Sequential Approach for Water Purification Using Seashell-Derived Calcium Oxide through Disinfection and Flocculation with Polyphosphate for Chemical Pollutant Removal.

Safe water supply is usually inadequate in areas without water treatment plants and even in a city under emergency conditions due to a disaster, even though safe water is essential for drinking and other various purposes. The purification of surface water from a river, lake, or pond requires disinfection and removal of chemical pollutants. In this study, we report a water purification strategy using seashell-derived calcium oxide (CaO) via disinfection and subsequent flocculation with polyphosphate for chemical pollutant removal. Seashell-derived CaO at a concentration (2 g L-1) higher than its saturation concentration caused the >99.999% inactivation of bacteria, mainly due to the alkalinity of calcium hydroxide (Ca(OH)2) produced by hydration. After the disinfection, the addition of sodium polyphosphate at 2 g L-1 allowed for the flocculation of CaO/Ca(OH)2 particles with adsorbing chemical pollutants, such as Congo red, dichlorodiphenyltrichloroethane, di(2-ethylhexyl)phthalate, and polychlorinated biphenyls, for removing these pollutants; purified water was obtained through filtration. Although this purified water was initially highly alkaline (pH ∼ 12.5), its pH decreased into a weak alkaline region (pH ∼ 9) during exposure to ambient air by absorbing carbon dioxide from the air with the precipitating calcium carbonate. The advantages of this water purification strategy include the fact that the saturation of CaO/Ca(OH)2 potentially serves as a visual indicator of disinfection, that the flocculation by polyphosphate removes excessive CaO/Ca(OH)2 as well as chemical pollutants, and that the high pH and Ca2+ concentrations in the resulting purified water are readily decreased. Our findings suggest the usability of seashell-derived material-polymer assemblies for water purification, especially under emergency conditions due to disasters.

Nanospiked paper: Microfibrous cellulose materials nanostructured via partial hydrolysis and self-assembly.

Nanocelluloses, such as cellulose nanofibers and nanocrystals, are sustainable nanomaterials that are generally extracted from natural raw materials in a top-down manner. These nanomaterials and their assemblies are facilitating new applications of biopolymers. However, creating nanostructures from conventional cellulosic materials including paper and cloth remains challenging. Herein, we report an approach for bottom-up nanostructuring of conventional microfibrous cellulose materials via a molecular self-assembly strategy. As a precursor cellulose material, paper was allowed to swell with aqueous phosphoric acid for the partial dissolution and hydrolysis of cellulose while maintaining its microfibrous structure. The generated cello-oligosaccharides in a dissolved state started to self-assemble upon adding water as a coagulant, resulting in nanospike-like assemblies on the microfiber surfaces. The resultant nanospiked papers were found to serve as a precursor for synthesizing silver nanoparticle-cellulose composites with bactericidal activities. Our findings promote the development of cellulose-based functional materials with nanostructures designed via molecular self-assembly.

Biofilm Degradation by Seashell-Derived Calcium Hydroxide and Hydrogen Peroxide.

Microbial cells and self-produced extracellular polymeric substances assembled to form biofilms that are difficult to remove from surfaces, causing problems in various fields. Seashell-derived calcium hydroxide, a sustainable inorganic material, has shown high bactericidal activity even for biofilms due to its alkalinity. However, its biofilm removal efficacy is relatively low. Herein, we report a biofilm degradation strategy that includes two environmentally friendly reagents: seashell-derived calcium hydroxide and hydrogen peroxide. A biofilm model of Escherichia coli was prepared in vitro, treated with calcium hydroxide-hydrogen peroxide solutions, and semi-quantified by the crystal violet stain method. The treatment significantly improved biofilm removal efficacy compared with treatments by calcium hydroxide alone and hydrogen peroxide alone. The mechanism was elucidated from calcium hydroxide-hydrogen peroxide solutions, which suggested that perhydroxyl anion and hydroxyl radical generated from hydrogen peroxide, as well as the alkalinity of calcium hydroxide, enhanced biofilm degradation. This study showed that concurrent use of other reagents, such as hydrogen peroxide, is a promising strategy for improving the biofilm degradation activity of seashell-derived calcium hydroxide and will contribute to developing efficient biofilm removal methods.

Recent Progress in the Development of Disinfectants from Scallop Shell-Derived Calcium Oxide for Clinical and Daily Use.

The current pandemic of novel coronavirus disease (COVID-19) has highlighted the importance of disinfectants. As a raw material for next-generation disinfectants, scallop shell-derived calcium oxide (CaO) has been revealed to exhibit significant virucidal and microbicidal activities and is compatible with living tissues and the environment. This minireview summarizes recent progress in the development of disinfectants from scallop shell-CaO, focusing especially on studies of clinical and daily use applications. We describe the preparation, basic characteristics, and virucidal and microbicidal activities of scallop shell-CaO disinfectants. Furthermore, their applications in the disinfection of contaminated masks and the treatment of infected wounds are briefly introduced.

Efficacy of Bioshell Calcium Oxide Water as Disinfectants to Enable Face Mask Reuse.

Bioshell calcium oxide (BiSCaO) is derived from scallop shells and after heat treatment exhibits broad microbicidal activity. BiSCaO Water is a disinfectant prepared by collecting the aqueous layer after adding BiSCaO powder to water, is colorless and transparent, and has a pH of 12.8. We compared the utility of commercially available BiSCaO Water, ethanol, sodium hypochlorite, hypochlorous acid and hydrogen peroxide solutions as sterilization agents to enable the reuse of surgical and N95 face masks. The microbicidal efficacy of each disinfectant was evaluated using pieces of surgical and N95 face masks contaminated with normal bacterial flora. The results suggest that BiSCaO Water has excellent disinfection activity toward contaminated polypropylene masks and has minimal adverse effect on the structure of non-woven masks.

Safety of Concentrated Bioshell Calcium Oxide Water Application for Surface and Skin Disinfections against Pathogenic Microbes.

Immediately post-production, commercially available bioshell calcium oxide (BiSCaO) water is colorless, transparent, and strongly alkaline (pH 12.8), and is known to possess deodorizing properties and broad microbicidal activity. However, BiSCaO Water may represent a serious safety risk to the living body, given the strong alkalinity. This study aimed to investigate the safety of BiSCaO Water for use as an antiseptic/disinfectant despite concerns regarding its high alkalinity. The change over time in pH of BiSCaO Water was measured during air contact (stirring BiSCaO Water in ambient air). When sprayed on metal, plastic, wood piece, paper, and skin surfaces, the pH of BiSCaO Water decreased rapidly, providing a white powder coating upon drying. Scanning electron microscopy images, energy dispersive X-ray elemental mapping, and X-ray diffractograms showed that the dried powder residues of BiSCaO Water were composed primarily of calcium carbonate. These results suggested that BiSCaO Water is a potent reagent that may overcome the obstacles of being strongly alkaline, making this material appropriate for use in disinfection against pathogenic microbes.

Concentrated Bioshell Calcium Oxide (BiSCaO) Water Kills Pathogenic Microbes: Characterization and Activity.

Bioshell calcium oxide (BiSCaO) exhibits deodorizing properties and broad microbicidal activity. In this study, we examined possible utility of BiSCaO Water for that purpose. BiSCaO Water was prepared by adding 10 wt% BiSCaO to clean water and gently collecting the supernatant in a bottle. The same volume of clean water was gently poured onto the BiSCaO precipitate and the supernatant was gently collected in a bottle; this process was repeated fifty times. The produced BiSCaO Water contained nanoparticles (about 400-800 nm) composed of smaller nanoparticles (100-200 nm), and was colorless and transparent, with a pH > 12.7. In vitro assays demonstrated that BiSCaO Water eliminated more than 99.9% of influenza A (H1N1) and Feline calicivirus, Escherichia coli such as NBRC 3972 and O-157:H7, Pseudomonas aeruginosa, Salmonella, and Staphylococcus aureus within 15 min. We compared BiSCaO Water with the other microbicidal reagents such as ethanol, BiSCaO, BiSCa(OH)2 suspensions, povidone iodine, NaClO, BiSCaO dispersion and colloidal dispersion with respect to deodorization activity and microbicidal efficacy. The results showed that BiSCaO Water was a potent reagent with excellent deodorization and disinfection activities against pathogenic bacteria and viruses (including both enveloped and nonenveloped viruses).

Bioshell Calcium Oxide-Containing Liquids as a Sanitizer for the Reduction of Histamine Production in Raw Japanese Pilchard, Japanese Horse Mackerel, and Chub Mackerel.

Recently, there has been an increase in the number of food poisoning cases associated with histamine in food, mainly in relation to histamine in fish. Here, we investigated methods to decrease histamine levels in Japanese pilchard, Japanese horse mackerel, and chub Mackerel, stored at 10 °C using various concentrations of heated scallop bioshell calcium oxide (BiSCaO) suspension, dispersion (BiSCaO + Na2HPO4), colloidal dispersion (BiSCaO + NapolyPO4), scallop shell powder (SSP) Ca(OH)2 in pure water (PW) or saline, and BiSCaO water. BiSCaO in a high alkaline pH solution chemically decomposes histamine poorly, but the partial flocculation/precipitation of histamine was observed with 1 and 0.2 wt.% BiSCaO dispersion and BiSCaO colloidal dispersion, respectively. Cleaning fish samples with BiSCaO suspension, dispersion, colloidal dispersion, or BiSCaO water remarkably reduced histamine levels and normal bacterial flora (coliform bacteria (CF) and total viable bacterial cells (TC)) after storage for four days at 10 °C, while much higher histamine levels were observed after cleaning with saline. These results suggest that cleaning fish with BiSCaO dispersion, colloidal dispersion, or BiSCaO water can significantly reduce histamine levels through their bactericidal activity against histamine-producing bacteria.

Bioshell Calcium Oxide (BiSCaO) Ointment for the Disinfection and Healing of Pseudomonas aeruginosa-Infected Wounds in Hairless Rats.

Bioshell calcium oxide (BiSCaO) possesses deodorizing properties and broad microbicidal activity. This study aimed to investigate the application of BiSCaO ointment for the prevention and treatment of infection in chronic wounds in healing-impaired patients, without delaying wound healing. The bactericidal activities of 0.04, 0.2, 1, and 5 wt% BiSCaO ointment, 3 wt% povidone iodine ointment, and control (ointment only) were compared to evaluate the in vivo disinfection and healing of Pseudomonas aeruginosa-infected wounds in hairless rats. Treatment of the infected wounds with 0.2 wt% BiSCaO ointment daily for 3 days significantly enhanced wound healing and reduced the in vivo bacterial counts compared with povidone iodine ointment and control (no wound cleaning). Although 5 wt% BiSCaO ointment provided the lowest bacterial counts during 3 days' treatment, it delayed wound healing. Histological examinations showed significantly advanced granulation tissue and capillary formation in wounds treated with 0.2 wt% BiSCaO ointment for 3 days compared to wounds treated with the other ointments. This study suggested that using 0.2 wt% BiSCaO ointment as a disinfectant for infected wounds and limiting disinfection to 3 days may be sufficient to avoid the negative effects of BiSCaO on wound repair.

Relationship between shape of peripheral initial lymphatics and efficiency of mechanical stimulation-induced lymph formation.

OBJECTIVE: The present study aimed to combine the physiological significance of irregularly shaped initial lymphatics and mechanisms of mechanical stimulation-induced lymph formation. METHODS: To confirm stretch-induced expansion of initial lymphatics, a finite element model that simulated morphological changes on a computer and fluorescent image and immunohistochemical analyses in mouse skin were adopted. Next, to quantitatively analyze the stretch-induced expansion, a simulation study was performed using a viscoelastic model of the tissue including initial lymphatics. RESULTS: On the finite element model, when the tissue was stretched, irregularly shaped lymphatics were confirmed to increase luminal volume compared with round-shaped lymphatics. Stretch-induced expansion of the real initial lymphatics was demonstrated by fluorescent images and histological studies. Thereafter, with the application of a viscoelastic model of the tissue, the relationship between the lymph formation rate (Q) and massage frequency (f) could be obtained using the following equation: Q=2Af(1-e-1/2τf) , where A and τ are constants. Excellent agreement was found between the previous data and the results of the present equation. CONCLUSIONS: We conclude that irregularly shaped initial lymphatics may lead to efficient lymph formation induced by mechanical stimulation of the tissue.

Application of Colloidal Dispersions of Bioshell Calcium Oxide (BiSCaO) for Disinfection.

Bioshell calcium oxide (BiSCaO) is a scallop-shell powder heated at a high temperature. BiSCaO is composed mainly of calcium oxide and exhibits broad microbicidal properties. The aim of this study is to evaluate the disinfection and decontamination abilities of BiSCaO colloidal dispersions with that of commercially available bioshell calcium hydroxide (BiSCa(OH)2) following the formation of flocculants/precipitates under strongly alkaline conditions (pH 11.5-12.2). Various concentrations of BiSCaO and BiSCa(OH)2 colloidal dispersions were prepared by mixing with Na-polyPO4 (PP) and Na-triPO4 (TP) as flocculating agents. The microbicidal activities, and the degree of flocculation/precipitation of trypan blue, albumin, chondroitin sulfate, heparin, non-anticoagulant heparin carrying polystyrene (NAC-HCPS), and low-molecular-weight heparin/protamine nanoparticles (LMWH/P NPs) were dependent on the pH, the average particle diameter, and the concentration of BiSCaO or BiSCa(OH)2 and of the phosphate compound. BiSCaO (average particle diameter: 6 µm) colloidal dispersions (0.2 wt.%) containing 0.15 wt.% PP or TP exhibited substantially stronger microbicidal activity and flocculation/precipitation under strongly alkaline conditions. These results suggest that BiSCaO colloidal dispersions together with phosphate compounds have practical applicability for disinfection.

Preparation and Application of Bioshell Calcium Oxide (BiSCaO) Nanoparticle-Dispersions with Bactericidal Activity.

Scallop-shell powder (SSP) heated at high temperature exhibits high pH and broad antimicrobial activity. Bioshell calcium oxide (BiSCaO) is an SSP composed mainly of calcium oxide. It is poorly water-soluble under alkaline conditions and the generated precipitate can plug spray nozzles. The aim of this study was to establish that BiSCaO dispersion caused no significant CaO loss and plugging of spray nozzles, and to evaluate its deodorization and microbicidal abilities and its ability to reduce the concentrations of NO2- and NO3-. BiSCaO dispersions were prepared by mixing various concentrations of BiSCaO suspension, while phosphate compounds such as Na3PO4, Na2HPO4 or NaH2PO4 and the pH, average diameter, zeta potential, and form of the compounds with cryo-SEM were evaluated. We evaluated deodorization using tainted pork meat and microbicidal efficacy using contaminated suspension with normal bacterial flora. The concentration of NO2- and NO3- after mixing BiSCaO dispersion and pure water containing a high proportion of NO2- and NO3- were measured. BiSCaO dispersion formed with Na2HPO4, whose ratio to BiSCaO was 60%, showed a high pH (>12), a small particle diameter (>181 nm) and was stable for seven days. The BiSCaO dispersion showed higher deodorization and microbicidal activities than SSP-Ca(OH)2, which was mainly composed of Ca(OH)2. BiSCaO, but not SSP-Ca(OH)2, could reduce the concentration of NO2- and NO3- by more than 90% within 15 min. We developed a stable BiSCaO dispersion, and it had high deodorization and microbicidal efficacy. These activities of BiSCaO might result from the high pH caused by CaO hydration and a reduction activity causing active radical species.

Comparison of Various Disinfectants on Bactericidal Activity Under Organic Matter Contaminated Environments.

The bactericidal activity of heated bio-shell calcium oxide (BiSCaO) powder suspension (pH 12.4) , hypochlorous acid (HClO; pH 6), sodium hypochlorite (NaClO; pH 8) , povidone-iodine (Isodine solution®) , and chlorhexidine gluconate (Hibiscrub®) under organic matter contaminated environments were compared for tests conducted on wood scraps and pig skin pieces that were incubated with normal bacterial flora (total viable counts and coliform bacteria) . The test results showed that BiSCaO suspension had higher bactericidal activity than HClO and NaClO. Furthermore, more than 10-fold higher concentrations of antiseptics such as povidone-iodine and chlorhexidine gluconate were required to achieve bactericidal activity comparable to that of BiSCaO suspension. Our results demonstrate the possibility of using BiSCaO suspension under organic matter contaminated environments as a disinfectant for environmental and food hygiene applications.

FGF-2-containing dalteparin/protamine nanoparticles (FGF-2&D/P NPs) ameliorate UV-induced skin photoaging in hairless mice.

UVB exposure penetrates deeply into the dermis to alter skin barrier function, which is a primary factor in skin photoaging. We previously reported that dalteparin and protamine nanoparticles (D/P NPs) are effective carriers of FGF-2. This study aimed to examine the ability of FGF-2-containing D/P NPs (FGF-2&D/P NPs) to ameliorate UVB-induced skin photoaging in hairless mice. Dorsal skin of HR-1 hairless mice were exposed to UVB irradiation 5 days/week for 8 weeks (UV (+): final total, 2700 mJ/cm2). Mice were divided into four groups: Non-UVB (UV (-)) + saline, UV (+) + saline, UV (+) + FGF-2&D/P NPs, UV (+) + FGF-2, and UV (+) + D/P NPs, and following UVB irradiation, FGF-2&D/P NPs, FGF-2, and D/P NPs were applied to the groups of mice just after each UVB irradiation. Each group was subjected to evaluation of skin changes (elasticity), and histological examination using hematoxylin & eosin and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) staining. UVB irradiation of mice significantly induced a decline in elasticity and acanthosis, which was alleviated by application of FGF-2&D/P NPs. Furthermore, TUNEL-staining showed the proportions of apoptotic dermal fibroblast cells (DFCs) and epidermal keratinocyte cells (EKCs) in the UV (+) + FGF-2&D/P NPs group were significantly lower than those in the UV (+) + saline, UV (+) + FGF-2, and UV (+) + D/P NPs groups. Thus, FGF-2&D/P NPs may be effective in preventing skin photoaging accelerated by UVB irradiation such as declining elasticity, acanthosis, and apoptosis of DFCs and EKCs.