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To update the accommodation mechanisms and propose a dual-wavelength, dual-function laser system for presbyopia and glaucoma treatments.Study Design: Laser sclera softening (LSS) for increased accommodation of presbyopic eyes.Place and Duration of Study: New Taipei City, Taiwan, between Jan., 2023 and Feb., 2023.Methodology: Accommodation gain (AG) can be improved by: (i) thermal shrinkage of the scleral stroma and ciliary body, or (ii) softening of the scleral stroma (with temperature range of 700C to 900C), such that the the lens front and back curvature change (or lens thickening), leading to the thickening of ciliary body and its apex, and the increase of the space of ciliary body and lens equation (SCL), and the length of the posterior vitreal zonules (PVZ) increases.Results: A novel dual-color laser system having wavelength A and B, acting on the front-zone and back-zone of the sclera, respectively, where laser-A has a deep thermal penetration the sclera and ciliary body (CB) (0.5 to 1.0 mm); and laser-B has a shallow penetration depth in the sclera (0.3 to 0.5 mm), based on the optical property of the sclera. Laser-A (having a wavelength about0.8 to 0.98 um) leads to thermal shrinkage of the ciliary body such that the CLS is increased for accommodation gain which is much more effective than the prior art.Conclusion: The increase of AG can be achieved by scleral softening and ciliary body shrinkage which increase the SCL. A proposed novel dual-color laser system acting on the front-zone and back-zone of the sclera, respectively, could provide higher AG than that of single wavelength, or prior arts using scleral ablation. However, further clinical studies are required to justified the proposed novel system with predicted advantages and efficacy based on the optical properties of sclera.
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Purpose: To derive and provide, for the first time, comprehensive analytic formulas for scleral softening volume efficacy (SVE) for accommodative gain (AG) via the increased space between ciliary body and lens (SCL) and mobility of the posterior vitreous zonules (PVZ).Study Design: To increase the AG of presbyopic eye by a new procedure, laser scleral softening (LSS).Place and Duration of Study: New Taipei City, Taiwan, between June 2022 and July 2022.Methodology: The SVE is calculated based on the time and spatial integral of the scleral temperature profiles, T(z,t), solutions of a heat diffusion equation. Analytic formulas for SVE is derived based on the covered area given by a triangle area. The SVE of a 3-D model is governed by the "volume" covered by the laser beam, or its spot size area, the effective penetration depth (z"), which is an increasing function of laser dose, but a decreasing function of the absorption coefficient (A), due to the Beer's law of laser intensity, I(z)=I0exp(-Az). The efficacy depth-range (dZ) and time-ranges (dT) are defined for efficient softening with T(z,t)>T*, where T* is the scleral softening threshold temperature.Results: The accommodative gain is proportional to the 3-D SVE given by: SEV(3D) = SEV(1D) x laser beam spot (2-D area) x total number of spots (N) acting on the sclera, which is proportional to the efficacy ranges dZ and dT, in which dZ is an increasing of laser irradiation time, whereas dT is a decreasing function of depth. Softening of the scleral tissue after a thermal laser leading to the increase of PVZ mobility and SCL. However, the actual relation of SVE and the PVZ and SCL changes require measured data.Conclusion: Safety and efficacy of scleral softening for presbyopia treatment depend upon the laser parameters (intensity, dose, spot size, wavelength) and the effective depths. The SVE is proportional to the efficacy depth-range (dZ) and time-range (dT), in which dZ is an increasing of laser irradiation time and dT is a decreasing function of depth. The AG is proportional to the SVE(in 3-D).
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Purpose: To derive and provide analytic formulas and proposed protocol for accommodative gain of presbyopia eyes via laser scleral softening, which causes increased space between ciliary body and lens (SCL) and mobility of the posterior vitreal zonules (PVZ).Study Design: To increase the accommodation of presbyopia by laser scleral heating/softening.Place and Duration of Study: New Taipei City, Taiwan, between April 2022 and June 2022.Purpose: To analyze the safety and efficacy of presbyopia treatment via scleral softening.Methodology: The scleral softening efficacy is calculated based on the rate equation of scleral tissue with a rate coefficient given by an Arrhenius formula, Temperature spatial and temporal profiles are given by the numerical solutions of a heat diffusion equation with a volume heating source. Various effective depths including tissue damage depth, temperature penetration depth and conversion depth, governed by tissue absorption coefficient, light intensity and dose (or irradiation time), and the related threshold values, are introduced in replacing the conventional penetration depth based on a Beer's law.Results: Given the the temperature spatial and temporal profiles, scleral softening efficacy can be calculated. Scleral surface damage can be prevented by cooling window. The suggested protocol for scleral softening treatments include: a diode laser at about 1.45 to 1.5 祄 or about 1.86 to1.9 祄, or about 2.0 to 2.15 祄, wavelength (with absorption coefficient about 20 to 100 cm-1); laser power about 0.2 to 0.8 W per spot, having a total of 4 to 16 spots; and irradiation time of 100 to 600 ms. Results of corneal thermal shrinkage are demonstrated by the topography changes of pig eyes, in which the scleral softening does not affect the corneal shapes. The accommodative gain is proportional to the softening efficacy (Seff) of the scleral tissue after a thermal laser leading to the increase of PVZ mobility and SCL. However, the actual relation of Seff and the PVZ and SCL changes require measured data.Conclusion: Safety and efficacy of scleral softening for presbyopia treatment depend upon the laser parameters (intensity, dose, spot size, wavelength) and the effective depths. By choosing the laser treated areas, a dual function treatment using scleral softening for presbyopia, and cornea stromal shrinkage for hyperopia is proposed and demonstrated by topography of pig eyes.
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Clinically， the incidence and mortality of malignant tumors are relatively high， especially in underdeveloped regions or countries. In recent years， with the continuous improvement of people's health awareness， living standards and medical standards， the incidence and mortality of malignant tumors have been declining. At present， malignant tumors are mostly treated by western medicine therapies in clinic， such as surgical resection or radiation therapy， chemical drug therapy， targeted drug therapy and immunotherapy. However， patients with postoperative tumors are prone to relapse and metastasis， with severe adverse reactions and a poor prognosis. And drug resistance and other issues have a serious impact on clinical efficacy and the quality of life of patients. Traditional Chinese medicine believes that malignant tumors belong to the "accumulation" and "abdominal mass"， with both internal and external etiologies. The internal etiology is mainly the insufficient anti-pathogenic energy. The external etiology is mainly six exogenous pathogenic factors to the body seven emotional stimulations. Pathogenic factors， such as deficiencies of Qi and blood， imbalance of Yin and Yang and visceral dysfunction， which lead to the occurrence of malignant tumors. The pathogenesis is mostly based on the asthenia in origin and access in superficiality. The asthenia in origin is mainly due to the insufficient anti-pathogenic energy， with Qi stagnation， blood stasis， phlegm coagulation， and toxic knot as the symptoms. For malignant tumors， like modules， the method for softening hardness to dissipate stagnation is the first choice. Chinese herbal medicine for softening hardness to dissipate stagnation is widely used for malignant tumors in clinic， with a remarkable clinical efficacy. Therefore， in recent years， anti-tumor mechanism and clinical studies of Chinese herbal medicine for softening hardness to dissipate stagnation have become a hotspot at home and abroad. This paper combines the domestic and foreign literatures of the effect of Chinese herbal medicine for softening hardness to dissipate stagnation in treating malignant tumors in both pharmacological trials and clinical research over the past cade. The progress of the studies is reviewed， in the expectation of providing a reference for the clinical anti-tumor application of Chinese herbal medicine for softening hardness to dissipate stagnation.
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BACKGROUND: Postoperative sore throat (POST) is a common adverse event after general anesthesia. The aim of this study was to evaluate the effectiveness of 2% lidocaine jelly applied on the single-lumen endotracheal tube (ETT) and thermal softening of the ETT, and a combination of both interventions on the development of POST. METHODS: Patients (n = 144) undergoing general anesthesia were randomly assigned to one of four groups: Control group (un-softened ETT lubricated with saline); Lidocaine group (un-softened ETT lubricated with 2% lidocaine jelly); Softened group (thermally softened ETT lubricated with saline); and Combined group (thermally softened ETT lubricated with 2% lidocaine jelly). Sore throat was evaluated at 0, 1, 6, 24, and 48 h after extubation. The occurrence of any postoperative complication was also assessed including hoarseness and coughing. RESULTS: No significant difference was observed in the severity of POST at all time points. However, the incidences of POST for overall (0–48 h) and the immediately following period (0 h) were significantly lower in the Combined group (52.9% and 47.1%) than in the Control group (79.4% and 76.5%), Lidocaine group (81.8% and 78.8%), and Softened group (82.9% and 74.3%). The overall incidence of hoarseness did not differ among the groups. No other postoperative complication was observed in any of the patients. CONCLUSIONS: No differences were observed in the severity of POST. However, 2% lidocaine jelly applied on thermally softened ETT reduced the overall incidence of POST. Therefore, this combined intervention could be considered as an alleviating strategy for POST.
Subject(s)
Humans , Anesthesia, General , Cough , Hoarseness , Incidence , Lidocaine , Pharyngitis , Postoperative ComplicationsABSTRACT
Decoction pieces are important raw materials in the production of traditional Chinese medicine( TCM),and their quality could directly affect the clinical efficacy and medication safety. Research on the production and processing technology of TCM is the basis for the normalization and standardization of Chinese medicine decoction pieces. At present,the production and processing standards for Scutellaria baicalensis pieces are non-regulated,lacking data foundation. In this study,with baicalin,baicalein,wogonoside and wogonin contents as evaluation indicators,single factor experiment was designed to optimize the softening,drying and cutting processes of S. baicalensis,providing a basis for the standardization of their production and processing. The effects of different softening,drying and cutting processes on the contents of the main components in S. baicalensis were comprehensively analyzed by the summation of relative differences. RESULTS:: showed that the contents of the four components and comprehensive indexes were affected by different softening methods and drying temperatures. The content of wogonin in boiling method was higher than that in boiling with cold water,and the content of glycosides in 70 ℃ drying condition was higher than that in other groups. The content of baicalin was significantly affected by different cutting thicknesses,but not by comprehensive index. Eventually,the optimal preparation process for S. baicalensis was determined as follows: boiled in boiling water for 20 min,cut into thin slices( 1-2 mm),and then dried at 70 ℃ in blast drier. This process was close to the actual production,practical and feasible and meanwhile,it was of great significance to improve the quality of S. baicalensis pieces.
Subject(s)
Desiccation , Drugs, Chinese Herbal , Reference Standards , Flavonoids , Medicine, Chinese Traditional , Quality Control , Scutellaria baicalensis , ChemistryABSTRACT
Background: Expansins play an important role in cell wall metabolism and fruit softening. Determination of expansin activity is a challenging problem since it depends on measuring cell wall properties by using ad hoc extensometers, a fact that has strongly restricted its study. Then, the objective of the work was to adapt a methodology to measure cell wall creep and expansin activity using a commercial texture meter, equipped with miniature tensile grips and an ad hoc cuvette of easy construction. Results: It was possible to measure hypocotyls acid growth and expansin activity in a reliable and reproducible way, using a commercial texture meter, common equipment found in laboratories of food science or postharvest technology. Expansin activity was detected in protein extracts from cucumber hypocotyls, tomato and strawberry fruits, and statistical differences in expansin activity were found in both fruit models at different ripening stages. Conclusions: The possibility of measuring expansin activity following this adapted protocol with a commercial texture meter could contribute to ease and increase the analysis of expansin in different systems, leading to a better understanding of the properties of these proteins under different experimental conditions.
Subject(s)
Plant Proteins/metabolism , Solanum lycopersicum/metabolism , Cucumis sativus/metabolism , Fragaria/metabolism , Plant Proteins/analysis , Cell Wall/metabolism , Hypocotyl/growth & development , Elasticity , Fruit/metabolismABSTRACT
Study on the standardization of Chinese materia medica is an important action for modernization and globalization for traditional Chinese medicine. Standardization on the processing of Chinese herbal pieces is an important part in the study on standardization of Chinese materia medica, so it is of great significance to establish the technical processing standards of Angelicae Sinensis Radix pieces for improving its quality. In this study, single factor experiment was designed to optimize the softening, cutting and drying processes of Angelicae Sinensis Radix. With ferulic acid, Angelicae Sinensis Radix polysaccharide, volatile oil and extracts (water and ethanol) content as the quality index, the effects of different softening, cutting and drying processes on the contents of the five components in Angelicae Sinensis Radix were analyzed, and the normalized distance evaluation method was used to analyze the experimental data. The results showed that the content of five components in Angelicae Sinensis Radix was affected by different softening methods and drying temperature, but the thickness of slice had little effect on the content. The best preparation process for Angelicae Sinensis Radix was as follows: Non-medicinal parts were removed; mildewed and rot as well as moth-eaten parts were removed; washed by the flowing drinking water; stacked in the drug pool; moistening method was used for softening, where 125 mL water was sprayed for every 1 kg of herbs every 2.5 h; upper part of herbs covered with clean and moist cotton, and cut into thin slices (1-2 mm) after 15 h moistening until appropriate softness, with disk thickness of 1-2 cm, then received blast drying for 6 h at 55 ℃, and turned over for 2 times during the drying.
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Notopterol, isoimperatorin, volatile oil and extract （water and ethanol) were used as the research objects in this study to investigate the effects of different softening method, slice thickness and drying methods on the quality of Notopterygii Rhizoma et Radix slices, and the experimental data were analyzed by homogeneous distance evaluation method. The results showed that different softening, cutting and drying processes could affect the content of five components in Notopterygii Rhizoma et Radix incisum. The best processing technology of Notopterygii Rhizoma et Radix slices was as follows: non-medicinal parts were removed; mildewed and rot as well as moth-eaten parts were removed; washed by the flowing drinking water; stacked in the drug pool; moistening method was used for softening, where 1/8 volume of water was sprayed for every 1 kg of herbs every 2 h; upper part of herbs covered with clean and moist cotton, and cut into thick slices (2-4 mm) after 12 h moistening until appropriate softness, then received blast drying for 4 h at 50 ℃, and turned over for 2 times during the drying. The process is practical and provides the experimental basis for the standardization of the processing of Notopterygii Rhizoma et Radix, with great significance to improve the quality of Notopterygii Rhizoma et Radix slices.
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This study was aimed to observe the effect of Softening Liver and Reducing Enzyme Mixed Agent (SLREXA) in the prevention of acute liver injury rats induced by carbon tetrachloride (CCl4). A total of 60 male SD rats were randomly divided into 6 groups, which were the SLREXA low-, middle-, high-dose group, glucurolactone group, normal group and model group. Intraperitoneal injection of CCl4 was used to induce acute liver injury rat mod-el. Intragastric administration of SLREXA was given to each Chinese medicine group. Intragastric administration of distilled water was given to the normal group and the model group. Intragastric administration of glucurolactone aque-ous solution was given to the glucurolactone group. On the 12th day of the experiment, after 16-hour fasting, rats were killed. Pathological changes in liver tissues were examined. Blood serum was determined for alanine aminotrans-ferase (ALT) and aspartate aminotransferase (AST). The liver homogenate was determined for superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), glutathione (GSH), catalase (CAT) and malondialdehyde (MDA) in liver tis-sues of rats. RT-PCR was used to detect the expression level of mRNA in liver heme oxygenase-1 (HO-1). The re-sults showed that in the microscopic examination of liver tissues, compared with the model group, different doses of SLREXA can alleviate pathological damages of liver in varying degrees. Levels of blood serum ALT and AST content in different doses of SLREXA groups and glucurolactone group were significantly lower than those of the model group (P < 0.05 or P < 0.01). Compared with the model group, contents of GSH-Px, GSH, SOD, CAT in the liver ho-mogenate were significantly increased, and MDA content was decreased significantly (P< 0.05 or P< 0.01) in differ-ent doses of SLREXA groups and glucurolactone group; compared with the model group, the HO-1 mRNA relative expression quantity in the normal group and each treatment group increased obviously, with statistical significance (P< 0.05 or P< 0.01). It was concluded that SLREXA can prevent CCl4-induced liver injury rats with definite thera-peutic effect.
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The aim of the present study was determine the O2 and CO2 levels for active modified atmosphere (MAP), besides evaluate 1-MCP effect about pulp softening delaying and skin browning during shelf life of persimmon fruit after storage in controlled atmosphere (CA) at the temperature -0.5°C. The experiment was carried in factorial arrangement (2x5) with three replications with eight fruit each. After storage plus shelf life was not found significant interaction on pulp softening, but fruit submitted to 1.0 kPa O2 during shelf life in MAP showed lower softening. However, for skin browning was observed significant interaction. The use of highly CO2 levels during storage at -0.5°C promotes higher skin browning in all the shelf life MAP conditions, except on fruit of control treatment, after six days of shelf life at 20°C. Partial pressure of 1.0 kPa O2 during MAP shelf life is the best condition for reduction pulp softening and skin browning of 'Fuyu' persimmon. Partial pressure of 6.0 kPa CO2 during storage in CA with 0.15 kPa O2, in cold storage (-0.5°C) keep higher skin browning during MAP shelf life. The use of 1-MCP no brink effect in low O2 level during MAP shelf life at 20°C.
O objetivo do presente trabalho foi determinar os níveis de O2 e CO2 para armazenamento em atmosfera modificada ativa (AMA), além de verificar o efeito do 1-MCP sobre o retardamento do amaciamento da polpa e escurecimento da epiderme durante a vida de prateleira de caquis armazenados em AC na temperatura de -0,5°C durante 14 semanas. O experimento foi conduzido em fatorial (2x5) com três repetições com oito frutos cada. Após o armazenamento mais o período de vida de prateleira não se verificou interação entre os fatores para amaciamento de polpa, sendo que os frutos submetidos a 1,0 kPa de O2 durante a vida de prateleira em AMA apresentaram menor amaciamento. Para incidência de escurecimento da epiderme ocorreu interação. O uso de alto CO2 durante o armazenamento a -0,5°C causou maior escurecimento da epiderme em todas as condições de AMA na vida de prateleira, exceto na testemunha, após seis dias de exposição a 20°C. Pressão parcial de 1,0 kPa de O2 durante a vida de prateleira é a melhor condição de AMA para manter o caqui 'Fuyu' com menor amaciamento e escurecimento da epiderme. Pressão parcial de 6,0 kPa CO2 durante o armazenamento em AC com 0,15 kPa O2, na temperatura de -0,5°C causa maior escurecimento da epiderme durante a vida de prateleira. O 1-MCP não tem efeito em baixa concentração de O2 durante a vida de prateleira em AMA a 20°C.
Subject(s)
Crop Production , DiospyrosABSTRACT
Objective To develop a equipment of vehicular multifunctional water,and the problem of water in disaster medical rescue was solved. Methods According to the features of the field water in earthquakes,floods and other disasters,a kind of disaster relief in the wild integrated production,supplying softening water and water purification technology was created,and a system by the pretreatment system, ion exchange,reverse osmosis systems,and storage and transportation system and other parts of the small truck-mounted multifunctional water treatment equipment was developed. Results The vehicular prototypes of multifunctional water treatment machine was developed,the pond water and river water were tested to prove that the throughput of the equipment in production and supply in the land of disaster relief,the puri-fied water and softening water reached 2 080 L/day and 12 100 L/day,which could meet the conducted large-scale medical treatment de-mand. Conclusion The successful design of the vehicular multifunctional water processor carried out the medical rescue with water prob-lem,and provided a new method and new equipment,which was worthy of popularization and application.
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Una de las mayores causas de pérdidas poscosecha de frutos de pitaya amarilla es su ablandamiento excesivo, el cual ha sido documentado previamente cuando la fruta es almacenada a temperaturas de cosecha o después de refrigeración. Además, tratamientos de choque térmico antes del almacenamiento refrigerado ofrecen control en el ablandamiento de estos frutos. Diferentes experimentos fueron llevados a cabo para evaluar el papel de algunas enzimas degradadoras de pared celular en el ablandamiento de frutos de pitaya amarilla: almacenamiento a 18 °C (TA) y refrigeración con choque térmico previo (ChT-R). Se incluyó también un tratamiento refrigerado control, sin choque térmico (control-R). Si midió el color de la corteza, la firmeza y las actividades de poligalacturonasa (PG), celulasa (CEL) y xilanasa (XIL). La evaluación del color indicó que los frutos almacenados a TA alcanzaron su madurez comercial luego de seis días. Luego de 12 días de almacenamiento a TA el pardeamiento y ablandamiento excesivo afectaron negativamente la calidad de los frutos. El pardeamiento y ablandamiento excesivo fueron detectados también en los frutos control-R cuando se movieron de 2 a 18 °C. Un ligero pardeamieno fue observado en los frutos ChT-R. Estos frutos alcanzaron su madurez comercial luego de 24 días de almacenamiento (nueve días luego de terminado el almacenamiento refrigerado). La actividad de XIL se asoció al ablandamiento en los frutos almacenados a TA y ChT-R. No se observó una clara correlación entre las actividades de PG y el ablandamiento, como tampoco entre CEL y el ablandamiento.
One of the major causes of yellow pitaya fruit loss during its marketing is its excessive softening, which has been previously documented when the fruit is stored at harvest temperature or after refrigeration. Furthermore, its excessive softening has been controlled by the application of heat shock treatments before refrigeration. Different experiments were carried out to evaluate the role of cell wall degrading enzymes on yellow pitaya fruit softening: storage at 18 °C (RT) as well as refrigeration with previous heat shock treatment (HS-R). A refrigerated control, without heat shock, was included (control-R). Peel color, firmness, poligalacturonase (PG), celulase (CEL) and xilanase (XIL) activities were measured. RT fruits reached the commercial ripeness after six days, as indicated by the color evaluation. After 12 days of storage at RT browning and excessive softening negatively affected the fruit quality. Browning and excessive softening were also detected in the control-R fruit when moving from 2 to 18°C. Minor browning was found in the HS- R fruit. HS-R fruit was full ripe 24 days of storage (nine days after finishing the refrigerated storage). XIL activity was associated to the softening in the RT and HS-R fruits. No clear correlation was observed between PG and softening neither between CEL and softening.
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Durante el periodo de poscosecha el principal problema de deterioro del lulo (Solanum quitoense Lam) es el ablandamiento que es generado principalmente por actividad de enzimas pécticas que atacan la red estructural de la pared celular. Esta investigación se basó en la búsqueda de las mejores condiciones de extracción y medida de actividad de las enzimas pectinesterasa, poligalacturonasa y pectato liasa; herramientas necesarias para estudiar posteriormente el rol de estas enzimas en el deterioro por ablandamiento sufrido por el fruto debido a diversos cambios metabólicos. Se encontró que las dos primeras enzimas pueden ser extraídas simultáneamente con buffer fosfatos 20 mM pH 7,0 + NaCl 0,06 M y 60 min de extracción, relación 1:2 (material vegetal: buffer de extracción), a su vez, pectato liasa se extrajo con buffer fosfatos 20 mM pH 7,0 + cisteína 20 mM y 30 min de extracción, relación 1:3. Para la cuantificación de la actividad pectinesterasa es necesario incubar 15 min a 42 °C 2.500 µL de extracto enzimático crudo (EE) en buffer fosfatos 20 mM pH 7,0 + NaCl 0,15 M y 1,6% de pectina cítrica como sustrato, con valores de Km aparente de 3,78% de PC y Vmax 17,95 µmolH+/min*mg prot. Para la cuantificación de la actividad poligalacturonasa es necesario incubar 15 min a 37 °C 30 µL (EE) en buffer acetatos 200 mM pH 4,5 + NaCl 0,25 M y 1,0% de APG como sustrato, con valores de Km aparente 0,141% de APG y Vmax 28,46 nKat/s*mg prot. Para la cuantificación de la actividad pectato liasa es necesario incubar 2 min a 17 °C 100 µL (EE) en buffer TRIS:HCl 50 Mm pH 8,5 + CaCl2 4 mM y 0,1% de APG como sustrato, con valores de Km aparente 0,0865% de APG y Vmax 82,75 µg/s*mg prot.
The main problem of post-harvest deterioration of lulo (Solanum quitoense Lam) is the softening is the main problem of post-harvest deteriorarion of Lulo, that is generated mainly by the activity of pectic enzymes, which attack the structural network of the cell wall. This research was based on finding the best conditions structural cell wall network for extraction and measurement of enzyme activity pectinesterase (PE), polygalacturonase (PG) and pectato liasa (PL); tools needed to study the further role of these enzymes in the deterioration of pectatelyase fruit softening, due to various metabolic changes. It was found that the first two enzymes can be extracted simultaneously with 20 mM phosphate buffer pH 7.0, 0.06 M NaCl and 60 minutes of extraction, ratio 1:2 (plant material: extraction buffer), pectatelyase extracted with 20 mM phosphate buffer pH 7.0, 20 mM cysteine and 30 minutes of extraction, ratio 1:3. For quantification of pectinesterase activity is necessary to incubate 15 minutes at 42 ° C, 2500 µL of crude enzyme extract (EE) in 20 mM phosphate buffer pH 7.0, to 0.15 M NaCl and 1.6% citrus pectin as (CP) substrate with apparent Km values of 3.78% CP and Vmax 17.95 mol H+/min * mg prot. For the quantification of pectinesterase activity is necessary to incubate 15 minutes to 42 °C 2500 µL of crude enzyme extract (EE) in 20 mM phosphate buffer pH 7.0, 0.15 M NaCl and 1.6% citrus pectin as substrate with apparent Km values of 3.78% CP and 17.95 µ Vmax mol H+/min*mg prot. For the quantification of polygalacturonase activity is necessary to incubate 15 minutes to 37 °C 30 µL (EE) in 200 mM Acetate buffer pH 4.5, 0.25 M NaCl and 1.0% of APG as substrate, with apparent Km values 0.141% of APG and Vmax 28.46 nKat/s*mg prot. For the quantification of the pectatelyase activity is necessary to incubate 2 minutes to 17 °C, 100 µL (EE) in buffer TRIS: HCl pH 8.5, 50 mM 4 mM CaCl2 and 0.1% PGA as substrate, with apparent Km values 0.0865% of APG and Vmax 82.75 µg/s*mg prot.
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En diversas técnicas aplicadas para la conservación en fresco de la pitaya amarilla (Acanthocereus pitajaya) se ha encontrado que el ablandamiento excesivo de su corteza contribuye al deterioro de su calidad. Puesto que pectinmetilestearasa (PME) se ha vinculado con el ablandamiento de frutos este estudio se desarrolló con el objeto de determinar el efecto de la incorporación de los aditivos tritón X-100, NaCl y cisteína en buffer fosfatos 20 mM pH 7,0 sobre la cantidad de proteína extraída y sobre la actividad de PME. También se evaluó la necesidad de recurrir al proceso de diálisis en buffer fosfatos 20 mM pH 7,0. En la medida de actividad se pusieron a punto el tiempo de incubación, la concentración del cofactor NaCl, pH, temperatura y concentración de sustrato (pectina cítrica). Se encontró que el mejor sistema de extracción fue el compuesto por buffer fosfato 20 mM, pH 7,0 con concentraciones de NaCl que pueden estar entre 0,0 a 1,0 M. La medida de actividad se puede realizar empleando pectina cítrica entre 0,40 a 0,75%, a valores de pH entre 5,0 a 8,0, con incubación a una temperatura entre 40 a 45 °C, durante 2,5 min.
Using diverse techniques applied to keep the freshness of yellow pitaya (Acanthocereus pitajaya) fruit it has been found that excessive softening of its crust leads to quality deterioration. Since pectinmethyl esterase (PME) has been related to fruit softening in this study we evaluated the protein levels and the PME activity after the addition of Triton X-100 1% and NaCl in concentrations from 0 to 2 M in buffer 20 mM phosphate pH 7.0. Effects of cysteine addition and dialysis were also evaluated for the extraction processes. Factors that can affect the activity of PME such as incubation time, different NaCl concentration, as value level of pH during the incubation, temperature and pectin (citric pectin) concentration were evaluated. The best system found in this study for PME extraction was buffer phosphate 20 mM, pH 7.0 and NaCl from 0.0 to 1.0 M. The best system for the activity measurement is to use pectin from 0.40 to 0.75%, keep the pH between 5 and 8 and incubate from 40 to 45 °C during 2.5 min.
ABSTRACT
Para pitaya amarilla (Acanthocereus pitajaya) se ha encontrado que el ablandamiento excesivo de la cáscara contribuye al deterioro del fruto, al aplicar diferentes técnicas de conservación en fresco. Dado que tanto la celulasa como la xilanasa se han vinculado con el ablandamiento de la cáscara de frutos, este trabajo se basó en la búsqueda de las mejores condiciones de extracción y medida de actividad de celulasa y xilanasa. El mejor sistema de extracción fue buffer fosfato 20 mM, NaCl 0,5 M, pH 7,0. Para la medida de actividad de celulasa es necesario incubar durante 60 min a 37 ºC, con un volumen de extracto enzimático crudo de 30 µL, empleando buffer acetato 100 mM a pH 5,0; los valores de constante aparente de Michaelis Menten (K M aparente) y velocidad máxima (V MÁX) fueron 0,279 mg/mL y 0,00014 nmol glucosa/min, respectivamente. Para determinar la actividad de xilanasa se establecieron 15 min de tiempo de incuba-ción, a 50 ºC, empleando 30 µL de extracto enzimático crudo a pH 4,0 (buffer acetato 100 mM); los valores de K M aparente y V MÁX para xilanasa fueron 0,073 mg/mL y 0,0011 nmol glucosa/min, respectivamente.
By applying different conservation techniques on yellow pitaya fruit (Acanthocereus pitajaya) it has been found that excessive softening of the peel contributes to the deterioration of the fruit. Due to that both cellulase and xylanase have been related to the softening of the fruit's peel; this work was based on the search of the best conditions not only for the extraction, but also for the activity measurement of both cellulase and xylanase. The best extraction system for both enzymes was 20 mM buffer phosphate, 0.5 M NaCl, pH 7.0. For the cellulase activity measurement it was necessary to incubate during 60 min at 37 ºC, with a volume of raw enzymatic extract of 30 µL, using buffer acetate 100 mM at pH 5,0; the values of apparent K M and V MÁX were 0.279 mg/mL and 0.00014 nmol glucose/min, respectively. To determine the xylanase activity it was necessary to incubate during 15 min, at 50 ºC, using 30 µL of raw enzymatic extract at pH 4.0 with 100 mM buffer acetate; the values of apparent K M and V MÁX for this enzyme were 0.073 mg/mL and 0.0011 nmol glucose/min, respectively.
ABSTRACT
OBJECTIVE:To give an account of the preparation process and quality control of a scar softening ointment METHODS:The scar softening ointment was prepared with zine oxide,Chinese gall,centiped and camphor as crude materals,and all major ingredients were identified and quantitatively determined according to the methods in CHINESE PHAMACOPOEIA Besides,stabitity test and skin irritation test were performed The therapeutic effect was observed in 50 patients RESULTS:The scar softening ointment was reliable in quality and non-irritative in clinical application with a total effective rate of 92 0% CONCLUSION:The preparation is simple in quality control,good in stability and valid in therapeutic effects,and can be used for clinical therapy