Acidic Environment Induces Pyroptosis of NLRP3 Inflammatory Vesicle-Mediated Nucleus Pulposus Cells Through Up-Regulation of POSTN Expression

SUMMARY: Intervertebral disc degeneration (IVDD) is induced by nucleus pulposus (NP) dysfunction as a result of massive loss of NP cells. It has been reported that the acidic microenvironment of the intervertebral disc (IVD) can induce NP cell pyroptosis, and that up-regulation of periostin (POSTN) expression has a negative effect on NP cell survival. However, the relationship between the acidic environment, POSTN expression level and NP cell pyroptosis is unclear. Therefore, the aim of this study was to explore the relationship between acidic environment and POSTN expression level in NP cells, as well as the effect of POSTN in acidic environment on NP cell pyroptosis. NP cells were obtained from the lumbar vertebrae of Sprague Dawley (SD) male rats. These cells were divided into normal and acidic groups according to whether they were exposed to 6 mM lactic acid solution. And NP cells in the acidic group were additionally divided into three groups: (1) Blank group: no transfection; (2) NC group: cells transfected with empty vector plasmid; (3) sh-POSTN group: cells transfected with sh-POSTN plasmid to knock down the expression level of POSTN. Quantitative real-time PCR (qRT-PCR) and western blot was performed to assess the expression of POSTN at the mRNAand protein levels. CCK8 was used to evaluate cell survival. Western blot, in addition, was performed to examine acid-sensing ion channels (ASIC)-related proteins. And pyroptosis was detected by ELISA and western blot. The expression level of POSTN was significantly increased in NP cells in acidic environment. Knockdown of POSTN expression promoted the survival of NP cells in acidic environment and reduced the protein levels of ASIC3 and ASIC1a in NP cells. Moreover, knockdown of POSTN expression decreased the pyroptosis proportion of NP cells and the levels of pro-inflammatory cytokines interleukin (IL)-1β and IL-18. The levels of pyroptosis-related proteins NLRP3, ASC, cleaved-Caspase-1, and cleaved-GSDMD were also affected by the decreased POSTN expression. The extracellular acidic environment created by lactic acid solution activated NLRP3 inflammatory vesicle-induced caspase-1 to get involved in NP cell pyroptosis by up-regulating POSTN expression.

La degeneración del disco intervertebral (DDIV) es inducida por una disfunción del núcleo pulposo (NP) como resultado de una pérdida masiva de células NP. Se ha informado que el microambiente ácido del disco intervertebral (DIV) puede inducir la piroptosis de las células NP y que la regulación positiva de la expresión de periostina (POSTN) tiene un efecto negativo en la supervivencia de las células NP. Sin embargo, la relación entre el ambiente ácido, el nivel de expresión de POSTN y la piroptosis de las células NP es poco clara. Por lo tanto, el objetivo de este estudio fue explorar la relación entre el ambiente ácido y el nivel de expresión de POSTN en células NP, así como el efecto de POSTN en ambiente ácido sobre la piroptosis de las células NP. Las células NP se obtuvieron de las vertebras lumbares de ratas macho Sprague Dawley (SD). Estas células se dividieron en grupos normales y ácidos según se expusieron a una solución de ácido láctico 6 mM. Las células NP en el grupo ácido se dividieron adicionalmente en tres grupos: (1) Grupo en blanco: sin transfección; (2) grupo NC: células transfectadas con plásmido vector vacío; (3) grupo sh-POSTN: células transfectadas con plásmido sh-POSTN para reducir el nivel de expresión de POSTN. Se realizó una PCR cuantitativa en tiempo real (qRT-PCR) y una transferencia Western para evaluar la expresión de POSTN en los niveles de ARNm y proteína. Se utilizó CCK8 para evaluar la supervivencia celular. Además, se realizó una transferencia Western para examinar las proteínas relacionadas con los canales iónicos sensibles al ácido (ASIC). La piroptosis se detectó mediante ELISA y Western blot. El nivel de expresión de POSTN aumentó significativamente en células NP en ambiente ácido. La eliminación de la expresión de POSTN promovió la supervivencia de las células NP en un ambiente ácido y redujo los niveles de proteína de ASIC3 y ASIC1a en las células NP. Además, la eliminación de la expresión de POSTN disminuyó la proporción de piroptosis de las células NP y los niveles de citocinas proinflamatorias interleucina (IL) - 1β e IL-18. Los niveles de proteínas relacionadas con la piroptosis NLRP3, ASC, Caspasa-1 escindida y GSDMD escindida también se vieron afectados por la disminución de la expresión de POSTN. El ambiente ácido extracelular creado por la solución de ácido láctico activó la caspasa-1 inducida por vesículas inflamatorias NLRP3 para involucrarse en la piroptosis de las células NP mediante la regulación positiva de la expresión de POSTN.

Advances on the production of organic acids by yeast / 生物工程学报

Organic acids are organic compounds that can be synthesized using biological systems. They often contain one or more low molecular weight acidic groups, such as carboxyl group and sulphonic group. Organic acids are widely used in food, agriculture, medicine, bio-based materials industry and other fields. Yeast has unique advantages of biosafety, strong stress resistance, wide substrate spectrum, convenient genetic transformation, and mature large-scale culture technology. Therefore, it is appealing to produce organic acids by yeast. However, challenges such as low concentration, many by-products and low fermentation efficiency still exist. With the development of yeast metabolic engineering and synthetic biology technology, rapid progress has been made in this field recently. Here we summarize the progress of biosynthesis of 11 organic acids by yeast. These organic acids include bulk carboxylic acids and high-value organic acids that can be produced naturally or heterologously. Finally, future prospects in this field were proposed.

Effectiveness of ultrasonic activation associated to glycolic acid on smear layer removal, dentin structure and bond strength to root dentin

This study evaluated the effects of ultrasonic activation (US) associated to glycolic acid (GA) on smear layer, dentin structure and bond strength (BS) of filling/restorative material to root dentin. The roots were used for antimicrobial activity, dentin structure and BS evaluation, being distributed into seven groups, according to irrigation protocols: G1:DW+US; G2:17% EDTA; G3:QMix; G4:17% GA; G5:17% EDTA+US; G6:QMix+US; G7:17% GA+US. Scanning electronic microscopy, transmission electronic microscopy and push-out were performed, with specific statistical analysis for each evaluation. The highest smear layer removal occured in Groups 6 and 7 (p<0.05), and the largest collagen dispersion in Group 7, being similar to Group 2 and 5 (p>0.05). The highest BS of filling and restorative material occurred in Groups 6 and 7, and Groups 5, 6 and 7, respectively, being similars between them (p>0.05). The use of GA+US promoted effective smear layer removal and dentin structure preservation, improving the BS of filling/restorative material to root dentin.

Effect of Lactobacillus coryniformis FZU63 on the flavor quality of black tea beverage / 生物工程学报

The tea beverages will be endowed with distinct aroma and taste, as well as various biologically active compounds including probiotic factors, when fermented with lactic acid bacteria (LAB). However, at present, few studies on the dynamics of flavors in tea soup at different fermentation stages were conducted. In this study, the composition of monosaccharides, aromatic components, free amino acids, and organic acids were measured, when the black tea beverages were fermented with Lactobacillus coryniformis FZU63 which was isolated from Chinese traditional kimchi. The results indicated that monosaccharides including glucose, fructose, mannose and xylose in black tea beverages are the main carbon sources for fermentation. In addition, the abundance of aromatic compounds in black tea soup are increased significantly at different fermentation stages, which endow the fermented black tea soup with fruit aroma on the basis of flowery and nutty aroma. Moreover, some bitter amino acids are reduced, whereas the content of sweet and tasty amino acids is elevated. Furthermore, the levels of lactic acid, malic acid, citric acid and other organic acids are accumulated during the fermentation. Additionally, sensory evaluation displays that black tea beverage is acquired with comprehensive high-quality after being fermented for 48 h. This study provides a theoretical basis to steer and control the flavor formation and quality of the fermented tea beverages during LAB fermentation.

Bioavailability augmentation of silymarin using natural bioenhancers: An in vivo pharmacokinetic study

Abstract Pharmacokinetic studies were carried out in male and female rats to quantify silymarin as silybin (A+B) after the oral administration of various silymarin formulations combined with three bioenhancers, namely, lysergol, piperine, and fulvic acid, and compared with plain silymarin formulation (control). A non-compartmental analysis, model independent analysis, was utilized, and various pharmacokinetic parameters (C max, T max, and AUC 0-t) were calculated individually for each treatment group, and the values were expressed as mean ± SEM (n = 6). Plasma samples obtained from the rats were analyzed for the concentration of silymarin through a validated RP-HPLC method and on the basis of data generated from the pharmacokinetic studies. Results indicated that the bioenhancers augmented pharmacokinetic parameters and bioavailability increased 2.4-14.5-fold in all the formulations compared with the control. The current work envisages the development of an industrially viable product that can be further subjected to clinical trials and scientifically supports the development of silymarin as a contemporary therapeutic agent with enhanced bioavailability and medicinal values.

HPLC quantitative analysis of protocatechuic acid contents in 11 Phellinus mushroom species collected in Thailand

Abstract Phellinus mushrooms have been traditionally used for various medicinal purposes. Protocatechuic acid, which was previously reported to be a component in some Phellinus mushrooms, has some pharmacological effects. This study aimed to validate a HPLC method for the quantitative analysis of the protocatechuic acid contents in the extracts from different Phellinus mushroom species collected in Thailand. HPLC was carried out using a C18 column and the gradient mobile phases of 0.1% formic acid in water and 0.1% formic acid in acetonitrile. Method validation was performed to assure the linearity, accuracy, precision, limit of detection and limit of quantitation of the analytical method. The linearity range of protocatechuic acid was 1 - 10 µg/ml. The average recovery was 104.16%. The method was shown to be precise with the RSD of repeatability and intermediate precision at less than 3%. The protocatechuic contents in 11 Phellinus mushrooms were in the range of less than 0.0099 - 0.4121 %w/w of the extract. The developed HPLC method was reliable and suitable for the quantitative analysis of protocatechuic acid content in Phellinus mushrooms.

Biopelícula cariogénica de Streptococcus mutans y efecto hemostático de ácido tánico en la hemorragia post-exodoncia / Streptococcus mutans cariogenic biofilm and hemostatic effect of tannic acid in post-extraction hemorrhage

La caries dental y la placa dental se encuentran entre las enfermedades más comunes en todo el mundo y son causadas por una mezcla de microorganismos y restos de alimentos. Tipos específicos de bacterias productoras de ácido, especialmente Streptococcus mutans, colonizan la superficie dental y causan daño a la estructura dental dura en presencia de carbohidratos fermentables, por ejemplo, sacarosa y fructosa. Por otro lado, el sangrado posterior a la extracción es una complicación reconocida y frecuente en la práctica dental, que se define como la pérdida de sangre que continúa más allá de las 8 a 12 horas después de la exodoncia. Existe una amplia gama de técnicas sugeridas y sustancias para el tratamiento del sangrado post-extracción, que incluyen intervenciones dirigidas tanto a causas locales como sistémicas. El ácido tánico es una de las sustancias astringente que precipitan proteínas, pero no penetran en las células, por lo que inciden solo en la capa superficial. Su objetivo se enfoca a robustecer la superficie para acrecentar su resistencia mecánica y reducir la exudación. El objetivo de este estudio fue determinar la presencia de S. mutans en las biopelículas dentales y al mismo tiempo evaluar la actividad del ácido tánico como inhibidor del sangrado profuso en las extracciones dentales. S. mutans se aisló en el 92,5% de los pacientes evaluados. Los tiempos de hemostasia post-exodoncia fue significativamente menor en el grupo de pacientes a los que se les aplicó el ácido tánico en comparación a los que no se les aplicó(AU)

Tooth decay and dental plaque are among the most common diseases worldwide and are caused by a mix of microorganisms and food debris. Specific types of acid-producing bacteria, especially Streptococcus mutans, colonize the tooth surface and cause damage to hard tooth structure in the presence of fermentable carbohydrates, for example, sucrose and fructose. On the other hand, post-extraction bleeding is a recognized and frequent complication in dental practice, defined as blood loss that continues beyond 8 to 12 hours after extraction. There is a wide range of suggested techniques and substances for the treatment of post-extraction bleeding, including interventions targeting both local and systemic causes. Tannic acid is one of the astringent substances that precipitate proteins, but does not penetrate the cells, so it affects only the superficial layer. Its objective is focused on strengthening the surface to increase its mechanical resistance and reduce exudation. The objective of this study was to determine the presence of S. mutans in dental biofilms and at the same time to evaluate the activity of tannic acid as an inhibitor of profuse bleeding in dental extractions. S. mutans was isolated in 92.5% of the patients evaluated. Post-extraction hemostasis times were significantly shorter in the group of patients who received tannic acid compared to those who did not(AU)

Spectrophotometric determination of acid dissociation constants of some arylpropionic acids and arylacetic acids in acetonitrile-water binary mixtures at 25ºC

Abstract The acid dissociation constant of drug active compounds (arylpropionic and aryl acetic acids) were determined in acetonitrile and water binary mixtures (corresponding volume fractions of 0.40, 0.45, 0.50, and 0.55) by using a multi-wavelength spectrophotometric method. Drug active compounds, which were slightly soluble in water, were studied in these binary mixtures. The dissociation constants of drug active compounds are important in drug design studies and in any research of the biopharmaceutical and physicochemical properties of drugs. The STAR program was used for the determination of dissociation constants. The acidity constants of arylpropionic and aryl acetic acids were correlated with the Kamlet and Taft solvaatochromic parameters. Aqueous pKa values of these arylpropionic and aryl acetic acids were determined from pKa values obtained from acetonitrile and water binary mixtures with varying volume fractions. The studied drugs had a pKa value corresponding to base functional group. Results showed that the acid dissociation constant values of the drug active compounds increased with an increase in acetonitrile content in the medium.

New and potential properties, characteristics, and analytical methods of ferulic acid: A review

Abstract Phenolic compounds are widely distributed in the plant kingdom and in the microorganisms. Cinnamic acid and its hydroxylated derivative-ferulic acid, are phenolic compounds. Ferulic acid possesses antioxidant potential, as well as anti-cancer, anti-inflammatory, and antimicrobial properties. It prevents the harmful effects of radiation both as an ultraviolet absorber and as a free radical scavenger; it is not cytotoxic. Although ferulic acid has beneficial properties, it is hardly used in cosmetic preparations and has been rarely studied in the literature. Herein, we review the literature on ferulic acid, to provide information which can contribute to further research on the compound.

Craniofacial and neurological manifestations in patients with Guillain-Barré syndrome / Manifestaciones neurológicas craneofaciales en pacientes con síndrome de Guillain-Barré

Introducción: Among the broad range of symptoms of the Guillain-Barré Syndrome (GBS), patients can present craniofacial manifestations. Consequently, the participation of the dental surgeon in the evaluation of the stomatognathic system using a multidisciplinary approach model is very important. Objective: To identify the craniofacial and neurological manifestations of the Guillain-Barré Syndrome in patients from three hospitals in the district of Lambayeque, Peru, in 2019. Material and Methods: A descriptive, prospective, and cross-sectional study was carried out in 59 patients diagnosed with GBS by means of a clinical evaluation carried out by previously calibrated neurologists (k=0.911). Additionally, an oral evaluation of the oral hygiene index, the periodontal status, and the oral pH was carried out. Data were processed using frequency distribution tables. Results: 4 4.07% of the patients presented at least one functional alteration, such as unilateral limitation for facial expressions (25.42%), dysphagia (18.64%), alteration in phonation (11.86%), and loss of the sense of taste (6.78%), showing involvement of the corresponding cranial nerves. In addition, unfavorable conditions of oral hygiene and acid pH were observed in some cases, which proved adverse for the periodontal structures of the patients. Conclusion: The study found craniofacial manifestations of GBS, such as cases of unilateral facial palsy, dysphagia, difficulty in phonation, and loss of the sense of taste, due to the alteration of the cranial nerves that command these functions. In addition, poor oral hygiene, and the presence of acidic pH in some patients created an adverse environment for the integrity of the periodontal structures

Introducción: El Síndrome de Guillain-Barré (SGB) dentro de su amplia sintomatología puede presentar manifestaciones craneofaciales, por lo que resulta importante la participación del cirujano dentista en la evaluación del sistema estomatognático desde un modelo de abordaje multidisciplinario. Objetivo: Identificar las manifestaciones neurológicas craneofaciales del Síndrome de Guillain-Barré en pacientes de tres establecimientos de salud de Lambayeque, Perú en el año 2019. Materiales y Metodos: Se realizó un estudio descriptivo, prospectivo y transversal, con 59 pacientes diagnosticados con SGB, mediante una evaluación clínica realizada por médicos neurólogos previamente calibrados (k=0,911). Adicionalmente se realizó una evaluación bucodental del índice de higiene oral, del estado periodontal y del pH bucal, siendo procesados los datos mediante tablas de distribución de frecuencias. Resultados: El 44,07% de pacientes presentaron por lo menos una alteración funcional, identificando limitación unilateral para expresiones faciales (25.42%), disfagia (18.64%), alteración en la fonación (11.86%) y pérdida del sentido del gusto (6.78%), evidenciando compromiso de los pares craneales correspondientes. Además se observaron condiciones desfavorables de higiene oral y de pH ácido en algunos casos, que resultaron adversas para las estructuras periodontales de los pacientes. Conclusión: Fueron encontradas manifestaciones craneofaciales del SGB, identificando casos con parálisis facial unilateral, disfagia, dificultad para la fonación y pérdida del sentido del gusto, debido a la alteración de los pares craneales que comandan esas funciones. Además las condiciones desfavorables de higiene oral y la presencia de pH ácido en algunos casos, propiciaron un ambiente adverso para la integridad de las estructuras periodontales.

Acrocarpus fraxinifolius Wight and Arn. Bark; phenolics, toxicity studies, antioxidant and anti-inflammatory activities

The purpose of the survey was to determine acute & chronic toxicity; in vivo antioxidant and anti-inflammatory actions of the different extracts of A. fraxinifolius Wight and Arn bark; along with estimation of the phenolic, flavonoidal contents and investigation of phenolic metabolites that may attribute to the activities. LD50 of the total ethanol extract (TEE) was 7.1 g/kg b. wt, the radical scavenging activity of DPPH showed 60.31% inhibition, FRAP ability and ABTS+ activity showed 55.024 and 67.217 µmol Trolox/100 g dry weight, respectively. TEE followed by ethyl acetate extract (EAE) at 100 mg/kg b.w exhibited the highest in vivo antioxidant activity (94.51% and 91.08% potency, respectively) compared with Vit E (100%). The TEE & EAE exhibited the highest anti-inflammatory activity (3.81±0.08 & 3.79±.0.04) respectively in comparison with indomethacin 3.83±0.01 measured as edema diameter after 4 hours of extract administration. The total phenolic and total flavonoid contents in the total ethanol extract (TEE) estimated as gallic acid and catechin equivalents were 61.06± 0.08 µg eq GA/g, 40.33± 0.20 µg CE/g extract respectively. EAE revealed five phenolic acids and eight flavonoid compounds isolated for the first time from the plant

Evaluation of Sealing Ability of Biodentine to Root-End Cavities Irrigated either with Maleic Acid or Irritrol Using Glucose Filtration Model

ABSTRACT Objective: To compare the effect of Maleic acid and Irritrol (combination of Chlorhexidine & Ethylenediaminetetraacetic acid) irrigation on the sealing ability of Biodentine when used as root-end filling material. Material and Methods: Thirty single-rooted human premolars were subjected to standardized root canal instrumentation and were irrigated with 2.5% Sodium Hypochlorite (NaOCl) after each instrument change. Apical root resections were done by removing 3 mm of the apex at a 90° angle to the long axis of the root with a diamond bur. The root end cavity preparation was done using ultrasonic tips at a low power setting under water coolant. They were randomly divided into 3 groups (n=10). Group 1 was irrigated with 7% Maleic acid, Group 2 was irrigated with Irritrol, and Group 3 was irrigated with 0.9% Saline (Control). Then, the root end cavities of all the samples were restored with Biodentine and were subjected to leakage analysis using a glucose filtration test. The mean values for each group were calculated and the obtained data was statistically analysed using ANOVA with post-hoc Tukey's test, ANOVA with post-hoc Games-Howell test and repeated measures ANOVA with post-hoc Bonferroni test. Results: The specimens irrigated with Irritrol showed least microleakage as compared to 7% Maleic acid and 0.9% Saline. Conclusion: A regimen involving irrigation of the root-end cavity with Irritrol followed by root-end filling with Biodentine can improve the apical seal.

Advances in the metabolic engineering for the production of tetracarbon organic acids / 生物工程学报

Tetracarbon organic acids are important platform chemicals that are widely used in the food, chemical, medicine, material industries and agriculture. Compared with the traditional petrochemical process, the production of tetracarbon organic acids by microbial fermentation is more promising due to milder reaction conditions, greener process and better environmental compatibility. This review summarizes the biosynthetic pathways and metabolic mechanisms for the production of tetracarbon organic acids, and illustrates recent advances, challenges, and future perspectives in the production of tetracarbon organic acids by naturally selected or purposefully engineered strains.

Evaluation of the action of precipitating agents to obtain allergenic proteins from crude mite extracts / Avaliação da ação de agentes precipitantes na obtenção de proteínas alergênicas presentes em extratos brutos de ácaros

Introduction: Allergen proteins found in dust mite extracts, such as Dermatophagoides farinae (DF), Dermatophagoides pteronyssinus (DP) and Tyrophagus putrescentiae (TP), are relevant for scientific studies in the allergy and immunotherapy fields. The precipitation/concentration of protein extracts may favor the aggregation of the allergens in homogenates. Objective and method: This paper investigates the precipitation process by submitting crude mite extracts to compounds such as ammonium sulfate (NH4)2SO4, trichloroacetic acid (TCA) and acetone. Results: The best results were obtained by fractionation with (NH4)2SO4 at 80% (w/v) saturation (~0° C), observing the protein markings on the electrophoresis gel. Major allergens were identified by immunoblot at 25 kDa (cysteine protease) for Der f and Der p; and 25 kDa, 30 kDa (tropomyosin) and Try p 3, near 26 kDa. For this percentage the total protein contents were 12.83 mg mL-1 for Der f, 24.78 mg mL-1 for Der p and 27.35 mg mL-1 for Try. Conclusion: An advantage of precipitation with (NH4)2SO4 over precipitation with acetone was the possibility of gradually obtaining protein fractions, which does not happen when using the latter. The addition of 80% (v/v) acetone to the mite extracts favored total protein precipitation in the concentrations 16.42 mg mL-1, 28.47 mg mL-1 and 13.41 mg mL-1. The use of TCA in concentrations above 20% (w/v) forms peptides that are not retained in the gel under the established experimental conditions, and dilute solutions of this acid are more efficient.

Introdução: As proteínas alergênicas presentes nos extratos dos ácaros de poeira, tais como Dermatofagoides farinae (DF), Dermatofagoides pteronyssinus (DP) e Tyrophagus putrescentiae (TP) são relevantes para estudos científicos na área de alergias e aplicação em imunoterapias. A precipitação/concentração desses extratos proteicos pode favorecer a agregação de alérgenos nos homogenatos. Objetivo e método: O trabalho investiga o processo de precipitação, submetendo os extratos brutos de ácaros de poeira a compostos como sulfato de amônio (NH4)2SO4, ácido tricloroacético (ATC) e acetona. Resultados: Os melhores resultados foram obtidos por fracionamento com (NH4)2SO4 em 80% (m/v) de saturação (~ 0°C), observando as marcações proteicas no gel de eletroforese. Os alérgenos principais foram identificados por immunoblot em 25 kDa (cisteína protease) para Der f 1 e Der p 1; e 25 kDa, 33 kDa (tropomyosin), 11 kDa para Tyr. Para esse percentual, os teores de proteína total foram de 12.83 mg mL-1 para DF; 24,78 mg mL-1 para DP; e 27,35 mg mL-1 para TP. Conclusão: A vantagem da precipitação com (NH4)2SO4 frente à precipitação com acetona foi a possibilidade de gradativamente se obter frações proteicas, o que não acontece quando utilizado esse solvente. A adição de 80% (v/v) de acetona aos extratos de ácaros favoreceu a precipitação total de proteína nas concentrações 16,42 mg mL-1; 28,47 mg mL-1; e 13,41 mg mL-1. O uso de ATC em concentrações acima de 20% (m/v) forma peptídeos que não são retidos no gel nas condições experimentais estabelecidas, sendo eficiente soluções mais diluídas desse ácido.

Oxidative foliar photo-necrosis produced by the bacteria Pseudomonas cedrina

BACKGROUND: Although bioactive metabolites capable of causing oxidative photo-necrosis in plant tissues have been identified in fungi, little is known about this type of mechanism in bacteria. These metabolites act as photosensitizers that generate reactive oxygen species (ROS) capable of causing damage to cells. In addition, these metabolites can pass into an energetically excited state when they receive some luminous stimulus, a condition in which they interact with other molecules present in the environment, such as molecular oxygen (O2), also known as triplet oxygen (3 O2), generating ROS. RESULTS: The suspension of the bacterial culture of Pseudomonas cedrina was shown to produce foliar necrosis in papaya leaves (Carica papaya L.) only in the presence of sunlight, which is evidence of photosensitizing mechanisms that generate singlet oxygen (1 O2). From the chemical study of extracts obtained from this bacteria, 3-(4-(2-carboxipropyl) phenyl) but-2-enoic acid (1) was isolated. This compound, in the presence of light and triplet oxygen (3 O2), was able to oxidize ergosterol to its peroxide, since it acted as a photosensitizer producing 1 O2, with which it was corroborated that a photosensitization reaction occurs, mechanism by which this bacterium could prove to cause oxidative foliar photo-necrosis. CONCLUSIONS: P. cedrina was able to induce oxidative foliar photo-necrosis because of its potential ability to produce photosensitizing metabolites that generate singlet oxygen in the plants it colonizes. Based on the above, it can be proposed that some bacteria can cause oxidative foliar photo-necrosis as an important mechanism in the pathogenesis of host species.

Optimization of cosmetic formulations development using Box-Behnken design with response surface methodology: physical, sensory and moisturizing properties

Considering the importance of an adequate composition of the formulation in the development of stable, safe and effective cosmetic products, experimental design techniques are tools that can optimize the formulation development process. The objective of this study was to develop topical formulations using the Box-Behnken design with response surface methodology and evaluate its physical, sensory and moisturizing properties. The experimental design used in the first step allowed to identify and to quantify the influence of raw materials, as well as the interaction between them. In the second step, the analysis identified the influence of soy lecithin, the phytantriol and capric acid triglyceride and caprylic on the consistency index, stickiness and greasiness and skin hydration. Cetearyl alcohol, dicetyl phosphate and cetyl phosphate 10EO and acrylates/C10-30 alkylacrylate crosspolymer showed effects in rheological parameters. The addition of soy lecithin had significant effects in terms of consistency index, stickiness, oiliness and immediate moisturizing effects. Phytantriol showed effects on increasing consistency index and oiliness sensation. Thus, the experimental design was shown to be an effective tool for research and development of cosmetics, since it allowed the assessment of the individual and interaction effects of raw materials in the responses: rheological parameters, sensory and clinical efficacy.

Antitumor efficacy of EDTA co-treatment with cisplatin in tumor-bearing mice

Ethylenediamine tetraacetic acid (EDTA) is used in various medical applications. The aim of this study is to investigate the antitumor efficacy of EDTA alone or with cisplatin (Cis). Fifty male albino mice were used to assess the median lethal dose (LD50) of EDTA via intraperitoneal (i.p) injection. To determine the antitumor activity, fifty female albino mice were divided into five groups as the following; Group 1 (Gp1) was negative control; (Gp2-5) inoculated i.p with 2×106 Ehrlich Ascites Carcinoma (EAC) cells/mouse. After one day, Gp3, Gp4 and Gp5 injected with Cis (2 mg/kg), EDTA (25 mg/kg) and Cis (2 mg/kg)/EDTA (25 mg/kg) for six days, respectively. At day 14, all groups were sacrificed to assess the tumor profile, liver enzymes (alanine transaminases and aspartate transaminases), kidney function (urea and creatinine) and electrolytes (Na+, K+ and Ca2+). The results showed that the i.p LD50 of EDTA was 250 mg/kg. Treatment with EDTA alone did not show any antitumor activity and did not interfere with the antitumor efficacy of Cis. Biochemical findings revealed that EDTA had mild toxicity on liver and kidneys functions. In summary, EDTA had no antitumor effect and did not alter the Cis efficacy.

Surface and mechanical properties of adhesives with calcium phosphates challenged to different storage media

Aim: To evaluate the behavior of experimental dental adhesives with hydroxyapatite (HAp), alpha-tricalcium phosphate (α-TCP) or octacalcium phosphate (OCP) after storing them in three different media: dry storage, distilled water, or lactic acid. Methods: An experimental adhesive resin was formulated with bisphenol A glycol dimethacrylate, 2-hydroxyethyl methacrylate, and photoiniciator/co-initiator system. HAp(GHAp), α-TCP (Gα-TCP), or OCP (GOCP) were added to the adhesive resin at 2 wt.%, and one group remained without calcium phosphates to be used as a control (GCtrl). The adhesives were evaluated for surface roughness, scanning electron microscopy (SEM), and ultimate tensile strength (UTS) after storing in distilled water (pH=5.8), lactic acid (pH=4) or dry medium. Results: The initial surface roughness was not different among groups (p>0.05). GHAp showed increased values after immersion in water (p<0.05) or lactic acid (p<0.05). SEM analysis showed a surface variation of the filled adhesives, mainly for Gα-TCP and GHAp. GHApshowed the highest UTS in dry medium (p<0.05), and its value decreased after lactic acid storage (p<0.05). Conclusions: The findings of this study showed that HAp, OCP, and α-TCP affected the physical behavior of the experimental adhesive resins in different ways. HAp was the calcium phosphate that most adversely affected the surface roughness and the mechanical property of the material, mainly when exposed to an acid medium

Study on components efficacy of Salviae Miltiorrhizae Radix et Rhizoma based on systematic traditional Chinese medicine / 中国中药杂志

This paper aimed to establish efficacy systems of tanshinones and salvianolic acids, two representative substances in Salviae Miltiorrhizae Radix et Rhizoma by using literature mining and biological network construction, based on systematic traditional Chinese medicine theory. The systematic study on the efficacy of traditional Chinese medicine was carried out from the basic unit, the structure and relationship between the basic units, the boundary of the research object and the function of the system, so as to explain the overall efficacy of the two kinds of components at the molecular level. Firstly, we collected the elements of the efficacy systems of these two kinds of components by literature mining, and defined their boundaries based on biological processes. After that, the structure of the efficacy systems was clarified according to the relationship in the KEGG database. Finally, the function of the efficacy systems was analyzed from the level of pharmacology, pharmacodynamics, and efficacy, revealing the scientific connotation of traditional Chinese medicine efficacy system. The results showed that there were 201 targets(elements), 12 target sets(boundary), and 12 pathway networks(structure) in salvianolic acids' efficacy system. Meanwhile, there were 189 targets(elements), 11 target sets(boundary), and 11 pathway networks(structure) in tanshinones' efficacy system. The results suggested that the functions of salvia-nolic acids' and tanshinones' efficacy systems were different in pharmacology and pharmacodynamics from aspects of elements, boundary, relationship and structure, but they were same in functional level as both of them could promote blood circulation, remove blood stasis, clear away heart-fire, relieve restlessness, and soothe the nerves. Based on systematic traditional Chinese medicine, we constructed the efficacy system of two representative components in Salviae Miltiorrhizae Radix et Rhizoma in this paper, elucidated the overall efficacy and builded the bridge between reductionism and holism in traditional Chinese medicine.