Increase In Calcium Concentration Research Articles

Development of an adrenocortical cell model of calcium signaling modulation to decipher the molecular mechanisms responsible for primary aldosteronism

Primary aldosteronism (PA) is the most frequent form of secondary hypertension. The identification of germline or somatic mutations in different genes coding for ion channels and defines PA as a channelopathy. These mutations promote activation of calcium signaling, the main trigger for aldosterone biosynthesis. The objective of our work was to elucidate, using chemogenetic tools, the molecular mechanisms underlying the development of PA by modulating sodium entry into the cells, mimicking some of known mutations identified in PA. We have developed an adrenocortical H295R-S2 cell line stably expressing a chimeric ion channel receptor formed by the extracellular ligand-binding domain of the α7 nicotinic acetylcholine receptor fused to the ion pore domain of the serotonin receptor 5HT3α and named α7-5HT3. Mutations have been introduced in the ligand-binding domain to allow only synthetic drugs to activate this channel receptor. Activation of α7-5HT3 by a specific drug, PSEM-817 leads to sodium entry into the cells. This cell line was characterized in terms of intracellular calcium concentrations, cell proliferation, aldosterone production, steroidogenic expression and electrophysiological properties. Treatment of α7-5HT3 expressing cells with increasing concentration of PSEM-817 (from 10-9 to 10-5 M) induced a significant increase in intracellular calcium concentrations, similarly to potassium (12 mM) or angiotensin II (10-8 M). This stimulation of calcium signaling did not affect cell proliferation, but was responsible for an increase in CYP11B2 expression and aldosterone production after 24 h of treatment. However, while increased intracellular calcium concentrations were observed starting from 10-8 M of PSEM-817, CYP11B2 expression and aldosterone production were only affected starting from 10-7 M, suggesting a dose-dependent effect. Finally, whereas cells were hyperpolarized in absence of stimulation (around −60 mV), PSEM-817 induced a strong depolarization, cells rising to a membrane potential around −10 mV. This cell line, in which we can modulate the intracellular calcium concentration “on demand”, is a useful tool for a better understanding of the alterations of intracellular ion balance and calcium signaling in the pathophysiology of PA.

Dual-specificity Phosphatase 1 Suppresses Vascular Smooth Muscle Cell Calcification by Optical Atrophy 1-related Pathway

Objective To investigate the effect of dual-specificity phosphatase 1/optical atrophy 1 (DUSP1/OPA1) signaling pathway on vascular smooth muscle cell (VSMC) calcification.Methods An in vitro model of VSMC calcification was induced by exposure to β-glycerophosphate and calcium chloride.VSMC calcification was assessed by Alizarin Red S staining and calcium content by ELISA.Apoptosis was detected by TUNEL.Western blotting was employed to determine the protein levels of DUSP1,OPA1,Runt-related transcription factor 2 (Runx-2),bone morphogenetic protein 2 (BMP-2),and cysteinyl aspartate-specific proteinase-3 (Caspase-3).The effects of DUSP1 overexpression and OPA1 knockdown on cell calcification were investigated.Results Calcium chloride and β-glycerolphosphate induced VSMC calcification and down-regulated the expression levels of DUSP1 (t=11.951,P<0.001) and OPA1 (t=8.487,P<0.001).DUSP1 overexpression promoted OPA1 expression (t=-8.921,P<0.001),attenuated VSMC calcification,reduced calcium content and apoptosis rate,and down-regulated the expression of Runx-2,BMP-2,and active Caspase-3 (all P<0.001).OPA1 knockdown increased calcium content and apoptosis rate,up-regulated the expression of Runx-2,BMP-2,and active Caspase-3,and promoted VSMC calcification (all P<0.001).Conclusion DUSP1 may inhibit the VSMC calcification through the OPA1 signaling pathway.

Optimization of vacuum drying and determination of functional properties of Kadam ( Neolamarckia cadamba ) fruit powder

In the present study, the vacuum drying of the Kadam fruit was performed, and the functional properties of the Kadam fruit powder were estimated. The drying conditions were optimized using the Response Surface Methodology for minimum color change and maximum vitamin C retention. The functional properties of the Kadam fruit powder dried at the optimized conditions (bed thickness 3.67 mm, drying temperature 65°C, drying pressure 160 mmHg, and blanching temperature 100°C) were observed as; tapping density 4.23 ml/g, oil retention capacity 5.7 ml/g, water retention capacity 16 ml/g, water-solubility 0.5%, emulsion activity 0.5%, and emulsion stability 0.85%. The optimum dried fruit powder had vitamin C content (508 mg/100 g), color change (6.32), and effective moisture diffusivity (2.16 × 10−7 m2/min). This study showed that the vacuum drying of the Kadam pulp minimized the thermal degradation of the color and vitamin C. Practical applications Kadam fruit is a good source of minerals and bioactive compounds but is highly under-utilized due to its taste and lack of awareness in the population. Preparation of fruit powder can add value to under-utilized Kadam fruit. Kadam fruit powder can be used in many food products as functional ingredients. Also, incorporating Kadam powder in food formulations can increase calcium content and dietary fiber. The vacuum drying method helped minimize the thermal degradation of the color and Vitamin C of the Kadam fruit powder.

Enhanced H2S gas sensing performance of Ca-doped Bismuth Ferrite thick films

In the present work, gas sensing properties of Calcium-doped Bismuth Ferrite (BFO) nano-particulate films have been investigated as a function of doping concentration, operating temperature and nature of the exposure gases. Pure Bismuth ferrite films were found to be sensitive towards H2S gas at 350 °C. With increase in calcium concentration, besides an increase in sensor response, from ∼4.5 in BFO to ∼226 in Bi0·7Ca0·3FeO3 on exposure to 50 ppm of H2S, a non-ideal p-type behavior is observed in response curves. Such a large enhancement has not been observed earlier in a p-type semiconductor. In BFO and Bi0·9Ca0·1FeO3, resistance of films is observed to increase in accordance to p-type behavior, followed by a small decrease before saturation in gas ambience. In samples with higher calcium content, on exposure to gas, resistance of the sample is observed to increase initially followed by a decrease in resistance while still in gas ambience (saturation in gas ambience is not at all observed). In order to investigate the reason for (i) the large enhancement in sensor response usually not observed in a p-type sensor, and (ii) origin of non-ideal p-type behavior, X-ray photoelectron spectroscopy analysis of Bi0·7Ca0·3FeO3 films was carried out, after gas exposure. Based on analysis of XPS results, an additional sensing mechanism involving chemical reaction between the cations of sensing material and H2S, was found; which could satisfactorily explain the non-ideal behavior as well as enhancement in sensor reponse.

Understanding Contributions of Divalent Cations in Mineral Carbonation Using Artificial Neural Network

The roles played by divalent cations (calcium, magnesium and iron) of rock minerals in the efficiency of mineral carbonation have been investigated. Statistical modeling with Artificial Neural Network (ANN) having configuration ANN[17-4-1] shows that carbonation efficiency largely increases as the quantity of calcium content increases. Averagely,there is approximately 5% rise in the original efficiency for 10% increase in the quantity of calcium. This changes to 3.4% and 1.6% increases in efficiency, relative to the original efficiency for 20% and 30% increases in calcium content, respectively. Iron content of minerals offers clear positive correlation to the carbonation efficiency. From the global average, there is approximately 17% rise in the original efficiency for 10% increase in the quantity of iron. This increases to 29% and 41% over the original efficiency for 20% and 30% increases in iron content, respectively.. The influence of magnesium was found to be mainly negatively correlated to carbonation efficiency, after exceeding an unknown threshold. The global average of the efficiency changes with magnesium content results in original efficiency rising by 2% at 10% quantity increase and then reduces by 3% and 9% for 20% and 30% increase in magnesium quantity, respectively, relative to the original efficiency. Thus, iron compounds are found to be most potent of the divalent cations in carbonation reaction while calcium and magnesium content should maintain a threshold ratio with silica content for improved efficiency.

Calcium Regulates Growth and Nutrient Absorption in Poplar Seedlings.

As a crucial element for plants, calcium (Ca) is involved in photosynthesis and nutrient absorption, and affects the growth of plants. Poplar is an important economic forest and shelter forest species in China. However, the optimum calcium concentration for its growth is still unclear. Herein, we investigated the growth, biomass, photosynthetic pigments, photosynthetic parameters and products, chlorophyll fluorescence parameters, water use efficiency (iWUE), and antioxidant enzyme activity of “Liao Hu NO.1” poplar (P. simonii × P. euphratica) seedlings at 0, 2.5, 5, 10, and 20 mmol·L−1 concentrations of Ca2+, and further studied the absorption, distribution, and utilization of nutrient elements (C, N, P, K, and Ca) in plants. We found that with increasing calcium gradient, plant height and diameter; root, stem, leaf, and total biomasses; net photosynthetic rate (Pn); stomatal conductance (Gs); intercellular carbon dioxide (Ci) level; transpiration rate (Tr); Fv/Fm ratio; Fv/F0 ratio; chlorophyll-a; chlorophyll-b; soluble sugar and starch content; superoxide dismutase (SOD), catalase (CAT), and peroxidase (POD) levels; and long-term water use efficiency (iWUE) of poplar seedlings first increased and then decreased. These parameters attained maximum values when the calcium concentration was 5 mmol·L−1, which was significantly different from the other treatments (P < 0.05). Moreover, a suitable Ca2+ level promoted the absorption of C, N, P, K, and Ca by various organs of poplar seedlings. The absorption of C, N, P, and K increased first and then decreased with the increased calcium concentration, but the optimum calcium concentrations for the absorption of different elements by different organs were different, and the calcium concentration in leaves, stems, and roots increased gradually. Furthermore, the increase in exogenous calcium content led to a decreasing trend in the C/N ratio in different organs of poplar seedlings. C/P and N/P ratios showed different results in different parts, and only the N/P ratio in leaves showed a significant positive correlation with Ca2+ concentration. In conclusion, the results of this study indicate that 5 mmol·L−1 concentration of Ca2+ is the optimal level, as it increased growth by enhancing photosynthesis, stress resistance, and nutrient absorption.

Nutrient composition, starch microstructure and thermal properties, and in vitro availability of selected minerals of nixtamalized Philippine quality protein maize variety IPB Var 6 and the production of healthy loaf bread using nixtamalized corn–wheat flour blends

Corn is one of the most important food crops globally providing at least 30% of the food calories in developing countries. This study aimed to determine the nutritional content and starch properties of nixtamalized Philippine quality protein maize (QPM) variety and its potential as a flour substitute in loaf bread production. The nixtamalized Philippine QPM flour had a significant increase in moisture and calcium contents. In vitro availability of calcium, iron, and zinc was relatively lower and associated with phytate content. The developed loaf bread utilizing the optimum nixtamalized corn–wheat flour blend had improved nutritional value and had no significant difference in sensory characteristics compared with the control loaf bread made from wheat flour. The use of nixtamalized IPB Var 6 flour as an ingredient in loaf bread making improved its crude fiber, calcium, and iron levels and can be a nutritious potential substitute ingredient for baked products. Novelty impact statement The process of nixtamalization can improve the nutritional content of corn and has the potential to improve the quality of a food staple such as loaf bread. Incorporating nixtamalized QPM flour into loaf bread resulted in increased fiber, protein, and calcium contents, and it was also acceptable to the consumers in terms of sensory characteristics and general acceptability. Data on the nutritional content and availability of selected minerals of nixtamalized QPM flour can be used as basis for future researches to optimize the method to maximize its potential effects in the improvement of the nutritional value of QPM, thereby providing a healthier alternative staple for the consumers.

Oxidative stress impairs the meat quality of broiler by damaging mitochondrial function, affecting calcium metabolism and leading to ferroptosis.

ObjectiveThis work was conducted to investigate the effects of oxidative stress on meat quality, mitochondrial function, calcium metabolism and ferroptosis of broilers.MethodsIn this study, a total of 144 one-day-old male Ross 308 chicks were divided into 3 groups (control group, saline group, and hydrogen peroxide [H2O2] group) with 6 replicates of 8 broilers each. The study lasted for 42 d. The broilers in the saline and H2O2 groups were intraperitoneally injected with 0.75% saline and 10.0% H2O2 on the 16th and 37th day of the experimental period respectively, the injection volumes were 1.0 mL/kg of broiler body weight. On the 42nd day of the experimental period, two chicks were randomly selected from each cage, a total of thirty-six chicks were stunned by electric shock and slaughtered to collect breast muscle samples.ResultsThe H2O2 exposure reduced pH value, increased drip loss and shear force of breast meat (p<0.05), impaired the ultrastructure and function of mitochondria. The H2O2 exposure damaged the antioxidant system in mitochondria, excessive reactive oxygen species carbonylation modified calcium channels on mitochondria, which impaired the activities of key enzymes on calcium channel, resulted in the increased calcium concentration in cytoplasm and mitochondria (p<0.05). In addition, the H2O2 exposure increased the iron content and lipid peroxidation (p<0.05), which induced ferroptosis.ConclusionOxidative stress could impair meat quality by causing mitochondrial dysfunction, resulting in calcium metabolism disorder and ferroptosis.

The Sodium-Calcium Exchanger Controls the Membrane Potential of AFT024: A Mesenchymal Stem Cell Hematopoietic Niche Forming Line.

The aim of this study was to characterize the functional expression of sodium-calcium exchangers on AFT024 cell line, a murine model of mesenchymal stem/stromal cells (MSCs) supporting human primitive hematopoiesis. All current-clamp and voltage-clamp experiments were performed using the perforated patch whole-cell recording technique with amphotericin B. The membrane potential of -14 mV shifted to -35 mV when lowering the external sodium concentration to 0.33 mM and an increase of cytosolic calcium concentration was observed. KB-R7943, a selective blocker of cardiac sodium-calcium exchangers, also named NCX1, induced a hyperpolarization at physiological sodium concentration while it blocked the hyperpolarization observed at low sodium concentration. This demonstrates for the first time the presence of the sodium-calcium exchangers in AFT024 cells and provides initial evidence that the membrane potential of these stromal cells is maintained depolarized by this exchanger. Lowering external sodium concentration and KB-R7943 had no effect on the membrane potential of 2018 cells, a nonhematopoietic-supportive cell line. Since NCX1 is differentially expressed in AFT024 cells as compared with nonhematopoietic supportive cells with more restricted differentiation potential, this study suggests a potential role of this sodium-calcium exchanger, in the differentiation process or hematopoietic support of MSCs.

Modulation of Purinergic Signaling in Keratinocytes in Spared Nerve Injury Model of Neuropathic Pain

Epidermal keratinocytes express various purinergic 2 receptors that play an essential role in cell growth, differentiation, and proliferation. In the conditions of injury, concentrations of extracellular adenosine triphosphate (ATP) may dramatically increase due to cell damage and inflammatory processes. In this situation activation of purinergic signaling in keratinocytes could act as a double-edged sword contributing to skin regeneration or cell apoptosis. As the role of keratinocytes in transducing and modulating nociceptive stimuli has been increasingly appreciated in recent years, the aim of the present study was to evaluate whether peripheral nerve injury affects purinergic signaling in keratinocytes. Spared nerve injury (SNI), a classical model of peripheral neuropathic pain, was induced in mice. The injury was induced by sparing of the tibial nerve, and ligation and cut of the sural and common peroneal nerves. Keratinocytes were isolated and cultured on Days 2-4 post-injury and ATP-mediated calcium responses in keratinocytes were examined by confocal imaging. On average, the number of keratinocytes that responded to ATP with an increase in intracellular calcium gradient as well as the magnitude of the peak response was not significantly different between sham and SNI groups. However, significantly less delay in ATP-induced increase in intracellular calcium concentration was observed in keratinocytes in SNI group compared to sham. Selective pharmacological inhibition of keratinocyte response to ATP indicated a major role of P2 × 4 receptors in the modulation of calcium homeostasis in SNI. Our results indicate that epidermal purinergic signaling undergoes dramatic changes following peripheral nerve injury that may contribute to injury-induced mechanical hypersensitivity.

APPLICATION OF MINERAL PLUS FERTILIZER TO INCREASE TOMATO PRODUCTION AND QUALITY IN BATURITI DISTRICT, TABANAN REGENCY, BALI PROVINCE, INDONESIA

Environmentally friendly agriculture and food health are needed to support tourism in Bali. Meanwhile, it is alleged that agricultural products do not meet the requirements, especially the quality that does not meet consumer requirements. This research refers to the Udayana University Research Master Plan, a leading field of food security. Food Security Research Roadmap, with the aim of producing appropriate technology. The research products can be implemented in activities to increase production, quality of horticultural crops and food health to support tourism. The research method was through testing the productivity and quality of tomato plants through the addition of organic fertilizers, mineral fertilizers, chemical fertilizers, and their combination. The highest production was obtained from the treatment of P6 = mineral plus fertilizer (5 tons of hill lime + 5 tons of organic fertilizer products organic fertilizer product simantri + 100 kg urea + 100 kg phonska) per ha, can increase production (34.46%). Treatment of mineral fertilization (P2) =mineral fertilizer 5 tons of hill limestone, contains the highest vitamin C (286.14 mg/kg), P2 is also able to neutralize heavy metals Cu and Mo, increase calcium content, increase shelf life, thicken tomato skin walls which Calcium sticks to tomato skin.

Investigations on alkali-silica reaction products using Raman spectroscopy

The alkali-silica reaction (ASR) remains a major challenge regarding the durability of concrete structures. The reaction mechanism is not sufficiently understood owing to the difficulty in characterizing the structure of the alkali-silica reaction gel (ASR gel) in concrete. Synthetic ASR gels with different compositions, i.e. Na/Si and Ca/Si molar ratios, were synthesized and analysed by Raman spectroscopy and, for comparison, 29Si NMR spectroscopy. The results show that higher Na/Si ratios increase the number of non-bridging oxygens in the gel structure, thus leading to a decrease in the degree of cross-linking. With increasing calcium content of the sodium-calcium silica gel, the structure tends towards that of C-S-H phases. Raman spectroscopy is a promising method to characterise synthetic ASR gels and provide new information on the effect of alkalis on the gel structure.

Trade-offs in the genetic control of functional and nutritional quality traits in UK winter wheat.

A complex network of trade-offs exists between wheat quality and nutritional traits. We investigated the correlated relationships among several milling and baking traits as well as mineral density in refined white and whole grain flour. Our aim was to determine their pleiotropic genetic control in a multi-parent population over two trial years with direct application to practical breeding. Co-location of major quantitative trait loci (QTL) and principal component based multi-trait QTL mapping increased the power to detect QTL and revealed pleiotropic effects explaining many complementary and antagonistic trait relationships. High molecular weight glutenin subunit genes explained much of the heritable variation in important dough rheology traits, although additional QTL were detected. Several QTL, including one linked to the TaGW2 gene, controlled grain size and increased flour extraction rate. The semi-dwarf Rht-D1b allele had a positive effect on Hagberg falling number, but reduced grain size, specific weight, grain protein content and flour water absorption. Mineral nutrient concentrations were lower in Rht-D1b lines for many elements, in wholemeal and white flour, but potassium concentration was higher in Rht-D1b lines. The presence of awns increased calcium content without decreasing extraction rate, despite the negative correlation between these traits. QTL were also found that affect the relative concentrations of key mineral nutrients compared to phosphorus which may help increase bioavailability without associated anti-nutritional effects of phytic acid. Taken together these results demonstrate the potential for marker-based selection to optimise trait trade-offs and enhance wheat nutritional value by considering pleiotropic genetic effects across multiple traits.

Melatonin-Induced Postconditioning Suppresses NMDA Receptor through Opening of the Mitochondrial Permeability Transition Pore via Melatonin Receptor in Mouse Neurons.

Mitochondrial membrane potential regulation through the mitochondrial permeability transition pore (mPTP) is reportedly involved in the ischemic postconditioning (PostC) phenomenon. Melatonin is an endogenous hormone that regulates circadian rhythms. Its neuroprotective effects via mitochondrial melatonin receptors (MTs) have recently attracted attention. However, details of the neuroprotective mechanisms associated with PostC have not been clarified. Using hippocampal CA1 pyramidal cells from C57BL mice, we studied the involvement of MTs and the mPTP in melatonin-induced PostC mechanisms similar to those of ischemic PostC. We measured changes in spontaneous excitatory postsynaptic currents (sEPSCs), intracellular calcium concentration, mitochondrial membrane potential, and N-methyl-D-aspartate receptor (NMDAR) currents after ischemic challenge, using the whole-cell patch-clamp technique. Melatonin significantly suppressed increases in sEPSCs and intracellular calcium concentrations. The NMDAR currents were significantly suppressed by melatonin and the MT agonist, ramelteon. However, this suppressive effect was abolished by the mPTP inhibitor, cyclosporine A, and the MT antagonist, luzindole. Furthermore, both melatonin and ramelteon potentiated depolarization of mitochondrial membrane potentials, and luzindole suppressed depolarization of mitochondrial membrane potentials. This study suggests that melatonin-induced PostC via MTs suppressed the NMDAR that was induced by partial depolarization of mitochondrial membrane potential by opening the mPTP, reducing excessive release of glutamate and inducing neuroprotection against ischemia-reperfusion injury.

Role of endoplasmic reticulum stress in apoptosis induced by HK2 inhibitor and its potential as a new drug combination strategy.

Compared with normal cells, tumor cells mainly obtain energy through aerobic glycolysis. Hexokinase 2 (HK2) plays a key role in the regulation of tumor cell aerobic glycolysis, and targeting HK2 has become a new strategy for cancer treatment. However, little is known about the role of HK2 in colon cancer and the regulation of its targeted inhibitors. In this study, we found that the expression of HK2 in colorectal cancer tissues was significantly higher than that in adjacent tissues, and the expression level of HK2 in metastatic colorectal cancer was further increased. Meanwhile, the expression level of HK2 was closely related to clinical TNM stage and outcome of colorectal cancer patients. We provide here evidence that HK2 inhibitor 3-Bromopyruvate acid (3-BP) can significantly inhibit the survival and proliferation of colon cancer cells, and induce apoptosis through mitochondrial apoptosis signaling pathway. In addition, we found that 3-BP can also induce endoplasmic reticulum stress in colon cancer cells, the mechanism may be through the increase of intracellular calcium concentration. In vitro and in vivo experiments showed that inhibition of endoplasmic reticulum stress could further increase the proliferation inhibition and apoptosis induced by 3-BP. Collectively, our results show that HK2 is highly expressed in colorectal cancer. 3-BP, an inhibitor of HK2, can induce apoptosis and endoplasmic reticulum stress in colon cancer cells. Endoplasmic reticulum stress plays a protective role in cell death induced by 3-BP. This result suggested that targeting HK2 and endoplasmic reticulum stress may be a valuable strategy in targeted and combination therapy of colon cancer.

دور الأكسيناتيد-ناهض مستقبل الببتيد الشبيه بالغلوكاغون1- في العلاج والوقاية من هشاشة العظام المستحدثة باستئصال المبايض عند إناث جرذان ويستر

هدفت هذه الدراسة إلى تقييم التأثير الوقائي والعلاجي للأكسيناتيد على هشاشة العظام المستحدثة باستئصال المبايض عند إناث جرذان ويستر. بينت نتائج هذه الدراسة أن إعطاء الأكسيناتيد بجرعة 10 ميكروغرام/كغ يومياً لمدة 4 أشهر أدت إلى الحد من أعراض هشاشة العظام عند الجرذان مستأصلة المبايض وتجلى ذلك من خلال التغيرات في المستويات المصلية لواسمات الاستقلاب العظمي (زيادة معنوية في التركيز المصلي لكل من الكالسيوم والفوسفور، مصحوباً بانخفاض معنوي في فعالية إنزيم الفوسفاتاز القاعدي)، وتحسن ملحوظ في البنية النسيجية لعظم الفخذ، بالمقارنة مع أفراد المجموعة مستأصلة المبايض. نستخلص من هذه الدراسة فعالية الأكسيناتيد في الحد من التأثيرات الضارة لنقص هرمون المبيض على كل من بنية ووظيفة العظم عند إناث جرذان ويستر.

Anti-phospholipase A2 receptor antibodies directly induced podocyte damage in vitro

Background The pathogenesis of primary membranous nephropathy (MN) involves the antibodies against antigens on the cell surface of podocytes, with the majority of M-type phospholipase A2 receptor (PLA2R), and a profound podocyte dysfunction. The effects of anti-PLA2R antibodies directly to the podocytes remain unclear. Methods Anti-PLA2R antibodies from patients with PLA2R-associated MN were affinity-purified using a column coupled with recombinant human PLA2R protein. Their effects on conditionally immortalized human podocytes were assessed by apoptosis assays, cellular calcium detection, wound healing assay, and immunofluorescent staining. Proteomics analysis was performed by LC-MS/MS and on PANTHER database. Results The stimulation by anti-PLA2R antibodies could induce early-stage apoptosis of podocytes (MFI of Annexin V = 104.3 ± 19.2 vs. 36.7 ± 7.6, p = 0.004). The increase of calcium concentration in podocytes (MFI = 3309.3 ± 363.6 vs. 1776.3 ± 212.7, p = 0.015) might attribute to the endoplasmic reticulum calcium efflux. The expression of calcium/calmodulin-dependent protein kinase IV (CaMK4) was also increased (MFI = 134.4 ± 9.8 vs. 105.3 ± 10.1, p = 0.011). Proteomics results suggested that anti-PLA2R antibody treatment led to damage on cellular structure, and produced functional disorders on protein binding, actin filament binding, and microtubule motor activity. The staining of F-actin on foot process was reduced (MFI = 27.3 ± 2.8 vs. 47.5 ± 1.0, p = 0.001) and the motility and adherence capacity of podocytes were reduced (number of migrated cells = 44.7 ± 3.1 vs. 53.3 ± 4.9, p = 0.001) after incubation with anti-PLA2R antibodies. Conclusion These data indicate that anti-PLA2R antibodies may directly induce podocyte damage independent of the complement system, which expands the mechanism of anti-PLA2R antibodies on MN.

Melatonin Mediates Osteoblast Proliferation Through the STIM1/ORAI1 Pathway.

Based on the positive correlation between bone mineral density and melatonin levels in blood, this study confirmed that melatonin supplementation prevents postmenopausal osteoporosis. We further confirmed that melatonin promotes an increase in intracellular calcium concentrations through the STIM1/ORAI1 pathway, thereby inducing the proliferation of osteoblasts. Introduction: Osteoporosis (OP) is a progressive, systemic bone disease that is one of the main causes of disability and death in elderly female patients. As an amine hormone produced by the human pineal gland, melatonin plays an important role in regulating bone metabolism. This study intends to investigate the relationship between melatonin levels in human blood and bone density and to suggest the efficacy of melatonin in treating osteoporosis by performing in vivo and in vitro experiments. Methods: We used liquid chromatography-tandem mass spectrometry to determine the serum melatonin levels in postmenopausal women with osteoporosis and young women with a normal bone mass. The bone density, BV/TV, Tb.Th, Tb.Sp and other indicators of postmenopausal osteoporosis and mice with a normal bone mass were detected by measuring bone density and micro-CT. The intracellular calcium ion concentration was detected using fluorescence microscopy and a full-wavelength multifunctional microplate reader, and the expression of SOCE-related genes and STIM1/ORAI1 proteins was detected using PCR and WB. Results: This study confirmed that bone density positively correlates with the melatonin level in human blood. In the animal model, melatonin supplementation reverses postmenopausal osteoporosis. We explored the internal mechanism of melatonin treatment of osteoporosis. Melatonin promotes an increase in intracellular calcium ion concentrations through the STIM1/ORAI1 pathway to induce osteoblast proliferation. Conclusions: This study provides an important theoretical basis for the clinical application of melatonin in patients with osteoporosis and helps to optimize the diagnosis and treatment of postmenopausal osteoporosis.

Discovery of calcium-binding peptides derived from defatted lemon basil seeds with enhanced calcium uptake in human intestinal epithelial cells, Caco-2

It is anticipated that calcium-chelating peptides may serve to enhance the absorption of calcium. This research examined defatted lemon basil seeds (DLBS) which had been treated with Alcalase under optimized parameters for the degree of hydrolysis for proteolysis, discovering that the activity for calcium-binding in a competitive condition with phosphate ion was 60.39 ± 1.545%. The purification of the hydrolysates was performed via ultrafiltration along with reversed-phase high performance liquid chromatography (RP-HPLC). Determination of the purified peptide amino acid sequence was confirmed for both peptides and reported as Ala-Phe-Asn-Arg-Ala-Lys-Ser-Lys-Ala-Leu-Asn-Glu-Asn (AFNRAKSKALNEN; Basil-1), and Tyr-Asp-Ser-Ser-Gly-Gly-Pro-Thr-Pro-Trp-Leu-Ser-Pro-Tyr (YDSSGGPTPWLSPY; Basil-2). The respective activities for calcium-binding were 38.62 ± 1.33%, and 42.19 ± 2.27%. Fluorescence spectroscopy, and fourier transform infrared spectroscopy were employed in order to assess the chelating mechanism between calcium and the peptides. It was found that the calcium ions took place through the activity of the amino nitrogen atoms and the oxygen atoms on the carboxyl group. Moreover, both of these peptides served to improve calcium transport and absorption in Caco-2 cell monolayers, depending on the concentration involved. It was revealed that the peptide-calcium complexes offered an increased calcium absorption percentage when compared to free calcium at similar concentrations. It might be concluded that the peptide within the peptide-calcium complex can promote calcium absorption through both active and passive transport pathways by increasing calcium concentration and promoting cell membrane interaction. Accordingly, DLBS protein can be considered a strong potential source of protein which can be used to produce calcium-binding peptides and might therefore play a role in the production of nutraceutical foods as a bioactive ingredient.

Food to food fortification of whole wheat flour biscuit using moringa leaves powder

The purpose of the research was to use food to food fortification of staples, as a tool to deliver micronutrients especially minerals to the population. In this research moringa leaves powder (MLP), a local nutrient dense source, was utilized for increasing the micronutrients in whole wheat flour biscuit. Microstructural and mineral analysis of whole wheat flour (WWF) and MLP was carried out. WWF biscuit were prepared by incorporating MLP in different percentages (0, 2, 4, 6, 8 and 10%). Mineral profile, phytic acid content and microstructure of the biscuit were analyzed. The microstructural analysis showed that the gluten network was infered by inculsion of MLP, creating some cavities, but the entire protein matrix was not disrupted. The results of the mineral analysis indicated a significant increase in magnesium (42.34%), calcium(84.91%), potassium(49.82%), phosphorus (32.32%) and iron content(50.92%) of the cookies fortified with MLP. Phytic acid of the biscuit decreased from 0.66 to 0.39%. It can be concluded that MLP can be a tool to alleviate malnutrition and solve food insecurity especially in the developing counteries.