Degreasing and bleaching bones using light sources as a tool to increase the safety of teaching osteology at the University of Veterinary Sciences Brno.

The key part of creating bone material for teaching is degreasing and whitening it. However, the substances used are often dangerous and toxic. We tested and compared safer methods based on two physical variables. These are light and heat. The material for our study was 45 femurs from 23 adult domestic dogs (Canis lupus f. familiaris). The bones were divided into three groups of 15 pieces according to the method used to remove muscles and ligaments from their surface. Five femurs from each group were exposed to three different light sources for 28 days-sunlight, warm light from a classical incandescent light bulb and cold light by a LED bulb. At regular intervals, the change in the colour of the bone surface and the amount of fat loss from the medullary cavity was also monitored. The best degreasing and bleaching results were achieved in macerated bones exposed to sunlight. They achieved the required condition as early as 21 days after the start of sun exposure. The biggest problem was haemoglobin, which permeated through the Haversian canals and discoloured the bone tissue. The results showed that the use of light and heat is a suitable and safe alternative to chemical methods of degreasing and bleaching bones. The disadvantage is the length of time, especially for native material.

Optimization of Enzymatic Degreasing of Sheep Leather for an Efficient Approach and Leather Quality Improvement Using Fractional Experimental Design.

Leather industry is making significant contributions to economic development. However, it is notably leading to a serious environmental pollution. Recently, the enzyme technology developments offer new opportunities for enzymatic application in leather making. In the present investigation, microbial lipases were studied and used in degreasing process of sheep leathers. In order to optimize degreasing efficiency, a fractional experimental design with four parameters (enzyme source, processing stage, lipase amount, and degreasing duration) was used. Lipases A from Aspergillus niger, F from Rhizopus oryzae, R from Penicillium roqueforti, and AY from Candida rugosa were selected for leather degreasing. Enzymatic treatment of sheep skin was carried out during two stages of beamhouse operations: deliming-bating and pickling. Obtained results showed that enzymatic degreasing efficiency is higher than those obtained with the conventional process. Lipase F from Rhizopus oryzae demonstrated the most interesting hydrolysis with yields of 58.3% and 37.2% for delimed and pickled skins, respectively. An enzymatic degreasing process on pickled leather using 0.125% (w/v) of lipase F during 3.5 h is the most promising for an industrial application with a 76.03 of degreasing efficiency. Results of the physico-mechanical tests of leathers having undergone enzymatic treatment complied with industry requirement. The enzymatic treatment may be carried out in the same conditions as employed in leather manufacturing process. Results suggested that the enzymatic degreasing improves the leather quality and reduces the use of chemical compounds and surfactant.

Cleaner degreasing of sheepskins by the Yarrowia lipolytica LIP2 lipase as a chemical-free alternative in the leather industry.

Conventional degreasing of skins and hides in the leather industry requires high amounts of organic solvents and detergents that cause environmental issues. In this study, the LIP2 lipase from the yeast Yarrowia lipolytica (YLLIP2) was shown to be effective in degreasing sheepskins, thus reducing the amount of harmful chemicals. Using 6 mg of lipase/kg of raw skin, successful degreasing was achieved in only 15 min at pH 8 and 30°C. ToF-SIMS mass spectra of chemically and enzymatically treated sheepskins are consistent with a selective elimination process for the enzymatic treatment. Comparative SEM microscopy, ATR-FTIR spectroscopy and physicochemical analyses showed better properties of the enzymatically treated leather than those of the chemical treatment. Effluent physicochemical parameters showed that the enzymatic treatment is a cleaner degreasing operation. Altogether, this work opens new horizons to use the YLLIP2 lipase as a more efficient alternative in the leather industry.

Degreasing cotton used as pore-creating agent to prepare hydrophobic and porous carbon cathode for the electro-Fenton system: enhanced H2O2 generation and RhB degradation.

A porous carbon cathode was prepared using graphite, polytetrafluoroethylene (PTFE), and degreasing cotton (DC) through sintering treatment. The carbonization of DC by heat treatment played an ideal role in pore-creating, which weakened the mass transfer resistance of O2, and as a result, the adoption of degreasing cotton significantly improved the performance of H2O2 electro-generation. The optimized cathode was able to generate 567 mg L-1 H2O2 with a current efficiency (CE) of 86.7% by the electrochemical reaction of 60 min in a divided reactor. Furthermore, the degradation of rhodamine B (RhB) was carried out by an electro-Fenton system using the optimal cathode selected. The developed electro-Fenton system exhibited an excellent RhB degradation performance. The RhB solution of 50 mg L-1 was decolorized completely by the treatment of 10 min. Moreover, the degradation of 50~90 mg L-1 RhB solution presented over 90% TOC removal by the treatment of 120 min, indicating the ideal mineralization of organic pollutants. In addition, it was found that •OH was the major oxidizing specie responsible for the organics degradation. Finally, the possible pathway of RhB degradation in the electro-Fenton system was proposed by GC-MS analysis. The adoption of natural fibers for pore-creating provides an innovative and low-cost method to prepare porous cathode, which may promote the application of electro-Fenton oxidation in wastewater treatment.

Aqueous ethyl acetate as a novel solvent for the degreasing of black soldier fly (Hermetia illucens L.) larvae: degreasing rate, nutritional value evaluation of the degreased meal, and thermal properties.

BACKGROUND: The aim of this study was to select appropriate low-toxicity degreasing solvents to degrease black soldier fly (BSF, Hermetia illucens L.) larvae to prepare high-quality protein. Aqueous ethyl acetate was chosen as the solvent to extract BSF protein, and traditional solvents, such as petroleum ether, n-hexane, and isopropanol, were chosen as controls. RESULTS: The meal degreased by aqueous ethyl acetate (the volume ratio of ethyl acetate to water is 90 to 10, EA + W10) shows a high degreasing rate (29.04%), crude protein content (562.3 g kg-1 ), essential amino acid index (EAAI, 95.57), and digestible indispensable amino acid score (DIAAS, 85). The digestibility of the degreased meal samples in the simulated in vitro intestine can reach 76.52%. Thermodynamic analysis and the apparent morphology of the protein fragments showed that the meal degreased by EA + W10 exhibited thermodynamic stability, which suggests that using aqueous ethyl acetate as the degreasing solvent did not affect the nutritional value of the degreased meal. CONCLUSION: The results suggest that aqueous ethyl acetate (EA + W10) can be used as a novel solvent in the degreasing of BSF larvae meal to prepare high-quality protein with high EAAI and DIAAS and good digestibility. © 2019 Society of Chemical Industry.

Extraction of porcine cancellous bone by supercritical carbon dioxide / 中国组织工程研究

BACKGROUND: In recent years, there have been many studies on the use of supercritical fluids for biological material treatment in countries outside China. However, little is reported on application of supercritical fluids to bone tissue extraction, in particular in China. OBJECTIVE: To evaluate the effectiveness of supercritical carbon dioxide extraction technology in the treatment of porcine femur cancellous bone and its effect on bone biological properties. METHODS: Porcine femoral bone blocks that were subjected to supercritical carbon dioxide extraction (study group) or not (control group) were prepared to determine bone mineral density, microstructure, maximum compressive strength, elastic modulus, bone tissue composition, collagen content and perform histological analysis. Bone marrow mesenchymal stem cells (BMSCs) were inoculated into two groups of bone blocks, and cultured for 1 day. The microporous structure of trabecular bone and cell adhesion and growth in bone material-cell composite were observed by scanning electron microscopy. The two groups of bone blocks were implanted subcutaneously in SD rats. The inflammatory reaction of subcutaneous tissue was observed histologically at 1, 2 and 4 weeks after surgery. The experimental protocol had been approved by the Animal Ethics Committee of Chinese PLA General Hospital, China. RESULTS AND CONCLUSION: There were no significant differences in pore size, bone mineral density, maximum compressive strength, elastic modulus and collagen content between the study and control groups (P>0.05). Scanning electron microscopy showed that in the control group, the material pores had poor connectivity and there was soft tissue residue; in the study group, material pores were connected to each other and the structure was intact. Fourier transform infrared spectroscopy and X-ray diffraction analysis showed that the two groups of bone tissue materials had similar absorption and diffraction peaks. Thermogravimetric analysis showed that supercritical carbon dioxide extraction could reduce water content in bone tissue. Hematoxylin-eosin staining showed that there were no soft tissue residues in the bone, and the cell residues in the bone pit were significantly reduced in the study group, while soft tissue and cell residues were observed in the control group. Sirius red staining and modified Masson staining showed that the structure of bone collagen in the study group was intact, the cytoplasmic components reduced, and the cytoplasmic components in the control group remained significantly. Scanning electron microscopy showed that there was no obvious cell adhesion in the control group, but cell adhesion growth was obvious in the study group. Perivascular inflammatory response in the bone tissue implantation region was obviously weaker in the study group than in the study group. These results suggest that supercritical carbon dioxide extraction technology is an effective and environment-friendly bone tissue processing technology. It can effectively remove porcine cancellous bone cells and soft tissue without affecting its collagen structure and content and mechanical properties, retaining intact bone pore structure, increasing cell adhesion and growth, and effectively reducing inflammatory rejection.

Study on the Mechanism of Periplaneta americana Extract Reversing Multi-drug Resistance of Human Hepato- cellular Carcinoma HepG 2/ADM Cells / 中国药房

OBJECTIVE：To study the mechanism of Periplaneta americana extract degreasing cream and CⅡ-3（shorted for “degreasing cream ”and“CⅡ-3”）reversing the multi-drug resistance of human HepG 2/ADM cells. METHODS ：MTT assay was used to investigate the toxicity effects of different concentrations of sorafenib （positive control ），degreasing cream and C Ⅱ-3 on HepG2/ADM cells ，then IC 20 was calculated. The experiment was divided into sensitivity drug ，drug-resistance group ，sorafenib group，degreasing cream group and C Ⅱ-3 group. HepG 2 cells were included in sensitivity group ，and HepG 2/ADM cells were included in the latter 4 groups. Sensitivity group and drug-resistance group were treated with routine medium ，and other 3 groups were treated with relevant medicine （IC20 as drug concentration ）. The content of ADM in HepG 2/ADM cells was determined by Laser scanning confocal microscopy. The expression of apoptosis-related protein as Bcl- 2 and Cleaved-Caspase- 9 p37 were detected by Western blotting assay. RT-qPCR and immunocytochemistry were adopted to detect mRNA and protein expressions that related to multidrug resistance [P-gp （expression produce of MDR1 gene），LRP，BCRP] and that related to enzyme-mediated multidrug resistance pathway （GST-π and Topo Ⅱ）. RESULTS ：The IC 20 of degreasing cream ，CⅡ-3 and sorafenib were （2.40±0.16）， （200.44±27.52），（18.00±1.82）μg/mL，respectively. Compared with sensitivity group ，the protein expressions of Bcl- 2，P-gp， LRP，BCRP and Topo Ⅱ，the mRNA expressions of MDR 1， LRP，BCRP and GST-π were increased significantly in drug resistance group （P＜0.05 or P＜0.01）. Compared with @qq.com drug-resistance group ，the mRNA and protein expression of MDR1 mRNA and LRP ，BCRP，GST-π were significantly decreased in degreasing cream group and C Ⅱ-3 group（P＜ 0.05 or P＜0.01）；the protein expression of Bcl- 2 and the mRNA expression of Topo Ⅱ were significantly decreased （P＜0.01）， while the protein expression level of Cleaved-Caspase- 9 p37 was significantly increased in C Ⅱ -3 group （P＜0.05）. CONCLUSIONS：Degreasing cream and C Ⅱ-3 can reverse multidrug resistance of HepG 2/ADM cells by reducing drug efflux ， promoting cell apoptosis ，reducing the mRNA and protein expression of multi-drug resistance gene as well as gene in enzyme-mediated multi-drug resistance pathway. The effect of C Ⅱ-3 is better than that of degreasing cream.

Room temperature/Corcoran/Dow Corning™-Silicone plastination process.

For 20 years, the cold temperature/S10/von Hagens' plastination technique was used to preserve biological specimens without challenge. It became the "gold standard" for preservation of beautiful, dry biological specimens. Near the end of the 21st century, a group from the University of Michigan and environs and Dow Corning™, USA, combined silicone ingredients, similar to the von Hagens' plastination products, however in a different sequence. The new polymer (Cor-tech) was combined with the cross-linker to design the "impregnation mix" which would invade the cellular structure of the specimen and yet was stable at room temperature. Later, curing would be by application of the catalyst onto the impregnated specimen. This unique sequencing of products would become the "Room temperature/Dow Corning™/Corcoran-Silicone plastination technique." The results of this room temperature technique provided similar plastinates, beautiful and practical for demonstration, containing no toxic chemical residues and forever preserved. As the name implies, impregnation of this silicone mix could be done at room temperature, without having to be kept cold. Both processes (cold and room temperature) required the same four basic steps for plastination. As well, both processes used similar basic polymers and additives to produce plastinates. However, they were combined in a different sequence. Cold temperature combines polymer and catalyst/chain extender, which is not stable and therefore must be kept colder than -15°C, while room temperature combines polymer with cross-linker which is stable, and likely forever.

Effect of a botanical cleansing lotion on skin sebum and erythema of the face: A randomized controlled blinded half-side comparison.

BACKGROUND: Elevated levels of skin sebum are associated with the growth of Propionibacterium acnes. Intensive degreasing of the skin reduces Propionibacterium acnes but also may cause skin irritation. AIMS: We assessed the degreasing effect and skin tolerability of a botanical face cleanser with hops and willow bark extract and disodium cocoyl glutamate as mild cleansing agent compared to a standard face cleanser with sodium laureth sulfate (SLES). MATERIALS AND METHODS: A total of 21 healthy volunteers with normal to oily skin were enrolled in this study. Both cleansers were applied twice a day on the left or right side of the forehead for 15 days in a standardized manner. Bioengineering measurements were performed on day 8 and 15 and on day 17 after an application break of 48 hours. The sebum level was determined using a Sebumeter® , and skin redness was measured using a Mexameter® . RESULTS: The botanical face cleanser significantly reduced the sebum level (P < .01) in the test area on day 17. The SLES containing cleanser showed a statistically relevant degreasing effect already on day 15, but after the application break the sebum level increased again on day 17. None of the cleansers caused skin irritation as determined by skin redness measurements. CONCLUSIONS: In contrast to the SLES containing cleanser, the botanical skin cleanser with hops and willow bark extract had a continuous degreasing effect without reactive seborrhoe after the treatment break. Skin cleansing without SLES might be advantageous for sensitive skin.

Assessment of workers' exposure to microorganisms when using biological degreasing stations.

Biological degreasing stations (BDSs) are used by mechanics. These BDSs use a water-based solution with a microbial degradation process. Occupational exposure during the use of BDSs has not been reported and few studies have identified the bacteria present. The objectives were to measure the concentration of microorganisms during BDSs' use and monitor the bacterial community in the liquid over time. Five mechanical workshops were studied. Six 30-min samples were taken at each workshop over one year. Bioaerosols in the ambient air samples were collected with Andersen impactors near the BDS Bioaerosols in the workers' breathing zone (WBZ) were collected on filters. Fresh bio-degreasing fluids were collected from unopened containers, and used bio-degreasing fluids were collected in the BDS. The results show that the use of BDSs does not seem to increase bioaerosols concentrations in the WBZ (concentrations lower than 480 CFU/m3) and that the bacterial communities (mainly yeasts, Bacillus subtilis and Pseudomonas aeruginosa) in the bio-degreasing fluids change through time and differ from the original community (B. subtilis). This study established that workers using BDSs were exposed to low levels of bioaerosols. No respiratory protection is recommended based on bioaerosols concentrations, but gloves and strict personal hygiene practices are essential.

Effects of degreasing-frosting on reducing toxicity and enhancing efficacy of Periplaneta americana / 中成药

AIM To study the effects of degreasing-frosting on reducing toxicity and enhancing efficacy of Periplaneta americana L..METHODS The 95％ ethanol extracts from P.americana raw powder and processed products were extracted by petroleum ether,ethyl acetate,n-butanol and water.TLC was applied to comparing the component changes in various fractions.GC-MS was used for analyzing the kinds and contents of liposoluble constituents before and after processing.Shaking flask method combined with UV spectrophotometry method was adopted in the determination of Papp values in octanol-water and different pH values (2.23,3.28,4.22,5.23,6.12,7.24,8.23,9.10,10.15 and pure water) of buffer solutions.RESULTS After the petroleum ether fraction was processed,the colors of TLC spots became much shallower,which changed more significantly with the prolonging of processing time and increase of pressure.The TLC spots in the n-butanol and water fractions were clear without obvious changes.And no TLC spots were found in the ethyl acetate fraction.The kinds of liposoluble constituents were the same before and after processing,whose absolute contents were decreased after processing.Compared with raw powder,the Papp values of minor polypeptides in various processed products at different pH values exhibited significant differences (P ＜0.05,P ＜0.01),which also showed certain differences at the same pH values.CONCLUSION Degreasing-frosting can affect the contents of liposoluble constituents in P.americana and Papp values of minor polypeptides,which may be the mechanism of reducing toxicity and enhancing efficacy.

Empleo del ultrasonido para la extracción de fracción apolar en hojas de Mangifera indica L. (árbol del mango) / Use of ultrasound for extraction of the apolar fraction in leaves of Mangifera indica L. (mango tree)

Introducción: la mangiferina es una glucosil xantona natural presente en varias partes de Mangifera indica L. (árbol del mango), que posee gran variedad de efectos farmacológicos. Su aislamiento, a partir de las hojas, es de vital importancia teniendo en cuenta las propiedades farmacológicas que se le atribuyen por sí sola. En Cuba, se ha desarrollado un procedimiento nacional de obtención de mangiferina a partir de hojas de M. indica, empleando un método tradicional en tanque agitado, que incluye una etapa de desengrase, sin embargo continúan los estudios encaminados al uso de métodos no convencionales de extracción como el ultrasonido, teniendo en cuenta la rapidez y eficiencia en la obtención de fitoconstituyentes. Objetivo: evaluar la etapa de extracción de la fracción apolar en el proceso de extracción de mangiferina mediante ultrasonido a partir de hojas de M. indica. Métodos: se trabajó con hojas de M. indica, molidas y almacenadas en bolsas de nylon a temperatura y humedad ambientes. Se evaluó la influencia de los parámetros de operación (tiempo y relación disolvente material vegetal) en la etapa de desengrase, mediante un diseño factorial de superficie de respuesta, utilizando el ultrasonido. Resultados: el mejor valor experimental (29,03 ± 1,10 mg/g) se alcanza para un tiempo de 90 min y una relación volumen de disolvente/ material vegetal de 47 mL/g, empleando n-hexano como disolvente. Conclusiones: el método de ultrasonido es adecuado para la extracción de la fracción apolar en el proceso de obtención de mangiferina a partir de hojas de M. indica(AU)

Introduction: mangiferin is a natural glucosyl xanthone found in several parts of Mangifera indica L. (mango tree) which has a great variety of pharmacological effects. Its isolation from leaves is of great importance, considering the pharmacological properties attributed to it. A national procedure has been developed in Cuba to obtain mangiferin from M. indica leaves using a traditional stirred tank procedure which includes a degreasing stage. However, studies continue to be conducted about the use of unconventional extraction techniques such as ultrasound, due to the speed and efficacy with which phytoconstituents may be obtained. Objective: evaluate the apolar fraction extraction stage of the process of ultrasound extraction of mangiferin from M. indica leaves. Methods: M. indica leaves were ground and stored in plastic bags at ambient temperature and humidity. Operating parameters (time and solvent / plant material ratio) were evaluated at the degreasing stage with a response surface factorial design, using ultrasound. Results: the best experimental value (29.03 ± 1.10 mg/g) is obtained for a time of 90 min and a solvent volume / plant material ratio of 47 ml/g, using n-hexane as solvent. Conclusions: the ultrasound technique is appropriate for apolar fraction extraction of mangiferin from M. indica leaves(AU)

Use of electrochemically activated aqueous solutions in the manufacture of fur materials.

The influence of characteristics of electrochemically activated aqueous processing mediums in the treatment of fur skins with different contents of fatty substances was investigated. The use of electroactive water, namely anolytes and catholytes, forgoing antiseptics or surface-active materials, helped to restore the hydration of fur skins and to remove from them soluble proteins, carbohydrates and fatty substances. The activating effect of anolyte and catholyte in solutions of water on the processes of treating raw furs is explained by their special physical and chemical properties, namely the presence of free radicals, ions and molecules of water which easily penetrate cells' membranes and into the structure of non-collagen components and microfiber structure of dermic collagen. The stage of lengthy acid and salt treatment is excluded from the technical treatment as a result of using electroactivated water with high oxidizing power. A low-cost technology of processing different kinds of fur with the use of electroactivated water provides for substantial economy of water and chemical reagents, a two to threefold acceleration of the soaking and tanning processes and creation of highly elastic fur materials with a specified set of physical and chemical properties. At the same time the technology of preparatory processes of fur treatment excludes the use of such toxic antiseptics as formalin and sodium silicofluoride, which gives grounds to regard it as ecologically safe.