Alkaline Phosphatase Enzyme Activity Research Articles

Evaluation of dental pulp stem cells response to flowable nano-hybrid dental composites: A comparative analysis.

Flowable resin composites (FRC) are tooth-colored restorative materials that contain a lower filler particle content, and lower viscosity than their bulk counterparts, making them useful for specific clinical applications. Yet, their chemical makeup may impact the cellular population of the tooth pulp. This in-vitro study assessed the cytocompatibility and odontogenic differentiation capacity of dental pulp stem cells (DPSCs) in response to two recent FRC material extracts. Extracts of the FRC Aura easyflow (AEF) and Polofil NHT Flow (PNF) were applied to DPSCs isolated from extracted human teeth. Cell viability of DPSCs was assessed using MTT assay on days 1, 3 and 7. Cell migration was assessed using the wound healing assay. DPSCs' capacity for osteo/odontogenic differentiation was assessed by measuring the degree of mineralization by Alizarin Red S staining, alkaline phosphatase enzyme (ALP) activity, and monitoring the expression of osteoprotegerin (OPG), RUNX Family Transcription Factor 2 (RUNX2), and the odontogenic marker dentin sialophosphoprotein (DSPP) by RT-PCR. Monomer release from the FRC was also assessed by High-performance liquid chromatography analysis (HPLC). DPSCs exposed to PNF extracts showed significantly higher cell viability, faster wound closure, and superior odontogenic differentiation. This was apparent through Alizarin Red staining of calcified nodules, elevated alkaline phosphatase activity, and increased expression of osteo/odontogenic markers. Moreover, HPLC analysis revealed a higher release of TEDGMA, UDMA, and BISGMA from AEF. PNF showed better cytocompatibility and enhancement of odontogenic differentiation than AEF.

Exploring the driving forces and digital mapping of soil biological properties in semi-arid regions

Soil biological properties (SBPSs) are crucial for soil health and ecosystem functioning. Mapping these properties can provide insights into soil ecosystems and help develop management and conservation strategies. The objective of this study was to map five important indicators of SBPSs, namely urease (Ur), acid phosphatase (ACP), alkaline phosphatase enzyme (ALP) activity, metabolic quotient (qCO2), and soil microbial biomass to soil organic carbon ratio (MBC: SOC) using machine learning algorithms (MLA) and to identify the most important driving factors of these properties. The study was conducted in Honam sub-basin, Lorestan, Iran, on an area of 14,000 ha. 90 surface soil samples (0–30 cm) were collected and analyzed in the laboratory. 60 soil and environmental covariates, including soil, topography, and remote sensing data (RS), were used to represent influencing factors. The most relevant covariates were selected based on correlation and recursive feature elimination (RFE) analysis using two state-of-the-art MLAs, Random Forest (RF) and Extreme Gradient Boosting (XGBoost) trees, were used to predict SBPSs based on the selected covariates. Relative importance analysis and partial dependence plots were used to evaluate the effects of different factors on the prediction of SBPSs. The results showed that 28 soil and environmental covariates were selected as relevant factors. Both RF and XGBoost models performed well and achieved acceptable accuracy (as measured by R2 value). XGBoost outperformed RF on qCO2 (R2 = 0.61), ALP (R2 = 0.73) and Ur (R2 = 0.75), while RF performed better on ACP (R2 = 0.65) and the MBC: SOC (R2 = 0.81). Relative importance analysis showed that ACP and Ur were most strongly associated with the soil variables at 79 % and 42 %, respectively. ALP and qCO2 were strongly correlated with topographic attributes at 56 % and 52 %, respectively. MBS: SOC was about 40 % associated with the RS indices. The influence of each SBPS varied depending on the specific types of covariates. These results highlight the potential of targeted strategies for effective management and protection of soil ecosystem leading to improved soil health and ecosystem function. Further research should focus on developing methods that encompass a broader range of indices that influence soil microbial activity in drylands.

Soil properties and quinoa yield as affected by irrigation and vermicompost application under a drip-tape irrigation system

IntroductionQuinoa a crop with high nutritional value has the widespread adaptability to different climates and there is an increased demand for this product in the world. Characterized by roots with extensive penetration capabilities and wide, alternate leaves giving rise to diverse inflorescences, quinoa plants exhibit variable heights ranging from 20 to 300 cm. The duration of their growth phase spans 95 to 125 days’ contingent upon cultivar selection and prevailing climatic conditions.Materials and methodsIn order to reduce chemical fertilizers consumption through using organic fertilizers a two-year study was conducted at the Research Farm of the University of Kurdistan, Iran, during the 2021–2022 growing seasons. In this study the effects of varying levels of irrigation and vermicompost application on soil characteristics and quinoa (Chenopodium quinoa) yield were investigated. The experiments were arranged as split plots within a randomized complete block design. The main factor consisted of four irrigation levels (50%, 75%, 100%, and 125% of quinoa’s water requirement), while the sub-factor encompassed four vermicompost application rates (0, 5, 10, and 15 tons per hectare). The irrigation method employed was a drip-tape irrigation system with a seven-day interval between irrigations, and water quantities were measured using a volumetric meter on the main pipeline.ResultsThe results demonstrated that the highest levels of available soil phosphorus (21.83 mg kg−1) and soil ammonium (36.08 mg kg−1) were observed in the treatment receiving 50% of quinoa’s water requirement combined with 15 tons per hectare of vermicompost. Additionally, the application of 15 tons per hectare of vermicompost led to the highest concentrations of soil nitrate (14.16 mg kg−1), available potassium (144.62 mg kg−1), and quinoa seed yield (1784.01 kg ha−1)). Over both years, the greatest activity of alkaline and acid phosphatase enzymes was noted in the treatment receiving 125% of quinoa’s water requirement in combination with 15 tons per hectare of vermicompost. Vermicompost application was found to enhance yield and ameliorate drought stress by enhancing soil physical and chemical properties and improving soil moisture retention.

Long-term dietary exposure to the non-steroidal anti-inflammatory drugs diclofenac and ibuprofen can affect the physiology of common carp (Cyprinus carpio) on multiple levels, even at “environmentally relevant” concentrations

The aim of the study was to evaluate the effects of emerging environmental contaminants, the non-steroidal anti-inflammatory drugs (NSAIDs) diclofenac (DCF) and ibuprofen (IBP), on physiological functions in juvenile common carp (Cyprinus carpio). Fish were exposed for 6 weeks, and for the first time, NSAIDs were administered through diet. Either substance was tested at two concentrations, 20 or 2000 μg/kg, resulting in four different treatments (DCF 20, DCF 2000, IBP 20, IBP 2000). The effects on haematological and biochemical profiles, the biomarkers of oxidative stress, and endocrine disruption were studied, and changes in RNA transcription were also monitored to obtain a comprehensive picture of toxicity. Fish exposure to high concentrations of NSAIDs (DCF 2000, IBP 2000) elicited numerous statistically significant changes (p < 0.05) in the endpoints investigated, with DCF being almost always more efficient than IBP. Compared to control fish, a decrease in total leukocyte count attributed to relative lymphopenia was observed. Plasma concentrations of total proteins, ammonia, and thyroxine, and enzyme activities of alanine aminotransferase (ALT), aspartate aminotransferase, and alkaline phosphatase (ALP) were significantly elevated in either group, as were the activities of certain hepatic antioxidant enzymes (superoxide dismutase, glutathione-S-transferase) in the DCF 2000 group. The transcriptomic profile of selected genes in the tissues of exposed fish was affected as well. Significant changes in plasma total proteins, ammonia, ALT, and ALP, as well as in the transcription of genes related to thyroid function and the antioxidant defense of the organism, were found even in fish exposed to the lower DCF concentration (DCF 20). As it was chosen to match DCF concentrations commonly detected in aquatic invertebrates (i.e., the potential feed source of fish), it can be considered “environmentally relevant”. Future research is necessary to shed more light on the dietary NSAID toxicity to fish.

Antibacterial mechanism of slightly acidic electrolyzed water against Pseudomonas arsenicoxydans A1 from greater amberjack (Seriola dumerili)

This study aimed to identify the specific spoilage organism (SSO) from the metamorphosed greater amberjack (Seriola dumerili) fish. Pseudomonas arsenicoxydans A1 was identified as SSO by culture-depending method. Further, the antibacterial efficacy of slightly acidic electrolyzed water (SAEW) was explored and the mechanism of action on P. arsenicoxydans A1 was studied. SAEW with 20–60 mg/L of available chlorine concentration (ACC) showed a good antibacterial effect on P. arsenicoxydans A1. The total number of colonies, growth curve, OD value at 260 nm and 280 nm, DNA fluorescence intensity, alkaline phosphatase (AKP), lactate dehydrogenase (LDH), superoxide dismutase (SOD), and catalase (CAT) enzyme activity and SEM results showed that SAEW could destroy the permeability and integrity of P. arsenicoxydans A1 cell membrane, and affect bacterial cell activity at metabolic, enzymatic and DNA level, which caused bacterial cell death. This study provided the application of SAEW in sterilization and preservation of raw fish fillets, which showed a promising effect on the growth of P. arsenicoxydans A1 in food cold chain.

Enzymatically Driven Mineralization of a Calcium-Polyphosphate Bleaching Gel.

To examined alkaline phosphatase enzyme (ALP) activity and the effects of incorporating it in the thickener solution of a hydrogen-peroxide-based bleaching gel containing calcium-polyphosphate (CaPP) on the orthophosphate (PO43-) levels, bleaching effectiveness, and enamel microhardness. ALP activity was assessed at different pH levels and H2O2 concentrations, and in H2O- and Tris-based thickeners. Circular dichroism (CD) was used to examine the ALP secondary structure in water-, Tris-, or H2O2-based mediums. The PO43- levels were evaluated in thickeners with and without ALP. Enamel/dentin specimens were allocated into the following groups: control (without bleaching); commercial (Whiteness-HP-Maxx); Exp-H (H2O-based); CaPP-H; ALP-H (CaPP+ALP); Exp-T (Tris-based); CaPP-T; and ALP-T (CaPP+ALP). Color changes (ΔE/ΔE00) and the bleaching index (ΔWID) were calculated, and surface (SMH) and cross-sectional microhardness (CSMH) were assessed. The two-way ANOVA and Tukey's post-hoc tests were used to compare ALP and PO43- levels; generalized linear models were used to examine: ΔE/ΔE00/SMH/CSMH; and Kruskal-Wallis and Dunn's tests were used for ΔWID (α = 5%). The ALP activity was higher at pH 9, lower in H2O2-based mediums, and similar in both thickeners. The CD-spectra indicated denaturation of the enzyme upon contact with H2O2. The PO43- levels were higher after incorporating ALP, and the ΔE/ΔE00/ΔWID were comparable among bleached groups. SMH was lower after bleaching in Exp-H, while CSMH was highest in ALP-T.

How does warmer sea water change the sensitivity of a Mediterranean thermophilic coral after immune-stimulation?

Anthropogenic climate change is warming sea water worldwide, pushing the limits of tolerance for marine organisms and driving a decline in biodiversity. The risk of thermal anomalies has increased particularly in the Mediterranean region over the last 30 yrs, where intense warming has been identified as one of the main stressors in coastal regions. To determine the influence of warmer conditions on the immunity of an endemic Mediterranean coral species, different immune activity parameters were compared in response to elevated temperature (~ 28 °C) and the presence of a pathogen-associated molecular pattern—Escherichia coli lipopolysaccharide (LPS)—as an elicitor of the innate immune response of Astroides calycularis. Immune parameters, which included phenoloxidase-like, glutathione peroxidase, lysozyme-like, alkaline phosphatase, and esterase enzyme activity, were measured over time after LPS balneation (0-, 12-, 48-, and 120 h time point). All five enzymes demonstrated constant values under environmental conditions (~ 23 °C), indicating a constituent activity. LPS at environmental temperature induced significant upregulation immediately after exposure (0 h-time point), demonstrating an immune response to the pathogen elicitor. Under warmer conditions (~ 28 °C), constituent values increased over time, indicating a shift in the immune strategy to maintain homeostasis. However, warmer sea water, within the summer range experienced by this coral species, impaired the immune response to LPS, delaying it over time. These changes in immune strategy indicate that temperature affects coral immunity and, in thermophilic A. calycularis, results in an energy trade-off that could maintain its health-state through suboptimal conditions during multiple perturbations, such as summertime diseases.

Insight into the inhibitory activity and mechanism of bovine cathelicidin BMAP 27 against Salmonella Typhimurium

This study synthesized an antimicrobial peptide based on the bovine cathelicidin BMAP 27 sequence. It was found to have a broad spectrum of antibacterial activity, with exceptionally high activity against Salmonella. However, the antibacterial mechanism of BMAP 27 against Salmonella remains unclear. The minimum inhibitory concentrations (MIC) and minimum bactericidal concentrations (MBC) of BMAP 27 against Salmonella enterica serovar Typhimurium were determined to be 2 μM and 4 μM, respectively. After treatment with 2 MIC of BMAP 27, the absorbance of DNA in centrifugal supernatant increased from 0.244 to 1.464, and that of protein rose from 0.174 to 0.774, respectively. BMAP 27 has compromised the cell membrane as observed through field emission scanning electron microscope (FESEM) and transmission electron microscopy (TEM), and confirmed by the propidium iodide (PI) test. The alkaline phosphatase (AKP) enzyme activity in the supernatant of the 2 MIC treatment group was 2.15 times higher than the control group, indicating extracellular membrane damage. BMAP 27 treatment increased intracellular ROS levels as tested by dichlorofluorescein diacetate (DCFH) staining. DNA interaction analysis revealed that BMAP 27 has a binding affinity towards DNA, causing its characteristic bands to disappear and peak intensity at 260 nm to reduce. Molecular docking identified its potential binding mode with DNA. The crystal violet biofilm staining results demonstrated that BMAP 27 inhibited S. Typhimurium biofilm formation by 43.1 % and cleared mature biofilms by 53.62 %. Confocal Laser scanning electron microscopy (CLSM) observed that BMAP 27 could kill bacteria within the biofilm and dislodge bacteria from the surface of glasses. Swimming tests identified that the motor capacity of S. Typhimurium was diminished by BMAP 27. By counting the total bacteria, BMAP 27 was revealed to exert bacteriostatic effects in chilled pork and orange juice, which might provide a basis for its application in the inhibition of Salmonella.

Investigating the toxic effects of ethoprophos on Eisenia fetida: Integrating light microscopy, scanning electron microscopy, and biochemical analysis

This research investigated the ecological impact of exposing Eisenia fetida, an essential component of soil ecosystems, to the organophosphate pesticide ethoprophos, widely used in agriculture. With a focus on understanding the specific effects on earthworms, we employed three concentrations (7.5, 15, and 30 mg/kg) over 28 days, considering the pesticide's short half-life and existing data on environmental concentrations. We aimed to contribute to a broader understanding of how these pesticides affect soil health. Histological analysis, including staining with Hematoxylin-eosin, Mallory Trichrome, Periodic acid-Schiff, and Alcian blue methods, was conducted on control and treatment groups. The histological and histopathological results were evaluated using the light microscopy, revealing various degenerations in the epithelial and muscle layers. Scanning electron microscopy analysis detected concentration-related notable compaction of the body surface, asymmetry, and distortion in the body segments. In the exposed groups, especially those subjected to higher ethoprophos concentrations, the grid-like appearance of the clitellum was visibly disturbed. This disturbance in the grid-like pattern is indicative of structural changes and disruptions at the microscopic level. Furthermore, total protein, carbohydrate, lipid analyses, as well as acid phosphatase and alkaline phosphatase enzyme activities, were also evaluated for earthworms from each experimental group. The analyses showed a concentration-related decrease in all biochemical measurements, except acid phosphatase enzyme activity. In conclusion, our study reveals that the environmentally realistic concentrations of ethoprophos, an effective and widely used pesticide in pest control, have detrimental effects on the health and physiology of E. fetida. These effects are manifested through histological deformities, altered biochemical profiles, and observable physiological disturbances. These results shed light on the harmful effects of ethoprophos on earthworms, underlining the necessity to restrict its usage in agricultural practices and thereby support environmental sustainability.

Effects of natural and prolonged crude oil pollution on soil enzyme activities

In this study the effects of natural and long-term oil pollution levels (H: high, M: moderate and L: low) on the activity of lipase, β-glucosidase, urease, phosphatase, and dehydrogenase were analyzed. For this purpose, 120 oil-contaminated soil samples were collected from four locations located in oil-rich region of the west of Iran (Kermanshah province, Iran). After determining the physicochemical characteristics of soils, microbial counting and enzyme activities were measured. To determine the total bacterial population and bacteria involved in crude oil decomposition, bacterial counting was done in NA and CFMM culture media, respectively, which had a direct relationship with the increase in crude oil content. The average percentage of crude oil measured by Soxhlet method was 4.03%, 9.95% and 22.50% respectively for L, M and H levels. The results showed that the bacterial population increased with the increase of the contamination intensity. All the studied enzymes, except urease, had the highest activity in heavily polluted soils (H soil), and their lowest activity was observed in lightly polluted soils (L soils). Location 1 had the highest bacterial population as well as the highest activity of dehydrogenase (52.13 μg TPFg−1 h−1), acid phosphatase (35.86 μg PNPg−1 h−1), alkaline phosphatase (78.13 μg PNPg−1 h−1), lipase (92.58 μ MLA g−1 h−1) and β-glucosidase (21.67 μg PNPg−1 h−1) enzymes. In the soil samples of location 4, which had the lowest number of bacteria, the highest activity of urease enzyme (592.36 μg NH4g−1 h−1) was obtained. Principal components analysis (PCA) was also performed and 67% of the cumulative of the samples could be explained by the first two components (biochemical and physical). The findings of this research showed that natural and prolonged crude oil pollution caused the selection of oil-resistant microbial communities, and therefore we saw their positive response to the presence of oil compounds and increased enzyme activity other than urease activity.

Effect of the Substrate and Inoculation of Phosphate-Dissolving Bacterial and Fungal with Humic Acids in the Activity of the Alkaline Phosphatase Enzyme in Cadmium-Contaminated Soil

A laboratory experiment was carried out in the laboratories of the Department of Soil Sciences and Water Resources, College of Agriculture, University of Basrah, to identify the role of the bacteria Bacillus subtilis and the fungus Aspergillus niger, with humic acids extracted from fermented cow dung, the effectiveness of the alkaline phosphatase enzyme in soil contaminated with cadmium and compare it with uncontaminated soil. The soil was treated with a cadmium sulfate solution up to the critical limit (3 mg Cd L-1) and humic and fulvic acid were added at a level of 50 L ha-1 separately, leaving the treatment without addition for control. Then the soil was inoculated with bacterial B. subtilis and fungal A. niger isolates individually and with a mixture of isolates, leaving a treatment without inoculation for control, the treatments were incubated at a temperature of 28±2 °C. Different levels of substrate concentration were used (0.010, 0.025, 0.050, 0.075, and 0.100 M). The results showed that cadmium contamination decreased alkaline phosphatase enzyme activity by 12.28%. There was also an increase in the activity of the alkaline phosphatase enzyme at all inoculation treatments compared to the control treatment, with the bio-mixture treatment being superior to the rest of the treatments, reaching 436.944 µg P. nitrophenol gm-1 soil 1 hour-1 in uncontaminated soil and 402.558 µg P. nitrophenol gm-1 soil 1 hour-1 in cadmium contaminated soil. The humic acid treatment showed a similar increase in the activity of the alkaline phosphatase enzyme compared to the control treatment the highest increase was in the fulvic acid treatment. As for the effect of the substrate, the concentration of 0.075 M recorded the highest increase in alkaline phosphatase enzyme activity in unpolluted and cadmium-polluted soil for all treatments.

Assessment of the liver biochemical status of carpenters occupationally exposed to wood dust in Enugu metropolis South-East Nigeria.

Liver disease has a global distribution and ranks as the twelfth leading cause of mortality. This cross-sectional study carried out between June and November 2021 aimed to assess the liver biochemical status of carpenters occupationally exposed to wood dust in Enugu metropolis South-east Nigeria. The Ethical Committee of the University of Nigeria Teaching Hospital in Enugu gave its approval. A total of one hundred and twenty-four (124) participants; sixty-two (62) carpenters (exposed) and sixty-two (62) age and sex-matched (males) control (unexposed) were recruited for this study. Study participants were chosen by simple random sampling. Spectrophotometric method was used to test participants’ blood samples for liver enzymes alanine transaminase (ALT), aspartate transaminase (AST) and alkaline phosphatase (ALP). A statistically significant increase (p&lt;0.05) was observed in the liver biochemical enzymes of the carpenters (exposed) in comparison to the controls (unexposed). The carpenters do not use any form of personal protective equipment (90.32%) or have a good knowledge of any occupational risk (83.87%) they are exposed to. A statistically significant positive correlation (p&lt;0.05) was observed between ALT, AST and ALP enzyme activity and the duration of exposure to wood dust (in years) of the exposed. This study suggests that occupational exposure to wood dust may cause metabolic changes in the Liver. Constant use of personal protective equipment when at work, personal hygiene and regular health check is recommended.

The challenge of hypophosphatasia diagnosis in adults: results from the HPP International Working Group Literature Surveillance.

Hypophosphatasia (HPP) is an inborn error of metabolism caused by reduced or absent activity of the tissue non-specific alkaline phosphatase (TNSALP) enzyme, resulting from pathogenic variants in the ALPL gene. Clinical presentation of HPP is highly variable, including lethal and severe forms in neonates and infants, a benign perinatal form, mild forms manifesting in adulthood, and odonto-HPP. Diagnosis of HPP remains a challenge in adults, as signs and symptoms may be mild and non-specific. Disease presentation varies widely; there are no universal signs or symptoms, and the disease often remains underdiagnosed or misdiagnosed, particularly by clinicians who are not familiar with this rare disorder. The absence of diagnosis or a delayed diagnosis may prevent optimal management for patients with this condition. Formal guidelines for the diagnosis of adults with HPP do not exist, complicating efforts for consistent diagnosis. To address this issue, the HPP International Working Group selected 119 papers that explicitly address the diagnosis of HPP in adults through a Medline, Medline In-Process, and Embase search for the terms "hypophosphatasia" and "HPP," and evaluated the pooled prevalence of 17 diagnostic characteristics, initially selected by a group of HPP clinical experts, in eligible studies and in patients included in these studies. Six diagnostic findings showed a pooled prevalence value over 50% and were considered for inclusion as major diagnostic criteria. Based on these results and according to discussion and consideration among members of the Working Group, we finally defined four major diagnostic criteria and five minor diagnostic criteria for HPP in adults. Authors suggested the integrated use of the identified major and minor diagnostic criteria, which either includes two major criteria, or one major criterion and two minor criteria, for the diagnosis of HPP in adults.

Cell-Free Supernatant of Lactiplantibacillus plantarum 90: A Clean Label Strategy to Improve the Shelf Life of Ground Beef Gel and Its Bacteriostatic Mechanism.

Lactic acid bacteria metabolites can be used as a clean-label strategy for meat products due to their "natural" and antibacterial properties. In this study, the feasibility of using cell-free supernatant of Lactiplantibacillus plantarum 90 (LCFS) as a natural antibacterial agent in ground beef was investigated. The sensitivity of LCFS to pH, heat and protease, as well as the changes of enzyme activities of alkaline phosphatase (AKP) and Na+/K+-ATP together with the morphology of indicator bacteria after LCFS treatment, were analyzed to further explore the antibacterial mechanism of LCFS. The results showed that the addition of 0.5% LCFS inhibited the growth of microorganisms in the ground beef gel and extended its shelf-life without affecting the pH, cooking loss, color and texture characteristics of the product. In addition, the antibacterial effect of LCFS was the result of the interaction of organic acids and protein antibacterial substances in destroying cell structures (cell membrane, etc.) to achieve the purpose of bacteriostasis. This study provides a theoretical basis for the application of LCFS in meat products and a new clean-label strategy for the food industry.

Cranial Neural Crest Specific Deletion of Alpl (TNAP) via P0-Cre Causes Abnormal Chondrocyte Maturation and Deficient Cranial Base Growth.

Bone growth plate abnormalities and skull shape defects are seen in hypophosphatasia, a heritable disorder in humans that occurs due to the deficiency of tissue nonspecific alkaline phosphatase (TNAP, Alpl) enzyme activity. The abnormal development of the cranial base growth plates (synchondroses) and abnormal skull shapes have also been demonstrated in global Alpl-/- mice. To distinguish local vs. systemic effects of TNAP on skull development, we utilized P0-Cre to knockout Alpl only in cranial neural crest-derived tissues using Alpl flox mice. Here, we show that Alpl deficiency using P0-Cre in cranial neural crest leads to skull shape defects and the deficient growth of the intersphenoid synchondrosis (ISS). ISS chondrocyte abnormalities included increased proliferation in resting and proliferative zones with decreased apoptosis in hypertrophic zones. ColX expression was increased, which is indicative of premature differentiation in the absence of Alpl. Sox9 expression was increased in both the resting and prehypertrophic zones of mutant mice. The expression of Parathyroid hormone related protein (PTHrP) and Indian hedgehog homolog (IHH) were also increased. Finally, cranial base organ culture revealed that inorganic phosphate (Pi) and pyrophosphate (PPi) have specific effects on cell signaling and phenotype changes in the ISS. Together, these results demonstrate that the TNAP expression downstream of Alpl in growth plate chondrocytes is essential for normal development, and that the mechanism likely involves Sox9, PTHrP, IHH and PPi.

Results of clinical and hematological examination in dogs with congenital portosystemic shunts

The article presents the results of a clinical and hematological study of data obtained from dogs, mainly small ornamental breeds, with subsequently confirmed portosystemic shunts. As a result of the work, it was determined that all animals had a disorder of the nervous system, which was the reason for their owners to contact the veterinary clinic to examine the pets. Thus, according to the results of the analysis of the anamnesis and during the full-time clinical examination, tremor, ataxia, impaired consciousness, apathy, play movements, auto-aggression and causeless aggression or vocalization, mydriasis were observed in sick dogs. A laboratory blood test revealed a number of signs indicating the possible presence of portosystemic vascular anomalies in the examined dogs, namely, hypoproteinemia (48.4±3.094 g/l) with dysproteinemia, an increase in the activity of ALT, AST and alkaline phosphatase enzymes up to 188.2±85.573 IU/l, 121.5±20.061 IU/l and 514.6±110.851 IU/l, respectively. A pronounced increase in the concentration of bile acids in the blood before and after feeding was also noted, up to 82.5±16.57 µmol/l and 155.4±26.103 µmol/l, respectively. An analysis of the results of a morphological blood test showed the presence of such nonspecific changes as hypochromic anemia (hemoglobin concentration was 129.3 ± 5.631 g/l) and leukocytosis (the number of leukocytes in the blood of sick dogs was 17.7 ± 2.586*109 /l). Thus, we can conclude that the complex of clinical and laboratory studies carried out may be sufficient for the differential diagnosis of portosystemic vascular anomalies in dogs, but, nevertheless, the final diagnosis is possible only with the use of instrumental diagnostic methods.

The Effect of Calcium hydroxide, Glass Ionomer and light cured resin modified calcium silicate on viability, proliferation and differentiation of stem cells from human exfoliated deciduous teeth

BackgroundVital pulp therapy, based on the use of stem cells, has promising research and therapeutic applications in dentistry. It is essential to understand the direct effect of capping materials on the dental pulp stem cells of primary teeth, which contribute to the healing powers of the tooth. The aim of this study is to evaluate the effect of different capping materials (Calcium Hydroxide (DyCal®) – Glass Ionomer (Fuji IX®) and light-cured resin modified calcium silicate (TheraCal LC®)) on the viability, proliferation, and differentiation of stem cells from human exfoliated deciduous teeth (SHEDs).MethodsSHEDs were isolated from extracted primary teeth, then divided into four groups and each of the capping materials were applied to the stem cells as follows: group I the controls, group II with Ca(OH)2, group III with the GIC, and group IV with the Theracal LC. For all groups assessment of viability and proliferation rate was done using the MTT cell proliferation assay. Also, Differentiation was evaluated by measuring the gene expression of Alkaline phosphatase enzyme activity (ALP) and Dentin matrix protein-1 (DMP1) through quantitative real-time PCR. Morphological assessment was conducted using Alizarin Red S staining. All evaluations were performed after 7 and 14 days of culture.ResultsTheraCal LC showed the highest values of proliferation, which was significant only compared to the control group after 2 weeks (p = 0.012). After one week, TheraCal LC showed the highest significant values of ALP and DMP1 compared to all other groups (p < 0.001).ConclusionThe three materials under study are biocompatible, maintain viability, and stimulate the proliferation and differentiation of SHEDs. However, TheraCal LC allows better proliferation of SHEDs than Dycal Ca(OH)2 and Fuji IX GIC.

THU457 Variable Presentations Of Hypophosphatasia In Two Patients In The Outpatient Setting

Abstract Disclosure: A. Elmaaz: None. S. Batool: None. P. Dandona: None. Introduction: Hypophosphatasia is a rare disorder characterized by defective mineralization of bone and teeth which is caused by an autosomal inherited mutation in the tissue nonspecific alkaline phosphatase (ALP) encoding genes. Here we present two cases of multiple dental and bony defects secondary to low ALP enzyme activity due to hypophosphatasia. Case 1: A 27-year-old female with history of mitochondrial disease, gastroparesis, malabsorption syndrome, and autonomic dysfunction was referred to Endocrinology clinic because of persistently low serum ALP levels associated with recurrent muscle, joint, and bone pain since the age of 13. She had history of multiple tooth extractions because of dental developmental abnormalities and fractures. She also reported history of fractures of three of her fingers in both hands during her childhood and teenage years. The patient was adopted and was unaware of any biological family history. Physical examination was remarkable for short-statured female, 4'8". Notably, the middle phalanges in little fingers bilaterally were shorter than the other fingers. Workup showed vitamin D level of 21 (30-80) ng/ml, despite being on 50,000 IU twice a week for 6 months with normal PTH and ionized calcium levels. The repeated serum level of ALP was 24 (31-125) U/L with bone and liver isoenzymes levels of 42% and 58 % respectively. Vitamin B6 level was 13.2 (2.1-21.7) ng/ml. Bone mineral density was unremarkable. Case 2: A 53-year-old female with history of congenital left hip dysplasia, hypertension, and dyslipidemia was seen by Endocrinology service for evaluation of decreased ALP levels. She denied any history of falls or fractures. However, she reported chronic bone pain, especially in bilateral hip joints with history of multiple surgeries in her left hip joint. She had required teeth braces and multiple dental surgeries during her childhood due to dental maldevelopment. Her height was 4'8". Laboratory workup was remarkable for low serum ALP level of 30 U/L with bone isoenzyme of 24 %. Vitamin B6 level was 206.7 (2.1-21.7) ng/ml. She had elevated serum phosphorus levels with normal calcium, PTH, and vitamin D levels. Pyridoxal-5- Phosphate level was high. Bone mineral density revealed osteopenia. Conclusion: Hypophosphatasia is a rare disease, hence it could be easily missed due to high variability in clinical presentations and severity of symptoms. Low serum levels of ALP with history of bone pain, recurrent fractures, premature loss of deciduous teeth, or loss of adult dentition should raise the suspicion of hypophosphatasia. Resistance to vitamin D supplementation is another clue to the diagnosis. A multidisciplinary approach is required for appropriate management. Presentation: Thursday, June 15, 2023

Panicum miliaceum L. and Bone Healing Properties in Rat Model of Femur Fracture by Activating Phosphate Stimulating Macrophages via BMP2 and RANK Signaling Pathway

Objectives The aim of this study was to investigate the effect of Panicum miliaceum involved in the repair of femur bone fracture in rats. Materials and Methods In this study, 40 Wistar rats in five groups were studied as healthy, rats with lesion induced by surgery in femur (FX), FX rats treated with P. miliaceum extract (FX +100 and 200 P. miliaceum) and FX rats treated with Osteocare syrup (FX + OC). After 40 days, the serum levels of blood calcium (Ca), phosphorus (P), and alkaline phosphatase (ALP) along with activity of glutathione peroxidase (GPx), superoxide dismutase (SOD), and catalase (CAT) enzymes, and nitric oxide (NO) level were measured. The serum level of cytokines (IL-6, C-reactive protein [CRP], IL-10, IL-1β, and tumor necrosis factor-α [TNF-α]) was also measured by ELISA. The expression of OPG, RANK, RANKL, and BMP-2 genes and proteins were evaluated by real-time PCR and Western blotting techniques, respectively. Results The results showed that P. miliaceum was able to decrease the activity of ALP enzyme in addition to increasing serum Ca and P levels and GPx, SOD, and CAT activity. P. miliaceum reduces serum IL-6, IL-1β, and TNF-α, and NO levels. Also, the extract of this plant could increase the expression of OPG/RANK/BMP-2 pathway genes and proteins. Conclusion P. miliaceum can be used as a prodrug in traditional medicine to repair bone fractures and osteoporosis.

Effects of Different Proportions of Amaranthus hypochondriacus Stem and Leaf Powder Inclusions on Growth Performance, Carcass Traits, and Blood Biochemical Parameters of Broilers.

This experiment aimed to study the effects of different proportions of Amaranthus hypochondriacus stem and leaf powder (AHSL) on the growth performance, apparent nutrient digestibility, carcass traits, meat quality, and blood biochemical parameters of broilers from day 1 to day 42. The experiment utilized a single-factor experimental design, with a total of 216 one-day-old male broilers (Ross 308) randomly assigned to three dietary treatment groups (eight replicate cages of nine birds per cage). The dietary treatments included a control diet, a 3% AHSL diet and a 6% AHSL diet for days 0-21. Then, the 3% and 6% AHSL diets were changed to 5% and 10% AHSL for days 22-42. The results showed that the inclusion levels of AHSL did not affect growth performance, carcass traits, or meat quality on days 21 and 42 (p > 0.05). However, the inclusion levels of AHSL decreased the apparent nutrient digestibility (AND) of dry matter (DM) (p ˂ 0.001) and neutral detergent fiber (NDF) (p ˂ 0.001) and increased the serum concentration of phosphorus (p ˂ 0.001) on day 21. On day 42, the inclusion levels of AHSL decreased the AND of DM (p = 0.025) and NDF content (p ˂ 0.001), but increased the AND of crude protein (CP) (p = 0.004). In particular, the diet containing 10% AHSL significantly increased the serum enzyme activity of alkaline phosphatase (ALP) (p = 0.046) and the serum concentration of total protein (TP) (p ˂ 0.001) on day 42. Overall, AHSL can be used as a new and effective feed ingredient in broiler diets. It can replace part of the corn-soybean meal diet without any adverse effects, which is beneficial for conserving feed resources. Additionally, AHSL can be included at a level of up to 10% during the broiler growth period.