Ferrous Ferric Chloride Research Articles

Elastin Peptides with Ferrous Ferric Chloride Activate Human Melanocytes and Elastin Fibers

Background: Elastin peptides stimulate the development of mouse melanocytes in neural crest culture. Ferrous ferric chloride (FFC) promotes mammalian melanocyte growth in culture. However, it is unclear whether elastin peptides in the presence of FFC can stimulate human melanocyte growth in situ. Objectives: This study aimed to investigate the mechanism of human melanocyte growth for skin and stem cell science since melanocytes control human skin color. Methods: In this clinical trial study, a lotion containing elastin peptides and/or FFC was applied to the normal skin of 6 volunteers twice a day for 1 to 3 months. Punch biopsies were taken from treated skin and surveyed by histochemical methods using the dopa reaction (detect melanocytes) and dopa-premelanin reaction (detect melanocytes and melanoblasts). Elastin fibers were detected by Victoria blue staining. Results: Only the combined treatment of elastin peptides and FFC increased melanocyte populations in addition to dopa reactivity, melanogenesis, dendritogenesis, and epidermal melanin pigmentation. Mitotic divisions of melanocytes were also observed. However, the melanoblast population failed to increase, and no mitotic melanoblasts were observed. In the dermis, elastin fibers became thicker and denser after the treatment. The data of statistical analyses were performed by tabulation, mean, and SD on Microsoft Excel for Macintosh OS Catalina 10 system. Conclusions: Our present study suggests that elastin peptides with FFC can promote melanocyte growth, melanin synthesis, skin pigmentation, and elastin fiber formation. Our study can be expected to contribute to advancing skin and stem cell science.

P020 Down-regulatory effect of Pairogen, a health drink, on inflammatory response to Rhodococcus aurantiacus infection in mice

Introduction The health drink Pairogen mainly consists of ferrous ferric chloride (FFC) water that appears to perform multiple biological functions by regulating intracellular redox reactions in living organisms (1). With respect to its contribution to host defense, FFC water reportedly protects plant against fungal infection (2). However, little is known regarding the capability of FFC water and Pairogen to regulate the defense mechanisms in animals or humans in order to respond properly to pathogenic organisms. Herein, we investigated whether Pairogen regulates inflammatory response to Rhodococcus aurantiacus ( R. aurantiacus ) infection in mice. Methods We assessed survival, organ bacterial load, and inflammation (cytokine production and histomorphological changes) in Pairogen-treated mice (F0) and first-generation mice (F1) following infection. Cytokine response to heat-killed R. aurantiacus was further explored in the peritoneal macrophages of Pairogen-treated F1 mice. Results Longitudinal ingestion of Pairogen improved survival of infected mice in a concentration-dependent manner. Rapid elimination of bacteria from the organs (such as livers and spleens), accompanied by decreased production of tumor necrosis factor-alpha and interleukin-6 and elevated release of interleukin-10, was observed in F0 mice treated with 10-fold-diluted Pairogen. Histological studies also exhibited a significant reduction in granuloma number and size as well as amelioration in the oxidative status in mouse livers at day 14 following infection. The effects of Pairogen were further pronounced in F1 mice treated with 10-fold-diluted Pairogen. Stimulating the peritoneal macrophages from these mice with heat-killed R. aurantiacus induced the production of three cytokines similar to those detected in vivo, whereas gene expression of these cytokines was down-regulated, suggesting an inhibitory role of Pairogen in cytokine expression and the mutual antagonistic relationship among the cytokines. Conclusion The present study reveals that longitudinal ingestion of Pairogen improves survival, accelerates bacterial clearance and suppresses the over-expression of inflammation-related cytokines by regulating cellular redox reactions, and consequently results in depression of granulomatous inflammation in R. aurantiacus -infected mice. Moreover, these effects of Pairogen were enhanced in their offspring when they continued to ingest Pairogen. Pairogen is thought to be a useful redox regulator that contributes to the regulation of inflammatory and immune responses. It may also be expected to improve the quality of human life by heightening the defense functions against infection.

Ferrous Ferric Chloride Downregulates the Inflammatory Response to Rhodococcus aurantiacus Infection in Mice

The healthy drink Pairogen is mainly composed of ferrous ferric chloride water that reportedly changes the status of intracellular water from oxidative to antioxidative. Here, we investigated whether Pairogen affects host immune function in a murine model of granulomatous inflammation in response to Rhodococcus aurantiacus (R. aurantiacus) infection. Longitudinal ingestion of Pairogen markedly improved the survival of infected mice in a concentration-dependent manner. Compared to mice received water, mice that ingested 10-fold-diluted Pairogen displayed rapid bacterial elimination, decreased production of tumor necrosis factor (TNF)-α and interleukin (IL)-6, and high levels of IL-10 in organs during the initial phase of infection. Moreover, histological studies showed significant reduction in the number and size of granulomas as well as amelioration of oxidative stress in the livers of mice ingested 10-fold-diluted Pairogen at 14 d post-infection. These characteristics were further pronounced in first-generation (F1) mice that also ingested 10-fold-diluted Pairogen. Following stimulation with heat-killed R. aurantiacus, the production of TNF-α, IL-6, and IL-10 by macrophages from F1 mice was similar to that detected in vivo, while their gene expression levels in these cells were significantly lower than the levels in macrophages from mice received water. Heat-killed R. aurantiacus also induced the expression of heme oxygenase-1 mRNA in the cells from F1 mice. Taken together, these results indicate that Pairogen contributes to the negative regulation of the immuno-inflammatory response to R. aurantiacus infection in mice by modulating the cellular redox state. Longitudinal ingestion of Pairogen further enhances the defense function in mouse progeny.

Stimulation of the Proliferation and Differentiation of Skin Cells by Ferrous Ferric Chloride from a Distance

Ferrous ferric chloride (FFC) is a distinct form of aqueous iron composed of a complex of ferrous chloride and ferric chloride that participates in both oxidation and reduction reactions, and stimulates the proliferation and differentiation of mammalian keratinocytes, melanocytes, and fibroblasts. However, it is not known whether FFC can stimulate their proliferation and differentiation without being added into culture media or painted on the skin. This study aims to clarify whether FFC can stimulate their proliferation and differentiation from a distance without being added to culture media. In this study, FFC-containing skin lotions were painted under the culture dishes (1 mm away from cells) or on the top of the covers of 1 to 5 polystyrene culture dishes (1 to 5 cm away) and tested for their proliferation- and differentiation-stimulating effects. FFC lotions stimulated the proliferation and differentiation of human keratinocytes, melanocytes, and fibroblasts from a distance of 1 mm to 1 cm. However, FFC lotions failed to stimulate the proliferation and differentiation of melanocytes from distances of 2 to 5 cm. Results using Teflon covers were similar to those of polystyrene covers. Moreover, the effects of FFC lotions painted on the top of the Teflon covers were completely lost by lead disks. These results suggest that FFC can stimulate the proliferation and differentiation of skin cells from a distance of 1 cm without being added into culture media through physical factors rather than chemical factors.

Ferrous Ferric Chloride Stimulates the Skin Cell Function and Hair Growth in Mice

Ferrous ferric chloride (FFC) is a distinct form of aqueous iron composed of a complex of ferrous chloride and ferric chloride that participates in both oxidation and reduction reactions. The author's previous study showed that the supplementation of culture medium with FFC stimulated the proliferation and differentiation of keratinocytes and melanocytes in newborn mice. FFC also stimulated the proliferation of cultured human keratinocytes, fibroblasts, and melanocytes. However, it is not known whether FFC stimulates the proliferation and differentiation of mammalian skin cells as well as hair growth in vivo. To answer this question, FFC-containing skin lotions (FFC Super Essence Plain and Moisture Type, Akatsuka Co.) were painted on the dorsal skin of newborn C57BL/10JHir (B10) mice and tested for their proliferation- and differentiation-stimulating effects on keratinocytes, fibroblasts, and melanocytes as well as for their stimulating effects on the hair growth. This treatment stimulated the proliferation and differentiation of epidermal keratinocytes, dermal fibroblasts, and epidermal and dermal melanocytes in the skin as well as hair growth. From 2 to 3 weeks after birth B10 mice generally lose their hairs except those on the head at the telogen stage of the hair growth cycle due to the expression of the alopecia. The treatment with FFC lotions markedly inhibited the alopecia hair-loss. These results suggest that FFC can stimulate the proliferation and differentiation of keratinocytes, fibroblasts, and melanocytes in the skin as well as the hair growth, and, in addition, can inhibit the alopecia hair-loss.

Ferrous Ferric Chloride Induces the Differentiation of Cultured Mouse Epidermal Melanocytes Additionally with Herbal Medicines

Ferrous ferric chloride (FFC) is a special form of aqueous iron that is a complex of ferrous chloride and ferric chloride and participates in oxidation and reduction reactions. My previous study showed that FFC stimulated the proliferation and differentiation of cultured epidermal melanoblasts or melanocytes derived from newborn mice. However, it is not known whether FFC stimulates the proliferation and differentiation of melanocytes in cooperation with natural factors, such as vinegars, vitamins, and herbal medicines. The drink Pairogen® (Akatsuka Co., Tsu, Japan) consists of FFC, vinegars, and vitamins, while Pairogen Gold® (Akatsuka Co.) contains herbal medicines in addition to FFC, vinegar, and vitamins. To clarify whether these natural factors supplemented to Pairogen or Pairogen Gold elicit stimulative effects on skin function, Pairogen and Pairogen Gold were added to the culture medium and tested for their proliferation- and differentiation-stimulating activity on melanocytes. Although Pairogen Gold failed to increase melanocyte proliferation beyond that elicited by FFC alone, it markedly increased melanocyte differentiation. In contrast, Pairogen possessed no such effect. The extracts of Chinese wolfberry (Lycium chinense) and Siberian ginseng (Eleutherococcus senticosus) that are included in Pairogen Gold stimulated melanocyte differentiation additionally with FFC. Therefore, these results suggest that FFC is involved in regulating the differentiation of melanocytes additionally with extracts of Chinese wolfberry and Siberian ginseng.

Ferrous Ferric Chloride Stimulates the Proliferation of Human Skin Keratinocytes, Melanocytes, and Fibroblasts in Culture

Ferrous ferric chloride (FFC®) is a distinct form of aqueous iron made up of a complex of ferrous chloride and ferric chloride that participates in both oxidation and reduction reactions. Although FFC is known to stimulate cellular function, its precise mechanism of action is largely unknown. My previous study showed that FFC stimulated the proliferation of cultured epidermal keratinocytes and melanocytes derived from newborn mice, suggesting that FFC stimulates their function of mice. However, it is not known whether FFC stimulates the proliferation of keratinocytes and melanocytes derived from human skin. Moreover, it is unknown whether FFC stimulates the proliferation of human skin fibroblasts. Pairogen® (Akatsuka Co., Tsu, Japan) is a drink comprising FFC, vinegars, and vitamins, Pairogen Gold® (Akatsuka) contains herbal medicines in addition to FFC, vinegars, and vitamins, while Noncalorie Pairogen® (Akatsuka) contains low molecular weight collagen and hyaluronic acid in addition to FFC, vinegars and vitamins. To clarify whether FFC in addition to the natural factors included in these drinks stimulates the proliferation of human skin cells, FFC and these different drinks were added to culture media and tested for their proliferation-stimulating activity on skin cells. FFC, Pairogen, and Pairogen Gold equally doubled the growth rate of cultured human keratinocytes, melanocytes, and fibroblasts. Non-calorie Pairogen tripled the growth rate of human fibroblasts, but Non-calorie Pairogen only doubled the growth rate of human keratinocytes and melanocytes. FFC plus low molecular weight collagen tripled the growth rate of human fibroblasts, whereas FFC plus low molecular weight hyaluronic acid only doubled the growth rate of human fibroblasts. Low molecular weight collagen, with or without hyaluronic acid failed to stimulate the proliferation of fibroblasts. These results suggest that FFC stimulates the proliferation of human skin keratinocytes and melanocytes. Moreover, FFC markedly stimulates the proliferation of human skin fibroblasts synergistically with low molecular weight collagen.

Ferrous Ferric Chloride Stimulates the Proliferation and Differentiation of Cultured Keratinocytes and Melanocytes in the Epidermis of Neonatal Mouse Skin

Ferrous ferric chloride (FFC) is a special aqueous iron which is ac omplex of ferrous chloride and ferric chloride and has a function in both oxidation and reduction. FFC is thought to stimulate the cellular function of living organisms. Although FFC is reported to stimulate the function of red blood cells, it has not been determined whether FFC stimulates the function of skin cells. To understand the role of FFC in the function of skin cells, we studied its effects on the proliferation and differentiation of keratinocytes and melanoblasts or melanocytes. FFC was added to a serum-free culture of neonatal mouse epidermal cells and its effects on the proliferation and differentiation of keratinocytes and melanoblasts or melanocytes were investigated. FFC stimulated the proliferation and differentiation of keratinocytes and melanoblasts or melanocytes. The proliferation of keratinocytes and that of melanoblasts or melanocytes was stimulatedto the same extent (atwofold increase), suggesting tha tt heproliferation of the two types of cells constituting the epidermis may be equally stimulated by FFC. These results suggest that FFC may activate skin function by promoting cell renewal via th es timulation of the proliferation and differentiation of keratinocytes and melanoblasts or melanocytes.

The use of simultaneous chemical precipitation in modified activated sludge systems exhibiting biological excess phosphate removal: Part 7: Application of the IAWQ model

The IAWQ Activated Sludge Model (ASM) No. 2 is a kinetic-based model and incorporates two simple processes for chemical precipitation and redissolution that are readily integrated with biological processes for carbon, nitrogen and phosphorus remov al. This model was applied to experimental data collected as part of this study from parallel pilot-scale 3-stage Phoredox systems with and without simultaneous dosing of chemical precipitant. The precipitants tested were alum, ferric chloride and ferrous-ferric chloride. The model was calibrated to the control unit (without precipitant addition) in order to match effluent phosphate (P) predictions (and hence P removal) as closely as possible. The same calibration was then applied to modelling the test unit (wit h precipitant addition). It was found that the default model input stoichiometry for the precipitation reaction (ideal 1:1 molar ratio of metal ion (Me) to P) was suitable for ferric chloride addition at a 20 d sludge age, but did not accurately reflect the test sy stem behaviour for all experimental periods. A lower stoichiometry (0.60 to 0.75) was required for alum at a 20 d sludge age, and fo r a blend of predominantly ferrous chloride at a 10 d sludge age. The input stoichiometry was further decreased under P-limiting conditions. A simple approach to, and possible reasons for, the manipulation of the model stoichiometry are discussed in the li ght of observed stoichiometry from system P removal and fractionation data collected as part of this study. Furthermore, an alterna tive approach based on manipulation of the precipitation (and hence redissolution) kinetic constant is suggested and evaluated using available experimental data. Model predictions and observed data in respect of polyphosphate (polyP) and suspended solids are a lso compared and discussed. It is concluded that the ASM No. 2 model provides a useful basis for modelling simultaneous P precipitation, provided certain minor modifications are made. Further investigation into the kinetics of the precipitation process(es) is recommended, particularly in relation to the effect of system sludge age. The model lends itself to further enha ncement by incorporating additional physico-chemical processes.