Effect of sheep bone protein hydrolysate on promoting calcium absorption and enhancing bone quality in low-calcium diet fed rats.

Calcium deficiency is prone to fractures, osteoporosis and other symptoms. In this study, sheep bone protein hydrolysates (SBPHs) were obtained by protease hydrolysis. A low-calcium-diet-induced calcium-deficiency rat model was established to investigate the effects of SBPHs on calcium absorption and intestinal flora composition. The results showed that an SBPHs + CaCl2 treatment significantly increased the bone calcium content, bone mineral density, trabecular bone volume, and trabecular thickness, and reduced trabecular separation, and changed the level of bone turnover markers (P < 0.05). Supplementation of SBPHs + CaCl2 can remarkably enhance the bone mechanical strength, and the microstructure of bone was improved, and the trabecular network was more continuous, complete, and thicker. Additionally, SBPHs + CaCl2 dietary increased the abundance of Firmicutes and reduced the abundance of Proteobacteria and Verrucomicrobiota, and promoted the production of short chain fatty acids. This study indicated that SBPHs promoted calcium absorption and could be applied to alleviate osteoporosis.

The effect of amorphous calcium carbonate as a culture media supplement on embryonic development of murine sibling embryos.

PURPOSE: The aim of this study was to compare the addition in culture media of stabilized amorphous calcium carbonate (ACC) versus calcium chloride (CaCl2) or calcium carbonate in crystalline form (CCC) on growth rates among sibling mouse embryos. METHODS: We evaluated the effect of different ACC concentrations on the rates of embryo compaction at 60 h, blastocyst rate at 84 h and percentage of fully hatched at 108 h following hCG injection. As ACC is stabilized by tripolyphosphate (TPP), we also evaluated the addition of TPP alone to the culture media. Finally, we compared supplemented ACC culture media to one-step SAGE and Irvine cleavage media. RESULTS: The results revealed that ACC accelerates the compaction and blastocyst rates, as well as the percentage of fully hatched embryos in a dose-dependent manner, with an increased positive effect at 2.5 mM. The magnitude of the effect for ACC-supplemented media on the embryo developmental rate was between 30 to 40% (p < 0.01) faster for each stage, compared to both SAGE and Irvine one-step standard media. Embryos cultured with SAGE or Irvine media with or without supplementation of CaCl2 or CCC, did not produce the same improvements as observed with ACC. CONCLUSION: In conclusion, the ACC demonstrates a rapid modulation effect for restoring media optimal pH. ACC can inhibit cathepsin B activity during in vitro culture of fibroblast cells. The beneficial impact of ACC on cleavage mouse embryos is likely due to an improved buffering effect causing slower pH media variations, which may enhance quality and implantation potential of embryos following in vitro culture.

Oral calcium and vitamin D supplements differentially alter exploratory, anxiety-like behaviors and memory in male rats.

Oral calcium and calcium plus vitamin D supplements are commonly prescribed to several groups of patients, e.g., osteoporosis, fracture, and calcium deficiency. Adequate and steady extracellular calcium levels are essential for neuronal activity, whereas certain forms of calcium supplement (e.g., CaCO3) probably interfere with memory function. However, it was unclear whether a long-term use of ionized calcium (calcium chloride in drinking water ad libitum), vitamin D supplement (oral gavage) or the combination of both affected anxiety and memory, the latter of which was probably dependent on the hippocampal neurogenesis. Here, we aimed to determine the effects of calcium and/or vitamin D supplement on the anxiety- and memory-related behaviors and the expression of doublecortin (DCX), an indirect proxy indicator of hippocampal neurogenesis. Eight-week-old male Wistar rats were divided into 4 groups, i.e., control, calcium chloride-, 400 UI/kg vitamin D3-, and calcium chloride plus vitamin D-treated groups. After 4 weeks of treatment, anxiety-, exploration- and recognition memory-related behaviors were evaluated by elevated pulse-maze (EPM), open field test (OFT), and novel object recognition (NOR), respectively. The hippocampi were investigated for the expression of DCX protein by Western blot analysis. We found that oral calcium supplement increased exploratory behavior as evaluated by OFT and the recognition index in NOR test without any effect on anxiety behavior in EPM. On the other hand, vitamin D supplement was found to reduce anxiety-like behaviors. Significant upregulation of DCX protein expression was observed in the hippocampus of both calcium- and vitamin D-treated rats, suggesting their positive effects on neurogenesis. In conclusion, oral calcium and vitamin D supplements positively affected exploratory, anxiety-like behaviors and/or memory in male rats. Thus, they potentially benefit on mood and memory in osteoporotic patients beyond bone metabolism.

Relaxation mechanisms of chloroform root extracts of Prangos heyniae and Prangos uechtritzii on mouse corpus cavernosum.

Erectile dysfunction (ED) is the inability to achieve/maintain an erection. Because of the side effects, interactions, or ineffectiveness of currently used drugs, novel drug discovery studies are ongoing. The roots of Turkish endemic plants Prangos uechtritzii and Prangos heyniae are traditionally used as aphrodisiacs in Anatolia and contain coumarin-like relaxant compounds. This study aims to reveal the relaxant effect mechanisms of chloroform root extracts of P. heyniae (Ph-CE) and P. uechtritzii (Pu-CE). Isolated organ bath experiments were performed on Swiss albino mouse corpus cavernosum by DMT strip myograph. Relaxant responses to extract (10-7 -10-4  g/ml) were obtained in the presence/absence of NO and H2 S synthesis inhibitors nitro-l-arginine methyl ester (l-NAME, 100 µM) and aminooxyacetic acid (AOAA, 10 mM) respectively. Sodium nitroprusside (SNP, 10-9 to 10-4  M) and Na2 S (10-6 to 3 × 10-3  M)-induced relaxations and CaCl2 (10-6 to 10-4  M), KCl (10-2.1 to 10-0.9  M) and phenylephrine (3 × 10-8 to 3 × 10-5  M)-induced contractions were taken in the presence/absence of the extracts (10-4  g/ml). Relaxations induced by Ph-CE but not by Pu-CE were inhibited in the presence of l-NAME and AOAA. Ph-CE increased Na2 S- and SNP-induced relaxations. Ph-CE and Pu-CE decreased the contractions of KCl, phenylephrine, and CaCl2 . It was concluded that NO and H2 S synthesis/downstream mechanisms play roles in relaxations of Ph-CE but not in Pu-CE-induced relaxations. Inhibition of calcium influx appears to be involved in the relaxant effect of Ph-CE and Pu-CE. Since the extracts act directly by relaxing smooth muscle or through H2 S as well as NO, they may be a potential therapeutic agent in diseases such as ED where the bioavailability of NO is impaired.

Antidiarrheal effect of ethanol extract from Lophatheri Herba and its effect on isolated jejunal smooth muscle in rabbits.

Lophatheri Herba is a traditional Chinese medicine, which is commonly used in the treatment of fever, stomatitis, urodynia. The aim of the study is to evaluate the antidiarrheal activity of the ethanol extract of Lophatheri Herba (Gramineae, ELH) and observe its effect on isolated jejunum smooth muscle in rabbits, so that we can provide a possible pharmacological basis for its clinical use. Methods: In vivo, the antidiarrheal activity of ELH (250, 500 and 1000 mg/kg; orally) in castor oil-induced Kun Ming mice was evaluated. In vitro, the effect of ELH (0.01-10 mg/mL) on the spontaneous and ACh (10µM)/K+ (60mM)-induced contraction of isolated rabbit jejunum smooth muscle was studied. The possible mechanism of spasmolytic effect of ELH (1, 3mg/mL) was explored by pretreatment of intestinal tract with CaCl2. Results: ELH (500 and 1000mg/kg) exhibited antidiarrheal effect and it (0.01-10 mg/mL) inhibited the spontaneous and ACh/K+-induced contraction with an EC50 value of 1.27 (0.89-1.34), 0.76 (0.54-1.02) and 0.34 (0.27-0.53), it also shifted the concentration-response curves of CaCl2 to right with decreased in max, similar to verapamil. ELH has significant antidiarrheal and spasmolytic effect, this provides the pharmacological basis for use in gastrointestinal disorders.

Cell wall stabilization and calcium absorption on mango fruit treated with a quarantine hot water treatment combined with calcium salts and stored at chilling temperature.

Hot water treatment (HT) induces chilling injury (CI) tolerance in mango, but prolonged exposure to HT causes softening. In this sense, calcium salts stabilize the cell wall. Nevertheless, there is little information on the effect of HT combined with calcium salts (HT-Ca) on calcium absorption and cell wall stability during storage of mango at CI temperature. We evaluated the effect of quarantine HT in combination with calcium chloride (CaCl2 ), calcium citrate (CaCit), or calcium lactate (CaLac) on calcium absorption, CI tolerance, and cell wall stabilization. HT and HT-CaCl2 had the lowest CI development. HT increased firmness loss and electrolyte leakage, and HT-Ca counteracted this effect. Overall, HT-Ca treatments had a similar effect on the cell wall degrading enzymes. HT-CaCl2 was the best treatment and did not present alterations on the epicuticular wax as observed on HT. HT-CaCl2 is a useful technology to stabilize cell wall and preserve mango during chilling storage. PRACTICAL APPLICATIONS: The addition of calcium salts in an established hot water quarantine procedure for mango exportation represents a viable alternative to counteract the negative effects of this thermal treatment upon cell microstructure, maintaining its positive effect of tolerance to chilling injury. In this sense, mango producers and packers can use a HT-CaCl2 treatment to reduce the presence of chilling injury and extent the fruit shelf life and improve its commercialization. Furthermore, technical and infrastructure changes are not necessary for the packaging chain.

Evaluation of phytochemical, anti-oxidant and cardiac depressant effect of Rumex dentatus by using Langendorff's isolated heart apparatus.

Rumex dentatus has been used traditionally for ailment of cardiovascular diseases. The aim of the present study was to assess cardiovascular effects in isolated perfused rabbit heart. Aqueous and n-butanolic fractions were assessed for their effect on perfusion pressure (PP), force of contraction (FC) and heart rate (HR) of rabbit heart using Langendorff's method. The possible mechanisms of action of extracts/fraction were assessed with and without application of different agonist/antagonist. Phytochemical, toxicity and anti-oxidant activities were also determined. Both extracts at 1mg/mL dose produced a highly significant decrease in FC and HR but PP remained unchanged. Moreover, aqueous fraction of Rumex dentatus at 0.001mg/mL dose produced a highly significant decrease in FC and HR but no significant change in PP was observed. Atropine 10-5 M did not inhibit the cardiac depressant response of both fractions. Furthermore, both fractions blocked the positive ionotropic and chronotropic effects of adrenaline and calcium chloride. Phytochemical studies have shown the presence of some phytochemicals. Acute and sub-chronic toxicity studies demonstrated that test extracts are safe and produced no significant changes in haematological and biochemical parameters. Crude extract showed significant antioxidant activity like ascorbic acid. This study revealed that this plant have good cardiac depressant effect.

Development of a thin-layer chromatography bioautographic assay for neuraminidase inhibitors hyphenated with electrostatic field induced spray ionisation-mass spectrometry for identification of active Isatis indigotica root compounds.

The identification of neuraminidase inhibitors from natural products is a promising strategy in the field of anti-influenza research. In this study, a new thin-layer chromatography (TLC) bioautographic assay for the screening of neuraminidase inhibitors from natural products was developed. This TLC bioassay is based on the one-step reaction of neuraminidase with the sodium salt of 5­bromo­4­chloro­3-indolyl-α-d-N-acetylneuraminic acid (substrate) and the subsequent formation of blue coloured products. Neuraminidase inhibitory activity was shown by the development of white spots against the blue TLC background. The key factors affecting the assay (such as enzyme concentration, substrate concentration, incubation time, reaction time, and pH) were investigated and optimised by a combination of a one-factor-at-a-time design and a Box-Behnken design/response surface method. The developed TLC bioautographic method was applied to identify neuraminidase inhibitory compounds in the roots of Isatis indigotica. Eleven active compounds including six alkaloids, three lignans, one sterol, and one fatty acid were identified in situ by direct coupling with an electrostatic field induced spray ionisation-mass spectrometry approach through analysis of their MSn (n = 4) data or comparison with reference substances. The developed TLC bioautographic assay is simple, rapid, and efficient for screening potential neuraminidase inhibitors from natural products.

Physiological and metabolic analysis of winter jujube after postharvest treatment with calcium chloride and a composite film.

BACKGROUND: Ziziphus jujuba Miller cv. Dongzao is extremely susceptible to reddening, browning, nutritional loss, and perishability after harvest. In this study, we evaluated the mechanisms of calcium chloride and chitosan/nano-silica composite film treatments on the quality, especially in reddening, by physiological and metabolomic assays. RESULTS: The treatment delayed the decline of phenylalanine ammonia-lyase (PAL), chalcone synthase (CHS), and chalcone isomerase (CHI) activities. Meanwhile, the treated groups retarded the increases in anthocyanin and quercetin contents by inhibiting the gene expressions of flavonol synthase (ZjFLS), dihydroflavonol 4-reductase (ZjDFR), and anthocyanidin synthase (ZjANS), while promoting leucoanthocyanidin reductase (ZjLAR) expression, which leads to retardation of fruit reddening. Anthocyanins were found to be responsible for post-harvest winter jujube reddening through principal component analysis. Results from the technique for order preference by similarity to an ideal solution indicated that the treated group delayed the decline of the quality of 'Dongzao' and extended its shelf life. CONCLUSION: The treatment induced the heightening of flavonoids metabolism. They enhanced the nutritional value and the ability to resist stress by delaying the decline of PAL, CHS, and CHI activities. Meanwhile, the treated groups retarded the increase in anthocyanin and quercetin contents by inhibiting the gene expressions of ZjFLS, ZjDFR, and ZjANS and promoting ZjLAR expression, which leads to retardation of fruit reddening. Anthocyanins are responsible for post-harvest winter jujube reddening. Coating treatment effectively delayed the decline of winter jujube quality. © 2020 Society of Chemical Industry.

Vineyard calcium sprays induce changes in grape berry skin, firmness, cell wall composition and expression of cell wall-related genes.

Having a central role in cell wall pectin cross-linking, calcium has been increasingly used as supplement to promote fruit firmness and extended shelf-life. However, the molecular rearrangements associated to increased fruit robustness are still a matter of debate. In this study, mechanical, histochemical and molecular assays were conducted to understand the mechanisms underlying the effects of Ca in fruit physical properties. In a two-year field trial, grapevines were sprayed with exogenous CaCl2 throughout the fruiting season. Results showed an increase in berry Ca concentration at harvest, associated to increased fruit consistency and skin resistance. Scanning electron microscopy showed that fruits from Ca-treated plants had smoother skin surfaces than control fruits, and that microcracks encircling the lenticels were less prominent. Histochemistry assays suggested higher deposition of pectin-like material in skin cell walls in grapes from Ca-treated vines, but no evident modifications in cellulose content were observed. Accordingly, the expression of cellulose synthase family gene CesA3 was not affected by exogenous Ca, while polygalacturonase-encoding genes PG1 and PG2 were downregulated, together with EXP6 belonging to expansin family, and CER9 and CYP15 involved in cuticle biosynthesis. These results suggested that Ca acts by inhibiting pectin degradation and cell wall loosening, while remodeling cuticle structure.

Endothelium-Independent Vasodilatory Effects of Isodillapiolglycol Isolated from Ostericum citriodorum.

Ostericum citriodorum is a plant with a native range in China used in herbal medicine for treating angina pectoris. In this study, we investigated the vasodilatory effects of isodillapiolglycol (IDG), which is one of the main ingredients isolated from O. citriodorum ethyl acetate extract, in Sprague-Dawley rat aortic rings, and measured intracellular Ca2+ ([Ca2+]in) using a molecular fluo-3/AM probe. The results show that IDG dose-dependently relaxed endothelium-intact or -denuded aortic rings pre-contracted with noradrenaline (NE) or potassium chloride (KCl), and inhibited CaCl2-induced contraction in high K+ depolarized aortic rings. Tetraethyl ammonium chloride (a Ca2+-activated K+ channel blocker) or verapamil (an L-type Ca2+ channel blocker) significantly reduced the relaxation of IDG in aortic rings pre-contracted with NE. In vascular smooth muscle cells, IDG inhibited the increase in [Ca2+]in stimulated by KCl in Krebs solution; likewise, IDG also attenuated the increase in [Ca2+]in induced by NE or subsequent supplementation of CaCl2. These findings demonstrate that IDG relaxes aortic rings in an endothelium-independent manner by reducing [Ca2+]in, likely through inhibition of the receptor-gated Ca2+ channel and the voltage-dependent Ca2+ channel, and through opening of the Ca2+-activated K+ channel.

Antifungal Activity of Lactic Acid Bacteria Isolates and Their Associations: The Effects of Ca and Mg Divalent Cations.

The study of effects of Ca2+ and Mg2+ on antifungal activity of lactic acid bacteria (LAB) isolates and their associations revealed inducing and inhibiting effects on antifungal activity. The addition of Ca2+ essentially inhibited the antifungal effect of L. rhamnosus MDC9661 but stimulated the activity of RIN-2003-Ls, MDC9632 and MDC9633 strains, as well as their associations. Mg2+ partly increased the inhibitory activity of LAB isolates, while the addition of ions combination did not cause changes of their antifungal activity. The supplementation of Ca2+ stimulated the antifungal effect of most associations against Penicillium sp., Trichoderma viride, Geotrichum candidum, and Aspergillus flavus compared with the native conditions. The addition of Mg2+ induced the antifungal activity of RIN-2003-Ls, MDC9632, MDC9633, and INR-2010-Tsov-G-St combinations. The antifungal effects of most associations were increased in the presence of ions mixture. The natural LAB associations including VKPM B-3386, MDC9632, and MDC9633 could not suppress the growth of any tested mold; however, the supplementation of ions combination revealed their antifungal effect against all kinds of molds. The finding of substantial stimulation of the most LAB associations antifungal effect by metal ions can be basis for creation of new effective antifungal preparations by the supplementation of ions combined mixture.

Effect of calcium chloride and sucrose on the composition of bioactive compounds and antioxidant activities in buckwheat sprouts.

In this study, we evaluated the effect of sucrose and CaCl2 on the growth profile, nutritional quality, and antioxidant capacity of sprouted buckwheat. Buckwheat seeds were germinated at 25 °C for 8 days and sprayed with four different solutions: distilled water, 3% sucrose, 7.5 mM CaCl2, and 3% sucrose plus 7.5 mM CaCl2. Our results showed that CaCl2 effectively improved sucrose-elicitation induced growth reduction in buckwheat sprouts. Elicitation with both sucrose and CaCl2 in buckwheat sprouts markedly enhanced the accumulation of bioactive compounds, such as polyphenols, flavonoids, γ-aminobutyric acid, vitamin C, and E, without negatively affecting sprout growth. Elicitation with both sucrose and CaCl2 not only significantly enhanced the antioxidant activities but also exerted cytoprotective effects against oxidative damage in HepG2 cells and fibroblasts. These findings suggested that simultaneous elicitation with 3% sucrose and 7.5 mM CaCl2 can potentially improve the nutritional value and potential health benefits of buckwheat sprouts.

Enhancing the hardness of potato slices after boiling by combined treatment with lactic acid and calcium chloride: Mechanism and optimization.

Potatoes usually suffer from greatly decrease of hardness after boiling, which limits their processing potential in food industry. Moreover, methods for enhancing the hardness of potatoes after boiling are underexplored. In this study, the hardness of potato slices after boiling were increased from 288 g to 2342 g by the combined treatment of lactic acid (LA) and calcium chloride (CC). Through the analysis of the microstructure of the potato cells, the molecular weight distribution and natural sugar ratio of different soluble pectin fractions, and the enzymatic activities (polygalacturonase, PG and pectin methylesterase, PME), the possible mechanism behind the hardness enhancement by LA and CC pretreatment, namely the direct link between pectin and potato structure was revealed. The obtained results confirmed the target spot for enhancing the hardness of potatoes after boiling lay in PG activity and gelation of the pectin, which also could be used to help other plants resist the heat process if pectin existed in their cell wall.

Understanding the role of calcium-mediated cell death in high-frequency irreversible electroporation.

High-frequency irreversible electroporation (H-FIRE) is an emerging electroporation-based therapy used to ablate cancerous tissue. Treatment consists of delivering short, bipolar pulses (1-10µs) in a series of 80-100 bursts (1 burst/s, 100µs on-time). Reducing pulse duration leads to reduced treatment volumes compared to traditional IRE, therefore larger voltages must be applied to generate ablations comparable in size. We show that adjuvant calcium enhances ablation area in vitro for H-FIRE treatments of several pulse durations (1, 2, 5, 10µs). Furthermore, H-FIRE treatment using 10µs pulses delivered with 1mM CaCl2 results in cell death thresholds (771±129V/cm) comparable to IRE thresholds without calcium (698±103V/cm). Quantifying the reversible electroporation threshold revealed that CaCl2 enhances the permeabilization of cells compared to a NaCl control. Gene expression analysis determined that CaCl2 upregulates expression of eIFB5 and 60S ribosomal subunit genes while downregulating NOX1/4, leading to increased signaling in pathways that may cause necroptosis. The opposite was found for control treatment without CaCl2 suggesting cells experience an increase in pro survival signaling. Our study is the first to identify key genes and signaling pathways responsible for differences in cell response to H-FIRE treatment with and without calcium.

Effect of NaCl stress and supplemental CaCl2 on carotenoid accumulation in germinated yellow maize kernels.

Germinated edible seeds and sprouts are becoming increasingly common in the human diet because they are rich in bioactive compounds and antioxidants and are highly nutritious. In this study, the effects of NaCl stress and supplemental CaCl2 on carotenoid accumulation, antioxidant capacity and expression of key enzymes in yellow maize kernels were investigated. The results showed that the lutein and zeaxanthin contents increased with NaCl treatment, and further increased with supplemental CaCl2. Additionally, germinated yellow maize kernels showed increased antioxidant capacity in response to NaCl and CaCl2. The transcript levels of carotenogenic genes ZmPSY and ZmCYP97C were upregulated and the expression levels of ZmLCYB and ZmBCH1 were downregulated under NaCl stress. The expression of all key carotenogenic genes was upregulated by CaCl2 supplementation. These results suggested that NaCl and CaCl2 contribute to carotenoid accumulation via increased expression of related carotenogenic genes and increased antioxidant capacity in germinated yellow maize kernels.

Calcium chloride and 1-methylcyclopropene treatments delay postharvest and reduce decay of New Queen melon.

In this study, newly harvested New Queen melons were treated with calcium chloride (CaCl2) and 1-methylcyclopropene (1-MCP) alone or in combination before storage. The results showed that the respiration rate, ethylene release, the activity and gene expression of pectinases such as polygalacturonase (PG), pectin methylesterase (PME) and pectate lyase (PL) in New Queen melons were dramatically decreased by treatments with 0.18 mol/L CaCl2 and/or 1 µL/L 1-MCP. Meanwhile, the climacteric behavior and flesh hardness reduction were inhibited. We also found that softer melon flesh was more conducive to the growth and reproduction of decay-causing microorganisms according to their growth curves in melons that were different in flesh hardness, suggesting inhibiting fruit softening can slow down the growth of microorganisms in fruit flesh, and thus reduce fruit decay rate. The combined use of CaCl2 and 1-MCP was more effective in suppressing respiration rate, ethylene release and protopectin hydrolysis, which could greatly delay the softening, reduce the decay rate, and extend the shelf life of New Queen melons.

Mesenchymal stem cells differentiate into keratinocytes and express epidermal kallikreins: Towards an in vitro model of human epidermis.

Epidermal differentiation is a complex process in which keratinocytes go through morphological and biochemical changes in approximately 15 to 30 days. Abnormal keratinocyte differentiation is involved in the pathophysiology of several skin diseases. In this scenario, mesenchymal stem cells (MSCs) emerge as a promising approach to study skin biology in both normal and pathological conditions. Herein, we have studied the differentiation of MSC from umbilical cord into keratinocytes. MSC were cultured in Dulbecco's modified Eagle's medium (DMEM) (proliferation medium) and, after characterization, differentiation was induced by culturing cells in a defined keratinocyte serum-free medium (KSFM) supplemented with epidermal growth factor (EGF) and calcium chloride ions. Cells cultivated in DMEM were used as control. Cultures were evaluated from day 1 to 23, based on the cell morphology, the expression of p63, involucrin and cytokeratins (KRTs) KRT5, KRT10 and KRT14, by quantitative polymerase chain reaction, Western blot analysis or immunofluorescence, and by the detection of epidermal kallikreins activity. In cells grown in keratinocyte serum-free medium with EGF and 1.8 mM calcium, KRT5 and KRT14 expression was shown at the first day, followed by the expression of p63 at the seventh day. KRT10 expression was detected from day seventh while involucrin was observed after this period. Data showed higher kallikrein (KLK) activity in KSFM-cultured cells from day 11th in comparison to control. These data indicate that MSC differentiated into keratinocytes similarly to that occurs in the human epidermis. KLK activity detection appears to be a good methodology for the monitoring the differentiation of MSC into the keratinocyte lineage, providing useful tools for the better understanding of the skin biology.

Calcium chloride treatment modifies cell wall metabolism and activates defense responses in strawberry fruit (Fragaria × ananassa, Duch).

BACKGROUND: Fruit dips in calcium ions solutions have been shown as an effective treatment to extend strawberries (Fragaria × ananassa, Duch) quality during storage. In the present work, strawberry fruit were treated with 10 g L-1 calcium chloride solution and treatment effects on cell wall enzymes activities and the expression of encoding genes, as well as enzymes involved in fruit defense responses were investigated. RESULTS: Calcium treatment enhanced pectin methylesterase activity while inhibited those corresponding to pectin hydrolases as polygalacturonase and ß-galactosidase. The expression of key genes for strawberry pectin metabolism was up-regulated (for FaPME1) and down-regulated (for FaPG1, FaPLB, FaPLC, FaßGal1 and FaAra1) by calcium dips. In agreement, a higher firmness level and ionically-bound pectins (IBPs) amount were detected in calcium-treated fruit compared with controls. The in vitro and in vivo growth rate of fungal pathogen Botrytis cinerea was limited by calcium treatment. Moreover, the activities of polyphenol oxidases, chitinases, peroxidases and ß-1,3-glucanases were enhanced by calcium ion dips. CONCLUSION: News insights concerning the biochemical and molecular basis of cell wall preservation and resistance to fungal pathogens on calcium-treated strawberries are provided. © 2019 Society of Chemical Industry.

Enhancement the Cellulase Activity Induced by Endophytic Bacteria Using Calcium Nanoparticles.

The huge applications of cellulosic and lignocellulosic materials in the various fields of life lead to accumulation of its wastes that became one of the major sources of environmental pollution. In this study, a Gram-positive cellulose-decomposing endophytic bacterium (Chi-04) was isolated from medicinal plant Chiliadenus montanus which inhabitant Saint Catherine (Sinai) region in Egypt. The bacterial strain was identified based on the sequence analysis of 16S rRNA genes as Lysinibacillus xylanilyticus. This isolate was capable of degrading 58% of cellulosic filter paper (100 g/l) within 15 days of incubation. The soluble and reduced sugars were spectrophotometrically determined as cellulose decomposition metabolites. The bacterial isolate exhibited an obvious activity toward cellulase enzyme production. The maximum cellulase activity (0.18 U/min) was detected after 12 days of incubation while the maximum release of soluble sugars (11.85 mg/ml) was detected after 15 days of incubation. CaCl2 nanoparticles (100 nm) were chemically prepared to enhance the activity of the enzyme. The optimum concentration of CaCl2 nanoparticles that showed the highest activity of cellulase (0.3 mg/ml reduced sugar) was 0.6%. The bacterial isolates showed potential convert of cellulose into reducing sugars which could be used in several applications.