Revisión de la displasia ectodéֹrmica hipohidrótica: diagnóstico, tratamientos y terapias génicas / Hypohidrotic ectodermal dysplasia: a review on diagnosis, treatment, and gene therapies

RESUMEN La displasia ectodérmica hipohidrótica (DEH) es una genodermatosis producida por un defecto monogenético que afecta la proteína ectodisplasina en los tejidos derivados del ectodermo. De acuerdo al mecanismo de transmisión, podemos distinguir tres tipos de DEH: autosómica recesiva, autosómica dominante, o ligada al cromosoma X (siendo ésta la más frecuente). Si bien la expresión fenotípica presenta diferencias de acuerdo al gen afectado, las tres características clínicas de la DEH son: hipodoncia, hipotricosis e hipohidrosis. El tratamiento convencional busca mejorar la calidad de vida del paciente, abarcando intervenciones dentales como ortodoncia e implantes dentales, rutinas dermatológicas para piel seca y escamosa, y el uso de sudor artificial. En los últimos años, la investigación científica desplazó el interés desde estos tratamientos hacia la ingeniería genética, surgiendo alternativas prometedoras que en ciertos ensayos clínicos lograron revertir la enfermedad, como lo es la terapia génica con EDA1 recombinante, o ER004, desarrollada por Schneider, H. A lo largo de la revisión se abordarán los síntomas de la enfermedad, las herramientas para el correcto diagnóstico y los tratamientos disponibles, así como otras condiciones a tener en cuenta en el diagnóstico diferencial.

ABSTRACT Hypohidrotic ectodermal dysplasia (HED) is a genodermatosis caused by a monogenic defect that affects the protein ectodysplasin in tissues derived from the ectoderm. Depending on the transmission mechanism, we can distinguish three types of HED: autosomal recessive, autosomal dominant and linked to the X chromosome (this being the most frequent). Although the phenotypic expression varies according to which gene is affected, all forms of HED share three clinical characteristics: hypodontia, hypohidrosis and hypothricosis. The conventional treatments have the objective of improving the patient's quality of life, and include dental interventions such as orthodontics and dental implants, dermatological routines for dry skin, and the use of artificial sweat. In recent years, scientific research has focused on genetic engineering to treat HED, developing new promising strategies such as gene therapy with recombinant EDA1, or ER004, developed by Schneider, H. and colleagues. In someclinical trials this strategy effectively reversed clinical manifestations of the disease. Throughout the review we will address HED's symptoms, tools for the correct diagnosis, available treatments and considerations for the differential diagnosis.

The protective role of IMM-H004 on hepatic ischemia-reperfusion injury in mice / 药学学报

This study investigates the protective role of IMM-H004, a novel coumarin derivative, on hepatic ischemia-reperfusion injury (HIRI) in mice. All animal experiments in this paper have been approved by the Ethics Committee of Institute of Materia Medica, Chinese Academy of Medical Sciences. The experimental animals were divided into three groups, including sham group, model group, and IMM-H004 treatment group. Serum biochemical indicators were detected and H&E staining was used to assess liver damage. Real-time quantitative PCR (qPCR) was performed to analysis the mRNA content of inflammatory factors. Immunohistochemistry and immunofluorescence were used to observe neutrophil infiltration. Western blot was used to examine the protein levels of NOD-like receptor protein 3 (NLRP3), apoptosis-associated speck-like protein (ASC), cysteinyl aspartate specific proteinase-1 (caspase-1), and interleukin-1β (IL-1β). The results showed that IMM-H004 could significantly reduce the serum levels of alanine transaminase (ALT), aspartate transaminase (AST), lactate dehydrogenase (LDH). H&E results showed IMM-H004 could alleviate liver damage caused by HIRI. The mRNA expression of tumor necrosis factor-α (TNF-α), IL-1β, and interleukin-6 (IL-6) were decreased by IMM-H004 administration. Meanwhile, IMM-H004 could markedly inhibit neutrophil infiltration. Furthermore, IMM-H004 could significantly down-regulate the protein expression of NLRP3, ASC, caspase-1, and IL-1β, inhibiting the activation of NLRP3 inflammasome pathway. Our results confirmed that IMM-H004 could protect mice from HIRI and provide a theoretical foundation for IMM-H004 application for treating HIRI.

Levantamento e comparação dos valores de multas dos principais grupos de infrações descritos no Decreto 15.004/14 da Bahia / Occurrence of fines and comparison of fee values in main group of infractions described in Decree 15.004/14 of Bahia

O Decreto Estadual 15.004/14 (BAHIA, 2014) instituiu diversas infrações com penalidade de multa conforme pode ser observado nos incisos e alíneas do artigo 163. Os valores são apresentados sempre em intervalos na moeda corrente. Nesse artigo foi realizado um estudo sobre a ocorrência dos principais grupos de infrações. A metodologia adotada foi observar os grupos que mais ocorreram, categorizando-os. Em seguida foi calculada uma média dos valores em Reais transformando-os em pontos na proporção de 1:1. Os resultados mostraram que os grupos mais encontrados foram produtos, anuência da ADAB, abate, matérias-primas, rótulos ou carimbos, embalagens, instalações ou equipamentos ou utensílios, registro, ingredientes, trânsito, contaminação e documentos. Somaram, respectivamente, 282.500, 190.000, 160.000, 137.500, 105.000, 75.000, 67.500, 67.500, 62.500, 60.000, 27.500 e 27.500 pontos. Assim, um amplo espectro de grupos de infrações foi observado no artigo 163 do citado Decreto.

Andrographis Paniculata HMPL-004, An Old Medicine with New Story / 世界科学技术-中医药现代化

In June 2004,the US Food and Drug Administration (FDA) issued Guidance for Industry:Botanical Drug Development,which opened a new path for the internationalization of traditional Chinese medicine (TCM).In October 2006,the first botanical drug product Veregen TM was approved by the FDA.The regulatory policy for botanical and herbal medicine coincides with the internationalization strategy of Hutchison MediPharma Ltd.(hereinafter referred to as Hutchison).In 2006,Hutchison's first herbal mixture medicine andrographis paniculata HMPL-004 successfully obtained Investigational New Drug (IND) from the US.From 2006 to 2014,there are eight clinical studies of HMPL-004 being conducted in Europe and the US,from Phase 1 safety study to proof-of-concept trials in ulcerative colitis (UC) and Crohn's disease.To strengthen HMPL-004 development on a global scale,Hutchison made business alliance with Nestle Health Science in clinical development as well as basic research.Due to futile outcome of an interim analysis,the Phase Ⅲ registration study in UC was terminated.However,valuable lessons were learned with the birth of the generation botanical drug product HMPL004-6599.

Estabilidad del D-004 ingrediente activo en frascos de polietileno de alta densidad / Stability of D-004 active ingredient in high density polyethylene flasks

Introducción: el D-004 es un ingrediente farmacéutico activo obtenido de los frutos de la palma real cubana (Roystonea regia), compuesto fundamentalmente por una mezcla de ácidos grasos libres entre 8 y 18 átomos de carbono, con potencialidades en el tratamiento de la hiperplasia prostática benigna. Objetivo: determinar la vida útil del D‒004 en frascos de polietileno de alta densidad en las zonas climáticas IV y II. Métodos: muestras de tres lotes pilotos de D-004, envasadas en frascos blancos de polietileno de alta densidad y protegidos por cajas de cartón, se colocaron en las condiciones de las zonas climáticas IV (30 ºC/70 % HR) y II (25 ºC/60 % HR). Se determinaron las características organolépticas y los contenidos de material insaponificable, ácidos grasos y microbiológicos desde el tiempo inicial hasta los 36 meses, y se determinaron gráficamente los tiempos de vida útil en ambas zonas. Resultados: en las dos zonas climáticas el D‒004 cumplió con los parámetros de estabilidad evaluados durante los primeros 33 meses, pero a los 36 meses el contenido de ácidos fue inferior al límite establecido en varias de las muestras. Los análisis de regresión permitieron determinar tiempos de vida útil del D‒004 en las zonas IV y II. Conclusiones: se demostró que el ingrediente activo D‒004 que se obtuvo a escala piloto, se envasa en frascos de polietileno de alta densidad y protegido por un envase secundario (caja de cartón), presenta una vida útil de 30 y 32 meses en las zonas climáticas IV y II, respectivamente(AU)

Introduction: D­004 is a pharmaceutical active ingredient obtained from the Cuban royal palm fruits (Roystonea regia), mainly composed by a mixture of free fatty acids between 8 and 18 carbon atoms, potentially useful to treat benign prostatic hyperplasia. Objective: to determine the shelf life of D­004 in high-density polyethylene flasks in climatic zones IV and II. Methods: samples of three pilot batches of D­004, packed in white high-density polyethylene flasks and protected by cardboard boxes, were placed under conditions of the climatic zones IV (30 ºC/70 % RH) and II (25 ºC / 60 % RH). Organoleptic characteristics and unsaponifiable matter, fatty acids and microbiological contents were determined from the initial time till 36 months, and shelf lives were graphically determined in both zones. Results: in the two climatic zones, the D­004 fulfilled the tested stability parameters during the first 33 months, but at the month 36th, the acid content in several samples was lower than the set limit. The regression analyses allowed determining the shelf lives of D­004 in climatic zones IV and II. Conclusions: it was demonstrated that the active ingredient D­004, obtained at pilot scale, packed in high density polyethylene flasks and protected by a secondary container (cardboard box), exhibits a shelf life of 30 and 32 months in the climatic zones IV and II, respectively(AU)

Telemedicine network Program for reperfusion of Myocardial infarction / Programa en red para la reperfusión del infarto con telemedicina

background: Early management of myocardial infarction in the area of public health requires the integration of specific programs for the coordination of healthcare services. Objective: The aim of this study was to evaluate the impact on delay times and reperfusion rate of a comprehensive program for the reperfusion of myocardial infarction in a hospital network of the Southern Greater Buenos Aires. Methods: The network consists of six low-mid-complexity hospitals and a third-level referral center with 24-hour cath-lab. Stage 1 of the program (2009-2010) evaluated the existing barriers to reperfusion; Stage 2 (2011-2013) implemented the progressive incorpora-tion of improvements and Stage 3 assessed the program (2013-2014) complemented with fellows in each hospital. Program impact was evaluated by the proportion of patients reperfused and time to its implementation. results: A total of 432 patients referred from the network were hospitalized with diagnosis of ST-segment elevation myocardial infarction. Mean age was 56±9 years and 83.3% were men. The proportion of reperfused patients progressively increased: S1 60.7%, S2 69% and S3 78%, p for trend=0.01. Time to reperfusion decreased significantly between S1 and S3, from 120 minutes (IQR 55-240) to 90 minutes (IQR 35-150), p=0.04, with a median reduction of 30 minutes in the door-to-balloon and door-to-needle times. Conclusions: The application of a program for myocardial reperfusion based on the diagnosis of barriers was associated with 28.5% increase in reperfusion, and a significant reduction in the implementation times. This public network model built on algorithms adapted to local barriers may contribute to improve the care of myocardial infarction in our country.

Carcinogenicidad del D-004 en ratas Srague Dawley: informe parcial a los 18 meses / Carcinogenicity of D-004 in Sprague Dawley rats: a partial report at 18 months

INTRODUCCIÓN. el D-004, extracto lipídico del fruto de Roystonea regia (palma real), constituido por una mezcla reproducible de ácidos grasos -principalmente oleico, palmítico, láurico y mirístico- ha mostrado ser efectivo en modelos experimentales de hiperplasia prostática. OBJETIVO. resumir los resultados parciales (18 meses) del estudio de carcinogenicidad por exposición repetida y prolongada del D-004 en ratas Sprague Dawley, el cual culminará al concluir el tiempo establecido (24 meses). MÉTODOS. ratas Sprague Dawley de uno y otro sexo fueron distribuidas aleatoriamente en cuatro grupos (50 ratas/sexo/grupo): uno control de vehículo y tres tratados con D-004 (500, 1000 y 1500 mg/kg, respectivamente). Los tratamientos se administraron oralmente por intubación gástrica. Este reporte incluye los resultados de peso corporal, observaciones clínicas, mortalidad y frecuencia de aparición de tumores hasta ese momento (18 meses). RESULTADOS. la frecuencia de mortalidad es baja: en los grupos tratados fue de un 8,3 % por un 9 % en los controles. No se evidencia una disminución en el tiempo de supervivencia y las observaciones diarias no revelaron signos indicativos de toxicidad atribuibles al tratamiento. Además, el momento de aparición de tumores y la frecuencia de estos en los animales tratados y en los controles fueron estadísticamente similares en ambos sexos. CONCLUSIONES. en conclusión, la administración oral de D-004 (500-1500 mg/kg) a las ratas no mostró, durante los primeros 18 meses del estudio, evidencias de toxicidad o carcinogenicidad temprana relacionadas con el tratamiento

INTRODUCTION: D-004, a lipid extract from royal palm (Roystonea regia) fruit constituted by a reproducible mixture of fatty acids, mainly oleic, palmitic, lauric and myristic, has shown to be effective in experimental models of prostate hyperplasia. OBJECTIVE: summarize the partial results (18 months) of the study on carcinogenicity by repeated and long-term exposure of Sprague Dawley rats to D-004, to be completed in the planned period of 24 months. METHODS: sprague Dawley rats of both sexes were randomly distributed into four groups (50 rats / sex / group): one group was the vehicle control and three were treated with D-004 (500, 1000 and 1500 mg/kg, respectively). Treatments were administered orally by gastric intubation. This report includes the results obtained so far (18 months) concerning body weight, clinical observations, mortality and frequency of appearance of tumors. RESULTS: mortality is low: 8.3% in treatment groups and 9% in controls. A reduction in survival time was not observed, and daily observations did not reveal any signs of toxicity attributable to the treatment. The time of appearance and the frequency of tumors in treated animals and controls were statistically similar in both sexes. CONCLUSIONS: oral administration of D-004 (500­1500 mg/kg) to rats during the first 18 months of the study did not show any evidence of early carcinogenicity or toxicity associated with the treatment.

A novel histone deacetylase inhibitor induces proliferation inhibition of human colon cancer cells / 军事医学

Objective To examine the anticancer effect of a novel histone deacetylase inhibitor (HDACi), JZ004, on colon cancer cells HCT-8 and HT-29, and to investigate the molecular mechanisms of proliferation inhibition and apoptosis induction of cancer cells treated by JZ 004.Methods Colon cancer cells were treated with a series of concentrations of JZ004 .MTT assay was used to detect the proliferation of cancer cells .The cell cycle distribution and apoptosis were deter-mined by flow cytometry .Rhodamine 123 and DCFH-DA were applied to detect the mitochondrial membrane potential (ΔΨm) and reactive oxygen species ( ROS) production.The protein expressions of acetyl-histone H3, p21, cyclin-dependent kinase(CDK)4, Bcl-2, Mcl-1 and Bax were assayed by Western blotting .Results JZ004 was found to inhibit proliferation and induce apoptosis of colon cancer cells in a time-and dose-dependent manner , accompanied by a dose-dependent hyperacetylation of histone H3.JZ004 induced the cancer cell arrest in G 0/G1 phase by increasing the expres-sion level of p21 while CDK4 was downregulated .JZ004 also increased cellular ROS production and reduced ΔΨm by regu-lating the expressions of Bcl-2 family proteins .Conclusion As a novel HDACi , JZ004 effectively inhibits proliferation and increases ROS production to induce apoptosis of colon cancer cells .The results indicate that JZ004 is a potential compound to be developed as an anti-colon cancer agent for clinic application .

Preparation of nanoparticles of ZL-004 and preliminary study on its pharmacokinetics / 中国药学杂志

OBJECTIVE: To prepare ZL-004 loaded PLGA nanoparticles (ZL-004-NP) and evaluate its release characteristics in vitro and pharmacokinetics in rats. METHODS: In order to determine physico-chemical property of ZL-004, saturation-constant temperature method, potentiometric method and shake flask method were employed to obtain relative parameters, respectively. After investigating single factors, the orthogonal design was used to gain optimal formulation. Then characteristic of ZL-004-NP prepared under condition of best formulation was determined by scanning electronmicroscopy, laser particle size analyzer, dialysis method, respectively. Afterward, amounts of ZL-004 in plasma were determined under UPLC-MS/MS and relative bioavailability between ZL-004-NP and its raw material was calculated. RESULTS: The feature of ZL-004-NP met aim of experiment, which contained smooth spheres, 121.34 nm of diameter, 0.16 of PDI, 89.63% of encapsulation efficiency, 7.65% of loading drug content, sustained release about 216 h in vitro and less burst in initiate stage. In vivo, cmax of ZL-004-NP was increased robustly 1.7 times, which reached to 125 μg · L-1 and tmax was cut down rapidly from (6.47 ± 0.51) h to (4.13 ± 0.48) h, compared with crude ZL-004. Relative bioavailability between ZL-004-NP and crude ZL-004 was 200.99%, which greatly improved effect of oral absorption. CONCLUSION: ZL-004-NP might be developed to improve water-insolube nature of ZL-004, which could increase oral relative bioavailability.

Carcinogenicidad del D-004 en ratas Sprague Dawley. Reporte parcial a los 18 meses / Carcinogenicity of D-004 in Sprague Dawley rats. A partial report at 18 months

Introducción: el D-004, un extracto lipídico del fruto de la palma real, Roystonea regia, constituido por una mezcla reproducible de ácidos grasos, principalmente oleico, palmítico, láurico y mirístico, ha mostrado prevenir la hiperplasia prostática (HP) inducida con T en roedores y actúa mediante un mecanismo multifactorial que implica la inhibición de 5a-reductasa prostática, el antagonismo de las respuestas mediadas por los a1-ADR, efectos anti-inflamatorios y antioxidantes. Objetivo: el presente trabajo tiene como propósito resumir los resultados parciales (18 meses) del estudio de carcinogenicidad a largo plazo del D-004 en ratas, el cual culminará con el tiempo establecido de 24 meses. Métodos: se distribuyeron ratas Sprague Dawley (SD) de ambos sexos aleatoriamente, en cuatro grupos (50 ratas/sexo/grupo): un grupo como control de vehículo, y tres tratados con D-004 (500, 1 000 y 1 500 mg/kg, respectivamente). Los tratamientos se administraron por intubación gástrica. Este reporte incluye los resultados de peso corporal, observaciones clínicas, mortalidad y frecuencia de aparición de tumores hasta ese momento. Resultados: la frecuencia de mortalidad es baja, en los grupos tratados fue de un 8.3 por ciento por un 9 por ciento en los controles, no se evidencia disminución en el tiempo de supervivencia y las observaciones diarias no revelaron signos indicativos de toxicidad atribuibles al tratamiento. Además, el momento de aparición y la frecuencia de tumores en animales tratados y controles fueron estadísticamente similares en ambos sexos. Conclusiones: en conclusión, la administración de D-004 (5001500 mg/kg) a ratas durante los primeros 18 meses del estudio, no mostró evidencias de toxicidad o carcinogenicidad temprana relacionadas con el tratamiento

Introduction: D-004, a lipid extract from royal palm (Roystonea regia) fruit made up of a reproducible mixture of fatty acids, mainly oleic, palmitic, lauric and myristic, has shown to prevent prostate hyperplasia (PH) induced by T in rodents. It acts through a multifactorial mechanism involving prostatic 5a-reductase inhibition, antagonism of a1-ADR-mediated responses, and antiinflammatory and antioxidant effects. Objective: the paper is aimed at summarizing the partial results (18 months) of the long-term carcinogenicity study of D-004 in rats, which is set to conclude in the established 24-month period. Method: Sprague Dawley (SD) rats of both sexes were randomly distributed into four groups (50 rats/sex/group): one group as vehicle control and three treated with D-004 (500, 1 000 and 1 500 mg/kg, respectively). Treatments were administered by gastric intubation. This report presents the results obtained so far concerning body weight, clinical observations, mortality and frequency of appearance of tumors. Results: mortality rate is low: 8.3 percent in the treated groups and 9 percent among controls. No evidence was found of survival time reduction, and daily observations did not reveal any signs of toxicity attributable to the treatment. The time of appearance and the frequency of tumors in treated animals and controls were statistically similar in both sexes. Conclusions: administration of D-004 (5001 500 mg/kg) to rats during the first 18 months of the study did not show any evidence of early carcinogenicity or toxicity associated with the treatment

Comparación de los efectos del D-004, imipramina y sertralina en el modelo de nado forzado en ratones / Comparative effects of D-004, Imipramine and Sertraline in the forced swimming test in mice

Introducción: La finasterida, inhibidor de la 5 a-reductasa, se emplea en el tratamiento de la hiperplasia prostática benigna, y ocasiona entre sus efectos adversos un aumento de cuadros de depresión. El D-004 es un extracto lipídico extraído a partir del fruto de la palma real (Roystonea regia), inhibe la 5 a-reductasa y previene la hiperplasia prostática benigna, y muestra un efecto antidepresivo moderado en el ensayo de nado forzado y suspensión por la cola. Objetivo: comparar el efecto del D-004 con la imipramina y la sertralina sobre la duración de las conductas de inmovilidad, nado y escalado en el ensayo de nado forzado. Métodos: se distribuyeron los ratones en ocho grupos: control (vehículo), tres tratados con D-004 (100, 250 y 500 mg/kg), dos con sertralina y dos con imipramina (30 y 50 mg/kg) respectivamente. Estos se colocaron en un cilindro de cristal que contenía agua a una altura de 6 cm y se cuantificaron las conductas. Resultados: la administración oral de D-004 (100, 250 y 500 mg/kg) durante 14 días redujo significativamente el tiempo de inmovilidad con respecto al grupo control (17, 22 y 25 porciento) y aumentó significativamente la conducta de nado en 1,58, 1,68 y 1,74 veces. Este efecto resulta moderado (25 porciento) comparado con las reducciones alcanzadas por la sertralina y la imipramina (³ 60 porciento). Las dosis mayores (250 y 500 mg/kg) ocasionaron incrementos de la conducta de escalado, 2,79 y 3,55 veces superiores a la del grupo control, lo que mostró semejanza con la imipramina, aunque con una menor eficacia. Conclusiones: el D-004 ejerce un moderado efecto antidepresivo, lo que pudiera contribuir al manejo de los pacientes con hiperplasia prostática benigna en los cuales se informa coincidencia de cuadros depresivos

Introduction: Finasteride is a 5 a-reductase inhibitor to treat benign prostatic hyperplasia (BPH) and one of the adverse effects is the increase of depressive symptoms. D-004 is a lipid extract from the real palm fruit (Roystonea regia) that is effective to prevent prostatic hyperplasia by inhibiting 5 a-reductase and shows moderate antidepressant effects in the forced swimming test (FST) and tail suspension test. Objective: to compare the effects of D-004, Imipramine and Sertraline on the duration of behaviours under conditions of immobility, swimming and climbing in the forced swimming test. Methods: mice were randomly distributed in 8 groups: control (vehicle), 3 treated with D-004 (100, 250 and 500 mg/kg), 2 with Sertraline and 2 with Imipramine (30 and 50 mg/kg) respectively. Mice were placed in a glass cylinder containing 6 cm high column of water and their behaviours were quantified. Results: oral administration of D-004 (100, 250 and 500 mg/kg) during 14 days reduced the length of time of immobility with respect to the control group (17, 22 and 25 percent), and significantly increased the behaviours at swimming by 1.58, 1.68 and 1.74 times. This is a moderate effect (25 percento) if compared with Sertraline and Imipramine (³ 60 percent) The doses of 250 and 500 mg/kg showed that climbing behaviours were 2.79 and 3.55 times higher than the control group. The results were similar to those of Imipramine but less effective. Conclusions: D-004 showed moderate antidepressant effect. This fact could help in the treatment of patients with benign prostatic hyperplasia, who reported similar depressive status

Determinación de alcoholes grasos en el ingrediente activo D004 por cromatografía de gases-espectrometría de masas / Determination of fatty alcohols in D-004 active ingredient by using gas chromatography-mass spectrometry

Introducción: el D004, nuevo extracto lipídico purificado a partir de los frutos de la palma real cubana (Roystonea regia [Kunth] F. Cook), ha mostrado ser efectivo en modelos experimentales de hiperplasia prostática, y ha mostrado efectos antioxidantes y antiiflamatorios. Objetivos: identificar los alcoholes que pudieran estar presentes en el ingrediente activo D004 utilizando la cromatografía de gases acoplada a espectrometría de masa. Métodos: muestras de 9 lotes de D004 se sometieron a saponificación con una disolución de KOH/EtOH y las fracciones insaponificables fueron extraídas con n-hexano. Los alcoholes se analizaron como derivados trimetilsilil, obtenidos por reacción con N-metil, N-trimetilsilil trifluoroacetamida. La cuantificación se basó en el empleo de 1-eicosanol como patrón interno. Resultados: el D004 presenta un contenido total de alcoholes entre 0,18 y 5,10 por ciento. Esta fracción está compuesta principalmente por 1-octacosanol (52,94 por ciento), 1-triacontanol (25,91 por ciento) y 1-hexacosanol (14,40 por ciento); y presenta como componentes minoritarios: 1-docosanol, 1-tricosanol, 1-tetracosanol, 1-pentacosanol, 1-heptacosanol, 1-nonacosanol, 1-dotriacontanol, 1-tetratriacontanol, 1-tetradecanol, 1-pentadecanol, 1-hexadecanol, 1-octadecanol y 1-eicosanol, los seis últimos en niveles de trazas. Conclusiones: se identificaron y se cuantificaron mediante cromatografía de gases acoplada a espectrometría de masa los alcoholes grasos en el ingrediente activo D004. Los componentes mayoritarios fueron 1-octacosanol, 1-triacontanol y 1-hexacosanol

Introduction: D004, a new lipid extract purified from the Cuban royal palm (Roystonea regia [Kunth] F. Cook) fruits, has been shown to be effective in experimental models of prostate hyperplasia, and to exhibit antioxidant and anti-inflammatory effects. Objectives: to identify the fatty alcohols that could be present in D004 active ingredient using gas chromatography-mass spectrometry. Methods: samples of nine batches of D004 were submitted to saponification with KOH/EtOH solution and the unsaponifiable fractions were extracted with n-hexane. The alcohols were analyzed as trimetilsilil derivatives, which were obtained by reaction with MSTFA. The quantification was carried out by the internal standard method using 1-eicosanol. Results: the total content of alcohols in D004 ranged from 0,18 to 5,10 percent. This fraction was mainly composed of 1-octacosanol (52,94 percent), 1-triacontanol (25,91 percent), and 1-hexacosanol (14,40 percent); whereas the minority components were 1-docosanol, 1-tricosanol, 1-tetracosanol, 1-pentacosanol, 1-heptacosanol, 1-nonacosanol, 1-dotriacontanol 1-tetratriacontanol, 1-tetradecanol, 1-pentadecanol, 1-hexadecanol, 1-octadecanol and 1-eicosanol were minor components, being the last six alcohols present in trace levels. Conclusions: fatty alcohols were identified and quantified in D004 active ingredient by gas chromatography-mass spectrometry. The major components were 1-octacosanol, 1-triacontanol and 1-hexacosanol

Effect of oleic, lauric and myristic acids on phenylephrine-induced contractions of isolated rat vas deferens.

-004, a lipid extract of Roystonea regia fruits that contains oleic, lauric and myristic acids as major components inhibits α1-adrenoreceptors-mediated contractile responses in isolated rat vas deferens and prostate trips; no study has demonstrated a similar effect for oleic, lauric or myristic acids individually. Therefore, the effects of D-004 (250 g/ml), oleic (100 g/ml), lauric (50 g/ml) or myristic (25 g/mL) acids and their combined effects on phenylephrine (PHE: 10-7- 10-4 mol/L) induced contractions has been studied. No treatment changed the basal tone of the preparations, but all inhibited PHE-induced contractions. D-004 produced the highest inhibition, followed by lauric acid, which was more effective than myristic and oleic acids against PHE-induced contractions of control group. D-004 and the mixture of the three acids produced similar inhibitions.

Estudio del D-004 sobre la defensa antioxidante endógena en ratas con hiperplasia prostática inducida por inyección de testosterona / Study of D-004 on the endogenous antioxidant defence in rats presenting with prostate hyperplasia induced by testosterone injection

La hiperplasia prostàtica benigna, enfermedad común en hombres mayores de 50 años de edad, se caracteriza por el crecimiento incontrolado de la glàndula prostàtica y la presencia de síntomas del tracto bajo urinario. El estrés oxidativo ha sido recientemente asociado con la causa de esta enfermedad. El D-004, extracto lipídico del fruto de la Roystonea regia, ha mostrado reducir la hiperplasia prostàtica inducida por testosterona en roedores y producir efectos antioxidantes in vitro e in vivo, pero sus efectos sobre las enzimas del sistema antioxidante endógeno no han sido estudiados. Este trabajo investigó los efectos del tratamiento oral con D-004, durante 14 días, sobre las enzimas superóxido dismutasa y catalasa en ratas con hiperplasia prostàtica inducida por testosterona. Los animales se distribuyeron en 4 grupos: un control negativo y tres inyectados con testosterona: uno tratado con el vehículo (control positivo) y dos con D-004 (400 y 800 mg/kg, respectivamente). Se determinó la capacidad antioxidante total del plasma y las actividades de las enzimas superóxido dismutasa y catalasa en eritocitos lisados y plasma, respectivamente. El tratamiento oral con D-004 (400 y 800 mg/kg) previno de modo marcado y significativo el agrandamiento de la próstata inducido con testosterona en ratas, y aumentó significativamente la capacidad antioxidante del plasma y la actividad de la catalasa, sin modificar la actividad de la superóxido dismutasa. Estos resultados sugieren que la actividad antioxidante del D-004 està relacionada, al menos parcialmente, con la estimulación de algunas enzimas del sistema antioxidante endógeno.

Benign prostatic hyperplasia, a common disease in men aged over 50 is characterized by uncontrolled growth of prostatic gland and the presence of low urinary tract symptoms. The oxidative stress has been recently associated with the disease cause. The D-004, a lipid extract from Roystonea regia, reduces the prostatic hyperplasia induced by testosterone in rodents and to produce in vitro and in vivo antioxidant effects nut its effects on endogenous antioxidant4e system enzymes haven't been studied. Present paper researched the effects of D-004 oral treatment over 14 days on dismutase and catalase superoxide enzymes in rats with prostatic hyperplasia induced by testosterone. Animals were randomized distributed into 4 groups: a negative control ant three-testosterone injected: one treated with the vehicle (positive control) and two with D-004 (400 and 800 mg/kg, respectively). The plasma total antioxidant ability was determined as well as the activity of catalase and dismutase superoxide enzymes in lysed erythrocyte and plasma, respectively. D-004 (400 and 800 mg/kg) oral treatment markedly and significantly prevented the prostate enlargement induced with testosterone in rats and increased very much the plasma antioxidant activity and of catalase, without modify the superoxide dismutase. These results suggest that D-004 antioxidant activity is partially related to stimulation of some enzymes of the endogenous antioxidant system.

Efecto del D-004, un extracto del fruto de Roystonea regia y omega-3 en el modelo de nado forzado en ratones / Effect of D-004, a fruit extact from Roystonea regia and Omega-3 in a model of forced swimming in mice

La depresión se encuentra entre las principales causas de morbilidad y mortalidad de la población adulta y su manejo farmacológico incluye, entre otras opciones alternativas, el uso de la medicina complementaria, como es el caso del aceite de pescado rico en ácidos grasos omega-3 (AGw3). El D-004, extracto lipídico de los frutos de la palma real (Roystonea regia) que consiste en una mezcla de ácidos grasos, ha mostrado poseer una moderada acción antidepresiva. El objetivo de este trabajo consistió en comparar los efectos del tratamiento oral con D-004, con los AGw3 y con su terapia combinada en el modelo del nado forzado en ratones. Para ello, ratones machos se distribuyeron en 8 grupos: uno control tratado con el vehículo, dos con D-004 (250 y 500 mg/kg ), dos con AGw3 (250 y 500 mg/kg), dos con la combinación y uno con imipramina (10 mg/kg i.p.). Todos los tratamientos disminuyeron significativamente el tiempo de inmovilidad con respecto al control, sin diferencias entre dosis similares de los respectivos tratamientos. En conclusión, el D-004, los AGw3 y su terapia combinada resultaron igualmente efectivos en reducir el tiempo de inmovilidad de los ratones, sin manifestarse efectos aditivos o sinérgicos con la administración conjunta de ambas sustancias.

Depression is among the major causes of morbidity and mortality of adult population and its pharmacologic management includes among other alternative options, the use of the complementary medicine, e.g. the fish oil rich in Omega-3 fatty acids (AGw3). The D-004, a lipid extract of Real Palm fruits (Roystonea regia) consisting of a fatty acid mixture, has a moderate antidepressant action. The aim of present paper was to compare the effects of oral treatment using D-004 with the AGw3 and with its combined therapy in forces swimming in mice. Thus, the male mice were distributed into 8 groups: one as control treated with vehicle, two with D-004 (250 and 500 mg.kg), two with AGw3 (250 and 500 mg/kg), two with the combination, and another with imipramine (10 mg/kg i.p.). All treatments decreased significantly the immobility time regarding the control, without differences among similar dose of respective treatments. We conclude that D-004, the AGw3 and its combined therapy were similary effective to reduce the immobility time in mices without additive or synergic effects with the combined administration of both substances.

Efecto de D-004, un extracto lipídico extraído de las frutas de la palma real (Roystonea regia) sobre las contracciones provocadas por la Fenilefrina sobre la próstata aislada de rata / Effect of D-004, a lipid extract from royal palm (Roystonea regia) fruits, on phenylephrine-induced contractions of isolated rat prostate

D-004, a lipid extract of Roystonea regia fruits, inhibits norepinephrine-elicited contraction in rat vas deferens. Nevertheless, the effect of D-004 on a1-adrenergic contractile responses in isolated prostate preparations had not been studied. Therefore, this study investigated the effects of D-004 on phenylephrine-induced contractions in isolated rat prostate strips. D-004 (125-500 mg/mL) was added to prostate preparations in which contractions were induced with phenylephrine (10-6-10-4 M) or with ClK (50 mM). D-004 (250-500 mg/mL) significantly (p < 0,01).and dose-dependently inhibited the phenylephrine-induced contractions through a non-competitive mechanism, since it reduced (12-60 percent) the maximal contraction to phenyle phrine with respect to control preparations. D-004 at 500 mg/mL inhibited similarly both ClK and phenylephrine-contractile responses. Tamsulosin (0.01 mg/mL) abolished (100 percent inhibition) the phenylephrine, but unaffected the ClK-induced contractions.

D-004, un extracto lipídico de las frutas de Roystonea regia, inhibe la contracción provocada por la norepinefrina en el vas deferens de las ratas. No obstante, el efecto de D-004 sobre las respuestas contráctiles adrenérgicas a1 en las preparaciones aisladas de la próstata no ha sido estudiado. Por lo tanto, este estudio investigó los efectos de D-004 sobre las contracciones provocadas por la fenilefrina en las extracciones aisladas de la próstata de la rata. D-004 (125-500 mg/mL) fue adicionado a las preparaciones a partir de la próstata en la que las contracciones fueron provocadas por la fenilefrina (10-6-10-4 M) o por CIK (50 mM). D-004 (250-500 mg/mL) en forma significativa y la dosis en forma dependiente inhibieron las contracciones provocadas por la fenilefrina mediante un mecanismo no competetitivo, reduciendo así (12-60 por ciento) la contracción máxima por la fenilefrina con respecto a las preparaciones controles. D-004 a 500 µg/mL inhibieron de la misma forma tanto CIK como las respuestas contráctiles-fenilefrina. Tamsulosin (0,01 mg/mL) eliminó la acción de la fenilefrina (100 por ciento de inhibición), pero no afectó las contracciones provocadas por CIK.

Longitud del ciclo estral en ratas Sprague Dawley tratadas in útero con extracto de Roystonea regia / Estrus cycle length in Sprague Dawley rats in uterus using Roystonea regia extract

El D-004 consiste en una mezcla de ácidos grasos que inhibe significativamente la hiperplasia prostática inducida por testosterona en roedores. El objetivo del presente estudio fue evidenciar los posibles efectos adversos sobre el ciclo estral de hembras F1 expuestas in útero al D-004. Se utilizaron ratas Sprague Dawley, distribuidas aleatoriamente en 4 grupos: un control y 3 tratados con D-004 a las dosis de 500, 750 y 1 000 mg/kg; las hembras recibieron la administración de la dosis por vía oral desde 15 días antes del apareo y hasta el fin de la lactancia. A una hembra por camada de la generación F1 se le estudió la citología vaginal y se calculó la longitud aproximada del ciclo, la cual no se vio afectada ya que no existieron diferencias significativas (p= 0,1537) entre los grupos tratados y el control. Estos resultados indican que el D-004 no reveló alteraciones del ciclo estral de las crías hembras expuestas in útero(.

D-004 is a mix of fatty acids inhibiting significantly Testosterone- induced prostatic hyperplasia in rodents. The aim of present paper was to demonstrate the potential side effects on estrus cycle of F1 female rats exposed in uterus to D-004. We used Sprague Dawley rats, distributed randomly in 4 groups: a control one and another three treated with D-004 at a dosage of 500, 750 and 1 000 mg/kg; in the female rats we administered the dose by mouth from the 15 days before mating, and up to breast feeding termination. In each female rat by litter of F1 generation, the vaginal cytology was studied, and we estimated the approximate length of cycle, remained un-affected since there were not significant differences (p= 0,1537) among treatment groups and the control one. These results show that D-004 fails to reveals alterations in the estrum cycle of female litters exposed in uterus.

Estudio de estabilidad acelerada del ingrediente activo D-004 en diferentes envases / Accelerated stability study of D-004 active ingredient in different containers

INTRODUCCIÓN: D-004 es un nuevo ingrediente activo en fase de investigación y desarrollo, obtenido de los frutos de Roystonea regia (Kunth) O.F. Cook. OBJETIVO: precisar la estabilidad de D-004 en condiciones aceleradas en frascos de vidrio ámbar, de tereftalato de polietileno y de polietileno de alta densidad. MÉTODOS: muestras de 3 lotes se almacenaron a (40 ± 2) ºC y (75 ± 5) porciento de humedad relativa durante 12 meses. Se determinaron las características organolépticas, la densidad relativa, el índice de refracción (USP 27) y el contenido de ácidos grasos mediante cromatografía de gases. RESULTADOS: los parámetros medidos se mantuvieron dentro de las especificaciones de calidad establecidas, con excepción del contenido de ácidos grasos, que en los frascos de tereftalato y polietileno disminuyó significativamente a los 3, 6 y 4,5 meses, respectivamente. CONCLUSIONES: el vidrio ámbar protege al D-004 de la degradación más que los envases plásticos estudiados.

INTRODUCTION: D-004 is a new active ingredient under research and development stage, which is obtained from Roystonea regia (Kunth) O. F. Cook fruits. OBJECTIVE: to precise over the D-004 stability under accelerated conditions in amber glass, polyethylene terephtalate and high density polyethylene containers. METHODS: samples from three batches were stored at (40 ± 2) ºC and (75 ± 5) percent relative humidity for 12 months. Organoleptic characteristics, relative density, refractive index (USP 27) and fatty acids content by gas chromatography, were determined. RESULTS: measured parameters were within the set quality specifications , except for fatty acids content that significantly decreased in polyethylene terephtalate and high density polyethylene containers after 3, 4 and a half and 6 months, respectively. CONCLUSIONS: the amber glass protects more D-004 from degradation than the analyzed plastic containers.

Evaluación genotóxica del D-004, extracto del fruto de Roystonea regia, mediante el ensayo de micronúcleos / Genotoxic assessment of D-004, extract from Roystonea regia fruit, by means of micronuclei assays

El D-004 es un extracto lipídico del fruto de la palma real (Roystonea regia), que ha demostrado ser eficaz en prevenir la hiperplasia prostßtica inducida por testosterona y por fenilefrina en modelos experimentales en ratas. El objetivo fue determinar si el D-004 induce cambios en la frecuencia de aparición de micronúcleos al realizar la administración oral a dosis repetida durante 8 semanas. Se formaron 5 grupos experimentales (7 animales/grupo): un grupo control solvente, tres tratados con D-004 (500, 1 000 y 1 500 mg/kg) y un control positivo tratado con ciclofosfamida. No ocurrieron muertes ni se detectaron signos clínicos de toxicidad. No hubo diferencias significativas entre controles y tratados en cuanto a la frecuencia de eritrocitos policromatófilos micronucleados y el índice de citotoxicidad. En conclusión, el D-004 administrado por vía oral a las dosis empleadas no presenta actividad clastogénica ni citotóxica en médula ósea de ratones OF-1 machos in vivo.

D-004 is a lipid extract from the real palm fruit (Roystonea regia), which has demonstrated that is effective to prevent prostatic hyperplasia from testosterone and from phenylephrine in rats' experimental models. Aim of this paper was to determine if D-004 provoke changes in appearance frequency of micronuclei during oral administration of repeated doses for 8 weeks. Five experimental groups were created (7animals/group): a solvent control-group, three treated with D-004(500, 1 000, and 1 500 mg/kg), and a positive control-group treated with cyclophosphamide. There were no deaths or clinical signs of toxicity neither significant differences among controls and treated ones as regards the frequency of micronucleated polychromatophils erythrocytes, and the cytotoxic index. In conclusion, D-004 administered per os at doses used has neither clastogenic nor cytotoxic activity in bone marrow male OF-1mice in vivo.

The diagnosis and method investigation of active tuberculosis situation among HIV/AIDS patients / 中华检验医学杂志

Objective To screen the active tuberculosis patients among HIV infected patients,and investigate the diagnostic methods for active tuberculosis among TB/HIV co-infected patients.Methods From August 2006 to March 2007,660 HIV/AIDS patients were enrolled.The study was conducted at 4 authorized hospitals for AIDS in Nanning and Liuzhou.Chest X-ray(CXR),acid-fast stain test of sputum smear and fast culture were applied if CD+4 T cell counts were below 350 cells/mm3 or the patients at least have one suspected symptom.Result The CD<;+>4 T cell count in 76.1% (502/660) of the patients was less than 200 cells/mm3.TB/HIV coinfection was found in 22.9% (151/660) of the HIV patients.Among them,74.8% (113/151) of them were pulmonary TB patients.One third of them were extra-pulmonary TB patients,and 68.1% of them involved lymph node.In 264 patients with negative sputum smear test and CXR,20.1% (53/264) of them showed positive results in fast culture tests.In addition,the non-tuberculosis mycobacterium (NTM) infection accounted for 38.5% culture positive cases.Conclusions The TB/HIV coinfection rate is 22.8%.Liquid rapid culture of sputum plays an import role in diagnosing of active tuberculosis among HIV patients.There are considerable proportions of NTM or extra-pulmonary TB coinfection in HIV patients.