Salmonella enterica subsp. enterica serovar Typhimurium and Lactobacillus spp. interactions in vitro elicit improved antimicrobial production

@#Antimicrobial resistance (AMR) is a global health crisis. Despite the drug discovery efforts, AMR is increasing, and discoveries are nearly nil. It is thus critical to design new strategies. Probiotics are tapped as alternatives to antibiotics for the treatment of gut-associated diseases. Lactobacillus species, common in food products, can inhibit the growth of gut pathogens. Here, we demonstrate the antimicrobial activities of Lactobacillus species – Lactobacillus paracasei, Lactobacillus casei, and Lactobacillus delbrueckii subsp. bulgaricus are enhanced when cocultured with Salmonella enterica subsp. enterica serovar Typhimurium. Cell-free culture supernatants (CFCS) from cocultures of Lactobacillus spp. and Salmonella enterica serovar Typhimurium more potently inhibit pathogen growth than their monoculture counterparts. Interestingly, we discovered that Salmonella enterica serovar Typhimurium could enhance the production of antimicrobials from Lactobacillus spp., most evident in L. delbrueckii subsp. bulgaricus. Also, L. delbrueckii subsp. bulgaricus CFCS upregulates key Salmonella virulence genes, hilA and sipA. Whether this increases Salmonella’s pathogenicity in vivo or reduces pathogen fitness and growth inhibition in vitro warrants further investigation. We propose that these probiotic isolates may be utilized for innovative natural food processing and preservation strategies to control Salmonella food contaminations. Importantly, our findings that Salmonella elicits an enhanced antimicrobial activity from Lactobacillus spp. provide evidence of a pathogen-mediated elicitation of antimicrobial production. Therefore, extending this phenomenon to other microbial interactions may help augment the strategies for drug discovery.

LATEMO-E: A Film Database to Elicit Discrete Emotions and Evaluate Emotional Dimensions in Latin-Americans / LATEMO-E: Banco de Dados de Filmes para Suscitar Emoções e Avaliar Dimensões Emocionais para Latino-Americanos / LATEMO-E: Set de Fragmentos de Películas para Inducir Emociones y Evaluar Dimensiones Emocionales en Latinoamericanos

Abstract Introduction: Film clips are the most used stimuli to induce different emotional states. Objective: Identify, select, and evaluate a set of film clips according to dimensionality and discreteness of emotions. Film experts suggested 437 fragments, but only 70 met the criteria to be included in the LATEMO-E film-clip database. Study 1: The 70 film clips were tested for their ability to induce disgust, anger, fear, sadness, amusement, tenderness, and neutral (n = 147 participants; mean age = 20.64 years, SD ±2.21 years). Results: A total of 28 film clips presented adequate scores on intensity and discreteness. Study 2: A total of 28 selected film clips were assessed again in a new sample of 106 participants (mean age = 20.96 years, SD ±2.45 years). Results: Anger and amusement film clips were the most extreme in the valence and activation dimensions. Anger and fear film clips were the most extreme categories in the intensity, certainty, and control dimensions. Conclusions: These stimuli were useful for inducing emotional states that can be judged from both the dimensional perspective of emotions and the basic emotions perspective. Evoked emotions could be differentiated according to cognitive patterns of evaluation that guide behavior in response to emotional stimulation.

Resumo Introdução: os clipes são os estímulos mais utilizados para induzir diferentes estados emocionais. Objetivo: identificar, selecionar e avaliar um conjunto de clipes de filme de acordo com a discrição e dimensionalidade da emoção. 437 fragmentos foram sugeridos por especialistas em filmes, mas 70 atenderam aos criterios de inclusão em um banco de dados de filmes. Estudo 1: os 70 clipes foram testados em sua capacidade de induzir: nojo, raiva, medo, tristeza, diversão, ternura e neutro (N = 147 participantes; M = 20,64; DP ±2,21). Resultados: 28 clipes apresentaram escores adequados de intensidade e discrição. Estudo 2: 28 clipes selecionados foram reavaliados em uma nova amostra de 106 participantes (M = 20,96; DP ±2,45). Resultados: os clipes de raiva e diversão foram os mais extremos nas dimensões de valência e ativação. Clips de raiva e medo foram as categorias mais extremas nos indicadores de intensidade, certeza e controle. Conclusões: os estímulos foram úteis para induzir estados emocionais que podem ser julgados tanto pela perspectiva dimensional das emoções quanto pela perspectiva básica das emoções. Além disso, as emoções evocadas são diferenciadas de acordo com os padrões cognitivos de avaliação que guiam o comportamento em resposta à estimulação emocional.

Resumen Introducción: los fragmentos de películas están entre los estímulos más usados para inducir distintos estados emocionales. Objetivo: identificar, seleccionar y evaluar un set de fragmentos de películas según la discreción y la dimensionalidad de la emoción. Expertos en cine sugirieron 437 fragmentos, pero solo 70 fueron identificados como aquellos que cumplieron con los criterios de inclusión en un set de fragmentos de películas. Estudio 1: los 70 fragmentos fueron evaluados en su capacidad para inducir: asco, ira, miedo, tristeza, diversión, ternura y neutro (N = 147 participantes; M = 20.64; DE ±2.21). Resultados: 28 fragmentos presentaron puntuaciones adecuadas en intensidad y discreción. Estudio 2: los 28 fragmentos fueron reevaluados en una nueva muestra de 106 participantes (M = 20.96, DE ±2.45). Resultados: los fragmentos de ira y diversión fueron los más extremos en las dimensiones valencia y activación. Ira y miedo fueron las categorías más extremas en los indicadores intensidad, certeza y control. Conclusiones: los estímulos resultaron útiles para inducir estados emocionales que pueden ser juzgados desde la perspectiva dimensional de las emociones, así como desde el enfoque de las emociones básicas. Además, las emociones provocadas se diferencian en función de patrones cognitivos de evaluación que guían la conducta en respuesta al estímulo emocional.

Effect of salicylic acid and silver nitrate on rutin production by Hyptis marrubioides cultured in vitro / Efeito do ácido salicílico e nitrato de prata na produção de rutina por Hyptis marrubioides cultivada in vitro

ABSTRACT: Hyptis marrubioides (Lamiaceae) is a medicinal plant that is native from Brazilian Cerrado. In vitro propagation techniques make use of elicitors to alter metabolic pathways, affecting how molecules are produced both qualitatively and quantitatively. This research aimed to evaluate how abiotic elicitors salicylic acid (SA) and silver nitrate (SN) at concentrations of 30µM or 60µM influence Hyptis marrubioides seedling growth by two different in vitro culture methods. The rutin content was quantified by HPLC-DAD. Compared to an untreated culture, the H. marrubioides methanolic extracts cultured in MS medium for 10 days followed by culture in MS medium containing SN (30µM) for 20 days had 1.28 times higher rutin content. In a second experiment, seedlings were cultured in MS medium for 20 days, and then the desired elicitor was added to the culture and allowed to remain in contact with the medium for three and six days. SA (30µM) gave the best results: rutin production was 16.56-foldhigher than the control after six days. SN (30µM) increased the rutin content by 1.17-fold. At the two concentrations evaluated during the elicitation experiments, neither SA nor SN altered the growth parameters shoot length, leaf number, and fresh and dry weight of H. marrubioides seedlings grown in vitro as compared to the control. Based on these results, the abiotic elicitors SA and SN successfully provide Hyptis marrubioides with increased rutin content in vitro.

RESUMO: Hyptis marrubioides (Lamiaceae) é uma planta medicinal nativa do Cerrado brasileiro. Técnicas de propagação in vitro fazem uso de elicitores para alterar as vias metabólicas, afetando a produção de moléculas qualitativa e quantitativamente. Este trabalho teve como objetivo avaliar como os elicitores abióticos ácido salicílico (SA) e nitrato de prata (SN) nas concentrações de 30µM ou 60µM influenciam no crescimento de plântulas de Hyptis marrubioides por dois diferentes métodos de cultivo in vitro. O teor de rutina foi quantificado por CLAE-DAD. Em comparação com uma cultura não tratada, os extratos metanólicos de H. marrubioides cultivados em meio MS por 10 dias, seguidos de cultura em meio MS contendo SN (30µM) por 20 dias, apresentaram 1,28 vezes maior teor de rutina. Em um segundo experimento, as plântulas foram cultivadas em meio MS por 20 dias, e então o elicitor desejado foi adicionado à cultura e deixado em contato com o meio por três e seis dias. SA (30µM) forneceu os melhores resultados na produção de rutina, sendo 16,56 vezes maior do que o controle após seis dias. SN (30µM) aumentou o teor de rutina em 1,17 vezes. Nas duas concentrações avaliadas durante os experimentos de elicitação, nem SA nem SN alteraram os parâmetros de crescimento, como comprimento da parte aérea, número de folhas ou peso fresco e seco das plântulas de H. marrubioides cultivadas in vitro em relação ao controle. Com base nestes resultados, os elicitores abióticos SA e SN forneceram com sucesso Hyptis marrubioides in vitro com maior conteúdo de rutina.

Micropropagation of Baccharis antioquensis (Asteraceae) and photoinduction of polyphenols by UV radiation / Micropropagación de Baccharis antioquensis (Asteraceae) y fotoinducción de polifenoles mediante radiación UV

Abstract Polyphenols are a large diversity of chemical types and interactions that can be responsible for a multiplicity of protective functions ranging from toxicity and light/UV shielding to signal transduction. Bacharis antioquensis has been described as a potential source of new photoprotective compounds with antioxidant capacity associated to polyphenols compounds. The aim of the present work was to develop a micropropagation protocol of B. antioquensis and evaluate the production of polyphenols by in vitro plants exposure to UVB radiation. Branches in juvenile stage of B. antioquensis were collected, desinfected and cultured on half strength Murashige and Skoog medium, supplemented or not with growth regulators (TDZ, BA or GA3) on light/darkness conditions and liquid/solid media. After UV treatments, the absorption coefficient in the UVA-UVB range, the antioxidant capacity and the total phenol content (TPC) from all tissue cultures and the wild tissue were evaluated. Growth regulators, light conditions and type of culture medium (solid or liquid) had a favorable effect on the response of explants. Treatments containing BA + GA3 regulators (2 and 0.5 mg/L respectively) and TDZ (0.5 mg/L) showed positive results in bud growth in liquid medium and darkness. Results showed that UVR exhibited promoting effects on the accumulation of polyphenols, enhancing the absorption coefficient in the UVA-UVB range, the antiradical capacity and the TPC of B. antioquensis in vitro plants. Rev. Biol. Trop. 66(2): 754-764. Epub 2018 June 01.

Resumen Los polifenoles son compuestos químicos con una diversidad de interacciones que pueden ser responsables de muchas funciones, que van desde la toxicidad hasta la protección y blindaje contra la luz/UV. Baccharis antioquensis es una fuente potencial de compuestos fotoprotectores con capacidad antioxidante. El objetivo del presente trabajo fue desarrollar un protocolo de micropropagación para B. antioquensis y evaluar la producción de polifenoles in vitro por exposición a la radiación UVB. Ramas juveniles fueron colectadas y cultivadas en medio de Murashige y Skoog (MS) y suplementadas o no con reguladores de crecimiento (TDZ, BA o GA3) en diferentes condiciones de luz/oscuridad y medios líquidos/sólidos. Después de los tratamientos UVR, se midió el espectro de absorción UV y se evaluó el coeficiente de absorción en la región UVA-UVB, la capacidad antioxidante y TPC tanto en las plantas in vitro como en las plantas silvestres. Los tratamientos que contenían reguladores BA + GA3 (2 y 0.5 mg / L respectivamente) y TDZ (0.5 mg / L) mostraron resultados positivos en el crecimiento del brote en medio líquido y en condiciones de oscuridad. Los resultados mostraron que la UVR tiene efectos promotores sobre la acumulación de metabolitos secundarios, aumentando el coeficiente de absorción en la región UVA-UVB, la capacidad antiradicalaria y TPC en las plantas in vitro.

Metabolic disturbance and phytochemical changes in Andrographis paniculata and possible action mode of andrographolide

Objective: To explore the effect of gibberellic acid (GA

Metabolic disturbance and phytochemical changes in Andrographis paniculata and possible action mode of andrographolide

Objective:To explore the effect of gibberellic acid (GA3) and its inhibitor paclobutrazol (PBZ)on chemical composition and their pharmacological effects on Andrographis paniculata (Burm.f.) Wall.ex Nees,and to clarify action mode of andrographolide.Methods:The chemical composition was extracted by sequential extraction with hexane,dichloromethane,ethyl acetate and methanol,respectively.Andrographolide and its derivatives were evaluated by HPLC.Moreover,the metabolic profiling was analyzed by GC-MS.Inhibitory effect of crude extracts was tested against Staphylococcus aureus using agar well diffusion method.Mode of action was tested against mutant yeast by spotting assay.Andrographolide were tested for their mode of action against eukaryotes.Results:Among different solvents,dichloromethane gave the highest yield of crude (3.58％ DW),with the highest andrographolide content (8.3 mg/g DW).The effect of plant hormone (10 mg/L GA3 or PBZ) on phytochemical variations and bioactivity of Andrographis paniculata was demonstrated.It was found that PBZ promoted sesquiterpene compounds about 3.5 times over than GA3 treatment.But inhibitory effect of extracts against Staphylococcus aureus was highest in GA3 treated plants;andrographolide and 14-deoxy-11,12-didehydroandrographolide contents were significantly higher than those of water or PBZ.It was found that there were 11 strains involving in ergosterol biosynthesis,V-ATPase activity and homeostasis,and superoxide detoxification process.In this regard,andrographollde might cause the damage on the lipid bilayer of yeast cell and plasma membrane by interfering ergosterol biosynthesis.Conclusions:It is found that GA3 promotes andrographolide and 14-deoxy-11,12-didehydroandrographolide content while PBZ promotes sesquiterpene content.Andrographolide might cause the damage on the lipid bilayer of yeast cell and plasma membrane by interfering ergosterol biosynthesis.It might also affect mitochondria electron transport chain,leading to the occurrence of ROS,which can further harm cell organelles.However,the library screening is the first step to investigate mode of action of andrographolide.

Elicitation Enhanced the Production of Phenolic Compounds and Biological Activities in Hairy Root Cultures of Bitter melon ( Momordica charantia L.)

ABSTRACT Momordica charantia (Cucurbitaceae) is an important vegetable and also medicinal crop which produces the bioactive compounds for various biological activities with potential uses in human health. The present investigation relates to elicitors of jasmonic acid (JA) and salicylic acid (SA) to enhance biomass accumulation and phenolic compound production in hairy root cultures of M. charantia. Hairy root cultures were elicited with JA and SA at 0, 25, 50 and 100 μM concentrations respectively. The adding of elicitation to the hairy root cultures on the 15th day of culture and the roots were harvested on day 25. Cultures supplemented with 100 μM JA and SA enhanced the phenolic compounds significantly compared to that of non-elicited hairy root cultures. The biomass of hairy root culture significantly increased by SA whereas decreased in JA elicitation at 100 μM. JA and SA-elicited hairy root cultures significantly produced a higher amount of phenolic compounds (12811.23 and 11939.37µg/g), total phenolic (4.1 and 3.7 mg/g) and flavonoid (3.5 and 3.2 mg/g) contents than non-elicited hairy root cultures (10964.25 µg/g, 2.8 and 2.5 mg/g). JA and SA-elicited hairy root cultures were significantly higher antioxidant activity of DPPH (84 and 78%), reducing potential (0.53 and 0.48), phosphomolybdenum (3.6 and 3.2 mg/g) and ferrous ion chelating assays (80 and 74%) than non-elicited hairy root cultures. The higher antimicrobial and anticancer activity were exhibited in JA and SA-elicited than non-elicited hairy root cultures. This protocol can be developed for the production of phenolic compounds from JA and SA-elicited hairy root cultures.

Estimación del valor predictivo positivo de la colangiopancreatografía magnética utilizando metodos de bayes / Estimation of the positive predictive value of the magnetic resonance cholangiopancreatography using bayes methods

Objetivo: este artículo presenta la manera de calcular, dentro de una prueba de diagnóstico clínico, la probabilidad de que un individuo, cuyo resultado para la prueba de tamizaje es positivo, esté realmente enfermo sabiendo que no se tiene su resultado con la prueba patrón de oro; probabilidad denominada Valor Predictivo Positivo (VPP). Método: se asumió el VPP como una cantidad desconocida para la que se puede construir una distribución de probabilidad. Dentro del paradigma Bayesiano de la estadística, se utilizó el método propuesto por Winkler and Smith para calcular VPP utilizando un conjunto de registros sobre coledocolitiaisis en pacientes con pancreatitis aguda. Resultados: los métodos bayesianos permitieron obtener regiones de credibilidad más estrechas que los intervalos de confianza clásicos. Discusión: el uso de métodos Bayesianos es una excelente alternativa para obtener estimaciones más precisas del VPP. Estimar los valores predictivos utilizando los datos de la tabla 2X2 conlleva a errores cuando el tamaño de la muestra no es tan grande como para asumirla cercana a la población.

Objetive: In this article we present different alternative to compute thePositive Predictive Value (PPV) for a diagnostic test. Method: we assumedtheVPP as a continuous and unknown quantity whose natural performanceis susceptible of model using a distribution of probability. We use themethod propose as Winkler and Smith to estimate the VPP using Bayesianprocedure. We illustrate our methodology with a data set of patients withacute pranceatitis taken the Magnetic Cholangiopancreatogrphy as test toscreening of Choledocoletise Results: The Bayesian approach allows to havea best estimates and the credibility regions were narrower than the confidenceintervals.Discussion:thje use of Bayesian methods is an excellent choise toobtain more accute estimates. To estimateVPP’s using the observed data intable 2X2 implies mistakes when the sample size is not large enough as to thinkthat the sample close the population size.

Preference elicitation approach for measuring the willingness to pay for liver cancer treatment in Korea

BACKGROUND/AIMS: The Korean government has expanded the coverage of the national insurance scheme for four major diseases: cancers, cardiovascular diseases, cerebrovascular diseases, and rare diseases. This policy may have a detrimental effect on the budget of the national health insurance agency. Like taxes, national insurance premiums are levied on the basis of the income or wealth of the insured. METHODS: Using a preference elicitation method, we attempted to estimate how much people are willing to pay for insurance premiums that would expand their coverage for liver cancer treatment. RESULTS: We calculated the marginal willingness to pay (MWTP) through the marginal rate of substitution between the two attributes of the insurance premium and the total annual treatment cost by adopting conditional logit and mixed logit models. CONCLUSIONS: The effects of various other terms that could interact with socioeconomic status were also estimated, such as gender, income level, educational attainment, age, employment status, and marital status. The estimated MWTP values of the monthly insurance premium for liver cancer treatment range from 4,130 KRW to 9,090 KRW.

Application of Elicitation Teaching Mode in the Pharmaceutical Standard Training / 中国药房

OBJECTIVE:To improve the teaching quality of pharmaceutical standard training. METHODS: By introducing the overview of adopting elicitation teaching in the pharmaceutical standard training in Beijing,the training effect was summarized and role and significance were studied. RESULTS:Supervisors applied five teaching modules in the elicitation teaching in the pharmaceu-tical standard training which are concluded“pharmaceutical education and rational use of medicine”“comments of prescription and rational use of medicine”“prescription transfers and rational use of medicine”“medicine counseling and rational use of medicine”“pharmaceutically communication and rational use of drugs medicine”,the result showed that participants had mastered more than 90%of what they had learned;with 92.5%of participants felt elicitation teaching could help improve students’interests in pharma-cy,and strengthen responsibility;80.0% of the participants believed that part of the modules strengthen the training of clinical think-ing;the training of satisfaction to 100%. CONCLUSIONS: The elicitation teaching mode can not only improve the teaching quality in pharmaceutical standard training,but also strengthen communication and feedback between teachers and students,students and students to make students more deeply understand and master knowledge and improve teaching quality.

Production of phenolic metabolites by Deschampsia antarctica shoots using UV-B treatments during cultivation in a photobioreactor

Deschampsia antarctica (DA), the only species in the Gramineae family endemic to the Antarctic territory, is characterized by a combination of high levels of free endogenous phenylpropanoid compounds under normal in situ and in vitro growth conditions. In this article, we describe the design and use of a specific temporary immersion photobioreactor to produce both increased DA biomass and secondary metabolite accumulation by UV-B elicitation during cultivation. Three min-long immersions in an induction medium applied every 4 hrs at 14ºC +/- 1 and 20/4 hrs light/darkness photoperiod increased DA biomass production over previous in vitro reports. Biomass duplication was obtained at day 10.7 of culturing, and maximum total phenolics and antioxidant activity were observed after 14 day of culturing. The addition of UV-B radiation pulses for 0.5 hrs at 6 hrs intervals increased total phenolics and antioxidant activity more than 3- and 1.5- fold, respectively, compared to controls with no UV-B. Significant accumulation of scopoletin, chlorogenic acid, gallic acid and rutin was found in these plantlets. This is the first bioreactor designed to optimize biomass and phenylpropanoid production in DA.

Biochemical and morphological responses to abiotc elicitor chitin in suspension-cultured sugarcane cells

Cells of Saccharum officinarum submitted to hydrolyzated chitin for 1 to 8h produced phenolic compounds. These alterations were observed through cytochemical methods using Toluidine Blue and Phloroglucinol/HCl. After 4 h, besides cell wall change, there was a change in nuclear pattern of chitin treated cells. There was a 96 percent increase in nuclear area in 6 h chitin treated material, as observed by Feulgen reaction. The treated cells showed chromatin compacted regions and a degeneration process of nucleoli. In the outer areas of cell wall, there was a polysaccharide desagregation, confirming results obtained for different plants with the use of other elicitors. Peroxidase activity was maximal after 4 h and decreased progressively. PAL activity started to increase at 4 h of incubation. These results showed that chitin hydrolyzate stimulated a defense response in sugarcane cells.

Células de Saccharum officinarum quando submetidas a quitina hidrolisada por 1 a 8h produziram material fenólico. Essas alterações foram observadas por meio de métodos citoquímicos como o Azul de Toluidina e Floroglucinol/HCl. Após 4 h, além das mudanças nas paredes celulares houve uma mudança no padrão nuclear das células tratadas com quitina. Por observação da reação de Feulgen, houve um aumento de 96 por cento na área nuclear no material em 6h. Para as células tratadas foram observadas regiões de cromatina compactada e um processo de degeneração do nucléolo. Nas áreas externas da parede celular existia uma desagregação dos polisacarídios confirmando os resultados obtidos para diferentes plantas com o uso de outros elicitores. A atividade da peroxidase foi maxima após 4 h e então decresceu progressivamente. A atividade da PAL aumentou a partir de 4 h de incubação. Estes resultados mostram que o hidrolisado de quitina estimula as respostas de defesa em células de cana.

Unrecognized intraneural injection as a possible cause of nerve injury associated with axillary block : A case report

A 71-year-old man who had a laceration on his right fifth finger underwent an emergency explorative operation under brachial plexus block that was performed by using transaxillary artery technique.During axillary block, no elicitation of paresthesia was presented.Postoperatively he complained numbness on his right arm along the dermatome of 7th cervical nerve distribution.Two weeks later the evidence of nerve injury was found on EMG and NCV carried by the department of neurology. Observing closely for another two weeks, the symptoms and signs were gradually improved.On 42nd postoperative day, the patient complained no neurologic problems and the findings of EMG and NCV returned to normal.Considering the method of anesthesia, operation time, tourniquet time and patient position, we conclude that the direct needle injury and intraneural injection during axillary block are the highly suggestive causes of nerve injury.

Unrecognized intraneural injection as a possible cause of nerve injury associated with axillary block : A case report

A 71-year-old man who had a laceration on his right fifth finger underwent an emergency explorative operation under brachial plexus block that was performed by using transaxillary artery technique.During axillary block, no elicitation of paresthesia was presented.Postoperatively he complained numbness on his right arm along the dermatome of 7th cervical nerve distribution.Two weeks later the evidence of nerve injury was found on EMG and NCV carried by the department of neurology. Observing closely for another two weeks, the symptoms and signs were gradually improved.On 42nd postoperative day, the patient complained no neurologic problems and the findings of EMG and NCV returned to normal.Considering the method of anesthesia, operation time, tourniquet time and patient position, we conclude that the direct needle injury and intraneural injection during axillary block are the highly suggestive causes of nerve injury.

Peroxidase production from hairy root cultures of red beet (Beta vulgaris)

The genetically transformed roots of red beet have been shown, for the first time, to produce very high levels of peroxidase (POD; EC 1.11.1.7) accounting for 1.21 x 10(6) Units L-1. Of the ten clones established using different strains of Agrobacterium rhizogenes, one was that from the strain LMG-150, three each from A 2/83, A 20/83 and A4. All the clones showed true integration of T-DNA when tested by PCR and Southern hybridization methods. Each clone differed significantly from the others in growth, hormone dependency and POD production where LMG-150 produced highest biomass (140 g FW L-1) as well as POD (ranging from 8000-9000 U g-1 FW and 1.18 x 10(6) U L-1 with a specific activity of 600 U mg-1 protein) on hormone-free medium, both in shake-flask as well as in bioreactor with a further enhancement to 1.21 x 10(6) U L-1 upon the addition of extra calcium chloride (5 mM). PAGE with active staining showed 4 distinct bands of Rm 0.06, 0.16, 0.25, 0.38 and 0.46 in the biomass and bands at Rm 0.06, 0.16, 0.25 and one extra band of Rm 0.575 in the spent medium where isozymes of Rm 0.38 and 0.46 were totally absent. The pH optima and other properties were grossly comparable with the standard horse-radish POD (HRP) with better thermal stability than HRP and therefore, the present source appears to offer a cheaper and additional alternative for the commercial production of POD.

Effect of three fungi of Helminthosporium on bioactive constituents in root of seedling Rumex gmelini / 中草药

Objective To study the effect of three fungi of Helminthosporium on the seven kinds of bioactive constituents in root of seedling Rumex gmelini. Methods Let the R. gmelini infect the fungi of Helminthosporium by dealing the leave surface of R. gmelini with fungal spore suspension. The content of seven kinds of bioactive constituents was determined by HPLC and the results were analyzed by variance analysis. Results During the treatment time, little effect of three fungi of Helminthosporium on content of polydatin, chrysophanol 1-glucoside, and physcion, but the obvious effect on resveratrol, musizin, emodin, and chrysophanol was found. Conclusion The effects of H. turcicum 001 on yield and content of resveratrol are significant in early-middle stage (0—40 d), but treatment time over 40 d is not suitable. The results of this study could provide the new idea and theoretical basis for the exploiting of natural medicines and the planting of Chinese herbal medicine.

Effect of Vehicle on Elicitation of DNCB Contact Allergy in Guinea Pig / 대한피부과학회지

This study was undertaken to investigate the optimal conditions for challenge testing with DNCB in 3 vehicles applied to guinea pigs in standardized amount/ area (pg/cm2) The results are sumrnarized as follows: 1) All test reactions were negative when tested in 10 unsensitized animals. 2) There were significant difference in positive rate between 0. 05%, DNCB in alcohol and 0.1% DNCB in alcohol, but there were no significant difference between 0. l%, DNCB in alcohol and 0, 2% DNCB in alcohol. Same results were obtaincd when DNCR was dissolved in acetone. 3) Acetone gives the significantly higher degree of positive reactions, indicating that alcohol may be the safer vehicle in eliciting a response than acetone. 4) With DNCB in olive oil, relatively few animals reacted compared to the results obtained with alcohol and acetone. Furthermore, olive oil is slippery and difficult to utilize when attempting to delineate surface area. 5) With alcohol, we obtained a good correlation between the amount applied and the resulting response, showing that this vehicle might be a superior vehicle in eliciting contact allergy.

Effects of protein deficiency on selective elicitation of lysosomal enzymes in rat peritoneal macrophages.

Young albino rats were fed ad libitum 4, 8 or 20 % (control) protein diet for 1–4 weeks. Total activities of some of the lysosomal enzymes, namely, acid phosphatase, aryl sulphatase, ß-glucuronidase and cathepsin D, were determined in resident and proteasepeptone elicited peritoneal macrophages. Total cell number, protein content and the lysosomal enzyme activities were increased significantly in protease-peptone elicited macrophages; though at a lower rate in 4 % protein-fed group compared to control ones. However, the rate of induction of the tested hydrolases was selective and their response to the stimulant varied widely. Similarly, response of each enzyme to low protein diet also varied. Thus, at 4 weeks, cathepsin D and ß-glucuronidase activities, expressed per total number of elicited macrophages were reduced by 45 and 60 %, respectively, in 4 % protein-fed animals. These results indicate that the metabolic events related to lysosomal function in macrophages, are affected by dietary restriction of proteins.