose group, and the β-glycerophosphate + high-glucose group when compared with the control group. High-glucose treatment of VSMCs can cause ERS and apoptosis and induces transdifferentiation of VSMCs into osteoblast-like cells, resulting in increased basal phosphatase activity, up-regulation of calcium content, and bone differentiation markers in VSMCs. The findings confirm that ERS plays an important role in the calcification of VSMCs.

A novel switchable fluorescent sensor for facile and highly sensitive detection of alkaline phosphatase activity in a water environment with gold/silver nanoclusters.

A novel fluorescent sensor based on bovine serum albumin stabilized gold/silver nanoclusters (BSA-Au/Ag NCs) was developed for sensitive and facile detection of alkaline phosphatase (ALP) activity. For this fluorescent sensor, ascorbic acid 2-phosphate (AAP) was decomposed into ascorbic acid (AA) and phosphate by catalysis with ALP. The initial red fluorescence of the BSA-Au/Ag NCs was effectively quenched by KMnO4 and then the fluorescence was recovered by addition of AA. The mechanism of interaction between BSA-Au/Ag NCs and KMnO4 and AA was studied with use of the fluorescence lifetime and UV-vis absorption spectra. The results indicated that the oxidation/reduction modulated by KMnO4/AA led to surface structure destruction/restoration of the BSA-Au/Ag NCs, resulting in fluorescence quenching/recovery. The proposed fluorescence-based method based on a dark background was used to detect ALP and had excellent sensitivity, with a detection limit of 0.00076 U/L. Moreover, the method was applied to the determination of added analytes, with satisfactory recoveries (97.0-105.0 %). In a simulated eutrophic water body, this method successfully detected ALP in actual water samples and could monitor the dynamic changes of ALP activity through visual observation. More importantly, the proposed fluorescent sensor not only has the advantages of simple operation and high sensitivity but has also been successfully used on filter paper to establish a rapid and visual test paper for ALP.

Change in alkaline phosphatase activity associated with intensive care unit and hospital length of stay in patients with septic acute kidney injury on continuous renal replacement therapy

BackgroundEvidence suggests that alkaline phosphatase attenuates inflammatory response in sepsis by lipopolysaccharide detoxification and adenosine triphosphate dephosphorylation. We sought to determine changes in alkaline phosphatase (AP) activity during septic acute kidney injury (AKI) and clinical parameters associated with AP activity.MethodsIn this retrospective study, we investigated baseline (when initiating CRRT) and follow-up AP activity on day 3, and associated outcomes in patients who underwent continuous renal replacement therapy (CRRT) due to septic AKI.ResultsWe analyzed the baseline AP activity of 155 patients and day 3 AP activity in 123 patients. Baseline AP activity was not associated with renal or inflammatory biomarkers, or outcomes. It did not significantly differ between the 75 survivors and 80 non-survivors (p = 0.155). AP activity was higher on day 3 than at baseline (105 U/L [interquartile range, 79–156] vs 90 U/L [interquartile range, 59–133]). In particular, liver and bone isoforms increased significantly (p < 0.05), but intestine isoforms did not reach statistical significance (p = 0.367). In addition, day 3 AP activity showed a weak correlation with length of ICU stay (r = 0.213, p = 0.018) and length of hospital stay (r = 0.216, p = 0.017), but not with survival (r = − 0.035, p = 0.698).ConclusionEndogenous AP activity significantly increased in patients with septic AKI. However, neither baseline nor follow-up AP activity was associated with survival.

Change in Alkaline Phosphatase Activity Induced By Tetracycline in Freshwater Mussel, Parreysia cylindrica (Annandale and Prashad)

Change in Alkaline Phosphatase Activity Induced By Tetracycline in Freshwater Mussel, Parreysia cylindrica (Annandale and Prashad)

Effects of Rainfall in Kunming on the Growth and Alkaline Phosphatase Activity of the Cyanobacterium Microcystis aeruginosa

The effects of rainwater on freshwater ecosystems have received increasing attention worldwide. Alkaline phosphatase activity (APA) affects the biochemical cycles of phosphorus in the water, thereby affecting the proliferation and outbreak of cyanobacteria blooms. However, it is still unclear whether the complex composition of rainwater has a significant effect on the alkaline phosphatase activity. In this study, the effects of rainfall on Microcystis aeruginosa in Kunming were evaluated based upon changes in alkaline phosphatase activity, algal growth, and chlorophyll a. The results showed that the addition of rainwater brought about different changes in alkaline phosphatase activity, algal growth, and chlorophyll a. The general trend showed that a certain amount of rainwater (equivalent to light, moderate rain, and heavy levels) had a positive effect on the alkaline phosphatase activity, algal growth, and chlorophyll a in the cultivation system. Therefore, the following preliminary inference can be drawn: in the Kunming area, a certain amount of rainwater may promote the growth of blue-green algae, exacerbating an outbreak of cyanobacteria blooms.

Pediatric reference intervals for alkaline phosphatase.

Interpretation of alkaline phosphatase activity in children is challenging due to extensive changes with growth and puberty leading to distinct sex- and age-specific dynamics. Continuous percentile charts from birth to adulthood allow accurate consideration of these dynamics and seem reasonable for an analyte as closely linked to growth as alkaline phosphatase. However, the ethical and practical challenges unique to pediatric reference intervals have restricted the creation of such percentile charts, resulting in limitations when clinical decisions are based on alkaline phosphatase activity. We applied an indirect method to generate percentile charts for alkaline phosphatase activity using clinical laboratory data collected during the clinical care of patients. A total of 361,405 samples from 124,440 patients from six German tertiary care centers and one German laboratory service provider measured between January 2004 and June 2015 were analyzed. Measurement of alkaline phosphatase activity was performed on Roche Cobas analyzers using the IFCC's photometric method. We created percentile charts for alkaline phosphatase activity in girls and boys from birth to 18 years which can be used as reference intervals. Additionally, data tables of age- and sex-specific percentile values allow the incorporation of these results into laboratory information systems. The percentile charts provided enable the appropriate differential diagnosis of changes in alkaline phosphatase activity due to disease and changes due to physiological development. After local validation, integration of the provided percentile charts into result reporting facilitates precise assessment of alkaline phosphatase dynamics in pediatrics.

Gingival crevicular fluid alkaline phosphate activity during the retention phase of maxillary expansion in prepubertal subjects: A split-mouth longitudinal study.

The aim of this study was to monitor the alveolar bone formation at the tension sites of teeth supporting the appliances for rapid maxillary expansion (RME) during the retention phase according to the local gingival crevicular fluid (GCF) alkaline phosphatase (ALP) activity. This split-mouth prospective study included 23 prepubertal subjects (15 girls, 8 boys; mean age, 9.0 ± 1.4 years) who had a constricted maxillary arch and were undergoing RME. Periodontal parameters, including probing depth, were recorded at 3 and 6 months after RME. Furthermore, the GCF ALP activity was measured at the tension sites of the supporting test teeth (TT) and at the antagonist control teeth (CT) sites. Periodontal parameters were generally similar between the TT and CT sites during the study, with the exception that probing depth underwent a slight increase at the TT sites. At baseline, the GCF ALP activity was similar between the TT and CT sites; however, at both 3 and 6 months, significantly greater enzymatic activity was seen at the TT sites. The overall probing depth changes were not significantly correlated with the corresponding GCF ALP activity changes for either the TT or the CT sites. Alveolar bone formation at the tension sites would last up to 6 months of retention after RME. These results warrant more comprehensive studies to assess whether the GCF ALP activity has potential as a diagnostic tool for bone formation during the retention phase of RME.