Protective effects of three kinds of borneol on different brain regions in acute cerebral ischemia/reperfusion model rats / 中国中药杂志

This study compared the ameliorating effects of L-borneol, natural borneol, and synthetic borneol on the injury of different brain regions in the rat model of acute phase of cerebral ischemia/reperfusion(I/R) for the first time, which provides a reference for guiding the rational application of borneol in the early treatment of ischemic stroke and has important academic and application values. Healthy specific pathogen-free(SPF)-grade SD male rats were randomly assigned into 13 groups: a sham-operation group, a model group, a Tween model group, a positive drug(nimodipine) group, and high-, medium-, and low-dose(0.2, 0.1, and 0.05 g·kg~(-1), respectively) groups of L-borneol, natural borneol, and synthetic borneol according to body weight. After 3 days of pre-administration, the rat model of I/R was established by suture-occluded method and confirmed by laser speckle imaging. The corresponding agents in different groups were then administered for 1 day. The body temperature was monitored regularly before pre-administration, days 1, 2, and 3 of pre-administration, 2 h after model awakening, and 1 d after model establishment. Neurological function was evaluated based on Zea-Longa score and modified neurological severity score(mNSS) 2 h and next day after awakening. The rats were anesthetized 30 min after the last administration, and blood was collected from the abdominal aorta. Enzyme-linked immunoassay assay(ELISA) was employed to determine the serum levels of tumor necrosis factor-alpha(TNF-α), interleukin-6(IL-6), IL-4, and transforming growth factor-beta1(TGF-β1). The brain tissues were stained with triphenyltetrazolium chloride(TTC) for the calculation of cerebral infarction rate, and hematoxylin-eosin(HE) staining was used for observing and semi-quantitatively evaluating the pathological damage in different brain regions. Immunohistochemistry was employed to detect the expression of ionized calcium binding adapter molecule 1(IBA1) in microglia. q-PCR was carried out to determine the mRNA levels of iNOS and arginase 1(Arg1), markers of polarization phenotype M1 and M2 in microglia. Compared with the sham-operation group, the model group and the Tween model group showed significantly elevated body temperature, Zea-Longa score, mNSS, and cerebral infarction rate, severely damaged cortex, hippocampus, and striatum, increased serum levels of IL-6 and TNF-α, and decreased serum levels of IL-4 and TGF-β1. The three borneol products had a tendency to reduce the body temperature of rats 1 day after modeling. Synthetic borneol at the doses of 0.2 and 0.05 g·kg~(-1), as well as L-borneol of 0.1 g·kg~(-1), significantly reduced Zea-Longa score and mNSS. The three borneol products at the dose of 0.2 g·kg~(-1) significantly reduced the cerebral infarction rate. L-borneol at the doses of 0.2 and 0.1 g·kg~(-1) and natural borneol at the dose of 0.1 g·kg~(-1) significantly reduced the pathological damage of the cortex. L-borneol and natural borneol at the dose of 0.1 g·kg~(-1) attenuated the pathological damage of hippocampus, and 0.2 g·kg~(-1) L-borneol attenuated the damage of striatum. The 0.2 g·kg~(-1) L-borneol and the three doses of natural borneol and synthetic borneol significantly reduced the serum level of TNF-α, and the 0.1 g·kg~(-1) synthetic borneol reduced the level of IL-6. L-borneol and synthetic borneol at the dose of 0.2 g·kg~(-1) significantly inhibited the activation of cortical microglia, and 0.2 g·kg~(-1) L-borneol up-regulated the expression of Arg1 and down-regulated the expression level of iNOS. In conclusion, the three borneol products may alleviate inflammation to ameliorate the pathological damage of brain regions of rats in the acute phase of I/R by inhibiting the activation of microglia and promoting the polarization of microglia from M1 type to M2 type. The protective effect on brain followed a trend of L-borneol > synthetic borneol > natural borneol. We suggest L-borneol the first choice for the treatment of I/R in the acute phase.

Expression of β-glucosidase An-bgl3 from Aspergillus niger for conversion of scopolin / 生物工程学报

Scopoletin is a coumarin compound with various biological activities including detumescence and analgesic, insecticidal, antibacterial and acaricidal effects. However, interference with scopolin and other components often leads to difficulties in purification of scopoletin with low extraction rates from plant resource. In this paper, heterologous expression of the gene encoding β-glucosidase An-bgl3 derived from Aspergillus niger were carried out. The expression product was purified and characterized with further structure-activity relationship between it and β-glucosidase analyzed. Subsequently, its ability for transforming scopolin from plant extract was studied. The results showed that the specific activity of the purified β-glucosidase An-bgl3 was 15.22 IU/mg, the apparent molecular weight was about 120 kDa. The optimum reaction temperature and pH were 55 ℃ and 4.0, respectively. Moreover, 10 mmol/L metal ions Fe2+ and Mn2+ increased the enzyme activity by 1.74-fold and 1.20-fold, respectively. A 10 mmol/L solution containing Tween-20, Tween-80 and Triton X-100 all inhibited the enzyme activity by 30%. The enzyme showed affinity towards scopolin and tolerated 10% methanol and 10% ethanol solution, respectively. The enzyme specifically hydrolyzed scopolin into scopoletin from the extract of Erycibe obtusifolia Benth with a 47.8% increase of scopoletin. This demonstrated that the β-glucosidase An-bgl3 from A. niger shows specificity on scopolin with good activities, thus providing an alternative method for increasing the extraction efficiency of scopoletin from plant material.

New topical microemulsions of etofenamate as sufficient management of osteoarthritis

Abstract In this study, microemulsions containing etofenamate were prepared and evaluated as dermal delivery carriers. The developed microemulsions consist of oleic acid, Span 80, Tween 20, Cremophor EL, Transcutol and ethanol. The percentage of etofenamate loading in the microemulsions was 5% (w/w). The characterization of formulations included droplet size, zeta potential, pH, conductivity, PDI, refractive index and viscosity. Moreover, ex vivo penetration study was carried out using mice abdominal skin. The developed formulations were analyzed for their cytotoxicity via MTT assay and tested for their anti-inflammatory properties opposed to LPS-stimulated nitrite prοduction in RAW 264.7 cells. As ideal formulation, M2ETF, was chosen due to its greater permeation, lower penetration as well as higher anti-inflammatory

Compatibility and efficiency of preservatives in emulsive cosmetics containing high surfactant content

Abstract The objective of this study was to determine the influence of nonionic surfactants on the effectiveness of preservatives used in emulsions containing high surfactant content. Mixtures of different concentrations were prepared between polyethoxylated (40) hydrogenated castor oil (PHCO) and polyoxyethylene sorbitan monooleate (PSO), with methylparaben, phenoxyethanol, methylparaben, ethylparaben, propylparaben, and isobutylparaben (PMEPBI) blend, phenoxyethanol and benzoic acid (BP) blend, and phenoxyethanol and caprylyl glycol (PC) blend. Subsequently, the compatibility of the formulation ingredients and the effectiveness of the preservatives were evaluated by the challenge test. It was found that PHCO and PSO inactivated the antimicrobial action of methylparaben and PMEPBI. Paraben-free preservatives BP and PC had less influence on surfactants than systems containing parabens. When incorporated into microemulsions and nanoemulsions containing 40% and 20% surfactants, methylparaben and BP 0.2% and 0.5% were only effective against Aspergillus niger. The PMEPBI 0.2% was effective as a preservative in nanoemulsified formulations against A. niger, Escherichia coli, Pseudomonas aeruginosa, and Staphylococcus aureus. The results demonstrate that the efficacy of the preservative system in formulations containing nonionic surfactant excipients depends on the type of excipient, the components of the formulation, the preservative systems composition, the excipient to preservative ratio, and the availability in the formulation.

Sordera neurosensorial congénita con malformación de oído interno ligada al X en una familia mexicana / Congenital sensorineural deafness X-linked, with inner ear malformation in a Mexican family

Introducción. Las sorderas o hipoacusias prelinguales son de etiología genética entre el 60 y el 68% de los casos; de estos, del 20 al 40% son malformaciones del oído interno. De los casos de hipoacusia no sindrómica ligada al X se han descrito siete tipos. De las malformaciones de oído interno, la partición coclear incompleta tipo III es la menos frecuente.Objetivo. Presentar el reporte clínico-genético de una familia mexicana, con indi-viduos varones afectados por sordera neurosensorial congénita con malformación de oído interno. Material y Métodos. Se realizó estudio de una familia en la que nueve miembros presentaban sordera. Se estudiaron cuatro de ellos y una madre sin manifestaciones, a través del estudio clínico general por médico genetista, el estudio audiológico (otos-copía y audiometría) por médico audiólogo y el estudio de tomografía computada (TC) por médico radiólogo.Resultados. Los pacientes estudiados presentaron sordera neurosensorial congéni-ta, de severa a profunda bilateral. A través de la TC, se evidenció malformación de oído interno. Tres pacientes presentaron partición coclear incompleta tipo III y un paciente partición incompleta tipo I. Debido al estudio clínico y al árbol genealógico, se definió diagnóstico de hipoacusia neurosensorial no sindrómica ligada al X. La TC de la madre sin manifestaciones no presentó evidencia de malformaciones en oído interno (MOI).Conclusión. El estudio de imagen es fundamental para definir presencia o no de MOI en todos los pacientes con hipoacusia y así poder guiar la terapéutica y el aseso-ramiento genético, así como realizar los estudios moleculares más adecuados

Introduction. The pre-lingual deafness or hearing loss are of genetic cause in be-tween 60% and 68% of cases, among these, between 20% and 40% are malforma-tion of the inner ear. From the non-syndromic hearing loss cases that are linked to the X chromosome, seven types have been described. Among these inner ear malforma-tions, incomplete cochlear partition type III is the less frequent.Objective. Present the clinical genetical report of a Mexican family, with male in-dividuals affected by congenital neurosensory deafness with inner ear malformation.Materials and methodology. A study on a family in which nine members were affected by deafness was done. Four of them, plus a mother without manifestation, were studied through a general clinical study by a geneticist, an audiological study (otoscopy and audiometry) by an audiologist, and a computed tomography (CT) scan by a radiologist.Results. The studied patients presented congenital neurosensory deafness, from se-vere to deep bilateral. Via the CT, the inner ear malformation was made clear. Three of the patients presented incomplete cochlear partition type III and one patient in-complete cochlear partition type I. Due to the clinical study and the family tree, it was diagnosed non-syndromic neurosensory deafness linked to X. The CT of the mother without manifestation did not show evidence of inner ear malformations.Conclusion. The study by image is fundamental to define whether there is or not a presence of inner ear malformations in any patient with heading loss to be able to guide the therapeutics and the genetic counseling, as well as to make more accurate molecular studies

Quantitative determination method and application of pharmaceutical adjuvant tween 80 / 中国中药杂志

Based on the fact that chromogenic reaction of blue complex, a reaction product which can be dissolved in organic solvents, can be realized by polyethoxy and ammonium thiocyanate in tween 80, a rapid and accurate way for the determination for tween 80 in pharmaceutical adjuvant was established in this study, providing reliable technical means for quality control of traditional Chinese medicine injections. Based on the study of reaction kinetics, chromogenic reaction temperature and time, as well as extraction of organic solvents and other key conditions were optimized, and Kumu injection was used as the test material for method validation and applicability investigation. It was finally determined that 3 mL ammonium thiocyanate solution was added in the sample solution, and the reaction was carried out in a boiling water bath for 2 h. After cooling to room temperature, 5 mL of dichloromethane was added to extract the chromogenic product. The absorbance value was measured at the wavelength of 623 nm to calculate the tween 80 content in the sample. Under optimized conditions, tween 80 solution showed a good linear relationship with the absorbance in the range from 0.8 mg to 3.0 mg. The linear regression equation was ＝0.258-0.047. The correlation coefficient was 0.999 6. Under the experimental conditions, the average recovery was 99.66%, and the precision RSD was less than 2.0%. The results showed that this method can quickly and accurately determine the content of tween 80 in Kumu injection, and it could be applicable to the quality control of traditional Chinese medicine injections.

Low But Continuous Occurrence of Microsporum gypseum Infection in the Study on 198 Cases in South Korea from 1979 to 2016

BACKGROUND: Microsporum gypseum has been isolated from South Korea since 1966. However, the incidence of M. gypseum infection is very low. OBJECTIVE: The aim of this study is to add massive data on M. gypseum to the literature and to provide useful information on clinical and mycological characteristics of M. gypseum. METHODS: We retrospectively analyzed data of 198 cases infected with M. gypseum from 1979 to 2016. The identification of M. gypseum was done with mycological examination using 15% KOH preparation and potato corn meal tween 80 agar culture media. RESULTS: The incidence of M. gypseum infection was very low in South Korea, showing a tendency to decrease. Out of 198 cases, men were 94 cases (47.5%) and women were 104 cases (52.5%). Mean age of all patients was 29.83 years old: 24.97 years old in men and 34.22 years old in women. M. gypseum infection occurred most frequently in September (16.7%) and August (16.2%). The most common clinical type of M. gypseum infection was tinea corporis (38.4%). CONCLUSION: M. gypseum infection shows very low incidence but still remains around us until recent years. We should keep in mind the characteristics of M. gypseum.

The influence of Tween 80 on laccase production by Pleurotus sajor-caju and the efficiency of crude enzyme broth in the removal of bisphenol-A / Influência do Tween 80 na produção de lacases de Pleurotus sajor-caju e eficiência do caldo enzimático bruto na remoção de bisfenol-A

Bisphenol-A is currently considered an environmental pollutant, capable of interfering in the endocrine system of organisms and causing alterations in its development and reproductive system. An alternative method to the chemical treatment of this pollutant has been the use of oxidative enzymes, especially laccases produced by fungi. In order to reduce production costs, agro-industrial waste can be used in the culture medium composition. Nonionic surfactants, which are only slightly toxic to biological membranes, can be applied, as well as Tween 80, to facilitate the excretion of these enzymes into the culture medium. The objectives of this work were: a) characterize the immersion water of banana straw used in the formulation of the culture medium; b) evaluate laccase production by Pleurotus sajor-caju in culture medium with and without addition of Tween 80, through shaken flasks; c) evaluate the efficiency of the crude enzyme broth in degrading bisphenol-A. The shaken flasks were incubated at 30°C for 12 days. The immersion water had a C:N ratio of 13.8, ash percentage of 28.6%, and pH close to neutrality. The addition of Tween 80 on the culture medium (7.5%, m/v) yielded laccase activity and productivity values equal to 3,016.47 U L-1 and 502.7 U L-1 day-1, respectively. These values were 50 and 33.5 times higher than those obtained in the culture medium without addition of Tween 80 for laccase activity and productivity, respectively. The crude enzyme broth degraded 100% of bisphenol-A after 48 hours, regardless of concentration (500, 750 and 1,000 mg L-1).(AU)

O bisfenol-A é considerado um poluente ambiental capaz de interferir no sistema endócrino dos organismos, ocasionando alterações em seu desenvolvimento e sistema de reprodução. Um método alternativo ao tratamento químico desse tipo de poluente tem sido a utilização de enzimas oxidativas, especialmente as lacases, produzidas por fungos. A fim de diminuir custos de produção, resíduos agroindustriais podem compor o meio de cultivo. Assim, surfactantes não iônicos e pouco tóxicos para as membranas biológicas, como o Tween 80, podem ser utilizados para facilitar a excreção dessas enzimas para o meio de cultivo. Os objetivos deste trabalho foram: caracterizar quimicamente o resíduo água de imersão de palha de bananeira, usado na formulação do meio de cultivo; avaliar a produção de lacase por Pleurotus sajor-caju em meio de cultivo líquido (frascos Erlenmeyer) com e sem adição de Tween 80; e avaliar a eficiência do caldo enzimático bruto em degradar bisfenol-A. Os frascos foram incubados a 30°C, por 12 dias. A água de imersão apresentou relação C:N 13,8, percentual de cinzas 28,6% e pH próximo da neutralidade. O cultivo adicionado de Tween 80 (7,5%, m/v) propiciou valores de atividade e produtividade em lacase iguais a 3.016,47 U L-1 e 502,7 U L-1 dia-1, respectivamente. Esses valores são 50 e 33,5 vezes maiores que os obtidos no cultivo sem adição de Tween 80, para atividade e produtividade em lacase, respectivamente. O caldo enzimático bruto degradou 100% do bisfenol-A após 48 horas, independentemente da concentração (500, 750 e 1.000 mg L-1).(AU)

Anti-bacterial effects of lavender and peppermint oils on Streptococcus mutans / 대한구강보건학회지

OBJECTIVES: The main objectives of this study were to verify the antibacterial activity of two essential oils, lavender and peppermint, against dental caries and to review their synergistic effect when used in combination. Our results provide basic data for the evaluation of the use of these two substances towards the prevention and cure of dental caries. METHODS: The sample solutions of lavender and peppermint oils were prepared in three different concentrations (30%, 50%, and 70% (v/v)) by diluting them with third-distilled water and Tween 20. Streptococcus mutans was selected as the bacterial species for testing. The disk diffusion method was used to measure the antibacterial activity of the sample solutions. For generating growth curves and measuring the number of clusters of the bacterial, the liquid medium-dilution method was used; the absorbance of the medium was measured at 600 nm after 3, 6, 12 and 24 hours. RESULTS: When the antibacterial activity of the oils was tested via the disk diffusion method, the activity improved with increasing concentrations of all the sample solutions of peppermint, lavender, and the blend, but there was no significant difference between them with respect to the type of oil. In the growth curves of S. mutans, growth inhibition was observed after 12 hours. The inhibitory effect of 30% lavender oil on growth was 64.9% and 80.1% after 12 and 24 hours of treatment, respectively whereas that of peppermint oil was 71.3% and 80.1% after 12 and 24 hours of treatment, respectively. The inhibitory effect of the blended oil was 71.9% and 81.0% after 12 and 24 hours of treatment, respectively. CONCLUSIONS: Further research is still required in order to determine the efficacy of lavender and peppermint oils, as well as other essential oils, for wider use in preventing dental caries.

Intravitreal injection of polysorbate 80: a functional and morphological study / Injeção intravítrea de polissorbato 80: estudo funcional e morfológico

ABSTRACT Objective : to determine the functional and morphological effects at rabbits retina of PS80 concentration used in the preparation of intravitreal drugs. Methods: eleven New Zealand rabbits received a intravitreal injection of 0.1ml of PS80. As control, the contralateral eye of each rabbit received the same volume of saline. Electroretinography was performed according to a modified protocol, as well as biomicroscopy and retina mapping before injection and seven and ten days after. Animals were euthanized in the 30th day and the retinas were analyzed by light microscopy. Results: eyes injected with PS80 did not present clinical signs of intraocular inflammation. Electroretinography did not show any alteration of extent and implicit time of a and b waves at scotopic and photopic conditions. There were no morphological alterations of retinas at light microscopy. Conclusion: intravitreal injection of PS80 in the used concentration for intravitreal drug preparations do not cause any functional or morphological alterations of rabbit retinas. These results suggest that PS80 is not toxic to rabbit retinas and may be safely used in the preparation of new lipophilic drugs for intravitreal injection.

RESUMO Objetivo: determinar os efeitos funcionais e morfológicos na retina de coelhos da concentração de PS80 utilizada na preparação de drogas intravítreas. Métodos: onze coelhos New Zealand receberam injeção intravítrea de 0,1ml de PS80. Como controle, o olho contralateral de cada coelho recebeu o mesmo volume de soro fisiológico. Foram realizados eletrorretinogramas de acordo com o protocolo modificado, biomicroscopia e mapeamento de retina antes da injeção, sete e dez dias depois. Os animais foram sacrificados no 30o dia e as retinas analisadas por microscopia de luz. Resultados: os olhos injetados com PS80 não apresentaram sinais clínicos de inflamação intraocular. O eletrorretinograma não apresentou alteração de amplitude e tempo implícito das ondas a e b nas condições escotópica e fotópica. Não houve alteração morfológica da retina na microscopia de luz. Conclusão: a injeção intravítrea de PS80 na concentração utilizada na preparação de drogas intravítreas não causa alterações funcionais e morfológicas na retina de coelhos. Esses resultados sugerem que o PS80 não é tóxico para a retina de coelhos e pode ser usado com segurança na preparação de novas drogas lipofílicas para injeção intravítrea.

Capacidad de formación de biopelículas en especies del género Candida de procedencia clínica / Capability of biofilm formation in Candida genus species of clinical origin

El objetivo de la investigación fue evaluar la capacidad de formación de biopelículas en especies del género Candida provenientes de muestras clínicas con procesos infecciosos. La muestra estuvo constituida por 70 aislados obtenidos de laboratorios clínicos privados. Para la identificación de las especies se utilizó la Prueba de tubo germinativo y la técnica de microcultivo en agar harina de maíz con tween 80, mientras que la cuantificación de la capacidad de formación de biopelículas se realizó por la técnica de microplaca de poliestireno. Entre los resultados obtenidos en la investigación las especies aisladas fueron: Candida glabrata (34%), Complejo Candida albicans (30%), Complejo Candida parapsilosis (27%) y Candida tropicalis (9%), de las cuales 41 aislados (59%) formaron biopelículas, resultando el Complejo Candida albicans y Candida glabrata las especies con mayor grado de formación de biopelículas con 29% cada una, seguida de Complejo Candida parapsilosis con 27% y Candida tropicalis con 15%. Las cepas que tuvieron mayor formación de biopelículas provenían de orina, seguidas de secreción ótica, secreción vaginal y sangre, sin embargo al aplicar la prueba de Chi-cuadrado (x2) de Pearson se determinó que no existe asociación estadísticamente significativa entre la capacidad de formación de biopelículas y el grado de formación con la procedencia clínica así como tampoco hay asociación entre la capacidad de formación de biopelículas y grado de formación con la especie involucrada (p > 0,05)

The aim of this research was to evaluate the capability of biofilm formation in Candida genus species from clinical samples with infectious conditions. The sample consisted of 70 isolates obtained from private clinical laboratories. Germ tube test and agar cornmeal with tween 80 microculture technique were used as species identification, meanwhile, quantification of biofilm formation capacity was performed by the polystyrene microplate technique. Among the results obtained in the research, isolated species were: Candida glabrata (34%), Candida albicans Complex (30%), Candidaparapsilosis Complex (27%) and Candida tropicalis (9%), of which 41 isolates (59%) formed biofilms, resulting Candida albicans and Candida glabrata Complex species with the highest degree of biofilms with 29% each, followed by Candida parapsilosis Complex with 27% and Candida tropicalis with 15%. Strains with more biofilm formation came from urine, followed by ear discharge, vaginal secretion and blood. However, by applying the Chi squaretest the Pearson, it was determined that there was no statistically significant association between thecapability of biofilm formation and the formation degree with clinical origin, neither there is association between the ability of biofilm formation and the formation degree with the species involved (p> 0.05).

Desenvolvimento e caracterização de nanopartículas de poli (n-butil cianoacrilato) contendo a associação lamivudina e zidovudina / Development and characterization of poly (n- butyl cyanoacrylate) nanoparticles containing the combination of lamivudine and zidovudine

A zidovudina (AZT), fármaco antirretroviral utilizado no tratamento da AIDS, apresenta biodisponibilidade oral em torno de 60% e seu uso prolongado pode ocasionar efeitos tóxicos e tolerância ao tratamento. A lamivudina (3TC), apesar de demonstrar menor citotoxicidade e menor resistência viral, é considerada também menos potente. A associação entre os dois fármacos é recomendável em função da boa resposta terapêutica e maior adesão ao tratamento. As nanopartículas são uma alternativa para melhorar a biodisponibilidade e o transporte de fármacos sobretudo através da BHE. Nesse sentido, as nanopartículas poliméricas de poli (n-butil cianoacrilato) (PBCA) apresentam grande potencial para melhoria das características farmacêuticas, além de possibilitar resultados terapêuticos mais eficazes por meio da modificação de sua superfície, direcionando o fármaco ao sítio alvo. Diante do exposto, foram desenvolvidas nanopartículas de PBCA contendo a associação lamivudina e zidovudina (3TC/AZT) revestidas com polissorbato 80 (Ps80). As nanopartículas obtidas foram caracterizadas e apresentaram resultados coerentes aos encontrados na literatura. Após a encapsulação dos fármacos e o revestimento com Ps80, notou-se um aumento no diâmetro médio e o potencial Zeta foi próximo de zero. Esses resultados juntamente com a análise de SAXS comprovam o revestimento das nanopartículas de PBCA. Os dados de DSC e TG/DTG mostram que a encapsulação foi eficiente para a estabilização térmica dos fármacos. Foi desenvolvido e validado o método analítico por CLAE, a fim de determinar a eficiência de encapsulação. A validação do método analítico para quantificação simultânea do 3TC e AZT, tanto nas nanopartículas de PBCA quanto nas nanopartículas revestidas, apresentou linearidade, especificidade, precisão e exatidão adequadas de acordo com as normativas. A porcentagem de encapsulação dos fármacos foi igual a 44,45% e 30,44%. As nanopartículas de PBCA e PBCAPs80, em concentrações abaixo de 100 µg/mL, apresentaram viabilidade celular superior a 70% em células Caco-2, comprovando que o sistema apresenta baixa citotoxicidade, o que representa uma alternativa promissora para a encapsulação de fármacos antirretrovirais e consequente progresso no tratamento da AIDS

Zidovudine (AZT), which is an anti-retroviral drug used in the treatment of AIDS, has oral bioavailability around 60% and its prolonged use can cause toxic effects and tolerance to the treatment. Lamivudine (3TC), although it has lower cytotoxicity and lower viral resistance, is also considered less potent. The association between these two drugs is recommended based on the good therapeutic response and greater adherence to treatment. Nanoparticles are an alternative to improve the bioavailability and the transport of drugs, particularly through the BBB. Thus, the polymeric nanoparticles of poly (n-butyl cyanoacrylate) (PBCA) have great potential for improving the pharmaceutical characteristics, besides enabling more effective therapeutic results through the modification of its surface, directing the drug to the target site. That being said, PBCA nanoparticles were developed containing the association of lamivudine and zidovudine (3TC/AZT) coated with polysorbate 80 (Ps80). Nanoparticles obtained were characterized and presented coherent results when compared to those found in the literature. After the encapsulation of pharmaceuticals and Ps80 coating, it was noted an increase in the average diameter and Zeta potential was close to zero. These results along with the SAXS analysis proved the coating of the PBCA nanoparticles. The data of DSC and TG/DTG show that encapsulation was efficient for thermal stabilization of pharmaceuticals. An analytical method by HPLC was developed and validated to determine the efficiency of encapsulation. The validation of the analytical method for simultaneous quantification of 3TC and AZT, in both the PBCA nanoparticles and coated nanoparticles, presented as in linearity, specificity, precision and accuracy according to the regulations. The percentage of drug encapsulation was equal to 44.45% and 30.44%. The nanoparticles of PBCA and PBCA-Ps80, at concentrations below 100 µg/ml, presented cell viability greater than 70% in Caco-2 cells, proving that the system has low cytotoxicity, which represents a promising alternative for the encapsulation of antiretroviral drugs and consequent progress in AIDS treatment

Improved Antitumor Efficacy of Hyaluronic Acid-Complexed Paclitaxel Nanoemulsions in Treating Non-Small Cell Lung Cancer

Paclitaxel (PTX) is a effectively chemotherapeutic agent which is extensively able to treat the non-small cell lung, pancreatic, breast and other cancers. But it is a practically insoluble drug with water solubility less than 1 μg/mL, which restricts its therapeutic application. To overcome the problem, hyaluronic acid-complexed paclitaxel nanoemulsions (HPNs) were prepared by ionic complexation of paclitaxel (PTX) nanoemulsions and hyaluronic acid (HA) to specifically target non-small cell lung cancer. HPNs were composed of dl-α-tocopheryl acetate, soybean oil, polysorbate 80, ferric chloride, and HA and fabricated by high-pressure homogenization. The HPNs were 85.2 ± 7.55 nm in diameter and had a zeta potential of −35.7 ± 0.25 mV. The encapsulation efficiency was almost 100%, and the PTX content was 3.0 mg/mL. We assessed the in vivo antitumor efficacy of the HPNs by measuring changes in tumor volume and body weight in nude mice transplanted with CD44-overexpressing NCI-H460 xenografts and treated with a bolus dose of saline, Taxol®, PTX nanoemulsions (PNs), or HPNs at a dose of 25 mg/kg. Suppression of cancer cell growth was higher in the PN- and HPN-treated groups than in the Taxol® group. In particular, HPN treatment dramatically inhibited tumor growth, likely because of the specific tumor-targeting affinity of HA for CD44-overexpressed cancer cells. The loss of body weight and organ weight did not vary significantly between the groups. It is suggest that HPNs should be used to effective nanocarrier system for targeting delivery of non-small cell lung cancer overexpressing CD44 and high solubilization of poorly soluble drug.

Enhanced production of erythritol and mannitol by Yarrowia lipolytica in media containing surfactants

Abstract Various chemical compounds, including surfactants, when introduced to culture media may increase the permeability of cellular membranes and thereby affect the quantity of metabolites excreted by cells. The aim of the present study was to evaluate the impact of detergents including Triton X-100, Span 20 and Tween 80 on erythritol production from glycerol by Yarrowia lipolytica Wratislavia K1 in a shake-flask experiment, batch and fed-batch cultures. When Span 20 was added to a fed-batch culture with glycerol as a carbon source (300 g L-1), erythritol production increased by 15% compared to the culture without the surfactant where it reached 142 g L-1 after 5 days, which corresponded to 0.47 g g-1 yield and productivity of 1.1 g L-1 h-1. Therefore, it was concluded that Span 20 considerably enhanced the production of this polyol from glycerol.

Optimization of growth and bacteriocin production by Lactobacillus sakei subsp. sakei2a

Lactobacillus sakei subsp. sakei 2a is a bacteriocinogenic lactic acid bacterium isolated from Brazilian pork sausage, capable of inhibiting the growth of microbial pathogens, mainly Listeria monocytogenes. In order to optimize bacteriocin production for industrial applications, this study evaluated the effect of supplementation of MRS broth with glucose, Tween 20, Tween 80, sodium citrate, potassium chloride and cysteine, and effect of the initial pH and temperature of incubation of the medium on production of bacteriocins by L. sakei 2a. Adding glucose and Tween 20 to the medium, an initial pH of 5.0 or 5.5, and incubation temperatures of 25 °C or 30 °C resulted to the highest bacteriocin yields. Thus, a 2⁴ factorial design with the four variables was performed, and statistical analysis showed that it was an adequate model (R² = 0.8296). In the studied range, the four parameters significantly influenced bacteriocin production, with the maximum yield produced at an initial pH between 5.5 and 7.0, a temperature between 25 and 30 °C and supplementation of the MRS broth with glucose from 3.25 to 6.0 g L⁻¹ and Tween 20 from 0.575 to 1.15% (v/v). Response Surface Methodology analysis indicated that the highest bacteriocin production (12800 AU mL⁻¹) occurred in the MRS broth supplemented with 5.5 g L⁻¹ glucose and 1.05% Tween 20 at an initial pH of 6.28 and an incubation temperature of 25 °C. The amount of bacteriocin produced in commercial MRS broths under the same conditions was only 5600AU mL⁻¹.

Intravenous tacrolimus and cyclosporine induced anaphylaxis: what is next?

Tacrolimus and cyclosporine have been used in various formulations, but their hypersensitivity reactions are rare in practice. Castor oil derivatives are nonionic surfactants used in aqueous preparations of hydrophobic active pharmaceutical ingredients. Castor oil derivatives that can be used as additives to tacrolimus and cyclosporine may play a role in the development of hypersensitivity reactions, especially anaphylaxis. Various immunologic and nonimmunologic mechanisms have been implicated in hypersensitivity reactions induced by castor oil derivatives. Physicians should be aware that not only the drug itself, but also its additives or metabolites could induce hypersensitivity reactions. We report a case of anaphylaxis caused by vitamin K (phytonadine), serotonin antagonist (granisetron), intravenous tacrolimus, and cyclosporine. Interestingly, the patient tolerated oral cyclosporine, which did not contain Cremophor EL or polysorbate 80.

From micelle to microemulsion: an investigation of structural changes using molecular dynamics

Objective: Although a huge number of experimental works may be observed in the literature for microemulsions, modelling reports on these nano-systems are rare. Additionally, no comprehensive work so far has detailed the changes occurring to a micellar droplet when oil molecules are introduced [i.e. obtaining microemulsions from micelles]. This work aimed to simulate a micelle and a microemulsion system containing polysorbate 80 - as surfactant - and study the structural changes in the two systems

Methods: Molecular dynamics simulation employing a coarse-grained model was used

Results: The results showed that the oil molecules located in the hydrophobic core of the micelle make its size larger, while the co-surfactant molecules distribute throughout the system and enter the hydrophilic shell of the micelle

Conclusion: It was shown that insertion of oil molecules makes the droplets more spherical, while co-surfactants tend to enlarge them

A Case of Alternaria alternata Keratitis Isolated from Corneal Tissue / 대한의진균학회지

A 71-year-old man presented with pain in the left eye that revealed a 3x3 mm deep corneal stromal infiltrate, with a 2x2 mm epithelial defect. The patient started topical moxifloxacin, voriconazole 2%, and natamycin for 2 weeks. However, the treatment was not effective and the corneal infiltration worsened. Subsequently, the patient underwent therapeutic penetrating keratoplasty. Thick brown/gray mold colonies on Potato Corn Meal Tween 80 agar was isolated from excised corneal tissue and on slide culture many septated, and club-shaped ascospores were revealed. Histological findings also showed numerous hyphae scattered in corneal tissue. A. alternata colonies were confirmed by 18S rRNA sequencing. Intracameral voriconazole was injected every other day for 2 weeks to eliminate remaining fungi on the deep corneal stroma. The remaining corneal infiltration was improved one month after the injection. During 5 months postoperative follow up, the infection did not recurred. In conclusion, deep corneal infection of A. alternata was effectively treated with intracameral voriconazole injection.

The Composition of Modified BHI Medium for Optimal Growth of Malassezia spp / 대한의진균학회지

BACKGROUND: There are several media for culture of Malassezia spp., such as Leeming & Notman (LN) medium, modified Leeming & Notman (mLN) medium, Dixon's medium and modified Dixon's medium etc. It is known that Malassezia spp. grow well in these media in general, but the kind and amounts of their ingredients are various and un-uniform according to researchers. Author propose the new and transparent BHI based medium for the optimal growth of Malassezia spp. OBJECTIVES: The purpose of this study was to design the simple and transparent BHI based medium and find essential ingredients for the growth of M. globosa and M. obtusa. METHODS: The colony size of eight standard strains (M. dermatis, M. furfur, M. globosa, M. japonica, M. obtusa, M. sloofiae, M. sympodialis, M. yamatoensis) on the modified BHI (mBHI) agar media with different ingredients was observed by naked eye after seven day culture. The compositions of mBHI medium were as follows; mBHI-1 was supplemented with 0.7% dextrose, 1.5% Tween 80, 1% glycerol to BHI medium, mBHI-2 was supplemented with 1.5% Tween 40 to mBHI-1 instead of Tween 80, mBHI-3 was supplemented with 1.5% Tween 60 to mBHI-1 instead of Tween 80, mBHI-4 was added with 0.8% bile salts to mBHI-1. mBHI-5 was supplemented with 1.5% Tween 60 to mBHI-4 instead of Tween 80, and mBHI-6 was supplemented with 1.5% Tween 40 to mBHI-4 instead of Tween 80. pH of six mBHI media was all adjusted to 6.5. RESULTS: M. furfur & M. japonica were grown well on mBHI-1 agar, but M. globosa & M. obtusa were not grown and others grown poorly. M. globosa & M. obtusa were not grown on mBHI-1 & mBHI-4 containing Tween 80 as lipid source, but others grown on all mBHI media. The media that all eight Malassezia strains grew well were slightly turbid mBHI-5 & transparent mBHI-6 medium. CONCLUSIONS: M. globosa & M. obtusa need glycerol and bile salts as well as Tween 60 or 40 instead of Tween 80 for growth. M. furfur & M. japonica need not bile salts for growth. Author proposes the transparent modified BHI medium supplemented with 0.7% dextrose, 1.5% Tween 40, 1% glycerol and 0.8% bile salts (mBHI-6) as new standard medium for culture of eight Malassezia species.

Cucurbita pepo oil as a drug microemulsion formulation: study of phase Diagram

Investigation of phase diagram of various drug formulations is effective to predict different phase region of drugs to detect final formula. The purpose of this research was to develop the ternary phase diagrams for a drug microemulsion system consisting of Cucurbita pepo [pumpkin] oil, surfactant [Tween 80] and deionized water. An electrical conductivity was used to study the properties of system. Particle size analysis of microemulsion system was performed by dynamic light scattering. The electrical conductivity of the microemulsions increases with increasing of aqueous phase content. Structural transitions from the oil-in-water to a bi-continuous phase then inversion to water-in-oil occured in the system. Diameter of particles was calculated 70 nm [for 75 percent of particles] and 35 nm [for 25 percent of particles]. Solubility results showed that microemulsion system of Cucurbita pepo oil can increase its solubility in aqueous medium due to droplet size reduction into nanometer size. Microemulsion technique can be used as a successful method in preparation of Cucurbita pepo oil drug formulation