Physicochemical and Active Properties of Gelatine-Based Composite Gels Loaded with Lysozyme and Green Tea Polyphenols.

RESEARCH BACKGROUND: The use of gel-based systems as a novel method for the delivery of natural antimicrobial, antioxidant and bioactive compounds is a developing innovative solution for the food industry. This research aims to develop multifunctional active edible gels based on gelatine and its composites with improved mechanical properties. EXPERIMENTAL APPROACH: Antilisterial and bioactive composite gels showing different physical and active properties from classical gelatine gel were developed by loading lysozyme and green tea extract into gelatine/starch and gelatine/wax composite gels. Mechanical properties, swelling profiles, colour, release profiles, and antimicrobial and bioactive properties of the gels were characterised. RESULTS AND CONCLUSIONS: Gelatine/wax gels showed 1.3- to 2.1-fold higher firmness and cutting strength than gelatine and gelatine/starch composite gels that had similar firmness and cutting strengths. Work to shear of both composite gels was 1.4- to 1.9-fold higher than that of gelatine gel. The gelatine/starch gel showed the highest water absorption capacity. Green tea extract reduced soluble lysozyme in all gels, but composite gels contained higher amount of soluble lysozyme than gelatine gel. All the gels with lysozyme inhibited Listeria innocua growth in the broth media, while green tea extract showed antilisterial activity only in gelatine/wax gels. Gels with green tea extract showed antioxidant, antidiabetic (α-glucosidase and α-amylase inhibition), antihypertensive (angiotensin-converting enzyme inhibition) and antiproliferative activities (on Caco-2 human colon carcinoma cells). However, gelatine and gelatine/wax gels showed the highest antioxidant and antidiabetic activity. The gelatine/wax gels prevented phenolic browning, while green tea extract in other gels showed moderate or extensive browning. NOVELTY AND SCIENTIFIC CONTRIBUTION: This work clearly showed the possibility of improving mechanical properties and modifying water absorption and controlled release profiles of gelatine gels using gelatine/starch and gelatine/wax composites. The novel composite gels reduced browning of incorporated polyphenols and showed antilisterial and bioactive properties.

Molecular mediators governing iron-copper interactions.

Given their similar physiochemical properties, it is a logical postulate that iron and copper metabolism are intertwined. Indeed, iron-copper interactions were first documented over a century ago, but the homeostatic effects of one on the other has not been elucidated at a molecular level to date. Recent experimental work has, however, begun to provide mechanistic insight into how copper influences iron metabolism. During iron deficiency, elevated copper levels are observed in the intestinal mucosa, liver, and blood. Copper accumulation and/or redistribution within enterocytes may influence iron transport, and high hepatic copper may enhance biosynthesis of a circulating ferroxidase, which potentiates iron release from stores. Moreover, emerging evidence has documented direct effects of copper on the expression and activity of the iron-regulatory hormone hepcidin. This review summarizes current experimental work in this field, with a focus on molecular aspects of iron-copper interplay and how these interactions relate to various disease states.

Mechanistic and regulatory aspects of intestinal iron absorption.

Iron is an essential trace mineral that plays a number of important physiological roles in humans, including oxygen transport, energy metabolism, and neurotransmitter synthesis. Iron absorption by the proximal small bowel is a critical checkpoint in the maintenance of whole-body iron levels since, unlike most other essential nutrients, no regulated excretory systems exist for iron in humans. Maintaining proper iron levels is critical to avoid the adverse physiological consequences of either low or high tissue iron concentrations, as commonly occurs in iron-deficiency anemia and hereditary hemochromatosis, respectively. Exquisite regulatory mechanisms have thus evolved to modulate how much iron is acquired from the diet. Systemic sensing of iron levels is accomplished by a network of molecules that regulate transcription of the HAMP gene in hepatocytes, thus modulating levels of the serum-borne, iron-regulatory hormone hepcidin. Hepcidin decreases intestinal iron absorption by binding to the iron exporter ferroportin 1 on the basolateral surface of duodenal enterocytes, causing its internalization and degradation. Mucosal regulation of iron transport also occurs during low-iron states, via transcriptional (by hypoxia-inducible factor 2α) and posttranscriptional (by the iron-sensing iron-regulatory protein/iron-responsive element system) mechanisms. Recent studies demonstrated that these regulatory loops function in tandem to control expression or activity of key modulators of iron homeostasis. In health, body iron levels are maintained at appropriate levels; however, in several inherited disorders and in other pathophysiological states, iron sensing is perturbed and intestinal iron absorption is dysregulated. The iron-related phenotypes of these diseases exemplify the necessity of precisely regulating iron absorption to meet body demands.

Silencing the Menkes copper-transporting ATPase (Atp7a) gene in rat intestinal epithelial (IEC-6) cells increases iron flux via transcriptional induction of ferroportin 1 (Fpn1).

The Menkes copper-transporting ATPase (Atp7a) gene is induced in rat duodenum during iron deficiency, consistent with copper accumulation in the intestinal mucosa and liver. To test the hypothesis that ATP7A influences intestinal iron metabolism, the Atp7a gene was silenced in rat intestinal epithelial (IEC-6) cells using short hairpin RNA (shRNA) technology. Perturbations in intracellular copper homeostasis were noted in knockdown cells, consistent with the dual roles of ATP7A in pumping copper into the trans-Golgi (for cuproenzyme synthesis) and exporting copper from cells. Intracellular iron concentrations were unaffected by Atp7a knockdown. Unexpectedly, however, vectorial iron ((59)Fe) transport increased (∼33%) in knockdown cells grown in bicameral inserts and increased further (∼70%) by iron deprivation (compared with negative control shRNA-transfected cells). Additional experiments were designed to elucidate the molecular mechanism of increased transepithelial iron flux. Enhanced iron uptake by knockdown cells was associated with increased expression of a ferrireductase (duodenal cytochrome b) and activity of a cell-surface ferrireductase. Increased iron efflux from knockdown cells was likely mediated via transcriptional activation of the ferroportin 1 gene (by an unknown mechanism). Moreover, Atp7a knockdown significantly attenuated expression of an iron oxidase [hephaestin (HEPH); by ∼80%] and membrane ferroxidase activity (by ∼50%). Cytosolic ferroxidase activity, however, was retained in knockdown cells (75% of control cells), perhaps compensating for diminished HEPH activity. This investigation has thus documented alterations in iron homeostasis associated with Atp7a knockdown in enterocyte-like cells. Alterations in copper transport, trafficking, or distribution may underlie the increase in transepithelial iron flux noted when ATP7A activity is diminished.

Golden thistle (Scolymus hispanicus L.) hydromethanolic extracts ameliorated glucose absorption and inflammatory markers in vitro.

Golden thistle (GT, Scolymus hispanicus L.) is an edible plant native to the Mediterranean. Several activities have been reported for the GT, as it is used for traditional medicinal purposes in some cultures. In this study, we aimed to investigate the effects of GT crude extract on phenolic bioavailability, antidiabetic, and anti-inflammatory activities by using colonic epithelium (CaCo-2) and murine macrophage (RAW 264.7) cell lines. The CaCo-2 cells were grown on the bicameral membrane system for intestinal bioavailability and glucose efflux. Lipopolysaccharide (LPS, 0.5 µg/mL) was used to induce systemic inflammation on RAW 264.7. The inflammatory medium of RAW 264.7 cells was given to Caco-2 cells to mimic colonic inflammation. Our results showed that 5-o-caffeoylquinic acid had an apparent permeability of (1.82 ± 0.07) × 10-6 cm/s after 6 h. The extract lowered the glucose efflux by 39.4%-42.6%, in addition to the reductions in relative GLUT2 mRNA expressions by 49%-66% in pre- and co-treatments (p < .05). Decreases in systemic inflammation markers of nitric oxide, tumor necrosis factor-alpha, and interleukin-6 (IL-6) were also detected in 30%-45% range after pre-treatments with the GT extract (p < .05). Lastly, colonic inflammation markers of IL-6 and IL-8 were reduced by 8.7%-19.5% as a result of GT pre-treatments (p < .05). Thus, an in vitro investigation of GT extract revealed promising results on antidiabetic and anti-inflammatory activities.

Formulation of Gluten-Free Cookies Utilizing Chickpea, Carob, and Hazelnut Flours through Mixture Design.

Legume flours, which offer high nutritional quality, present viable options for gluten-free bakery products. However, they may have an objectionable flavor and taste for some consumers. In this study, it was aimed to improve the gluten-free cookie formulation by incorporating carob and hazelnut flours to pre-cooked chickpea flour and to investigate the techno-functional properties of the formulated cookies. The flours used in the formulations were assessed for their chemical and physical properties. This study employed a mixture design (simplex-centroid) to obtain the proportions of the flours to be used in the cookie formulations. The rheological characteristics of the doughs and the technological attributes of the baked cookies were determined. The addition of the hazelnut and carob flours had the overall effect of reducing the rheological characteristics of the cookie doughs. Furthermore, the textural attribute of the hardness of the baked cookies decreased as the ratio of hazelnut flour in the formulations was raised. The analysed results and sensory evaluation pointed to a formulation consisting of 30% pre-cooked chickpea/30% carob/30% hazelnut flours, which exhibited improved taste and overall acceptability scores. A total of 16.82 g/100 g of rapidly digestible starch, 5.36 g/100 g of slowly digestible starch, and 8.30 g/100 g of resistant starch exist in this particular cookie. As a result, combinations of chickpea, hazelnut, and carob flours hold promise as good alternatives for gluten-free cookie ingredients and warrant further exploration in the development of similar products.

Techno-Functional and In Vitro Digestibility Properties of Gluten-Free Cookies Made from Raw, Pre-Cooked, and Germinated Chickpea Flours.

Chickpea flour, which is produced in various forms, has high protein and fiber content; therefore, it can be a good ingredient for gluten-free cookies. The objective of this study was to investigate and compare the properties of cookies formulated using raw (RCF), cooked (CCF), and germinated (GCF) chickpea flours. The techno-functional properties of these flours were determined, and scanning electron microscope images and mid-infrared spectra were obtained. The rheological properties of cookie doughs were measured along with their mid-infrared spectra. Baked cookies were analyzed for their technological properties as well as their in vitro digestion properties. Sensory analysis was also performed for all the cookies. The most significant difference among the flours was observed in their water retention capacity, and CCF had 119.7% higher water retention capacity compared to RCF. The dough made with CCF had quite different rheological properties from the others. The cookies baked with GCF had the highest baking loss and spread ratio. The CCF-containing cookies had the hardest structure. The cookies made from RCF had a higher resistant starch content followed by the cookies with GCF. All the cookies had similar scores in all aspects tested in the sensory analysis. The use of three different forms of chickpea flour in cookie formulations resulted in products with very different properties; however, their overall acceptability levels were close.

Natural and synthetic nanovectors for cancer therapy.

Nanomaterials have been extensively studied in cancer therapy as vectors that may improve drug delivery. Such vectors not only bring numerous advantages such as stability, biocompatibility, and cellular uptake but have also been shown to overcome some cancer-related resistances. Nanocarrier can deliver the drug more precisely to the specific organ while improving its pharmacokinetics, thereby avoiding secondary adverse effects on the not target tissue. Between these nanovectors, diverse material types can be discerned, such as liposomes, dendrimers, carbon nanostructures, nanoparticles, nanowires, etc., each of which offers different opportunities for cancer therapy. In this review, a broad spectrum of nanovectors is analyzed for application in multimodal cancer therapy and diagnostics in terms of mode of action and pharmacokinetics. Advantages and inconveniences of promising nanovectors, including gold nanostructures, SPIONs, semiconducting quantum dots, various nanostructures, phospholipid-based liposomes, dendrimers, polymeric micelles, extracellular and exome vesicles are summarized. The article is concluded with a future outlook on this promising field.

Sucrose-induced hyperglycemia dysregulates intestinal zinc metabolism and integrity: risk factors for chronic diseases.

Objective: Zinc is an essential micronutrient that is critical for many physiological processes, including glucose metabolism, regulation of inflammation, and intestinal barrier function. Further, zinc dysregulation is associated with an increased risk of chronic inflammatory diseases such as type II diabetes, obesity, and inflammatory bowel disease. However, whether altered zinc status is a symptom or cause of disease onset remains unclear. Common symptoms of these three chronic diseases include the onset of increased intestinal permeability and zinc dyshomeostasis. The specific focus of this work is to investigate how dietary sources of intestinal permeability, such as high sucrose consumption, impact transporter-mediated zinc homeostasis and subsequent zinc-dependent physiology contributing to disease development. Method: We used in vivo subchronic sucrose treatment, ex vivo intestinal organoid culture, and in vitro cell systems. We analyze the alterations in zinc metabolism and intestinal permeability and metabolic outcomes. Results: We found that subchronic sucrose treatment resulted in systemic changes in steady-state zinc distribution and increased 65Zn transport (blood-to-intestine) along with greater ZIP14 expression at the basolateral membrane of the intestine. Further, sucrose treatment enhanced cell survival of intestinal epithelial cells, activation of the EGFR-AKT-STAT3 pathway, and intestinal permeability. Conclusion: Our work suggests that subchronic high sucrose consumption alters systemic and intestinal zinc homeostasis linking diet-induced changes in zinc homeostasis to the intestinal permeability and onset of precursors for chronic disease.

The Dual Diverse Dynamic Reversible Effects of Ankaferd Blood Stopper on EPCR and PAI-1 Inside Vascular Endothelial Cells With and Without LPS Challenge.

OBJECTIVE: Ankaferd blood stopper (ABS) is comprised of a mixture of the plants Thymus vulgaris, Glycyrrhiza glabra, Vitis vinifera, Alpinia officinarum, and Urtica dioica. ABS is used as a topical hemostatic agent due to its antihemorrhagic effect, yet its hemostatic mechanism of action remains to be investigated. ABS does not affect the levels of coagulation factors II, V, VII, VIII, IX, X, XI and XII. The aim of this study was to investigate the effects of ABS on endothelium and immune response. As such, we evaluated changes in endothelial cell protein C receptor (EPCR) and plasminogen activator inhibitor type-1 (PAI-1) expression inside human umbilical vein endothelial cells (HUVECs) in the presence and absence of lipopolysaccharides (LPSs). MATERIAL AND METHODS: We exposed HUVECs to 10 µL and 100 µL of ABS for 5 min, 25 min, 50 min, 6 h, and 24 h. Additionally, 10 µg mL-1 of LPS was administered for 1 h to observe the effects of LPS challenge on HUVECs, and then the cells were treated with ABS for 5 min, 25 min, 50 min, and 6 h to observe the effects of ABS on HUVECs. Total RNA was isolated from HUVECs and then the level of expression of EPCR and PAI-1 mRNA was measured. RESULTS: Cells were microscopically observed to arise from the surface and adhere to each other following the administration of ABS to HUVECs. Additionally, after 24 h the cells had normal growth and physiology, which suggests that the adhesive cellular effects of ABS might be reversible. ABS had a negative effect on EPCR and PAI-1 expression; the effect in response to 100 µL was greater than that to 10 µL. EPCR and PAI-1 expression increased over time in response to LPS and 10 µL of ABS. EPCR and PAI-1 expression was very low during the first hour of exposure to LPS and 100 µL of ABS, but after 6 h increased to levels similar to those observed in response to LPS and 10 µL of ABS. CONCLUSION: It was observed that ABS had dual diverse dynamic reversible effects on EPCR and PAI-1 expression in HUVECs, which were dependent on dose and concentration. ABS might play a role in numerous cellular mechanisms, in addition to having hemostatic effects. CONFLICT OF INTEREST: None declared.

Comparison of apical and basolateral Cu treatment for iron-related gene regulation during deferoxamine induced iron deficiency.

BACKGROUND: Intestinal copper transporter (Atp7a) mutant-brindled mice with systemic Cu deficiency had elevated Cu levels in enterocyte cells without any perturbation of iron-regulating genes, suggesting that blood Cu level might be important for intestinal iron homeostasis during iron deficiency (ID). We hypothesized that the blood Cu level and polarization (apical and basolateral) of enterocyte cells might be important regulators for the compensatory response on the regulation of genes in enterocyte cells during iron deficiency. METHODS: We grew Caco-2 cells on a bicameral cell culture plate to mimic the human intestine system and on a regular tissue culture plate. Iron deficiency was induced by deferoxamine (DFO). The cells were treated with Cu and Cu with Fe following mRNA expressions of DMT1, FPN, TFR, and ANKRD37 were analyzed. RESULTS: Our main finding was that basolateral treatment of Cu significantly reduced mRNA expressions of iron-regulated genes, including DMT1, FPN, TFR, and ANKRD37, compared to DFO-treated and DFO with apical Cu-treated groups in both bicameral and regular tissue culture plates. CONCLUSIONS: Cu level in the basolateral side of Caco-2 cells significantly influenced the intracellular gene regulation in DFO-induced iron-deficient condition, and polarization of the cells might be important factor gene regulation in enterocyte cells.

Effects of Golden Thistle (Scolymus hispanicus L.) on Cytotoxic Activity: Cell Cycle Arrest and Apoptotic Properties on the CaCo-2 Cell Line.

Cancer is a global concern for many individuals with high mortality rates, with colon cancer being the third most common diagnosed cancer worldwide. A phytochemical-rich diet is often recommended in the prevention and during the treatment of cancer cases. Golden thistle (GT) plant (Scolymus hispanicus L.) is a wild edible plant widely consumed in the Mediterranean countries. In this study, we aimed to obtain a hydromethanolic extract from three parts of the GT plant and test its antiproliferative activity in the CaCo-2 human adenocarcinoma cell line. Concentrations of the golden thistle extract (GTE) were used to treat CaCo-2 cells and the most significant reduction was detected with 4 mg/mL GTE after 72 h, with 78.3% decrease in cell viability (P < .05). Additionally, 4 mg/mL GTE caused 7.8-fold higher release of lactate dehydrogenase enzyme, indicating cell death after treatment. Flow cytometric analyses concluded both 3.3-fold higher early and late apoptotic activity of the 4 mg/mL GTE compared with the nontreated control group (P < .05). Last, 4 mg/mL GTE showed 24.1% reduction in the G1 phase and 38.1% increase in the S phase of cell cycle distribution. The alteration of G1 and S phases in the cell cycle led to growth reduction of CaCo-2 cells and caused apoptosis.

Iron Absorption: Factors, Limitations, and Improvement Methods.

Iron is an essential element for human life since it participates in many functions in the human body, including oxygen transport, immunity, cell division and differentiation, and energy metabolism. Iron homeostasis is mainly controlled by intestinal absorption because iron does not have active excretory mechanisms for humans. Thus, efficient intestinal iron bioavailability is essential to reduce the risk of iron deficiency anemia. There are two forms of iron, heme and nonheme, found in foods. The average daily dietary iron intake is 10 to 15 mg in humans since only 1 to 2 mg is absorbed through the intestinal system. Nutrient-nutrient interactions may play a role in dietary intestinal iron absorption. Dietary inhibitors such as calcium, phytates, polyphenols and enhancers such as ascorbic acid and proteins mainly influence iron bioavailability. Numerous studies have been carried out for years to enhance iron bioavailability and combat iron deficiency. In addition to traditional methods, innovative techniques are being developed day by day to enhance iron bioavailability. This review will provide information about iron bioavailability, factors affecting absorption, iron deficiency, and recent studies on improving iron bioavailability.

The effects of Ankaferd® Blood Stopper on transcription factors in HUVEC and the erythrocyte protein profile.

OBJECTIVE: Ankaferd® Blood Stopper (ABS) is an herbal extract that has historically been used as a hemostatic agent in traditional Turkish medicine. ABS is comprised of a standardized herbal mixture of T. vulgaris, G. glabra, V. vinifera, A. officinarum, and U. dioica. ABS's basic mechanism of action is the formation of an encapsulated protein web, which represents the focal point for vital erythrocyte masses. The hemostatic effects of ABS have been observed in vitro and in vivo. ABS was registered as a hemostatic agent for external hemorrhages and dental bleeding following phase I randomized, double-blind crossover placebo-controlled clinical research, and safety and efficacy reports. In terms of the potential use of ABS, transcription factors may be novel factors that play a role in the hemostatic and other pleiotropic effects of ABS. METHODS: Hence, the present study aimed to investigate the effects of ABS on endothelium, and possible transcription factor changes in HUVEC (human umbilical vein endothelial cells) and the erythrocyte membrane profile. ABS (5 µL and 50 µL) was administered to HUVEC (in 75 cm2; ~75% fullness) for 5 min and 15 min. RESULTS: ABS caused significant increases in the level of activation of the following transcription factors; AP2, AR, CRE/ATF1, CREB, E2F1-5, E2F6, EGR, GATA, HNF-1, ISRE, Myc-Max, NF-1, NFkB, p53, PPAR, SMAD 2/3, SP1, TRE/AP1, and YY1. Following erythrocyte membrane isolation, protein complexes were undissolved, but denatured. The protein complex formed was resistant to heat and detergent. Trypsin and sonication were used in order to break this complex; the complex dissolved and erythrocyte membrane proteins were released in SDS-PAGE. CONCLUSION: ABS established a very fast and solid protein web, and increased the level of transcription factor activation. Therefore the cellular effects of ABS could be related to different intracellular biological pathways.

The Effect of Ankaferd Blood Stopper on Colonic Inflammation: An In Vitro Study in RAW 264.7 and Caco-2 Cells.

Ankaferd Blood Stopper (ABS) is a medicinal plant extract that has anti-inflammatory effect. Inflammatory bowel disease is a pathological condition that directly affects colon health and increases the risk of colon cancer. Especially inflammation is an important factor in the formation and progression of this disease. The aim of the study was to investigate the protective effect of ABS on colonic inflammation. Caco-2 and RAW 264.7 cells were used as a model of in vitro colonic inflammation. RAW 264.7 cells were treated with lipopolysaccharide for 12 h to induce inflammation, and an inflammatory medium (IM) was obtained. Caco-2 cells were treated with 15 µL/mL ABS for 4 h, then incubated with IM. The cells also were incubated with 15 µL/mL ABS and IM together for 12 h. Tumor necrosis factor alpha (TNF-α) protein levels were targeted in testing inflammatory condition and cyclooxygenase-2 (COX-2) mRNA level was used as a marker gene to show the possible anti-inflammatory effect of ABS in Caco-2 cells. TNF-α level was 26.1-fold higher than the control group. IM caused 3.2-fold increase in COX-2 expression in Caco-2 cells. Pretreatment of Caco-2 cells with ABS resulted in 3.3-fold decrease in COX-2 mRNA levels relative to IM group. Furthermore, COX-2 mRNA level reduced 4.7-fold when ABS and conditional medium were given at the same time. ABS has suppressive effect on COX-2 mRNA expression in Caco-2 cells. These results suggest that ABS might have protective and therapeutic effect for colonic inflammation.

Cross-species comparison of genomewide gene expression profiles reveals induction of hypoxia-inducible factor-responsive genes in iron-deprived intestinal epithelial cells.

Molecular mechanisms mediating the induction of metal ion homeostasis-related genes in the mammalian intestine during iron deficiency remain unknown. To elucidate relevant regulatory pathways, genomewide gene expression profiles were determined in fully differentiated human intestinal epithelial (Caco-2) cells. Cells were deprived of iron (or not) for 6 or 18 h, and Gene Chip analyses were subsequently performed (Affymetrix). More than 2,000 genes were differentially expressed; genes related to monosaccharide metabolism, regulation of gene expression, hypoxia, and cell death were upregulated, while those related to mitotic cell cycle were downregulated. A large proportion of induced genes are hypoxia responsive, and promoter enrichment analyses revealed a statistical overrepresentation of hypoxia response elements (HREs). Immunoblot experiments demonstrated a >60-fold increase in HIF2α protein abundance in iron-deprived cells; HIF1α levels were unchanged. Furthermore, comparison of the Caco-2 cell data set with a Gene Chip data set from iron-deficient rat intestine revealed 29 common upregulated genes; the majority are hypoxia responsive, and their promoters are enriched for HREs. We conclude that the compensatory response of the intestinal epithelium to iron deprivation relates to hypoxia and that stabilization of HIF2α may be the primary event mediating metabolic and morphological changes observed during iron deficiency.

The development of lentil derived protein-iron complexes and their effects on iron deficiency anemia in vitro.

Iron deficiency anemia (IDA) is the most common nutrient-dependent health problem in the world and could be reversed by commercially available iron supplementation. The form of iron supplement is important due to its toxicity on the gastrointestinal system (GI), so the development of new dietary strategies might be important for the prevention of IDA. It has been shown that plant-based proteins bind to iron and might decrease the free form of iron before absorption and increase iron bioavailability. Thus, we aimed to form lentil derived protein-iron complexes and to test the functional properties of hydrolysed protein-iron complexes in anemic Caco-2 cell line. Our main findings were that (i) lentil derived proteins had the capacity to chelate iron minerals and (ii) hydrolysed protein-iron complexes significantly reduced the mRNA levels of iron regulated divalent metal transporter-1 (DMT1), transferrin receptor (TFR), and ankyrin repeat domain 37 (ANKRD37) marker genes that were induced by iron deficiency anemia. The current findings suggest that hydrolysed protein-iron complexes might have functional properties in iron deficiency anemia in vitro. Further in vivo studies are necessary to show lentil derived proteins and iron might be used as supplements or food additives to reduce the risk of iron deficiency anemia.

Iron loading increases ferroportin heterogeneous nuclear RNA and mRNA levels in murine J774 macrophages.

The transmembrane protein ferroportin is highly expressed in tissue macrophages, where it mediates iron export into the bloodstream. Although ferroportin expression can be controlled post-transcriptionally through a 5' iron-responsive element in its mRNA, various studies have documented increased ferroportin mRNA levels in response to iron, suggesting transcriptional regulation. We studied the effect of iron loading on levels of macrophage ferroportin mRNA, as well as heterogeneous nuclear RNA (hnRNA), the immediate product of ferroportin gene transcription. J774 cells, a mouse macrophage cell line, were incubated for 0, 3, 6, 9, 12, and 24 h in medium supplemented or not with 200 mumol/L iron. Quantitative RT-PCR was used to measure steady-state levels of ferroportin mRNA and hnRNA. Ferroportin mRNA levels increased by 12 h after iron treatment, reaching 6 times the control levels after 24 h. Changes in ferroportin mRNA levels were paralleled by similar changes in the levels of ferroportin hnRNA. Time course studies of ferroportin mRNA and hnRNA abundance after incubating cells with the transcriptional inhibitor actinomycin D revealed that ferroportin mRNA has a half-life of approximately 4 h and that iron loading does not stabilize ferroportin mRNA or hnRNA. Collectively, these data are consistent with the hypothesis that iron increases macrophage ferroportin mRNA levels by inducing transcription of the ferroportin gene.

Increased iron content and RNA oxidative damage in skeletal muscle with aging and disuse atrophy.

Muscle atrophy with aging or disuse is associated with deregulated iron homeostasis and increased oxidative stress likely inflicting damage to nucleic acids. Therefore, we investigated RNA and DNA oxidation, and iron homeostasis in gastrocnemius muscles. Disuse atrophy was induced in 6- and 32-month old male Fischer 344/Brown Norway rats by 14 days of hind limb suspension (HS). We show that RNA, but not DNA, oxidative damage increased 85% with age and 36% with HS in aged muscle. Additionally, non-heme iron levels increased 233% with aging and 83% with HS at old age, while staining for free iron was strongest in the smallest fibers. Simultaneously, the mRNA abundance of transferrin receptor-1 decreased by 80% with age and 48% with HS for young animals, while that of the hepcidin regulator hemojuvelin decreased 37% with age, but increased about 44% with disuse, indicating a dysregulation of iron homeostasis favoring increased intracellular free iron in atrophied muscles. RNA and DNA concentrations increased with age and were negatively correlated with muscle mass, whereas protein concentrations decreased with aging, indicating a preferential loss of protein compared to nucleic acids. Furthermore, xanthine oxidase activity increased with age, but not with HS, while mRNA abundance of the Y box-binding protein-1, which has been suggested to bind oxidized RNA, did not change with age or HS. These results suggest that RNA oxidation, possibly mediated by increased non-heme iron, might contribute to muscle atrophy due to disuse particularly in aged muscle.

MEF2A sequence variants in Turkish population.

Myocyte enhancer factor 2 (MEF2) is present in skeletal, cardiac, and smooth muscles and in neurons. MEF2A gene encodes a transcription factor which was on 15q26. The objective was to study the MEF2A gene in patients with premature MI. The control group consisted of 87 subjects who were older than 45 years with no history of cardiovascular disease or MI and no family history of CAD. The premature MI group consisted of 69 patients with documented MI younger than 45 years. No abnormal bands with single strand conformation polymorphism were detected after screening exon 1 through exon 8. This is the first study that detected 145408: T>C polymorphism in intron 10. In both study groups, the rare polymorphism P279L in exon 7, T>C polymorphism in intron 10, and 21-bp deletion in exon 11 of the gene were not found. The data supported the previous studies indicating no association between MEF2A gene and premature MI.