Alkaline Phosphatase Enzyme Activity Research Articles

In situ synthesis of micro-plastics embedded sewage-sludge co-pyrolyzed biochar: Implications for the remediation of Cr and Pb availability and enzymatic activities from the contaminated soil

Careful designing of biochar (BC) employing biological and industrial waste materials has gained much attention to improve soil health via reducing the bioavailability of heavy metals (HMs) in the contaminated-soil. Herein, a forty-day pot experiment was conducted to explore the influence of micro-plastic (MP) embedded sewage-sludge (SS). The effects of SS and MP dosage ratios (0–15%) on the physicochemical properties of the modified-biochars were also studied with slow-pyrolyzed (SBC-500 °C) and flash-pyrolyzed biochar (FSBC-700 °C). Our investigations found remarkable positive synergistic effects for the SBC-15% involving increased soil pH, CEC, high carbon contents, and alleviation in Cr and Pb leaching than control and FSBC-700 °C. Besides, SBC-15% containing high carbon functional groups can effectively mitigate the Cr and Pb availability stress by intensifying the adsorption or passivation in the amended-soil, thereby, significantly reducing the Cr and Pb EDTA-extractable contents. Chemical fractionation analyses further confirmed that SBC-15% addition was more helpful for Cr and Pb immobilization and ultimately reducing transfer-rate, bioconcentration-factor, and translocation-factor as compared to FSBC-15% and control due to its higher alkalinity, surface area/porosity, and available carbon functional groups. The maize biomass (root and shoot) increased by more than 50%, and the activities of soil enzymes such as urease, alkaline phosphatase, and glucosidase enzyme activities were also enhanced. This ecologically feasible strategy would pave an efficient way to make full use of the SS and MP for the biochar synthesis with excellent soil remediation performance.

Seed Priming and Coating by Nano-Scale Zinc Oxide Particles Improved Vegetative Growth, Yield and Quality of Fodder Maize (Zea mays)

Nano-fertilizers of essential plant nutrients, including micronutrients, have the potential to improve nutrient use efficiency and productivity of field crops in deficient soils. The present study reports the comparative influence of zinc oxide nanoparticles (ZnONPs) and bulk Zn salt (ZnSO4) on the growth, yield, and quality of fodder maize (Zea mays) (var. J-1006) cultivated under field conditions in the year 2019. Three levels (0, 20, and 40 mg L−1) of Zn fertilizers were used for seed priming and coating in triplicate following the randomized complete block design model. An increase in vegetative and yield parameters (number of plants, plant height, stover yield, plant biomass), acid detergent fiber (ADF%), and hemicellulose contents and shoot zinc (Zn) content on treatment of seeds with ZnONPs (20 mg L−1) concentration as compared to bulk ZnSO4 and control treatments was observed. The application of ZnONPs (40 mg L−1) significantly enhanced the total chlorophyll content, available soil nitrogen and phosphorus, neutral detergent fiber (NDF%), and cellulose contents and improved the total soil microbial counts and soil enzyme activities (dehydrogenase, acid and alkaline phosphatase enzyme activities), whereas a significant increase in available soil potassium and zinc contents was recorded under ZnONPs (20 mg L−1) treatments. These findings suggest an encouraging effect on the growth and yield attributing characteristics of fodder maize after ZnONPs seed coating at low concentration. Furthermore, ZnONPs seed coating can also be considered an effective tool for the delivery of Zn micronutrient to fodder maize crop.

Effects of Co-Applications of Biochar and Solid Digestate on Enzyme Activities and Heavy Metals Bioavailability in Cd-Polluted Greenhouse Soil

To deal with the problems of increasing the heavy metal (HM) bioavailability and declining the soil biological properties resulting from a direct application of solid digestate (SD). A low-temperature fruit biochar and pig-SD (BSD-0, BSD-1, BSD-2, BSD-4, BSD-8) co-application experiment was performed to evaluate enzyme activities and HM bioavailability in Cd-polluted greenhouse soil. The advantage of BSD co-applications compared to SD application was maintained the stable of pH and electrical conductivity (EC) in soil and was more effective to improve soil organic matter (OM). BSD-8 treatment significantly promoted the uptake of available nitrogen, available phosphorus, and available potassium by plants. The immobilization effect of BSD co-applications on Cu, Zn, and Cd was better than SD application. BSD-8 treatment has the best immobilization effect on Cd and the contents of bioavailable Cd was 0.167 mg kg−1. The optimal enzyme activities of invertase, urease, and alkaline phosphatase were shown in BSD-8 treatment, which were 0.027 mg glucose g−1 soil h−1, 88.654 mg NH3-N g−1 soil h−1, and 15.766 μmol PNP g−1 soil h−1, respectively. The activities of enzymes also were influenced by soil physicochemical properties and HM bioavailability. BSD-8 treatment was suggested as an appropriate mixing proportion to alleviate soil acidification and salinization, decreasing HM bioavailability and stimulating enzyme activities in Cd-polluted soil.

Microbial Biomass Carbon, Activity of Soil Enzymes, Nutrient Availability, Root Growth, and Total Biomass Production in Wheat Cultivars under Variable Irrigation and Nutrient Management

Intensive mono-cropping without a balanced supply of nutrients and declining water resources are degrading soil health, and as a consequence, agriculture production is becoming unsustainable and causing environmental degradation. The field experiment was conducted during Rabi season to assess the effect of an irrigation schedule, nutrient management, and wheat (Triticum aestivum L.) varieties on soil microbial biomass carbon (SMBC) and soil enzymes activities. Two nutrient levels, recommended rate of chemical fertilizer (RDF) and 50% RDF + 50% recommended dose of nitrogen (RDN) through farmyard manure (FYM) designated as Integrated Nutrient Sources (INS), and three irrigations levels, one irrigation at crown root initiation (CRI), two irrigations at CRI and flowering stages, and five irrigations at all main stages of the crop (CRI, tillering, jointing, flowering, and grain filling) were allocated to main-plots while four varieties of wheat, HD 2967, WR 544, HD 2987, and HD 2932, were allocated to sub-plots. The results revealed that SMBC and activities of dehydrogenase, alkaline phosphatase enzymes, and acid phosphatase were higher under restricted irrigation (irrigation at CRI stage) than other irrigation schedules. SMBC, dehydrogenase, acid phosphatase, and alkaline phosphatase activities were 73.0 µg g soil−1, 86.0 µg TPF g soil−1d−1, 39.6 µg PNP g soil−1 h1, and 81.8 µg PNP g−1 soil h−1, respectively, with the use of INS that was higher than RDF. Root weight and root volume followed a similar pattern. Applying single irrigation at CRI left behind the maximum available nitrogen (166.4 kg ha−1) in soil compared to other irrigation schedules and it was highest (149.31 kg ha−1) with the use of INS. Moreover, total organic carbon (TOC) was 0.44 and 0.43% higher with irrigation at CRI stages and the use of INS, respectively. The INS with single irrigation at the CRI stage is important to improve the root growth, SMBC dehydrogenase, alkaline phosphatase, and acid phosphatase enzyme activity in the wheat production system.

Evaluation of BMP2/miRNA co-expression systems for potent therapeutic efficacy in bone-tissue regeneration.

Reconstruction of bone defects and compensation of deficient repair mechanisms represent important goals within the field of regenerative medicine and require novel safe strategies for translation into the clinic. A non-viral osteogenic gene therapeutic vector system ('hybrid vectors') was generated, combining an improved bone morphogenetic protein 2 (BMP2) gene cassette and single pro-osteogenic microRNAs (miR-148b-3p, miR-20-5p, miR-590b-5p), driven by the U6 promoter. The vectors were tested in vitro for their osteogenic differentiation potential in C2C12 and C3H/10T1/2 cell lines, using BMP2 alone as control. After confirming BMP2 expression and miRNA transcription, increased osteogenic differentiation was observed by all hybrid vectors, but most consistently by BMP2/miR-590-5p, using alkaline phosphatase enzyme activity assays and osteogenic marker mRNA quantitation, including runt-related transcription factor 2 (Runx2), collagen type 1 (Col1a1) and osteocalcin. To visualise target mRNAs of the respective miRNAs, next generation sequencing was performed, confirming down-regulation of mRNA targets of the hybrid vectors. Since the hybrid vector consisting of BMP2 and miR-590-5p showed the largest increase in osteogenic differentiation in vitro, this was tested in a mouse ectopic-bone model. Mineralisation was more than with BMP2 alone. The present study showed hybrid vectors as a novel non-viral gene therapeutic plasmid system for combining therapeutic effects of recombinant protein expression and miRNA transcription that did not add to the burden of the translation machinery, while improving the therapeutic efficacies. In vivo proof-of-principle in the context of bone regeneration suggested that such hybrid vectors will be applicable in a wide array of gene therapeutic strategies.

Copper Oxide Nanoparticles Alter Serum Biochemical Indices, Induce Histopathological Alterations, and Modulate Transcription of Cytokines, HSP70, and Oxidative Stress Genes in Oreochromis niloticus.

Simple SummaryCopper oxide nanoparticles (CuONPs) are increasingly manufactured because of their wide range of biomedical uses and industrial applications. Nonetheless, their release into the aquatic ecosystems is predictably increased, which will sequentially induce serious toxicological influences on the exposed aquatic biota. Several research studies have been published on CuONPs toxicity in fish; however, the mechanisms of their toxicity at the molecular levels in Nile tilapia (Oreochromis niloticus) are not completely described. The current study investigated the influences of sub-lethal CuONPs levels on serum biochemical indices, histopathological alterations, and transcriptomic responses in the hepatic and gill tissues of Nile tilapia juveniles.In the present study, fish were exposed to sub-lethal doses of CuONPs (68.92 ± 3.49 nm) (10 mg/L, 20 mg/L, and 50 mg/L) for a long exposure period (25 days). Compared to the control group (0.0 mg/L CuONPs), a significant dose-dependent elevation in blood urea and creatinine values, serum alanine transaminase, aspartate transaminase, and alkaline phosphatase enzyme activities were evident in CuONPs-exposed groups (p < 0.05). Fish exposure to 50 mg/L CuONPs significantly upregulated the transcription of pro-inflammatory cytokines (tumor necrosis factor-alpha, interleukin-1beta, interleukin 12, and interleukin 8), heat shock protein 70, apoptosis-related gene (caspase 3), and oxidative stress-related (superoxide dismutase, catalase, and glutathione peroxidase) genes in liver and gills of the exposed fish in comparison with those in the control group (p < 0.05). Moreover, varying histopathological injuries were noticed in the hepatopancreatic tissues, posterior kidneys, and gills of fish groups correlated to the tested exposure dose of CuONPs. In summary, our results provide new insights and helpful information for better understanding the mechanisms of CuONPs toxicity in Nile tilapia at hematological, molecular levels, and tissue levels.

Effect of TNF-α on the proliferation and osteogenesis of human periodontal mesenchymal stem cells.

The aim of the present study was to investigate the effect of tumor necrosis factor-α (TNF-α) on the proliferation and osteogenesis of human periodontal mesenchymal stem cells (hPDLSCs). Antigen expression in hPDLSCs was detected by flow cytometry. hPDLSCs were divided into four groups: A control group with no TNF-α treatment, and three experimental groups treated with 0.1, 1 and 10 ng/ml TNF-α, respectively. The effect of TNF-α on proliferation of hPDLSCs in vitro was detected using a Cell Counting Kit-8 assay. Differentiation into an osteogenic lineage was detected by alkaline phosphatase sand alizarin red staining, and the mRNA and protein expression levels of runt-related transcription factor 2 (Runx2), osteocalcin (OCN) and type I collagen (Col-I) were detected using reverse transcription-quantitative PCR and western blot respectively. Following treatment with 10 ng/ml TNF-α, proliferation was significantly increased compared with an untreated control group (P<0.01). Additionally, there was a significant inhibition of alkaline phosphatase enzyme activity, alizarin red mineralization node size, and in the gene and protein expression levels of osteogenic differentiation markers, including Runx2, OCN and COL-I (all, P<0.05). Taken together, the results indicated that treatment with 10 ng/ml TNF-α promoted the proliferation of hPDLSCs in vitro and inhibited osteogenic differentiation of hPDLSCs, providing an experimental basis for regulation of hPDLSC-mediated periodontal tissue regeneration.

Biological Properties of Experimental Methacrylate-Based Sealers Containing Calcium Phosphates.

This study aimed to evaluate, in vitro and in vivo, the biocompatibility of experimental methacrylate-based endodontic sealers containing α-tricalcium phosphate (α-TCP) or nanostructured hydroxyapatite (HAp). Experimental methacrylate-based dual-cure sealers with the addition of α-TCP or HAp, at 10%wt were formulated and compared to AH Plus (AHP). Cell viability was assessed by 3-(4,5-dimethyl-thiazoyl)-2,5-diphenyl-tetrazolium bromide (MTT), and sulforhodamine B (SRB). Sealers were implanted in rats' subcutaneous tissue and histologically evaluated. Bioactivity was assessed by alkaline phosphatase enzyme activity (ALP) and Alizarin Red (AR), using apical papillary cells (SCAPs), and by the bone deposition measured in surgical cavities on rats' femur filled with AH Plus or α-TCP. In both viability assays, HAp and AHP sealers were similar, and α-TCP presented lower viability compared to the others at MTT assay (p<0.05). A gradual decrease of the inflammatory response according to the periods was observed and AHP was the only that presented giant cells (7-day period). Collagen fibers condensation increased according to the periods, with no differences among sealers. There was an increase at ALP activity and mineralized nodules deposition according to periods. HAp and α-TCP presented higher values for ALP activity at 5 days and at 5, 10, and 15 days for AR and were different from AHP (p<0.05). α-TCP presented superior values at 10 and 15 days compared to HAp and AHP for AR (p<0.05). At 90 days, α-TCP and control (empty cavity) showed high bone deposition compared to AHP (p<0.05). α-TCP and HAp, in a methacrylate-based sealer, presented biocompatibility and bioactivity, with the potential to be used as endodontic sealers in clinical practice. Further investigations are required to gain information on the physicochemical properties of these sealers formulation before its clinical implementation.

Role of 1,25-Dihydroxyvitamin D3 on Osteogenic Differentiation and Mineralization of Chicken Mesenchymal Stem Cells.

1,25-dihydroxyvitamin D3 (1,25OHD) has been suggested to play an important role in osteogenic differentiation and mineralization. However, limited data have been reported in avian species. In the present study, the direct role of 1,25OHD on osteogenic differentiation and mineralization in chicken mesenchymal stem cells (cMSCs) derived from day-old broiler bones was investigated. cMSCs were treated with control media (C), osteogenesis media (OM), OM with 1, 5, 10, and 50 nM 1,25OHD, respectively. The messenger RNA (mRNA) samples were obtained at 24 and 48 h and 3 and 7 days to examine mRNA expression of key osteogenic genes [runt related transcription factor 2 (RUNX2), bone morphogenetic protein 2 (BMP2), collagen type I alpha 2 chain (COL1A2), bone gamma-carboxyglutamate protein (BGLAP), secreted phosphoprotein 1 (SPP1), and alkaline phosphatase (ALP)]. Cells were stained at 7, 14, and 21 days using Von Kossa (mineralization), Alizarin Red (AR; mineralization), and Alkaline Phosphatase (early marker) staining methods. From the mRNA expression results, we found a time-dependent manner of 1,25OHD on osteoblast differentiation and mineralization. In general, it showed an inhibitory effect on differentiation and mineralization during the early stage (24 and 48 h), and a stimulatory effect during the late cell stage (3 and 7 days). The staining showed 1,25OHD had an inhibitory effect on ALP enzyme activities and mineralization in a dosage-dependent manner up to 14 days. However, at 21 days, there was no difference between the treatments. This study provides a novel understanding of the effects of 1,25OHD on osteogenic differentiation and mineralization of cMSCs depending on cell stage and maturity.

Effects of plastic and straw mulching on soil microbial P limitations in maize fields: Dependency on soil organic carbon demonstrated by ecoenzymatic stoichiometry

Mulching rain-fed farmland ecosystems changes the soil physicochemical properties, especially soil organic carbon (SOC), but the metabolic limitations of soil microorganisms after these changes are unclear. We established a long-term experiment in 2012 with three treatments: no mulch (CK), straw mulch (SM), and plastic film mulch (FM). In 2019 the soil enzyme activities were measured in five maize growing periods: planting time, six-leaf period, silking period, milk period, and physiological maturity. Extracellular enzymatic stoichiometry models were used to examine microbial metabolic limitations. The vector length and angle were employed to determine the C and N/P limitations of soil microorganisms. Compared with CK, the average SOC and total nitrogen (TN) contents were 9.7% and 7.8% higher under SM, respectively, in each period. The SOC, TN, and total phosphorus (TP) contents were 5.6%, 4.8%, and 2.8% lower under FM, respectively. Compared with CK, the C- and N-acquiring enzyme activities were 20.5% and 5.2% lower under FM, respectively. The alkaline phosphatase enzyme activities were 2.7% and 13.5% higher under SM and FM, respectively, than CK. Soil nutrients, pH, and temperature influenced the C and P limitations of soil microorganisms. The different P limitation responses under SM and FM were mainly due to SOM. The decomposition of SOC was a key source of soil available P. The soil hydrothermal conditions under FM accelerated the decomposition of SOC in the early years, thereby increasing the P limitation. However, long-term SM increased the SOC due to the annual input of straw and its decomposition released available P to alleviate the P limitation for microorganisms. Thus, the temperature, water, pH, and SOC affect the P limitation for microbes under mulching conditions, but the SOC content of alkaline soil in arid farmland is the main factor that leads to microbial P limitation.

Microbial necromass formation, enzyme activities and community structure in two alpine elevation gradients with different bedrock types

Microbial necromass is an important part of soil organic carbon (SOC), but the changes in its contribution along elevational gradients and soil types are poorly understood. At two sites, the alkaline site Hochschwab and the acidic site Rauris, soil samples from three elevation levels and depths were analysed for amino sugars as microbial necromass indices, extracellular enzymes, and phospholipid fatty acids (PLFA) as microbial biomass indices. The three elevation levels at each site represented similar temperature and precipitation regimes. Microbial necromass contribution to SOC was lower (26%) at the alkaline site than at the acidic site (37%), indicating lower transformation of microbial biomass into stable SOC and support the increased accumulation of fungal necromass under N limitation at Hochschwab. Fungi generally dominate microbial necromass, with 88% at Hochschwab and 77% at Rauris. At the alkaline site Hochschwab, phenoloxidase, peroxidase and phosphatase enzymes activities were lower by 96%, 98% and 75%, respectively than at the acidic Rauris site, but not those of the hydrolytic enzymes β-glucosidase, exo-glucanase, exo-chitinase, and protease, indicating retarded decomposition of plant residues. Gram positive bacteria dominate bacterial biomass at both sites, with 70% at Hochschwab and 72% at Rauris. The higher fungal to bacterial PLFA ratio suggest increased contribution of fungi to total microbial PLFA at Hochschwab due to the better adaptability of fungi to the site-specific environmental conditions. Effects along alpine elevation gradients depend on vegetation induced changes in soil properties, which interact with bedrock properties.

Enzymatic Markers of Pulmonary Inflammation in Tracheal Wash Fluid in Racehorses: Alkaline Phosphatase and Lactate Dehydrogenase

Background: Diagnosis of the lower airway diseases (LAD) in horses relies on clinical signs, endoscopic examination and cytologic evaluation of lower respiratory tract flushing fluids as well. Specific enzyme activities in respiratory tract fluids of horses could be used as useful indicators in lower airway disorders. The main goal of the study was to evaluate the diagnostic usefulness of alkaline phosphatase (ALP) and lactate dehydrogenase (LDH) enzyme activites in tracheal wash (TW) fluids of horses with lower respiratory tract inflammation and/or damage and also in distinguishing of non-septic and septic inflammation in racehorses with LAD.Materials, Methods &amp; Results: A cross-sectional study was conducted in racehorses. TW fluid samples were obtained from 83 racehorses (71 with lower airway diseases and 12 healthy controls) for the study. The One-Way ANOVA Test was used for the significance analysis of the differences between the groups in showing the normal distribution of blood serum ALP and LDH properties. Kruskal Wallis H Test and Tamhane T2 test were used for the significance analysis of the differences between the groups of tracheal ALP and LDH properties, which were not normally distributed. The sampled horses were classified into 3 groups as healthy control, non septic neutrophilic inflammation (NS-LAD) and septic neutrophilic inflammation (S-LAD) according to the tracheal mucus scores determined during endoscopic examination, clinical signs, cytologic and bacteriologic examination of collected TW samples. According to cytology results normal neutrophil levels (&lt; 20%) were determined in the heatlhy control group (14.5%). However, increased neutrophil levels (≥ 20% and ≥ 80%) were encountered in the NS-LAD (61.4%) and S-LAD (24.1%) groups respectively. Also, the signs of degenerative changes and presence of intracellular bacteria in the neutrophil cells in the S-LAD group was observed. Bacteriologic analysis of the samples revealed Streptococcus equi subsp. zooepidemicus was the most prevalent agent (85.6%) followed by Escherichia coli (4.8%), Staphylococcus aureus (4.8%) and Klebsiella pneumoniae (4.8%). ALP and LDH enzyme activities of TW fluid were found to be significantly higher in racehorses with LAD compared with healthy controls (P &lt; 0.01). On the other hand, a significant difference was also determined for LDH levels between NS-LAD and S-LAD cases determining with cytological and bacteriological examination (P &lt; 0.01). No statistically significant difference was found between NS-LAD and S-LAD group. Serum ALP and LDH levels of racehorses revealed no significant difference between each study groups.Discussion: Respiratory system diseases have been known to be one of the main causes of training disruption and interruption of racing events in racehorses. Specifically, LAD is the most frequently diagnosed disorder in racehorses evaluated for poor performance. In the present study established for determining the usefulness of ALP and LDH enzyme levels in the TW samples of horses with lower respiratory tract inflammatory disorders. It was concluded that ALP and LDH enzyme activities of TW fluid could be considered as indicators of pulmonary inflammation and/or damage and LDH enzyme activity is thought to be useful in distinguishing neutrophilic non-septic and septic pulmonary inflammation in racehorses with LAD. On the other hand, data obtained from the study indicated that TW fluid ALP activity was not enough statistically to differenatiate horses with non-septic and septic lower airway inflammation.

A preamble study on ecophysiological impact of bamboo species for sustaining soil health

Bamboo, perennial flowering plant species of family Poaceae, was studied in present study to estimate the effect of different bamboo species on physico-biochemical properties of soil collected from rhizosphere of ten bamboos species viz, Bambusa balcooa, B. bamboos, B. smultiplex, B. nutans, B. tulda, B. vulgaris, Dendrocolomus asper, D. giganteus, Dendrocolomus hamiltoni, Dendrocolomus stricus. In the present study soil macronutrients and enzyme activities were analyzed that play role in soil physical, chemical and biological properties that in turn help in crop production, soil health improvement, biomass production with enhanced microbial activities. In the bamboo rhizosphere the pH and EC correspondingly ranged between6.56 to 8.10 and 18.80 to 45.23 milli S/m where highest pH and EC were observed in Dendrocolomus strictus and Bambusa balcooa respectively. While in the bamboo rhizosphere total organic carbon and total organic matter respectively ranged between 9.133% to 18.567% and 15.708% to 31.935%. The Highest total organic carbon and total organic matter was recorded in Bambusa bambos. The highest available Nitogen was found in the B. tulda(1.833 kgh-1) whereas highest available Phosphorus was observed to be 0.088 kgh-1 (D. asper and D. stricus). The highest available Pottassium was found in biosphere of, B. balcooa (33.167 kgh-1). The highest β glucosidase and alkaline phosphatase enzyme activity and its specific activity was reported in Dendrocolomus gigantus 115.17 (Catechol mg-1soil g-1 h -1), 355 (U mg-1) and 413.5 (mg PNP g-1soil h-1) and 200.10 (U mg-1) respectively. The inputs of organic matter through bamboos species increased crop roots density and better microbial and enzymatic activities. The results indicate that adoption of the agroforestry practices help to improved organic matter status of the soil, which is also reflected in the increased nutrient pool necessary for long-term productivity of the soil.

Cytotoxicity and Bioactivity of Calcium Silicate-based Cements in a Culture of Stem Cells from the Apical Papilla.

The present in vitro study evaluated the cytotoxicity and bioactivity of commonly-used calcium silicate-based cements in a culture of stem cells from the apical papilla (SCAPs). NeoMTA Plus (Avalon Biomed), BiodentineTM (Septodont) and MTA HP Repair (Angelus) cements were evaluated using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) and sulphorhodamine-B (SRB) viability assays. Cells were seeded (1*104 cells mL-1) in 96-well plates and exposed to 1:4 diluted extract in 24 h and 72 h. For the analysis of bioactivity, alkaline phosphatase (ALP) enzyme activity and Alizarin Red S (AZR) were assessed after 24 h of cell culture in 12-well plates (1*104 cells mL-1), where cells were exposed to 1:4 diluted extract on days 1 and 7. Minimum Essential Eagle's Medium alpha modification was used as control. ANOVA and Tukey's post hoc test were used to compare the different cements at each experimental time point. No significant differences were found between the cements and the control specimens on MTT at 24 h and 72 h (P>0.05); however, the calcium silicate-based cement materials showed higher cell viability compared to the control group (P<0.05). In the 24-h SRB, NeoMTA Plus showed lower cell viability than BiodentineTM and MTA HP Repair (P<0.05), with all groups similar to the control group (P>0.05). Compared to 24-h results, only NeoMTA Plus presented increased cell viability at 72 h (P<0.05). ALP activity was similar across the materials at 1 day (P>0.05). ALP activity was higher for BiodentineTM when compared to NeoMTA Plus (P<0.05), nevertheless, it was similar to MTA HP Repair and control groups (P>0.05) at 7 days. At 1- and 7-day periods of AZR assay, BiodentineTM presented higher levels of mineralized nodule formation (P<0.05). All evaluated calcium silicate-based cements demonstrated cell viability and bioactivity, suggesting that these (bio)materials may be indicated for use in regenerative dentine-pulp complex procedures.

Avaliação da conformidade de queijos industriais fiscalizados em Minas Gerais, Brasil

O objetivo do presente trabalho foi realizar uma análise histórica do perfil de conformidade de 2.580 amostras de queijos industriais sob fiscalização no estado de Minas Gerais, em relação aos parâmetros físico-químicos regulamentados Umidade, Matéria Gorda no Extrato Seco, Atividade da Enzima Fosfatase Alcalina Residual e Nitrato, no período de sete anos. Os parâmetros com maior prevalência de não conformidades foram Matéria Gorda no Extrato Seco e Atividade da Enzima Fosfatase Alcalina, alavancados pelas contribuições dos queijos Minas Frescal e Muçarela, respectivamente. Estabilidade no perfil de não conformidades foi evidenciada no período considerado. Concluiu-se que o investimento em ações voltadas para aqueles parâmetros identificados como críticos seria uma importante tática para promoção de uma mudança no patamar de qualidade e conformidade de queijos no estado. Ainda, observou-se que a aplicação de ferramentas estatísticas sobre as séries históricas de análises fiscais permite um melhor entendimento e monitoramento do sistema, com direcionamento das ações de melhoria, visando à promoção da segurança dos alimentos.

Influence of photobiomodulation and vitamin D on osteoblastic differentiation of human periodontal ligament stem cells and bone-like tissue formation through enzymatic activity and gene expression.

(1)Human periodontal ligament stem cells (HPDLSCs) are a unique population of mesenchymal stem cells (MSCs). Recently, the positive effects of photobiomodulation on the regulation of MSCs proliferation and osteogenic differentiation have gained significant attention. This study aimed to assess the effects of photobiomodulation and vitamin D (as an anabolic factor) on HPDLSCs for bone regeneration. (2)HPDLSCs were collected, isolated, and characterized and then divided into six groups: groups I and II, control and (10-7 Mol) vitamin D, respectively; group III, irradiation at 1 J/cm2 of 808-nm diode laser; group IV, irradiation at 1 J/cm2 and culture with vitamin D; group V, irradiation at 2 J/cm2, and group VI, irradiation at 2 J/cm2 and culture with vitamin D. Cell viability assay was measured through MTT assay and cell growth curve. Alkaline phosphatase (ALP) enzyme activity and mRNA levels of RUNX2, collagen 1 (Col-1), ALP, and osteonectin were also assessed. (3)Photobiomodulation at 1 and 2 J/cm2 combined with vitamin D significantly promoted HPDLSC proliferation (in MTT assay and cell growth curve results) and osteogenic differentiation (through the gene expression of RUNX2, Col-1, ALP, and osteonectin levels (p < 0.05). (4)Laser irradiation at 2 J/cm2 combined with vitamin D3 enhanced osteoblast differentiation and proliferation of cultured HPDLSCs and thus could further substitute bone grafting.

Bioaccumulation of heavy metals and their toxicity assessment in Mystus species.

This study was conducted on two native fish species namely Mystus vittatus and Mystus tengara inhabiting challenging environment of Yamuna River. The heavy metals concentrations in the river water were found to be as follows: Fe > Mn > Zn > Cu > Ni > Cr > Cd, all above the Bureau of Indian Standards (BIS) and World Health Organization (WHO) guidelines. The high metal pollution index in gill, liver, and kidney of M. vittatus was recorded compared to M. tengara. The pathology caused by the accumulation of heavy metals resulted significantly (p < 0.05) higher enzyme activities of alkaline phosphatase (ALP), aspartate aminotransferase (AST), alanine aminotransferase (ALT), and creatinine kinase (CK) in M. tengara as compared to M. vittatus. However, albumin: globulin ratio was found to be below 0.8 in both fishes. Higher total leukocyte (TLC) (48.5 × 103/mm3), lymphocytes (40%), respiratory burst activity (1.9), and nitric oxide synthase (NOS) activity (13.11 U/L) in M. vittatus reflect high immune response. In addition, chromosomal breakage study showed significantly (p < 0.05) low micronuclei frequency, lobed nuclei, and kidney-shaped nuclei (KSN) in M. vittatus. These results indicate that under the same challenging conditions M. vittatus have more capability of resistance and its continuous survival points towards its suitability to serve as a bioindicator than M. tengara.

АКТИВНОСТЬ ЩЕЛОЧНОЙ И КИСЛОЙ ФОСФАТАЗ В ТКАНЯХ ТОЩЕЙ КИШКИ У РАЗНОВОЗРАСТНЫХ КРОЛИКОВ

The article describes the nature and intensity of age-related changes in the activity of alkaline and acid phosphatases in the tissues of the mucous and muscle layers of the proximal, medial and distal parts of the jejunum in rabbits. The enzyme activity was determined spectrophotometrically in the scientific laboratory of the Federal State Budget Educational Institution of Higher Education Chuvash State Agricultural Academy. It was revealed that high metabolic processes in the tissues of the mucous and muscle layers of the jejunum in rabbits are found up to thirty days of age. The low-est activity of metabolic processes is detected from the age of 45 days. At the same time, by the age of four months of rabbits, stabilization of the activity of alkaline and acid phosphatase enzymes does not occur.

Concentrated Growth Factors (CGF) Induce Osteogenic Differentiation in Human Bone Marrow Stem Cells.

Simple SummaryOsteogenesis is a complex physiologic process that occurs during development as well as during damaged bone regeneration. This process requires several growth factors that act on stem cell populations, including Bone Marrow Stem Cells (BMSC). The present study fits into the research field for safe improvement of cell osteogenesis induction. In this context there is a great interest on an autologous and biocompatible blood derived product, named Concentrated Growth Factor (CGF). In particular, the ability of CGF to induce osteogenic differentiation of human BMSC (hBMSC) in vitro was here investigated. The osteogenic differentiation was evaluated measuring typical osteogenic markers such as alkaline phosphatase enzyme activity, matrix mineralization of hBMSC, and expression of some osteogenic-related genes. The results show that CGF alone is able to induce osteogenic differentiation of hBMSC. This finding opens up further, interesting perspectives in the biotechnological use of CGF in the tissue regeneration field.Bone regeneration is a complex process regulated by several factors that control overlapping biological processes, coordinating interactions among distinct cell populations. There is a great interest in identifying new strategies for inducing osteogenesis in a safe and efficient manner. Concentrated Growth Factor (CGF) is an autologous blood derived product obtained by centrifugation of venous blood following the procedure set on the Silfradent device. In this study the effects of CGF on osteogenic differentiation of human Bone Marrow Stem Cells (hBMSC) in vitro have been investigated; hBMSC were cultured with CGF or osteogenic medium, for 21 days. The osteogenic differentiation was evaluated measuring alkaline phosphatase (ALP) enzyme activity, matrix mineralization by alizarin red staining and through mRNA and protein quantification of osteogenic differentiation markers by Real-time PCR and Western blotting, respectively. The treatment with CGF stimulated ALP activity and promoted matrix mineralization compared to control and seems to be more effective than osteogenic medium. Also, hBMSC lost mesenchymal markers and showed other osteogenic features. Our study showed for the first time that CGF alone is able to induce osteogenic differentiation in hBMSC. The application of CGF on hBMSC osteoinduction might offer new clinical and biotechnological strategies in the tissue regeneration field.

The effects of hand-stripping on some epidermal mucus immune parameters in rainbow trout, Oncorhynchus mykiss

Fish epidermis functions as the first defense barrier against physical, chemical, and biological stressors. However, the effects of hand-stripping on fish mucosal immune responses have been hardly investigated. The present study investigated the effects of stripping procedures on skin mucosal immune responses of rainbow trout ( Oncorhynchus mykiss ) breeders. The skin mucus was sampled from six male and six female trout before and one week after the stripping handling. The results showed that stripping had significant effect on all parameters except protease activity, but gender had a significant effect only on the mucosal protease and alkaline phosphatase activity. The results did not show any effect of the interaction between stripping and gender on mucosal lysozyme activity. The data revealed that enzymatic activities of alkaline phosphatase, lysozyme, esterase, as well as the total immunoglobulin level and bactericidal activity were significantly reduced in the skin mucus of fish one week after the stripping. The reduction of immune parameters in the skin mucus could be related to immunosuppression caused by stripping stress which, in turn, might have made the fish more susceptible to microbial infections and diseases. Therefore, care should be taken during stripping to minimize the manipulation stress.