The inhibitory effects of live and UV-killed Akkermansia muciniphila and its derivatives on cytotoxicity and inflammatory response induced by Clostridioides difficile RT001 in vitro

Clostridioides difficile infection (CDI) is the leading cause of healthcare-acquired infections worldwide. Probiotics are widely recommended to prevent CDI and its recurrences. Akkermansia muciniphila, as a therapeutic symbiont colonizing the intestinal mucosal layer, is considered to be a promising next-generation probiotic. In this work, we assessed the inhibitory effects of A. muciniphila MucT and its derivatives on cytotoxicity and inflammatory response induced by C. difficile RT001 in Caco-2 cells. The results obtained from SEM revealed that the morphology of UV-killed A. muciniphila remained unchanged after UV inactivation. TEM analysis showed that A. muciniphila–isolated extracellular vesicles (EVs) were spherical and ranged from 50 to 200 nm in size. Toxigenic supernatant (Tox-S) of C. difficile RT001 (500 μg/ml) significantly (P <0.01) reduced the cell viability of Caco-2 cells. Caco-2 cells treated with live (MOI 10), UV-killed (MOI 10), cell-free supernatant (CFS, 106 cfu/ml), and EVs (20 μg/ml) of A. muciniphila exhibited over 90% viability in comparison to untreated control. The neutralized CFS preparation using A. muciniphila and its derivatives could notably reduce the expression level of inflammatory markers. Additionally, A. muciniphila and its derivatives modulated the production of IL-1β, TNF-α, and IL-10 in Tox-S stimulated Caco-2 cells. We demonstrated that A. muciniphila and its derivatives can modulate changes in the gut barrier–related genes and inflammatory response caused by C. difficile Tox-S in Caco-2 cells. (AU)

Anticancer Properties of Caulerpa racemosa: A Review Study

Introduction: Cancer is the leading cause of death in theworld, with approximately 10 million deaths expected by2020. Several approaches are used in cancer management.However, the cost is one of the main obstacles in cancer therapy as well as side effects in sufferers. Caulerpa racemosa is a type of seaweed that is naturally abundant in theIndonesian sea. Recently, there has been much research onthe anticancer effects of Caulerpa sp. This study aims to findthe potency of Sea grapes extract (Caulerpa racemosa) in thetreatment of cancer and its mechanisms.Method: A review of the literature was constructed on thepotential of the C. racemosa extract with the PICOS criteriaand the data were extracted from ‘PUBMED’, ‘ScienceDirect’and ‘SpringerLink’. The search method was using a booleanoperator with the main keywords ‘Caulerpa racemosa’, ‘cancer’, and ‘Management’.Results: The main results were 8 articles including in vitroand in vivo experimental studies based on inclusion criteria.Several studies (n=8) revealed the potency of C. racemosaextract as an anticancer agent through various activities, suchas antiproliferative, apoptotic, antioxidant, cytotoxic activity,and inhibition of tumor progression genes, DNMT, and upregulation of proapoptotic genes, including BAX, P53, Caspase-3, Caspase-8, and Caspase-9.Discussion: C. racemosa possesses several potent antioxidant substances, along with gene regulation activities and in hibition of cell line proliferation. Seaweeds has been usedwidely as functional food and showed minimal or no toxicitiesagainst human. With all these benefits, C. racemosa has thepotential to be commercialized as a promising diet for cancerpatients.Conclusion: Sea grapes extract (C. racemosa) has goodpotential as an anticancer agent through antiproliferationmechanisms, induction of apoptosis, cytotoxic and antioxidantactivity.(AU)

Cancer Immunotherapy associated with Interstitial Lung Disease

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Could formaldehyde induce mutagenic and cytotoxic effects in buccal epithelial cells during anatomy classes?

BACKGROUND: Due to increased formaldehyde exposure, carcinogenic to humans, several researches have been studying the potential toxicity and the safe levels for human beings. The aim of this study was to investigate mutagenicity and cytotoxicity in buccal epithelial exfoliated cells (BEC) of students subjected to formaldehyde (FA) during anatomy classes. MATERIAL AND METHODS: BEC were collected periodically from 17 volunteers of undergraduate programs, who had participated in practical anatomy classes, before and after FA exposure. Cells were stained according to Feulgen method and then micronucleus test was applied. A total of 1,500 cells were assessed per individual in this study for the micronucleus frequency and other parameters of cytotoxicity. RESULTS: There was statistically significant increase in number of micronucleated BEC after FA exposure (after 1 month p=.034 and after 3.5 months p=.017). However, FA exposure caused no significant increase in other nuclear alterations closely related to cytotoxicity (p≥.05). CONCLUSIONS: FA induced mutagenicity during anatomy classes. Cell death increased, but it was not statistically significant. Efforts have to be made to improve air quality and reduce exposures during anatomy classes

Evaluation of the in vitro ocular toxicity of the fortified antibiotic eye drops prepared at the Hospital Pharmacy Departments / Evaluación de la toxicidad ocular in vitro de los colirios fortificados antibióticos elaborados en los Servicios de Farmacia Hospitalaria

The use of parenteral antibiotic eye drop formulations with non-marketed compositions or concentrations, commonly called fortified antibiotic eye drops, is a common practice in Ophthalmology in the hospital setting. The aim of this study was to evaluate the in vitro ocular toxicity of the main fortified antibiotic eye drops prepared in the Hospital Pharmacy Departments. We have conducted an in vitro experimental study in order to test the toxicity of gentamicin, amikacin, cefazolin, ceftazidime, vancomycin, colistimethate sodium and imipenem-cilastatin eye drops; their cytotoxicity and acute tissue irritation have been evaluated. Cell-based assays were performed on human stromal keratocytes, using a cell-based impedance biosensor system [xCELLigence Real-Time System Cell Analyzer (RTCA)], and the Hen’s Egg Test for the ocular irritation tests. All the eye drops, except for vancomycin and imipenem, have shown a cytotoxic effect dependent on concentration and time; higher concentrations and longer exposure times will cause a steeper decline in the population of stromal keratocytes. Vancomycin showed a major initial cytotoxic effect, which was reverted over time; and imipenem appeared as a non-toxic compound for stromal cells. The eye drops with the highest irritating effect on the ocular surface were gentamicin and vancomycin. Those antibiotic eye drops prepared at the Hospital Pharmacy Departments included in this study were considered as compounds potentially cytotoxic for the ocular surface; this toxicity was dependent on the concentration used (AU)

El uso de reformulaciones de antibióticos parenterales en forma de colirios de composición o concentraciones no comercializadas, comúnmente denominados colirios antibióticos reforzados, es una práctica habitual en oftalmología a nivel hospitalario. El objetivo del presente trabajo ha consistido en evaluar la toxicidad ocular in vitro de los principales colirios antibióticos reforzados elaborados en los Servicios de Farmacia Hospitalaria. Hemos realizado un estudio experimental in vitro para evaluar la toxicidad de los colirios de gentamicina, amikacina, cefazolina, ceftazidima, vancomicina, colistimetato de sodio e imipenem-cilastatina en el que se ha evaluado su citotoxicidad y la irritación tisular aguda. Los ensayos celulares se realizan sobre queratocitos estromales humanos, mediante la utilización de un sistema biosensor de impedancia celular [(xCELLigence Real-Time System Cell Analyzer (RTCA)] y los ensayos de irritación ocular mediante el ensayo Hen´s Egg Test. Todos los colirios, excepto vancomicina e imipenem, han mostrado un efecto citotóxico de concentración y tiempo dependiente, siendo las concentraciones más altas y los tiempos más prolongados los que provocan un descenso más pronunciado en la población de queratocitos estromales. La vancomicina muestra un importante efecto citotóxico inicial que revierte con el transcurso del tiempo y el imipenem se muestra como un compuesto no tóxico para las células estromales. Los compuestos con mayor efecto irritante para la superficie ocular son la gentamicina y la vancomicina. Los colirios antiinfecciosos elaborados en los Servicios de Farmacia Hospitalaria estudiados se muestran como compuestos potencialmente citotóxicos para la superficie ocular, siendo esta toxicidad dependiente de la concentración utilizada (AU)

Investigation of twenty selected medicinal plants from Malaysia for anti-Chikungunya virus activity

Chikungunya virus is a reemerging arbovirus transmitted mainly by Aedes mosquitoes. As there are no specific treatments available, Chikungunya virus infection is a significant public health problem. This study investigated 120 extracts from selected medicinal plants for anti-Chikungunya virus activity. The plant materials were subjected to sequential solvent extraction to obtain six different extracts for each plant. The cytotoxicity and antiviral activity of each extract were examined using African monkey kidney epithelial (Vero) cells. The ethanol, methanol and chloroform extracts of Tradescantia spathacea (Commelinaceae) leaves showed the strongest cytopathic effect inhibition on Vero cells, resulting in cell viabilities of 92.6% ± 1.0% (512 µg/ml), 91.5% ± 1.7% (512 µg/ml) and 88.8% ± 2.4% (80 µg/ml) respectively. However, quantitative RT-PCR analysis revealed that the chloroform extract of Rhapis excelsa (Arecaceae) leaves resulted in the highest percentage of reduction of viral load (98.1%), followed by the ethyl acetate extract of Vernonia amygdalina (Compositae) leaves (95.5%). The corresponding 50% effective concentrations (EC50) and selectivity indices for these two extracts were 29.9 ± 0.9 and 32.4 ± 1.3 µg/ml, and 5.4 and 5.1 respectively. Rhapis excelsa and Vernonia amygdalina could be sources of anti-Chikungunya virus agents (AU)

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Cytotoxic activity and induction of inflammatory mediators of the methanol:chloroform extract of Fusarium moniliforme / Actividad citotóxica e inducción de mediadores inflamatorios del extracto metanol: cloroformo de Fusarium moniliforme

Background. Fusarium moniliforme is a phytopathogenic facultative fungus with a cosmopolitan distribution in all types of climates, and has a wide host range, including, among others, bean, rice, wheat and sorghum crops. There is a current lack of knowledge regarding the potential of these fungi, so it is considered to be of great importance to obtain information related to the biological activity of extracts and secondary metabolites. Aims. An evaluation of the role of methanol:chloroform extract of F. moniliforme in the production of inflammatory cytokines and their cytotoxic activity. Methods. The production of nitric oxide was analyzed by the Griess method, the production of cytokines using ELISA, and the effects of the extract on cell cycle and induction of apoptosis by flow cytometry. Results. The extract of F. moniliforme was seen to be able to stimulate nitric oxide (NO) production in J774A.1 cells, as well as to produce cytokines such as, IL-1β, IL-6, and TNF-α. It was also observed that the extract of F. moniliforme produces activity on cell cycle modulation and apoptosis when tested in carcinogenic cell lines. Conclusions. The results obtained from this study open the possibility of obtaining and identifying metabolites of the extract of F. moniliforme that can be evaluated for possible use in cancer therapy (AU)

Antecedentes. Fusarium moniliforme es un hongo fitopatógeno facultativo con distribución cosmopolita en todos los tipos de climas y con una gran variedad de huéspedes, como el frijol, el arroz, el trigo y el sorgo. Actualmente existe una falta de conocimiento sobre el potencial de este hongo, por lo que es de gran importancia obtener información relacionada con la actividad biológica de los extractos y sus metabolitos secundarios. Objetivos. En este trabajo se evaluó el papel del extracto metanol:cloroformo de F. moniliforme sobre la producción de citocinas y su actividad citotóxica sobre líneas celulares. Métodos. Se analizó la producción de óxido nítrico por el método de Griess, la producción de citocinas se evalúo por el método de ELISA y los efectos del extracto sobre el ciclo celular e inducción de apoptosis se analizó por citometría de flujo. Resultados. El extracto de F. moniliforme fue capaz de inducir la producción de óxido nítrico (ON) en células J774A.1, así como la producción de citocinas IL-1β, IL-6 y TNF-α. También se observó que el extracto de F. moniliforme posee actividad en la modulación del ciclo celular y en la inducción de apoptosis observada en líneas células cancerígenas. Conclusiones. Los resultados de este trabajo abren la posibilidad de obtener e identificar los metabolitos del extracto de F. moniliforme para que puedan ser evaluados en una posible terapia para el cáncer (AU)

¿Es posible curar la infección por VIH? / Is it possible to cure HIV infection?

El tratamiento antirretroviral ha logrado normalizar las expectativas de vida de las personas infectadas por VIH, pero no logra la curación de la enfermedad. Se han identificado obstáculos que impiden la curación con solo tratamiento antirretroviral, que incluyen la existencia de un reservorio de células latentemente infectadas, la replicación vírica persistente en tejidos y los santuarios anatómicos. Se persigue como principal estrategia de curación la administración de fármacos que reactiven el virus latente para de este modo eliminar el reservorio celular. Los ensayos clínicos en marcha han mostrado la prueba de concepto, pero aún no se ha demostrado la eficacia de estos fármacos en disminuir el tamaño del reservorio (AU)

Antiretroviral therapy has significantly improved the life expectancy in HIV-infected people, but it cannot cure the disease by itself. Several barriers have been identified for the cure of HIV infection, including a reservoir of latently infected cells, persistent viral replication in tissues, and anatomical sanctuaries. The main strategy proposed for the cure of HIV consists on the administration of drugs that, through the reactivation of latent HIV, would eliminate the cell reservoir. Ongoing clinical trials have shown the proof of concept, but the efficacy of these drugs in decreasing the reservoir size has not been proved so far (AU)

La actividad citotóxica de las células natural killer como herramienta diagnóstica en pacientes pediátricos críticos con sospecha de síndrome hemofagocítico / Natural killer cell cytotoxic activity in critical pediatric patients with suspected hemophagocytic syndrome

OBJETIVO: Determinar el papel de la citotoxicidad de las células natural killer (NK) en pacientes pediátricos críticos con sospecha de síndrome hemofagocítico (SH). DISEÑO: Estudio prospectivo realizado en el período comprendido entre septiembre de 2008 y febrero de 2014. ÁMBITO: Se ha realizado en el Laboratorio de Onco-Hematología del Hospital Infantil Universitario Niño Jesús de Madrid. PACIENTES: Se analizaron 30 muestras de sangre periférica de pacientes ingresados en la Unidad de Cuidados Intensivos de diferentes centros hospitalarios. Los pacientes, 18 niños y 12 niñas, tenían una media de edad de 4,7 años (rango 0,2-22). La citotoxicidad NK se comparó con un control sano del mismo sexo y similar edad. INTERVENCIÓN: Determinamos la citotoxicidad in vitro NK frente a la línea celular K562 mediante fluorescencia resuelta en el tiempo (Europium-TDA) en condiciones de reposo, tras estimulación con interleucina 15 y bloqueo con el anticuerpo Fas ligando. VARIABLE DE INTERÉS: La citotoxicidad NK. RESULTADOS: En 20 pacientes la citotoxicidad NK estaba disminuida (p = 0,001). De ellos, 9 fueron diagnosticados de SH primario y 11 de SH secundario. En 10 pacientes la citotoxicidad fue normal; ninguno fue diagnosticado de SH. La estimulación con interleucina 15 aumentó la citotoxicidad NK en los SH secundarios, y el bloqueo de Fas ligando la disminuyó en los pacientes con SH primarios (p = 0,001). CONCLUSIONES: En nuestra experiencia, la citotoxicidad NK constituye una prueba útil para el diagnóstico de los SH. La estimulación con interleucina 15 y el bloqueo con Fas ligando nos pueden ayudar al diagnóstico diferencial de los SH primarios y secundarios

AIM: To determine the role of natural killer (NK) cytotoxic activity in patients with suspected hemophagocytic lymphohistiocytosis syndrome (HLH). DESIGN: A prospective study was conducted from September 2008 to February 2014. SCOPE: The study was carried out in the Hematological Oncology Laboratory of Hospital Infantil Universitario Niño Jesús, Madrid (Spain). PATIENTS: We analyzed 30 peripheral blood samples from intensive care patients with suspected HLH. There were 18 males and 12 females, with a mean age of 4.7 years (range 0.2-22). NK cell cytotoxicity was compared with healthy controls according to age and sex. INTERVENTION: In vitro NK cell cytotoxicity against the K562 cell line was determined by time-resolved fluorescence (Europium-TDA) under resting conditions, after interleukin 15 stimulation, and following block with Fas ligand antibody. VARIABLE OF INTEREST: NK cell cytotoxicity. RESULTS: A total of 20 patients showed a significant decrease of NK cell activity compared with controls (P=.001). Nine of these patients were diagnosed with primary HLH. A total of 10 patients were diagnosed with secondary HLH. Cytotoxic activity was normal in 10 subjects. None of them were diagnosed with HLH. Interleukin 15 stimulation increased NK cell cytotoxicity in secondary HLH, and blocking Fas ligand on NK cells decreased cytotoxic activity in primary HLH patients (P = .001). CONCLUSIONS: In our experience, NK cell cytotoxic activity measured by time-resolved fluorescence is a simple and useful clinical diagnostic test for HLH. Interleukin 15 stimulation and Fas ligand blocking on NK cells could help differentiate between primary and secondary HLH

Dendritic cells transfected with hepatocellular carcinoma (HCC) total RNA induce specific immune responses against HCC in vitro and in vivo

BACKGROUND: Immunotherapy is an effective method for preventing metastasis and recurrence of carcinoma. Hepatocellular carcinoma (HCC) is a common malignancy with a high rate of recurrence, and has not successfully been introduced to immunotherapy. METHODS: Peripheral blood mononuclear cells were isolated from whole blood of HCC patients and stimulated to transform into dendritic cells (DCs). These DCs were then transfected with RNA extracted from HepG-2 hepatoma cells to induce expression of specific antigens. RESULTS: The transfected DCs stimulated T lymphocytes to produce cytotoxic T lymphocytes, which specifically attacked HepG-2 cells. Injection of T lymphocytes from HCC patients and transfected DCs into severe combined immunodeficiency mice limited the growth of HepG-2 tumors. CONCLUSION: A specific immune response against hepatoma can be generated in vivo by administering DCs transfected with RNA from a specific tumor. This method may have therapeutic application in humans to reduce recurrence of HCC (AU)

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Measuring human mast cell-mediated cytotoxicity against human tumour cells: three-colour flow cytometry using monoclonal antibody target staining with annexin V / propidium iodide co-labelling of death

Background: Chromium release assay is the standard method for evaluation of cell-mediated cytotoxicity, including that of mast cells. Although this is a reproducible method, it has more drawbacks than even radioactivity. In addition to the shortcoming of measuring only necrotic killing, some non-radioactive methods have not been widely used either. The numerous limitations of these methods led researchers to develop other techniques. This study describes a new flow cytometric approach that measures human mast cell-mediated cytotoxicity by marking target cells with monoclonal antibody alongside annexin V/ PI co-labelling. Methods: A colony forming unit - mast in vitro was developed from human bone marrow mononuclear cells in serum-free methylcellulose medium. Six-week-old human bone marrow-derived mast cells were used as effectors, and malignant B-lymphoblastoid cell lines like Daudi / Raji cells as targets. Effectors and targets were both co-incubated for short and long-term durations, and experiments were repeated several times. Cytotoxicity was calculated by flow cytometric mast cell-mediated cytotoxicity assay. Results: This method was able to clearly show mast cell-mediated cytotoxicity against human tumours. It is well-known that some lymphokine-activated killer-sensitive cells are resistant to mast cell-mediated cytotoxicity. However, a different type of lymphokine activated killer-sensitive cell in this study was found to be very sensitive to mast cell-mediated cytotoxicity. Moreover, this technique also allowed us to separate killing into different stages: early and late apoptosis. Conclusions: When compared to chromium release and non-radioactive methods, this method has the advantages of allowing evaluation of early apoptosis and short/long term mast cell-mediated cytotoxicity with specific target marking(AU)

Interleukin-15 enhances T-cell responses by stimulation with dendritic cells

INTRODUCTION: Cytokines play important roles in regulating immune responses. Interleukin-2 (IL-2) has usually been used as an adjuvant to enhance antitumour immune responses. However, its crucial role in activation-induced cell death, inhibition of homeostatic proliferation of CD8+ memory T cells and its notable biological side effects impair its prospect of application. IL-15 has several similar functions to IL-2 and shows potential advantages over IL-2, and is being investigated to enhance antitumour dendritic cell (DC) vaccine strategies in our ongoing studies. OBJECTIVE: In this preliminary study, we evaluated the ability of IL-15, compared with IL-2, to act as an adjuvant to enhance T-cell responses activated by DCs in vitro. MATERIALS AND METHODS: Bone marrow-derived DCs (BMDCs) were pulsed with tumour antigens and used to stimulate lymphocyte responses in the presence of IL-15 or IL-2. The activated T lymphocytes were examined by flow cytometric analysis, and interferon-γ (IFN-γ) enzyme-linked immunospot and cytotoxicity assays. RESULTS: IL-15 was observed to activate lymphocytes with comparable phenotype characteristics of activated/memory CD8+ lymphocytes, compared with IL-2. Both in primary and secondary stimulation with DCs, when using IL-15 as an adjuvant, activated lymphocytes showed higher proportions of IFN-γ-secreting subsets. In secondary stimulation with BMDCs in the presence of IL-15, the activated lymphocytes showed a stronger cytotoxicity to antigen-specific tumour target cells. CONCLUSIONS: Our study suggested that IL-15 might be a prospective adjuvant for a DC vaccine strategy against cancers. The further observation that IL-15 acts as an adjuvant for an antitumour DC vaccine strategy is worth investigating (AU)

Poria cocos, un hongo con propiedades antiinflamatorias e inmunoestimulantes / Poria cocos, a fungus with anti-inflammatory and immunoestimulant properties

El esclerocio de Poria cocos (Schw.) Wolf (Poliporáceas) denominado "fu-ling" (China) o "hoelen" (Japón) es una de las drogas más apreciadas en las medicinas tradicionales orientales por sus propiedades diuréticas, sedantes y tónicas. Se trata de un hongo saprofítico que crece sobre diferentes especies de Pinus, y cuyos principios activos más relevantes son los polisacáridos y los triterpenoides, los cuales han sido el principal objetivo de los estudios farmacológicos. De ellos, los triterpenos destacan por sus propiedades antiinflamatorias y los polisacáridos como inmunoestimulantes, mientras que en las propiedades citotóxicas están implicados ambos tipos de principios. Sin embargo, el número de ensayos clínicos realizados, para establecer su eficacia y seguridad, es limitado. En esta revisión se han recopilado todos los datos sobre química y farmacología de Poria cocos, con el fin de establecer su interés clínico para futuros empleos en patologías donde la inflamación y el sistema inmunológico estén implicados (AU)

The screrotium of Poria cocos (Schw.) Wolf (Polyporaceae), also called "fu-ling" (China) or "hoelen" (Japan), is a wellknown traditional Asian medicine used for its diuretic, sedative, and tonic effects. It is a saprophytic fungus growing in diverse species of Pinus. Polysaccharides and triterpenoids, the two relevant groups of phytochemicals present in Poria cocos, have been the principal subjects of pharmacological studies on this fungus. Whereas triterpenes exhibit marked anti-inflammatory activity, polysaccharides have immunostimulant effects, but both groups have been shown to possess anticancer properties. Unfortunately, the amount of published research on the clinical properties of Poria cocos is insufficient to establish its efficacy and safety from a scientific point of view. In this review we have compiled all the data on the plant’s chemistry and pharmacology in order to determine its clinical interest for future use in various pathologies in which inflammation and immunodepression are implicated (AU)

Os escleródios de Poria cocos (Schw.) Wolf (Polyporaceae) chamado "fu-ling" (China) ou "hoelen" (Japão) é uma das drogas mais populares na medicina oriental tradicional para a sua diurética, sedativa e tônica. É um fungo saprófita que cresce em várias espécies de Pinus, e cujos princípios são polissacarídeos e triterpenos, que têm sido o principal objetivo dos estudos farmacológicos. Destes, os triterpenos são conhecidos por suas propriedades antiinflamatórias e polissacarídeos como imunoestimulante, enquanto as propriedades citotóxicas envolvendo ambos os tipos de princípios activos. No entanto, o número de ensaios clínicos para determinar sua segurança e eficácia é limitada. Nesta revisão, reunimos todos os dados sobre a química e farmacologia de Poria cocos, a fim de estabelecer a sua relevância clínica para os trabalhos futuros em doenças onde a inflamação eo sistema imunológico estão envolvidos (AU)

Cytotoxic activity of Alfa-humulene and transcaryophyllene from Salvia officinalis in animal and human tumor cells / Actividad citotóxica del Alfa-humuleno y del tras-cariofileno de Salvia officinalis en dos líneas celulares tumorales animal y humana

Background: The purpose of the present work is two-fold: the fractionationof Salvia officinalis essential oil and the cytotoxic study ofthis oil with its fractions ¡°in vitro¡± tumor cell lines. Materials andMethods: S. officinalis essential oil was obtained by hydrodistillationand fractionated with column chromatography; the essential oil andits fractions were analyzed by gas chromatography (GC) coupled tomass spectrometry (MS). The cytotoxic activity was evaluated incellular lines of breast cancer MCF-7, colon cancer HCT-116, andmurine macrophage RAW264.7 cell lines by the MTT assay. Results:the sub-subfraction F1.1.1 of S. officinalis essential oil containing ¦Á-humulene present highest activity on RAW264.7 and HCT-116 withIC50 values of 41.9 and 77.3 ¦Ìg/ml, respectively. The sub-subfractionF1.2.1 of S. officinalis essential oil with trans-caryophyllene showedless activity on RAW246.7 and HCT-116 with IC50 values of 90.5 and145.8 ¦Ìg/ml. Conclusion: This paper suggests that the ¦Á-humulene andtrans-caryophyllene extracted from S.officinalis essential oil inhibit tumorcell growth (AU)

Antecedentes: Este trabajo tiene dos objetivos: el fraccionamiento del aceite esencial de la especie Salvia officinalis y la determinación de la citotoxicidad del mencionado aceite esencial con sus fracciones en líneas celulares tumorales “in vitro”. Material y Métodos: El aceite esencial de Salvia officinalis fue obtenido por hidrodestilación y fraccionado mediante cromatografía en columna; el aceite esencial y sus fracciones fueron analizadas mediante cromatografía de gases (GC) acoplada a espectrometría de masas (MS). La actividad citotóxica fue evaluada en líneas celulares de cáncer de mama MCF-7; cáncer de colon HCT-116 y en macrófago murino. RAW264.7 con el ensayo MTT. Resultados: La sub-subfracción F1.1.1 del aceite esencial de Salvia officinalis que contiene alfa-humuleno presenta la actividad mas acusada frente a las líneas celulares RAW264.7 y HCT-116, con valores de IC50 de 41,9 y de 77,3 μg/ml respectivamente. La sub-subfracción F1.2.1 del aceite esencial de Salvia officinalis con trans-cariofileno, muestra menor actividad sobre células RAW246.7 y HCT-116 con valores de IC50 de 90,5 y 145,8 μg/ml respectivamente. Conclusión: Estos resultados sugieren que el alfa-humuleno y el trans-cariofileno de los extractos del aceite esencial de Salvia officinalis inhiben el crecimiento de células tumorales (AU)

Características químicas, mecanismo de acción y actividad antiviral de darunavir / Chemical characteristics, mechanism of action and antiviral activity of darunavir

Darunavir es el resultado de una amplia y profundainvestigación en el campo de la farmacología del bloqueode la proteasa del virus de la inmunodeficiencia humana(VIH). Se trata de un inhibidor de la proteasa (IP) nopeptídico, con una estructura química distinta que,confiriéndole una mayor y mejor afinidad por la diana yuna mayor estabilidad frente a su disociación, lo hace máspotente que el resto de los IP conocidos.Sus características farmacocinéticas exigen lacoadministración de dosis bajas de ritonavir y permitenbien la administración oral (preferentemente con lascomidas), la administración una vez al día frente a cepasdel VIH no resistentes y una dosificación cómoda en muyvariadas situaciones, incluyendo las insuficiencias renal yhepática leves-moderadas.Su potencial de interacciones farmacológicas se asumefácilmente y puede administrarse sin ajuste de dosis concasi todos los antirretrovirales, excepto maraviroc,lopinavir, saquinavir y tipranavir. Tampoco hay problemasde antagonismo farmacodinámico con ninguno de ellos.Las dosis citotóxicas están muy por encima de las dosisterapéuticas, lo que le otorga un amplio margen deseguridad.Su espectro de acción es muy amplio, y resulta eficazfrente a todos los subtipos del VIH-1 y frente al VIH-2, yactúa bien en las líneas celulares mononucleares ymonociticomacrofágicas. Además, es activo frente a lamayoría de los VIH resistentes al resto de los IP, y surobustez frente a los mecanismos conocidos de resistenciadel VIH es también superior al del resto de los IPdisponibles, de tal manera que la inducción y selección demutaciones que confieren resistencia a este fármacoparece ser más lenta y difícil, lo que le podría permitirmantener su efecto antiviral inalterado durante largosperíodos(AU)

Darunavir is the result of wide and in-depth investigationinto HIV protease inhibitors (PIs). This drug is a nonpeptidePI, with a distinct chemical structure that, byconferring it drug with enhanced binding affinity and aslower dissociation rate, makes it more potent than theremaining PIs developed to date.Because of its pharmacokinetic characteristics, darunavirmust be coadministered with low doses of ritonavir.Furthermore, these characteristics allow oraladministration (preferably with meals), once-dailyadministration in non-resistant HIV strains, and a lesscomplicated treatment regimen with improvedconvenience in highly varied contexts, including mild-tomoderaterenal and hepatic impairment.The potential of darunavir for pharmacological interactionsis highly acceptable and this drug can be administeredwithout dose adjustments with almost all antiretroviralagents except maraviroc, lopinavir, saquinavir andtipranavir. There are no problems of pharmacodynamicantagonism with any of these drugs.Cytotoxic doses are well above therapeutic doses,providing a wide safety margin.The spectrum of action is very wide, and darunavir iseffective against all subtypes of HIV-1 and against HIV-2and acts well in mononuclear and monocyte/macrophagecell lines. Darunavir is also active against most HIV strainsresistant to the remaining PIs and the robustness of thisdrug against the known mechanisms of resistance of HIVis also superior to that of the other available PIs.Consequently, the induction and selection of mutationsconferring resistance to this drug may be slower and moredifficult, resulting in its antiviral effect remainingunchanged for prolonged periods(AU)

Actividades citotóxicas y antibacterianas de las raíces de Capparis zeylanica Linn / Antibacterial and Cytotoxic Activities of Capparis zeylanica Linn Roots

Se analizaron extractos crudos y un ácido graso, ácido octadec-7-en-5-ynoic (1), de la corteza de la raíz de Cappariszeylanica Linn. (familia de las Capparidaceae) para observar sus actividades antibacterianas frente a la bacteria Gram positiva y Gram negativa. Entre los extractos crudos, el extracto de cloroformo mostró una buena actividad frente a todos los organismos de prueba. El ácido graso (1) aislado del extracto de cloroformo mostró actividades antibacterianas frente a todos los organismos de prueba, a excepción de E. coli. Las actividades se compararon con un antibiótico estándar: la kanamicina. Las concentraciones inhibitorias mínimas (CIH) de 1, determinadas mediante la técnica de dilución en serie, fueron 64 mg/ml frente a Bacillus subtilis y Shigella dysenteriae. Las actividades citotóxicas del extracto crudo y del ácido graso (1) se observaron mediante el bioensayo de gambas en salmuera yel valor de LC50 del compuesto fue 6,27 mg/ml (AU)

Crude extracts and a fatty acid, octadec-7-en-5-ynoic acid (1), from the root bark of Capparis zeylanica Linn. (Fam. Capparidaceae) were screened for their antibacterial activities against Gram positive and Gram negative bacteria. Among the crude extracts, chloroform extract showed good activity against all test organisms. The fatty acid (1) isolated from chloroform extract exhibited antibacterial activities against test organisms except E. coli. The activities were compared to a standard antibiotic- kanamycin. The minimum inhibitory concentrations (MICs) of 1, determined byserial dilution technique, were found to be 64 mg/ml against Bacillus subtilis and Shigella dysenteriae. The cytotoxic activities of crude extract and fatty acid (1) were observed by brine shrimp biassay and LC50 value of the compound was found to be 6.27 mg/ml (AU)

Síntesis de nuevos antitumorales por manipulación de modelos naturales de origen marino / Synthesis of new antitumour agents through structure manipulation of

Se están estudiando diversas series de compuestos con estructura de pirazino[2,1-b]isoquinolina y 6,15-iminoisoquino[3,2-b]-3-benzazocina tomando comomodelos productos naturales de origen marino con actividad antitumoral. Hasta elmomento se han desarrollado diversas estrategias de síntesis, y se ha evaluado lacitotoxicidad in vitro de estos compuestos frente a tres líneas celulares de cáncerhumano. El paso clave para su obtención se basa fundamentalmente en la síntesisde pirazinoisoquinolina-1,4-dionas por un procedimiento «one-pot» N-alquilaciónciclaciónde 3-arilmetilpiperazina-2,5-dioxosilillactimas con dimetil acetales de diversosaldehídos, utilizándose también otras reacciones de α-amidoalquilación yciclación intramolecular tipo Pictet-Spengler. Se han obtenido diversas pirazino[2,1-b]isoquinolinas, pirazino[2,1,6-bc]isoquinolinas, 6,15-iminoisoquino[3,2-b]-3-benzazocinas y otros compuestos octacíclicos más complejos, y se están derivandoen algunos casos a precursores de especies iminio alquilantes. La actividadcitotóxica micromolar y submicromolar que se ha encontrado hasta el momentoparece no depender de su capacidad para generar intermedios que permitan elenlace covalente con el ADN

Several series of pyrazino[2,1-b]isoquinoline and 6,15-iminoisoquino[3,2-b]-3-benzazocine compounds related to marine antitumour natural products are beingsynthesized and their in vitro cytotoxic activities are being evaluated against threehuman cancer cell lines. The key synthetic step is based on the synthesis of pyrazinoisoquinoline-1,4-diones by the one-pot N-alkylation-cyclization of 3-arylmethylpiperazine-2,5-dioxosilyllactims with aldehyde dimethyl acetals or throughdifferent alpha-amidoalkylation reactions and intramolecular cyclizations of Pictet-Spengler type. Pyrazino[2,1-b]isoquinolines, pyrazino[2,1,6-bc]isoquinolines, 6,15-iminoisoquino[3,2-b]-3-benzazocines and other more complex octacyclic compoundshave been obtained and derived in some cases to precursors of alkylatingiminium ion species. The micromolar an submicromolar cytotoxic activity foundup to date is apparently uninfluenced by the ability to generate intermediateswhich would permit covalent bonding to DNA

Sistema inmune y enfermedad hepática por alcohol / Immune system and alcoholic liver disease

Es un hecho bien conocido que el alcoholismo se asocia con una respuesta inmune anormal. Hasta ahora, el hallazgo más importante ha sido un estado de inmunodepresión, lo que conlleva un mayor riesgo de infecciones graves en el paciente alcohólico. Sin embargo, recientemente se ha constatado que el consumo de etanol se acompaña de cambios en la síntesis y en las concentraciones circulantes de determinadas citocinas y de una activación de subpoblaciones linfocitarias citotóxicas que podrían contribuir a la aparición de la enfermedad hepática por alcohol. En el desarrollo de la lesión hepatocelular es relevante el papel del factor de necrosis tumoral alfa como inductor de apoptosis y necrosis; esta citocina, junto con las interleucinas 1, 6 y las quimiocinas, propicia la inflamación del parénquima hepático. Los factores transformantes de crecimiento y el derivado de las plaquetas actúan sobre las células estrelladas y favorecen la fibrogénesis hepática. El avance en los conocimientos de los mecanismos inmunológicos que intervienen en la enfermedad hepática alcohólica ha permitido abrir nuevas vías terapéuticas, que en el futuro podrían modificar su evolución

It is well established that alcoholism is associated with imbalanced immune responses. To date, most relevant finding reported is the existence of an immunodepressed state which leads to a higher risk of suffering from severe infections in alcoholic patients. However, recent studies have shown that ethanol intake is followed by changes involving the synthesis and serum levels of specific cytokines as well as the activation of several different subsets of cytotoxic lymphocytes, that could be involved in the development of alcoholic liver disease. Accordingly, tumor necrosis factor-alpha plays a key role in the development of alcoholic liver damage through the induction of both apoptosis and necrosis of hepatocytes. This cytokine, together with interleukin (IL) 1, IL6 and several chemokines, facilitate the development of inflammation of the liver. Additionally, both transforming growth factor-beta and platelet-derived growth factor, act over stellate cells favouring hepatic fibrogenesis. The advances in the knowledge of the immunological mechanisms involved in alcoholic liver disease may lead to the discovery of new potential therapeutic targets, which may modify disease outcome in the near future

Avances en la inmunopatología de la infección por el VIH / Advances in the immunopathology of HIV infection

La infección por el virus de la inmunodeficiencia humana (VIH) origina una destrucción progresiva e irreversible del sistema inmunitario. Aunque este proceso está directamente relacionado con la infección de los linfocitos CD4, la disminución de esta población celular a lo largo de la infección por el VIH no se debe únicamente al efecto citopático provocado por la replicación viral. La inmunopatogenia del sida es un proceso extraordinariamente complejo en el que se encuentran implicados mecanismos patogénicos muy diferentes, algunos de los cuales no son todavía completamente comprendidos. En este artículo se analizan los mecanismos mediante los cuales el VIH se adapta y destruye sus células diana, la respuesta del sistema inmunitario frente a la infección por el VIH y, por último, los mecanismos de escape que permiten al VIH eludir esta respuesta inmunitaria (AU)

Inhibición de la actividad citotóxica no restringida por MHC de células NK y de clones T CD8+alfa-beta por la agregación de moléculas MHC de clase I / Inhibition of non MHC-restricted cytotoxicity of human NK cells and a CD8+alfa-beta T cell clone by MHC class I cross-linking

La lista de ligandos del sistema inmune que se convierten en receptores es cada día más amplia. Nuestros datos experimentales muestran que la agregación de moléculas MHC de clase I humanas por anticuerpos monoclonales inhibe la lisis no específica de células Natural Killer (NK) y de linfocitos T citotóxicos. Estos resultados sugieren que las moléculas MHC de clase I humanas expresadas por células efectoras T citotóxicas (CTL) o células NK pueden transmitir señales negativas tras su interacción con ligandos expresados por las células que necesitan protegerse de la citotoxicidad no específica. En consecuencia, las moléculas MHC I no son únicamente ligandos del receptor de antígeno T o de receptores NK no reordenados, sino que también son moléculas transmisoras de señales por sí mismas probablemente tras co-localización en grupos de activación supramolecular o asociación con receptores con capacidad de transducción de señales. Además, nuestros resultados experimentales pueden tener consecuencias en el estudio de las señales inhibitorias que modulan la respuesta citotóxica de linfocitos T y células NK en donde está generalizado el uso de anticuerpos anti-MHC I. En resumen, nuestros datos sugieren una nueva función de las moléculas MHC de clase I humanas relevante en el estudio de las señales reguladoras que controlan la actividad de las células efectoras NK y citotóxicas T (AU)