Está en la página 1de 44

HOMEONEWS

Edicin N 4 Enero/Febrero de 2006 Director: Farm. Fernando Estevez Castillo

PUBLICACION BIMESTRAL DISTRIBUCION GRATUITA PARA PROFESIONALES DE LA SALUD

Homeonews
Edicin N 4 Enero-Febrero de 2006 Edicin n 4 Enero / Febrero de 2006 Registro de la Propiedad Intelectual n: 418380 Director: Fernando Oscar Estevez Castillo Propietario: Fernando Oscar Estevez Castillo (C.I.: 10.103.605)

Direccin postal: Pte. Quintana 414 Lans Oeste Pcia de Buenos Aires (B1824NVJ), Argentina Tel.: (54-11) 4241-4441 E.Mail: festevez_castillo@hotmail.com festevezcastillo@fullzero.com.ar

Director: Fernando Estevez Castillo

Homeonews
Edicin N 4 Enero-Febrero de 2006 INDICE 1- Editorial, pg. 4. 2- Fitoterapia:

-Efecto de los brotes del rbano japons (Raphanus sativus) sobre el metabolismo de los lpidos y carbohidratos en ratas normales o con diabetes inducida por estreptozotocina; pg. 5. -Actividad antifngica y mecanismos de accin de compuestos extraidos de Trbulus terrestris L.; pg. 6. 3- Alopata: Comparacin del cido azelaico y la antralina en el tratamiento de la alopecia areata circunscripta: estudio piloto; pg. 20. 4- Homeopata: Estimulacin de la actividad mitocondrial del esperma bovino mediante diluciones homeopticas de monensina; pg. 21. 5- Nutricin: Los cidos grasos Omega-3 disminuyen la irritabilidad de pacientes con desrdenes bipolares: estudio abierto; pg. 22. 6- Notas de inters: I Curso Internacional de Farmacia Homeoptica (I Diplomatura en Terapias Alternativas, Universidad Nacional Mayor de San Marcos, Lima, Per); pg. 32. 7- Novedades: CHIACAPSR: nuevo suplemento dietario; pg. 34. Homeodinamizador HiemusR; pg. 38. 8- Otros temas: Cursos; pg. 39. Libros; pg. 43. 10- Formulario de suscripcin, pg. 44. Foto de tapa: Salvia hispnica L. (Labiatae) Las opiniones vertidas en los artculos firmados son responsabilidad de sus autores. Director: Fernando Estevez Castillo

Homeonews
Edicin N 4 Enero-Febrero de 2006

EDITORIAL
Estimados colegas del equipo de salud: Nuevamente nos reencontramos con un nuevo nmero de mucha informacin y novedades.

Homeonews, con

Quiero aprovechar esta oportunidad para alertar a mis colegas sobre ciertos Cursos de Farmacia Homeoptica que una Farmacia est difundiendo en Argentina, con una larga enumeracin de antecedentes (Universidades Nacionales y Extranjeras) que toman como propios, omitiendo que los mismos corresponden al profesional (que ejerca la Direccin Tcnica) que los ha dictado durante aos por invitacin directa a su persona y no a esa institucin. Es por esto que a modo de consejo, y para no crear falsas expectativas, sugiero verificar siempre los antecedentes acadmicos de los docentes de los Cursos, as como tambin la validez de los certificados emitidos, fundamentalmente para la certificacin profesional. En este nmero podrn encontrar dos trabajos muy interesantes de Fitoterapia realizados con el Raphanus sativus y el Trbulus terrestris; la comparacin de dos sustancias (cido azelaico y antralina) para el tratamiento de la alopeca areata circunscripta; la estimulacin de la actividad mitocondrial del esperma bovino mediante diluciones homeopticas de monensina y un estudio que demuestra que los cidos grasos Omega-3 disminuyen la irritabilidad de pacientes con desrdenes bipolares. En el captulo NOTAS DE INTERES, las repercusiones del I Curso Internacional de Farmacia Homeoptica dictado en la Universidad Nacional Mayor de San Marcos de Per. Tambin dos NOVEDADES muy interesantes: Laboratorios Dr. Madaus & Co. present CHIACAPSR, el primer suplemento dietario aprobado en Argentina con aceite de Cha y la eleccin del HOMEODINAMIZADOR HIEMUS para la realizacin de los trabajos prcticos del Curso de Farmacia Homeoptica dictado en la UNMSM (Per). En el captulo de Cursos, dos actividades para recomendar, el Curso 2006 de Medicina Biolgica, inmunomodulacin, terapias complementarias y oculoanlisis y el Curso Anual de Fitomedicina y en referencia a Libros, dos textos del Dr. Jorge Alonso, que todo profesional que trabaja con plantas medicinales debera tener. Prximamente SAIDAH organizar un Taller y una Jornada que oportunamente les comunicar por este medio. Llego el momento esperado para recorrer Homeonews en toda su extensin, deseando que sea de vuestro agrado, los saludo hasta el prximo nmero. Farm. Fernando Estvez Castillo

Director: Fernando Estevez Castillo

Homeonews
Edicin N 4 Enero-Febrero de 2006

FITOTERAPIA
EFECTO DE LOS BROTES DEL RBANO JAPONS (RAPHANUS SATIVUS) SOBRE EL METABOLISMO DE LOS LPIDOS Y CARBOHIDRATOS EN RATAS NORMALES O CON DIABETES INDUCIDA POR ESTREPTOZOTOCINA Hironobu Taniguchi1, Kazuo Kobayashi-Hattori2 *, Chie Tenmyo2, Tomoko Kamei 2 , Yasushi Uda 3, Yoshiko Sugita-Konishi4, Yuichi Oishi5, Toshichika Takita2
1

Department of Nutritional Science and Culinary Arts, Toita Woman's College, 2-21-17 Shiba, Minato-ku, Tokyo 105-0014, Japan 2 Department of Nutritional Sciences, Faculty of Applied Bio-Science, Tokyo University of Agriculture, 1-1-1 Sakuragaoka, Setagaya-ku, Tokyo 156-8502, Japan 3 Department of Bioproductive Sciences, Faculty of Agriculture, Utsunomiya University, 350 Minemachi, Utsunomiya, Tochigi 321-8505, Japan 4 Division of Microbiology, National Institute of Health Sciences, 1-18-1 Kamiyoga, Setagaya-ku, Tokyo 158-8501, Japan 5 Department of Food and Nutritional Science, College of Bioscience and Biotechnology, Chubu University, 1200 Matsumoto-cho, Kasugai-City, Aichi 487-8501, Japan. [Phytotherapy Research, Vol 20, No 4 (Marzo 24), 2006: pp. 274-278] RESUMEN No existe informacin disponible sobre los efectos de los brotes del rbano japons (JRS) sobre la diabetes. Para aclarar el tema, la influencia del JRS sobre el metabolismo de los lpidos y carbohidratos fue investigada en ratas normales y diabticas inducidas por estreptozotocina. Estos animales fueron alimentados con una dieta conteniendo 0%, 2.5% o 5% de JRS ad libitum durante 21 das. Comparados con los correspondientes grupos controles, las ratas normales alimentadas con JRS mostraron niveles plasmticos ms bajos de colesterol total (TC), triglicridos (TG), fosfolpidos (PL), fructosamina, glucosa e insulina y niveles ms altos de LDL-Colesterol, mientras que las ratas diabticas alimentadas con JRS mostraron niveles plasmticos ms bajos de fructosamina, glucosa e insulina sin modificaciones en los parmetros lipdicos del plasma. El JRS tambin disminuy los niveles hepticos de TC, TG and PL en las ratas normales y los TG en las ratas diabticas. Estos resultados muestran que el JRS tuvo una actividad hipoglucemiante tanto en las ratas normales como diabticas y parcialmente mejoraron el metabolismo lipdico en las ratas normales. El JRS tiene el potencial de mejorar a los individuos con hiperglucemia en los casos donde la diabetes est presente y en la prevencin primaria de la diabetes mellitus.

Director: Fernando Estevez Castillo

Homeonews
Edicin N 4 Enero-Febrero de 2006

Palabras clave: Diabetes. Brotes de rbano japons. Metabolismo de carbohidratos. Metabolismo lipdico. Efecto hipoglucemiante. Estreptozotocina.

ACTIVIDAD ANTIFUNGICA Y MECANISMOS DE ACCION DE COMPUESTOS EXTRAIDOS DE TRIBULUS TERRESTRIS L. Jun-Dong Zhanga, Zheng Xua, Yong-Bing Caoa, Hai-Sheng Chenb, Lan Yana, Mao-Mao Ana, Ping-Hui Gaoa, Yan Wanga, Xin-Ming Jiaa and Yuan-Ying Jianga,
a

Department of Pharmacology, College of Pharmacy, Second Military Medical University, 325 Guo He Road, Shanghai 200433, PR China b Department of Natural Products Chemistry, College of Pharmacy, Second Military Medical University, 325 Guo He Road, Shanghai 200433, PR China [Journal of Ethnopharmacology, Volume 103, Issue 1 , 3 January 2006, Pages 36-42] RESUMEN La actividad antifngica de los productos naturales est siendo ampliamente estudiada. Las saponinas son conocidas por sus propiedades antifngicas y antibacterianas. Nosotros utilizamos un fraccionamiento tipo bioensayo guiado para aislar ocho saponinas esteroides a partir del Tribulus terrestris L., las cuales fueron identificadas como hecogenin-3-O--D-glucopyranosyl (1 4)-D-galactopyranoside (TTS-8), tigogenin-3-O--D-glucopyranosyl (1 4)--Dgalactopyranoside (TTS-9), hecogenin-3-O--D-glucopyranosyl (1 2)--Dglucopyranosyl (1 4)--D-galactopyranoside (TTS-10), hecogenin-3-O--Dxylopyranosyl (1 3)--D-glucopyranosyl (1 4)--D-galactopyranoside (TTS11), tigogenin-3-O--D-xylopyranosyl (1 2)-[-D-xylopyranosyl (1 3)]--Dglucopyranosyl (1 4)-[-L-rhamnopyranosyl (1 2)]--D-galactopyranoside (TTS-12), 3-O-{-D-xylopyranosyl (1 2)-[-D-xylopyranosyl (1 3)]--Dglucopyranosyl (1 4)-[-L-rhamnopyranosyl (1 2)]--D-galactopyranosyl}26-O--D-glucopyranosyl-22-methoxy-(3,5,25R)-furostan-3,26-diol (TTS-13), hecogenin-3-O--D-glucopyranosyl (1 2)-[-D-xylopyranosyl (1 3)]--Dglucopyranosyl (1 4)--D-galactopyranoside (TTS-14), tigogenin-3-O--Dglucopyranosyl (1 2)-[-D-xylopyranosyl (1 3)]--D-glucopyranosyl (1 4)-D-galactopyranoside (TTS-15). La actividad antifngica in vitro de las ocho saponinas contra cinco levaduras, Candida albicans, Candida glabrata, Candida parapsilosis, Candida tropicalis and Cryptococcus neoformans fueron estudiadas utilizando un ensayo de dilucin en microcaldo. Se estudi en particular, la actividad in vivo de TTS-12 en un modelo de infeccin vaginal con Candida albicans. Los resultados mostraron que TTS-12 y TTS-15 fueron muy efectivas in vitro contra muchas especies de cndidas patognicas y Cryptococcus neoformans. Es digno de destacar que TTS-12 y TTS-15 fueron muy activas contra Candida albicans (MIC80 = 10 y 2.3 g/mL) y Cryptococcus neoformans (MIC80 = 1.7 y 6.7 g/mL). La microscopa de contraste de fases Director: Fernando Estevez Castillo

Homeonews
Edicin N 4 Enero-Febrero de 2006

mostr que TTS-12 inhibe la formacin de hifas, un factor de virulencia importante de Candida albicans, mientras que la microscopa de transmisin electrnica que TTS-12 destruye la membrana celular de Candida albicans. Como conclusin, TTS-12 tiene una actividad antifngica in vitro e in vivo, debilitando la virulencia de Candida albicans y matando a los hongos a travs de la destruccin de la membrana celular. Palabras clave: Tribulus terrestris L.; Saponinas; Antifngicos; Candida albicans Abreviaturas: MICs, concentracin inhibitoria mnima; TEM: microscopa de transferencia electrnica.

Artculo original
1. Introduction In recent 20 years, the risk of opportunistic fungal infections has greatly increased in patients who are severely immunocompromised due to cancer chemotherapy, organ or bone marrow transplantation and human immunodeficiency virus infection (Wingard et al., 1979, Wingard et al., 1991 and Wingard et al., 1993). Candida albicans is an organism that is most often associated with serious fungal infections, and can cause fungal diseases in immunocompromised patients, including cancer patients, organ transplant patients, and those with human immunodeficiency virus infections (Fridkin and Jarvis, 1996). Candidal vaginitis is predominantly caused by strains of Candida albicans (90%) (Sobel et al., 1995, Sobel et al., 1998a, Sobel et al., 1998b and Sobel et al., 2001), and remains to be a common problem in immunocompetent or healthy women. Despite advances in antifungal therapies, many problems remain to be solved for most antifungal drugs available. For example, the use of amphotericin B, known as the gold standard, is limited because of its infusion-related reactions and nephrotoxicity (Grasela et al., 1990 and Fanos and Cataldi, 2000). The use of azoles, such as fluconazole, ketoconazole and miconazole, has resulted in clinically resistant strains of Candida spp. (Lyman and Walsh, 1992 and Sojakova et al., 2004). A 3.67.2% of vaginal isolates of Candida albicans from women with candidal vaginitis is resistant to fluconazole (Sobel et al., 2003). This situation highlights the need for advent of safe, novel and effective antifungal compounds. Plants provide abundant resources of antimicrobial compounds and have been used for centuries to inhibit microbial growth. Tribulus terrestris L. (Zygophyllaceae) is an annual creeping herb widely growing in China. It is also distributed in Japan, Korea, western Asia, southern Europe and Africa. In traditional Chinese pharmaceuticals, Tribulus terrestris L. is used for treating cutaneous pruritus, edema, inflammation and tracheitis (Jiangsu New Medical College, 1977). In our previous study, we isolated and identified 10 compounds from Tribulus terrestris L. (Xu et al., 2000). In the present report, eight of the 10

Director: Fernando Estevez Castillo

Homeonews
Edicin N 4 Enero-Febrero de 2006

saponins were tested to investigate their antifungal properties, especially against Candida albicans. 2. Materials and methods 2.1. Plant materials Tribulus terrestris L. was collected from Henan Province, China. It was identified by Prof. H.C. Zheng of the Department of Pharmacognosy, College of Pharmacy of the Second Military Medical University, Shanghai. A voucher specimen (950812) is available in the herbarium of this Department. 2.2. Extraction and purification The air-dried and powdered plant (10.7 kg) was extracted three times with an excess of 80% EtOH at room temperature. After removal of the solvent by evaporation, the residue was extracted with petrol, CHCl3 and n-butanol. Our previous preliminary study on Candida albicans showed that the n-butanol extract using the macrobroth dilution method had antifungal activity. The nbutanol layer was chromatographed over a macroporous resin column (10 cm 50 cm, 2 kg), and first eluted successively with water and then with 50%, 70% and 90% EtOH. The four fractions were separated by a combination of chromatography over silica gel, reversed phase RP-18 chromatography, sephadex G-25 and HPLC to yield pure compounds TTS-8 (33 mg), TTS-9 (21 mg), TTS-10 (46 mg), TTS-11 (27 mg), TTS-12 (1 g), TTS-13 (72 mg), TTS14 (43 mg), TTS-15 (52 mg), TTS-16 (56 mg) and TTS-18 (42 mg). 2.3. Organisms used A total of 69 American Type Culture Collection (ATCC) and clinical isolates of Candida species and Cryptococcus neoformans obtained from different hospitals in China, or commercially, or kindly donated were tested (Table 1). The collection included the following numbers of isolates: 51 isolates of Candida albicans (ATCC76625, ATCC64550 isolates and 49 clinical isolates), 4 isolates of Candida tropicalis (4 clinical isolates), 5 isolates of Candida glabrata (ATCC11006 isolate and 4 clinical isolates), 5 isolates of Candida parapsilosis (ATCC18062 isolates and 4 clinical isolates) and 3 isolates of Cryptococcus neoformans (ATCC32609 isolate and 2 clinical isolates). All isolates were identified by Shanghai Changhai Hospital. The isolates were stored as water suspensions until use. Prior to test, each isolate was passaged on potato dextrose agar (Sangon, Shanghai, China) to ensure purity and viability. Table 1. In vitro MIC80 values of eight compounds from Tribulus terrestris L. against different yeasts
No.

Compound TTS-8

Candida albicans >128.0

Candida glabrata >128.0

Candida parapsilosis >128.0

Candida tropicalis >128.0

Cryptococcus neoformans >128.0

Director: Fernando Estevez Castillo

Homeonews
Edicin N 4 Enero-Febrero de 2006
Compound TTS-9 TTS-10 TTS-11 TTS-12 TTS-13 TTS-14 TTS-15 FLC AMB ICZ Candida albicans >128.0 >128.0 >128.0 1.0 0.6 >128.0 41.7 20.5 2.3 1.0 1.3 0.7 0.29 0.15 0.12 0.06 Candida glabrata >128.0 >128.0 >128.0 8.8 4.4 >128.0 57.6 41.7 19.2 12.1 1.4 0.5 0.28 0.21 0.35 0.14 Candida parapsilosis >128.0 >128.0 >128.0 45.3 21.3 >128.0 >128.0 74.7 26.1 5 2 3.3 0.75 027 0.38 0.14 Candida tropicalis >128.0 >128.0 >128.0 21.3 8.3 >128.0 >128.0 106.7 33.0 1.5 0.6 0.63 0.25 0.38 0.14 Cryptococcus neoformans >128.0 >128.0 >128.0 1.7 0.6 >128.0 48.0 27.7 6.7 2.3 1.33 0.58 0.21 0.07 0.17 0.07

No.

2 3 4 5 6 7 8 9 10 11

Candida albicans SC5314, a strain most often used in the study of virulence and genetics of Candida albicans, was kindly donated by White TC from the University of Washington, and Spencer Redding from the University of Texas Health Science Center at San Antonio. 2.4. Media All strains used in this study were grown in two complete media consisting of a YEPD liquid medium (1% Bacto Peptone [Difco, USA], 0.5% yeast extract [Difco], 2% glucose [Sangon]), and a solid medium prepared by adding 2% agar (Sangon). 2.5. Laboratory animals Forty female SpragueDawley (SD) rats weighing 100120 g (Center of Experimental Animals, Second Military Medical University, Shanghai, China) were used for the study of vaginal infections with Candida albicans. This experiment was approved by the Bioethic Committee of the Second Military Medical University, and the procedures of the experiment were strictly according to generally accepted international rules and regulations. 2.6. Antifungal susceptibility test The in vitro minimal inhibitory concentrations (MICs) of the compounds were determined by the micro-broth dilution method according to the methods defined by the National Committee for Clinical Laboratory Standards (NCCLS, 2002). Candida krusei (ATCC6258) and Candida parapsilosis (ATCC22019) were quality controlled strains, and tested in each assay. Fluconazole (FLC), itraconazole (ICZ) and amphotericin B (AMB) obtained from their respective manufacturers served as the positive control. The drug MIC80 was defined as

Director: Fernando Estevez Castillo

Homeonews
Edicin N 4 Enero-Febrero de 2006

10

the first well with an approximate 80% reduction in growth compared to the growth of the drug-free well. The eight compounds to be tested were dissolved in dimethyl sulfoxide (DMSO), and the stock solutions of the serial two-fold dilutions were prepared in RPMI 1640 medium (Gibco, USA) with the final concentrations between 128.0 and 0.25 g/mL (111.300.220 mol/L), and the final concentrations of FLC, ICZ and AMB were 64.00.125 g/mL (209.150.410 mol/L), 2.00.004 g/mL (2.830.006 mol/L) and 2.00.004 g/mL (2.160.004 mol/L), respectively, depending on the MIC results from our preliminary study. 2.7. Growth curve study The effect of TTS-12 and TTS-15 exposure in relation to time and concentration on Candida albicans SC5314 was determined in YEPD liquid medium. TTS-12 and TTS-15 solutions (in DMSO) were added to the cultures to form an optical density of 0.1 (measured at a wavelength of 600 nm), the final concentrations of which were 0, 2, 4, 8 or 16 g/mL (0, 1.74, 3.48, 6.96, 13.91 mol/L). The growth was monitored by measuring the optical density (600 nm) of the cultures during the subsequent 48 h. 2.8. Vaginal infection model with Candida albicans The vaginal infection animal model was established based on modified models previously described by Sobel et al. (1998) to obtain a more chronic and homogeneous infection. Briefly, 40 female animals were ovariectomized, and estrus was induced with subcutaneous administration of estradiol at a dose of 10 mg/kg 3 days before infection and maintained by subcutaneous estradiol at a dose of 4 mg/kg weekly throughout the experiment. Candida albicans was inoculated intravaginally with 107 yeast cells per 0.1 mL of sterile saline and 0.1 mL per rat. Inoculation was performed using a micropipette with disposable tips. The 32 infected animals were equally randomized into four groups: Group 1, control; Group 2, miconazole (MCZ, 30 mg/kg); Group 3, TTS-12 (30 mg/kg); Group 4, TTS-12 (60 mg/kg). MCZ was served as the positive control. The vaginal Candida albicans load was evaluated at day 3 post-infection, and day 3, 7, 14 after initiation of drug administration. TTS-12 or MCZ was administered to the infection animals for 14 consecutive days. 2.9. Hyphal induction Candida albicans SC5314 cells were induced to form hyphae in medium 199 (10 M199, Gibco). The medium was pre-warmed to 37 C. The cells from a 48 h stationary-phase culture were transferred to 5 mL of 1 M199 to a final concentration of 3 106 cells/mL, and TTS-12 solution was added to the growth medium to final concentrations of 0, 4 and 16 g/mL (0, 3.48 and 13.91 mol/L), and the cultures were incubated for 6 h at 37 C, 5% CO2. The hyphal formation of Candida albicans SC5314 was seen with an inverted phase contrast microscope with the magnification of 400. FLC was used as a positive control with the final concentration of 4 g/mL.

Director: Fernando Estevez Castillo

Homeonews
Edicin N 4 Enero-Febrero de 2006 2.10. Ultrastructure analysis by transmission electron microscopy

11

Transmission electronic microscopy was performed to observe the effect of TTS-12 on cell ultrastructure. Candida albicans SC5314 cells (1 108 cells/mL) were collected after being treated with TTS-12 at 8 g/mL for 16 h, washed twice with PBS solution, centrifuged for 10 min at 3000 rpm, fixed in 2% glutaraldehyde at 4 C for 72 h, and then placed in 1% phosphotungstic acid. The cells were desiccated with gradients, and embedded with EPON-812. Ultrathin sections were prepared and observed after double staining with uranium and plumbum under a transmission electron microscope (HITACHI H800, Japan) with 2 104 magnification. At the same time, the untreated cells were used as control, and FLC (8 g/mL) was served as the positive control. 3. Results 3.1. Identification of 10 compounds Identification of the 10 compounds showed that they were hecogenin-3-O--Dglucopyranosyl (1 4)--D-galactopyranoside (TTS-8), tigogenin-3-O--Dglucopyranosyl (1 4)--D-galactopyranoside (TTS-9), hecogenin-3-O--Dglucopyranosyl (1 2)--D-glucopyranosyl (1 4)--D-galactopyranoside (TTS-10), hecogenin-3-O--D-xylopyranosyl (1 3)--D-glucopyranosyl (1 4)--D-galactopyranoside (TTS-11), tigogenin-3-O--D-xylopyranosyl (1 2)-[-D-xylopyranosyl (1 3)]--D-glucopyranosyl (1 4)-[-L(TTS-12), 3-O-{-Drhamnopyranosyl (1 2)]--D-galactopyranoside xylopyranosyl (1 2)-[-D-xylopyranosyl (1 3)]--D-glucopyranosyl (1 4)[-L-rhamnopyranosyl (1 2)]--D-galactopyranosyl}-26-O--D-glucopyranosyl22-methoxy-(3,5,25R)-furostan-3,26-diol (TTS-13), hecogenin-3-O--Dglucopyranosyl (1 2)-[-D-xylopyranosyl (1 3)]--D-glucopyranosyl (1 4)-D-galactopyranoside (TTS-14), tigogenin-3-O--D-glucopyranosyl (1 2)-[D-xylopyranosyl (1 3)]--D-glucopyranosyl (1 4)--D-galactopyranoside (TTS-15), 2S,3S,4S,5S-hexitol (TTS-16), inorganic salt (a mixture of NaNO3, KNO3 and K2NO2FHF in a ratio of 46.1:36.4:17.4) (TTS-18). The chemical structures of the eight compounds were shown in Fig. 1.

Fig. 1. Chemical structures of compounds 18. Director: Fernando Estevez Castillo

Homeonews
Edicin N 4 Enero-Febrero de 2006 3.2. Antifungal susceptibility results

12

Of the 10 compounds isolated from Tribulus terrestris L., 8 compounds were identified as steroid saponins, the in vitro activity of which were evaluated against five human pathogenic yeasts (Candida albicans, Candida tropicalis, Candida parapsilosis, Candida glabrata, Cryptococcus neoformans) which are often encountered clinically. The results were showed in Table 1. TTS-8, TTS-9, TTS-10, TTS-11 and TTS-13 were inactive against fungi tested, and compound TTS-14 had somewhat activities against Candida albicans, Candida glabrata, Cryptococcus neoformans, with MIC80 values of 41.7, 57.6, 48.0 g/mL, respectively. Especially, TTS-12 and TTS-15 had significant antifungal activities against the five yeasts tested, Candida albicans, Candida glabrata, Candida parapsilosis, Candida tropicalis, Cryptococcus neoformans. Importantly, TTS-12 and TTS-15 clearly inhibited the growth of Candida albicans, and the MIC80 value was determined to be 1.0 and 2.3 g/mL, respectively. They were also very effective against Cryptococcus neoformans at 1.7 and 6.7 g/mL. 3.3. Growth curve TTS-12 and TTS-15 activity showed dose and time dependency against the growth of Candida albicans SC5314 (Fig. 2). By 8 h post-incubation significant inhibition was observed at concentrations as low as 8.16 g/mL compared with the control.

Fig. 2. The effect of TTS-12 or TTS-15 on the growth of Candida albicans. Yeast cells were treated with TTS-12 (0, 2, 4, 8 and Director: Fernando Estevez Castillo

Homeonews
Edicin N 4 Enero-Febrero de 2006 16 g/mL) (A) or TTS-15 (0, 2, 4, 8 and 16 g/mL) (B) in the subsequent for 48 h. 3.4. The vaginal infection model

13

After completing the experiment with in vitro activity, we examined the activity of TTS-12 in vivo. For this purpose, an experimental vaginal infection model (oestrogen-dependent rat vaginitis) was established, where the animals were challenged with Candida albicans strain. All rats were infected with Candida albicans and assessed mycologically 3 days after infection. Fig. 3 shows that TTS-12 caused a rapid clearance of the strain from the vagina of the experimentally infected rats. In the vaginal infection model with Candida albicans, the number of the animals infected significantly decreased after TTS12 was administrated at the dose of 30 and 60 mg/kg, and there was a statistically significant difference between the control and the TTS-12 treatment groups at all time-points.

Fig. 3. The number of vaginal infection animals. The rats were infected vaginally with Candida albicans, and then treated with MCZ (30 mg/kg), TTS-12 (30 mg/kg) and TTS-12 (60 mg/kg) in various treatment days (0 days, 3 days, 7 days and 14 days). Significance: *P < 0.05, **P < 0.01, ***P < 0.001 compared to the control. 3.5. Hyphal induction Candida albicans SC5314 cells were incubated for 6 h in the presence of either DMSO (control), or 4 and 16 g/mL TTS-12, or 4 g/mL FLC, and then observed by phase contrast microscopy. In the absence of the drug, hyphal formation was observed in the isolate SC5314, while in the 4 and 16 g/mL TTS-12 or FLC groups, hyphal formation of Candida albicans was inhibited markedly (Fig. 4).

Director: Fernando Estevez Castillo

Homeonews
Edicin N 4 Enero-Febrero de 2006

14

Fig. 4. Hyphalformation of Candida albicans cells. Candida albicans SC5314 cells were induced to form hyphae in medium 199, and then were treated with TTS-12 (A, control; B, 4 g/mL; C, 16 g/mL), and FLC (D, 4 g/mL) as the positive control. Hyphal formation of Candida albicans cells was obviously inhibited by TTS-12. The white bar represents a length of 20 m (A). 3.6. Ultrastructure analysis In TEM photographs, the cell membrane and cell wall of Candida albicans SC5314 were clearly seen in normal Candida albicans (Fig. 5A). TTS-12 and FLC strongly destroyed the cell membrane of Candida albicans SC5314 (Fig. 5B and C).

Fig. 5. Ultrastructure of Candida albicans cell. Candida albicans

Director: Fernando Estevez Castillo

Homeonews
Edicin N 4 Enero-Febrero de 2006 cells were treated with TTS-12, and were observed by transmission electron microscopy. (A) Normal ultrastructure of Candida albicans cell; (B) ultrastructure of Candida albicans treated with TTS-12 g/mL); (C) ultrastructure of Candida albicans treated with FLC (8 g/mL) as the positive control. The cell membrane of Candida albicans was seriously destroyed by TTS12 or FLC (arrows indicate destroyed cell membrane). The white bar represents a length of 2 m (A). 4. Discussion and conclusion

15

In traditional Chinese Medicine, the plant Tribulus terrestris L. has long been used for the treatment of cutaneous pruritus, edema and inflammation, but no detailed studies concerning the related active components have been reported (Jiangsu New Medical College, 1977 and Chu et al., 2003). Earlier studies showed that Tribulus terrestris L. contained flavanoids, steroid saponins, alkaloids and polysaccharides (Bourke et al., 1992, Wu et al., 1996, Yan et al., 1996, Li et al., 1998, Liu et al., 2003 and Conrad et al., 2004). In our previous studies, we isolated from Tribulus terrestris L. eight steroid saponins, TTS-8, TTS-9, TTS-10, TTS-11, TTS-12, TTS-13, TTS-14 and TTS-15, but did not study their biological activities in detail (Xu et al., 2000). Many studies in the literature (Ekabo et al., 1996, Mshvildadze et al., 2000, Renault et al., 2003 and Sautour et al., 2004) reported that most steroid saponins have antifungal activities. In the present study, we therefore conducted a series of experiments to investigate antifungal activities of the eight steroid saponins we had isolated previously, and did some pioneer work concerning the effects of steroid saponins on the ultrastructure and the hypha, an important factor of fungal virulence. The results of our study showed that TTS-8, TTS-9, TTS-10, TTS-11 and TTS13 were inactive, and TTS-14 had insignificant activities against Candida albicans, Candida glabrata, Cryptococcus neoformans. It is noteworthy that TTS-12 and TTS-15 had significant antifungal activities against the five yeasts tested: Candida albicans, Candida glabrata, Candida parapsilosis, Candida tropicalis, Cryptococcus neoformans. Among these fungi, Candida albicans is the most common infection-causing fungus; about 45% of clinical fungal infections were caused by Candida albicans (Gupta et al., 2004) The present experiment showed that TTS-12 and TTS-15 had potent anti-Candida albicans activity, where MIC80 was 1.0 and 2.3 g/mL, respectively, by far lower than that of the saponins previously reported. According to the literature, MIC of most saponins against Candida albicans is greater than >50 g/mL and that of individual saponins is between 4 and 20.8 g/mL (Ekabo et al., 1996, Mshvildadze et al., 2000, Renault et al., 2003 and Sautour et al., 2004). The studies of Renault et al. (2003) and De Lucca et al. (2002) demonstrated that CAY-1, a steroid saponin, from the ground fruit of Capsicum frutescens, had antifungal activity on Candida albicans, and the IC50 and IC90 were determined to be 3.8 g/mL (3.1 mol/L) and 7.7 g/mL (6.2 mol/L), respectively. Extracts of Eriocephalus africanus L., Felicia erigeroides DC and Helichrysum crispum (L.) D. Don inhibited the growth of Candida albicans (Salie et al., 1996). Director: Fernando Estevez Castillo

Homeonews
Edicin N 4 Enero-Febrero de 2006

16

Prosapogenin A of dioscin exhibited antifungal activity against human pathogenic yeasts Candida albicans (MIC 20.8 g/mL), Candida glabrata (MIC 6.25 g/mL) and Candida tropicalis (MIC 25 g/mL) (Sautour et al., 2004). TTS12 and TTS-15 were also very active on Cryptococcus neoformans (MIC80 = 1.7 and 6.7 g/mL, respectively), similar to the antifungal activities of several other saponins reported with the MICs of 212.5 g/mL (Ekabo et al., 1996 and Mshvildadze et al., 2000). The time course study indicated that TTS-12 and TTS-15 reduced the fungal viability rapidly at a dose dependent rate (Fig. 2). Candidal vaginitis is predominantly caused by strains of Candida albicans (90%) (Sobel et al., 1995, Sobel et al., 1998a, Sobel et al., 1998b and Sobel et al., 2001), and remains to be a common problem in immunocompetent or healthy women. So we also observed the in vivo antifungal activity of TTS-12. According to the literature, the candidal vaginitis rat model is a stable model that can be used for study of drugs on candidal vaginitis. Our result showed that vaginal administration of TTS-12 had a marked therapeutic effect on candidal vaginitis. Above all, steroid saponin TTS-12 has marked in vitro and in vivo antifungal activities. The steroidal glycosides tested in the experiment are from the same chemical class, but only TTS-12 and TTS-15 exhibited significant antifungal activity against Candida albicans, Candida glabrata, Candida tropicalis, Candida parapsilosis and Cryptococcus neoformans. These results indicate that there are critical structural features that are responsible for the antifungal activity. The chemical difference between the aglycons of TTS-12, TTS-15 and compounds TTS-8, TTS-10, TTS-11, TTS-14 was the presence of a carbonyl group at C-12 for TTS-8, TTS-10, TTS-11 and TTS-14. Only TTS-14 showed some antifungal activity. Another chemical difference between TTS-12, TTS-14, TTS-15 (which connect more than four oligosaccharides) and TTS-8, TTS-9, TTS-10, TTS-11 (which connect three or fewer oligosaccharides) is the number of connecting sacchorides. So the absence of a carbonyl group at C-12 and the number of connecting sacchorides are probably related to the antifungal activity of compounds. That is probably the reason why TTS-9, a saponin without a carbonyl base at C-12, did not show antifungal activity. Our result also showed that TTS-13 was inactive against fungi, which confirmed the earlier observations that furostanol-type steroidal glycosides were fungally inactive (Hufford et al., 1988). Bedir et al. (2002) also studied the antifungal activities of seven steroid saponins from Tribulus terrestris L., and the chemical structure of spirostanol saponin 2 (Compound 2) in their study was identical with that of TTS-14. Their results showed that four furostanol-type steroidal glycosides 47 (Compound 4 7) were inactive against fungi, which was coincident with our results. Without a carbonyl base at C-12, spirostanol saponin 1 (Compound 1) in their study, which connects three oligosaccharides, failed to show antifungal activity. This finding is similar to the antifungal activity of TTS-9, which connects two oligosaccharides. In addition, spirostanol saponins 2 and 3 of their study with carbonyl base at C-12 had strong antifungal activities, their MIC values against Candida albicans were all 6.25 g/mL, and MIC values against Cryptococcus neoformans were 2.00 and 3.12 g/mL, respectively. However, our result failed to show that TTS-14 had strong activity against Candida albicans and

Director: Fernando Estevez Castillo

Homeonews
Edicin N 4 Enero-Febrero de 2006

17

Cryptococcus neoformans, with MIC being 41.7 and 48.0 g/mL, respectively. We do not know the reason causing the difference, which needs further experiments. Based on our results and the literature, spirostanol framework and the number of oligosaccharide residue attached at C-3 of aglycon seem closely related to antifungal effects of steroid saponins, but further studies are required to confirm the relation of carbonyl base at C-12 and strong antifungal activity. Hyphae are an important factor of fungal virulence. It is through hyphae that Candida albicans invades human tissues. There have been few studies reporting the effect of saponins on hyphae. The observations of phase contrast microscopy showed that TTS-12 clearly inhibited hyphal formation during the hyphal induction of Candida albicans (Fig. 4). Candida albicans is a dimorphic yeast. Its ability to switch from yeast cells to hyphae is considered to be important for the interactions of Candida albicans with its host (Cutler, 1991). Hyphae are long, slender, continuous tubules with septae that separate each of the nuclei without distinct indentation at the septae. Both yeast cells and hyphae are present in the host during commensal growth and during infection. Hyphae are thought to be an important virulence factor that promotes invasion of cells into the mucosa, allowing candidal cells to resist macrophage and neutrophil engulfment (Yang, 2003). The action mechanisms of saponins may lie in damage to the membrane and leakage of cellular materials, ultimately leading to cell death (Mshvildadze et al., 2000). This activity has been documented in a number of saponins, and the damaging effects have been shown against a variety of fungi, including Candida albicans, Saccharomyces cerevisiae, Trichodemta viride, Acremonium spp. and Cryptococcus neoformans (Lalitha and Venkataraman, 1991 and Polacheck et al., 1991). For example, medicagenic acid 3-O-beta-D-glucopyranoside, an antimycotic saponin from alfalfa root, formed stable complexes with ergosterol, causing lethal leakage of ions out of yeast cells (Polacheck et al., 1991). The present study clearly revealed the antifungal activity of the steroidal saponins. TTS-12 destroyed the yeast cell membrane through TEM (Fig. 5), so that the cytoplasm components leaked out of the cells and the yeast cells were killed by TTS-12. The results of the present study provide pharmacological reference for the traditional use of Tribulus terrestris, documenting that saponins exert antifungal activity by inhibiting fungal hyphae and destroying the ultra structure of fungi in particular. In conclusion, TTS-12 and TTS-15 are steroidal saponins with potent properties against a number of fungal pathogens, and identifying the mode of action and its in vivo toxicity warrants further study in the light of developing new antifungal drugs. Further studies are also required to confirm the structure-activity relation of steroid saponins we propose in the paper. Acknowledgements The authors are grateful to Professor Jun GU of Shanghai Changhai Hospital for donating the yeast isolates. This study was supported by Shanghai Key Director: Fernando Estevez Castillo

Homeonews
Edicin N 4 Enero-Febrero de 2006

18

Basic Research Projects (02DJ14016) and Shanghai R&D Key Fund (02DZ19120). References Bedir et al., 2002 E. Bedir, I.A. Khan and L.A. Walker, Biologically active steroidal glycosides from Tribulus terrestris, Pharmazie 57 (2002), pp. 491493. Bourke et al., 1992 C.A. Bourke, G.R. Stevens and M.J. Carrigan, Locomotor effects in sheep of alkaloids identified in Australian Tribulus terrestris, Australian Veterinary Journal 69 (1992), pp. 163165. Chu et al., 2003 S.D. Chu, W.J. Qu, M. Li and Q.H. Cao, Research advance on chemical component and pharmacological action of Tribulus terrestris, Chinese Wild Plant Resources 22 (2003), pp. 47. Conrad et al., 2004 J. Conrad, D. Dinchev, L. Klaiber, S. Mika, I. Kostova and W. Kraus, A novel furostanol saponin from Tribulus terrestris of Bulgarian origin, Fitoterapia 75 (2004), pp. 117122. Cutler, 1991 J.E. Cutler, Putative virulence factors of Candida albicans, Annual Review of Microbiology 45 (1991), pp. 187218. De Lucca et al., 2002 A.J. De Lucca, J.M. Bland, C.B. Vigo, M. Cushion, C.P. Selitrennikoff, J. Peter and T.J. Walsh, CAY-I, a fungicidal saponin from Capsicum sp. fruit, Medical Mycology 40 (2002), pp. 131137. Ekabo et al., 1996 O.A. Ekabo, N.R. Farnsworth, T.O. Henderson, G. Mao and R. Mukherjee, Antifiingal and molluscicidal saponins from Serjania salzmanniana, Journal of Natural Products 59 (1996), pp. 431435. Fanos and Cataldi, 2000 V. Fanos and L. Cataldi, Amphotericin B-induced nephrotoxicity: a review, Journal of Chemotherapy 12 (2000), pp. 463470. Fridkin and Jarvis, 1996 S.K. Fridkin and W.R. Jarvis, Epidemiology of nosocomial fungal infections, Clinical Microbiology Reviews 9 (1996), pp. 499 511. Grasela et al., 1990 T.H. Grasela, S.D. Goodwin and M.K. Walawander, Prospective surveillance of intravenous amphotericin B use patterns, Pharmacotherapy 10 (1990), pp. 341348. Gupta et al., 2004 N. Gupta, A. Haque, A.A. Lattif, R.P. Narayan, G. Mukhopadhyay and R. Prasad, Epidemiology and molecular typing of Candida isolates from burn patients, Mycopathologia 158 (2004), pp. 397405. Hufford et al., 1988 C.D. Hufford, S.C. Liu and A.M. Clark, Antifungal activity of Trillium grandiflorum constituents, Journal of Natural Products 51 (1988), pp. 9498. Jiangsu New Medical College, 1977 Jiangsu New Medical College, 1977. Dictionary of Chinese Herbal Medicine. Shanghai People's Publishing House, Shanghai, p. 1274. Lalitha and Venkataraman, 1991 T. Lalitha and L.V. Venkataraman, Antifungal activity and mode of action of saponins from Madhuca butyracea Macb, Indian Journal of Experimental Biology 29 (1991), pp. 558562. Director: Fernando Estevez Castillo

Homeonews
Edicin N 4 Enero-Febrero de 2006

19

Li et al., 1998 J.X. Li, Q. Shi, Q.B. Xiong, J.K. Prasain, Y. Tezuka, T. Hareyama, Z.T. Wang, K. Tanaka, T. Namba and S. Kadota, Tribulusamide A and B, new hepatoprotective lignanamides fromthe fruits of Tribulus terrestris: indications of cytoprotective activity in murine hepatocyte culture, Planta Medica 64 (1998), pp. 628631. Liu et al., 2003 J. Liu, H.S. Chen, Y.X. Xu, W.D. Zhang and W.Y. Liu, Studies on chemical constituents of Tribulus terrestris L, Di Er Jun Yi Da Xue Xue Bao 24 (2003), pp. 221222. Lyman and Walsh, 1992 C.A. Lyman and T.J. Walsh, Systemically administered antifungal agents. A review of their clinical pharmacology and therapeutic applications, Drugs 44 (1992), pp. 935. Mshvildadze et al., 2000 V. Mshvildadze, A. Favel, F. Delmas, R. Elias, R. Faure, Q. Decanosidze, E. Kemertelidze and G. Balansard, Antifungal and antiprotozoal activities of saponins from Hedera colchica, Pharmazie 55 (2000), pp. 325326. NCCLS, 2002 National Committee for Clinical Laboratory Standards, 2002. Reference method for broth dilution antifungal susceptibility testing of yeasts. Approved standard. Document M27-A2. National Committee for Clinical Laboratory Standards, Wayne, Pa. Polacheck et al., 1991 I. Polacheck, M. Levy, M. Guizie, U. Zehavi, M. Naim and R. Evron, Mode of action of the antimycotic agent G2 isolated from alfalfa roots, Zenfralbl Bakteriol 275 (1991), pp. 504512. Renault et al., 2003 S. Renault, A.J. De Lucca and S. Boue, CAY-1, a novel antifungal compound from cayenne pepper, Medical Mycology 41 (2003), pp. 7581. Salie et al., 1996 F. Salie, P.F. Eagles and H.M. Leng, Preliminary antimicrobial screening of four South African Asteraceae species, Journal of Ethnopharmacology 52 (1996), pp. 2733. Sautour et al., 2004 M. Sautour, A.C. Mitaine-Offer, T. Miyamoto, A. Dongmo and M.A. Lacaille-Dubois, Antifungal steroid saponins from Dioscorea cayenensis, Planta Medica 70 (2004), pp. 9092. Sobel et al., 1995 J.D. Sobel, D. Brooker, G.E. Stein, J.L. Thomason, D.P. Wermelmg, B. Bradley and L. Wemstem, Single oral dose fluconazole compared with conventional topical therapy of Candida vaginitis, American Journal of Obstetrics and Gynecology 172 (1995), pp. 12631268. Sobel et al., 1998a J.D. Sobel, S. Faro, R.W. Force, B. Foxman, W.J. Ledger, P.R. Nyirjesy, B.D. Reed and P.R. Summers, Vulvovaginal candidiasis: epidemiologic, diagnostic and therapeutic consideration, American Journal of Obstetrics and Gynecology 178 (1998), pp. 203211. Sobel et al., 1998b J.D. Sobel, A. Hasegawa, F. Debernardis, D. Adriani, G. Pellegrini, A. Cassone, P.L. Fidel, C. Haidaris, A.G. Gigliotti, D. Harmsen, S. Fujita, K. Yamamoto, K. Makimura, K. Shibuya, K. Uchida and H. Yamaguchi, Selected animal models: vaginal candidosis, pneumocystosis pneumonia, dermatophytosis and trichosporonosis, Medical Mycology 36 (1998), pp. 129 136. Director: Fernando Estevez Castillo

Homeonews
Edicin N 4 Enero-Febrero de 2006

20

Sobel et al., 2001 J.D. Sobel, P.S. Kapernick, M. Zervos, B.D. Reed, T. Hooton, D. Soper, P. Nyirjesy, M.W. Heine, J. Willems, H. Panzer and H. Wittes, Treatment of complicated Candida vaginitis: comparison of single and sequential doses of fluconazole, American Journal of Obstetrics and Gynecology 185 (2001), pp. 363369. Abstract | PDF (63 K) Sobel et al., 2003 J.D. Sobel, M. Zervos, B.D. Reed, T. Hooton, D. Soper, P. Nyirjesy, M.W. Heine, J. Willems and H. Panzer, Fluconazole susceptibility of vaginal isolates obtained from women with complicated Candida vaginitis: clinical implications, Antimicrobial Agents and Chemotherapy 47 (2003), pp. 34 38. Sojakova et al., 2004 M. Sojakova, D. Liptajova and M. Borovsky, Fluconazole and itraconazole susceptibility of vaginal yeast isolates from Slovakia, Mycopathologia 157 (2004), pp. 163169. Wingard et al., 1991 J.R. Wingard, W.G. Merz, M.G. Rinaldi, T.R. Johnson, J.E. Karp and R. Saral, Increase in Candida krusei infection among patients with bone marrow transplantation and neutropenia treated prophylactically with fluconazole, The New England Journal of Medicine 325 (1991), pp. 12741277. Wingard et al., 1993 J.R. Wingard, W.G. Merz, M.G. Rinaldi, C.B. Miller, J.E. Karp and R. Saral, Association of Torulopsis glabrata infections with fluconazole prophylaxis in neutropenic bone marrow transplant patients, Antimicrobial Agents and Chemotherapy 37 (1993), pp. 18471849. Wingard et al., 1979 J.R. Wingard, W.G. Merz and R. Saral, Candida tropicalis: a major pathogen in immunocompromised patients, Annals of Internal Medicine 91 (1979), pp. 539543. Wu et al., 1996 G. Wu, S. Jiang, F. Jiang, D. Zhu, H. Wu and S. Jiang, Steroidal glycosides from Tribulus terrestris, Phytochemistry 42 (1996), pp. 16771681. Xu et al., 2000 Y.X. Xu, H.S. Chen, H.Q. Liang, Z.B. Gu, W.Y. Liu, W.N. Leung and T.J. Li, Three new sapomns from Tribulus terrestris, Planta Medica 66 (2000), pp. 545550. Yan et al., 1996 W. Yan, K. Ohtani, R. Kasai and K. Yamasaki, Steroidal saponins from fruits of Tribulus terrestris, Phytochemistry 42 (1996), pp. 1417 1422. Yang, 2003 Y.L. Yang, Virulence factors of Candida species, Journal of Microbiology, Immunology, and Infection 36 (2003), pp. 223228.

ALOPATIA
COMPARACION DEL ACIDO AZELAICO Y LA ANTRALINA EN ELTRATAMIENTO DE LA ALOPECIA AREATA CIRCUNSCRIPTA: ESTUDIO PILOTO Sasmaz, Sezai1; Arican, Ozer1 Director: Fernando Estevez Castillo

Homeonews
Edicin N 4 Enero-Febrero de 2006
1

21

Department of Dermatology, School of Medicine, Sutcuimam University, Kahramanmaras, Turkey

Kahramanmaras

[American Journal of Clinical Dermatology,Volume 6, Number 6, 2005, pp.403406] RESUMEN Antecedentes: Aunque el cido azelaico ha sido utilizado en forma tpica para el tratamiento de la alopeca, no existen hasta la fecha ensayos controlados con esta droga para esta patologa. Objetivo: El objetivo de este estudio fue determinar la eficacia, tolerabilidad y seguridad del tratamiento de la ALOPECIA AREATA CIRCUNSCRIPTA (AA) con cido azelaico comparado con la antralina. Mtodos: Este estudio incluy 31 sujetos con AA circunscripta quienes no haban recibido ningn tratamiento por lo menos un ao antes del comienzo del estudio. Se registraron las caractersticas demogrficas y clnicas iniciales de todos los individuos que participaron de la experiencia. Los sujetos fueron elegidos al azar para aplicarle cido azelaico al 20% (15) o antralina al 0,5% (16) durante 12 semanas. Luego se continu con un perodo de seguimiento de 8 semanas sin ningn tipo de crema aplicada. Dos investigadores independientes realizaron, a las 20 semanas, una evaluacin de eficacia con examinacin clnica utilizando la escala de crecimiento de cabello terminal (RGS) con un rango desde 0 (respuesta inadecuada) a 2 (respuesta completa), teniendo la respuesta parcial un score igual a 1. Resultados: Todos los sujetos completaron el ensayo. A las 20 semanas el RGS fue 1.27 0.9 para el grupo del cido azelaico versus 1.37 0.8 del grupo de antralina (p > 0.05). Una respuesta completa fue observada en el 53,3% de los casos tratados con el cido azelaico (8 de 15) comparados con el 56.2% (9 de 16) del grupo de antralina (p > 0.05). Ningn efecto adverso serio fue observado en ambos grupos durante el estudio. Conclusin: el presente estudio piloto mostr que el uso de cido azelaico da resultados similares a la antralina con respecto al crecimiento capilar, y que puede ser una agente teraputico tpico efectivo para el tratamiento de la Alopeca Areata circunscripta. Sin embargo, se necesitan ms ensayos para poder llegar a una conclusin definitiva.

HOMEOPATIA
ESTIMULACION DE LA ACTIVIDAD MITOCONDRIAL DEL ESPERMA BOVINO MEDIANTE DILUCIONES HOMEOPATICAS DE MONENSINA

Director: Fernando Estevez Castillo

Homeonews
Edicin N 4 Enero-Febrero de 2006 DM Aziz1, and H Enbergs2
1

22

Department of Surgery and Obstetrics, College of Veterinary Medicine, University of Mosul, Mosul, Iraq 2 Institute of Anatomy, Physiology and Hygiene of Domestic Animals, University of Bonn, Bonn, Germany [Homeopathy, Volume 95, Issue 2, April 2006, Pages 94-97] RESUMEN La actividad mitocondrial es un parmetro importante de la viabilidad de los espermatozoides y se relaciona con la motilidad espermtica. La monensina es comnmente utilizada en laboratorio como un inhibidor de la actividad mitocondrial espermtica. Este estudio fue realizado para evaluar la influencia de algunas diluciones homeopticas de monensina sobre la actividad mitocondrial espermtica. Se utilizaron para el estudio, eyaculados frescos provenientes de seis toros adultos. Las muestras de semen fueron evaluadas usando un citmetro de flujo para determinar la actividad mitocondrial y la viabilidad espermtica, utilizando Rhodamina 123 y SYBR-14, respectivamente. La monensina 9X produjo un resultado muy altamente significativo (P<0.001) en la estimulacin de la actividad mitocondrial espermtica, mientras que la 5X, 7X, 8X y 13X fue altamente significativo (P<0.01). Otras diluciones homeopticas de monensina (6, 10, 11, 12 and 14) tambin tuvieron un efecto estimulante significativo (P<0.05). El uso de monensina no tuvo ningn efecto negativo sobre la viabilidad espermtica. Podemos concluir que algunas diluciones homeopticas de monensina incrementan la actividad mitocondrial de los espermatozoides bovinos sin efecto negativo con respecto a su viabilidad, siendo la 9X la dilucin ms efectiva. Es necesario realizar ms estudios in vivo para estimar el efecto de las diluciones homeopticas de monensina sobre la calidad del semen. Palabras clave: esperma; actividad mitocondrial, monensina, homeopata.

NUTRICION
LOS ACIDOS GRASOS OMEGA-3 DISMINUYEN LA IRRITABILIDAD DE PACIENTES CON DESORDENES BIPOLARES: ESTUDIO ABIERTO Kemal Sagduyu1, Mehmet E Dokucu2, Bruce A Eddy3, Gerald Craigen4, Claudia F Baldassano5 y Ayegl Yldz6
1

University of Missouri Kansas City, Missouri, 8801 West 148th Terrace, Overland Park, KS 66221, USA.

Director: Fernando Estevez Castillo

Homeonews
Edicin N 4 Enero-Febrero de 2006
2

23

Washington University, School of Medicine, Department of Psychiatry, Campus Box: 8134, 660 South Euclid Avenue, St. Louis, Missouri, 63110, USA. 3 Department of Psychiatry, School of Medicine, University of Missouri-Kansas City, Resource Development Institute, 601 Walnut Street, Kansas City, MO 64106, USA. 4 Mood Disorders Psychopharmacology Unit, University Health Network, Toronto Western Hospital, 399 Bathurst Street, ECW-3D-010, Toronto, Ontario M5T 2S8, Canada. 5 Mood and Anxiety Disorders Clinic, Department of Psychiatry, University of Pennsylvania, 3535 Market Street, 2nd floor, Philadelphia, PA 19104, USA 6 Dokuz Eyll Medical School, Department of Psychiatry, zmir, Turkey. [Nutrition Journal 2005, 4:6] RESUMEN Este es el reporte de la continuacin de un estudio abierto con 37 pacientes sobre cidos grasos Omega-3 (O-3FA). Los sujetos fueron los 19 pacientes originales, a los cuales se le agregaron 18 nuevos, elegidos y con seguimiento realizado de la misma forma que con los primeros 19. Los individuos tenan el diagnstico de desrdenes bipolares y visitaban regularmente una Clnica para estas patologas a lo largo de toda la experiencia. En cada visita, el estado de cada paciente fue seguido utilizando la planilla de monitoreo clnico. Los sujetos reportaban la frecuencia y severidad de los episodios de irritabilidad experimentados en los ltimos 10 das previos a la consulta; la frecuencia se midi por el porcentaje de das en los cuales experimentaban irritabilidad, mientras que para la severidad se utiliz la escala de Likert de 1 a 4 (si se presentaba). El componente de irritabilidad de acuerdo a la Escala de evaluacin de Mana de Young (YMRS) fue registrado en 13 de los 39 pacientes. Los pacientes tuvieron una persistente irritabilidad a pesar de su tratamiento continuo farmacolgico y psicoteraputico. La ingesta de cidos Omega-3 ayud con el componente de irritabilidad de pacientes sufriendo desrdenes bipolares con una presencia significativa de signos de irritabilidad. Bajas dosis de Omega- 3 (1 a 2 gramos por da), pueden ayudar tambin con el componente de irritabilidad de diferentes condiciones clnicas, tales como esquizofrenia, problemas de personalidad y otras condiciones psiquitricas con un signo comn presente de irritabilidad.

ARTICULO ORIGINAL INTRODUCTION

Director: Fernando Estevez Castillo

Homeonews
Edicin N 4 Enero-Febrero de 2006

24

According to the United States National Institute of Mental Health (NIMH), Bipolar Disorder (BPD), also known as manic-depressive illness, is a serious medical illness that causes shifts in a person's mood, energy, and ability to function. Different from the normal ups and downs that everyone goes through, the symptoms of bipolar disorder are severe. Bipolar disorder is a complex, chronic condition associated with considerable morbidity and mortality, including a high rate of suicide. Bipolar disorder causes dramatic mood swings from overly "high" and/or irritable to sad and hopeless, and then back again, often with periods of normal mood in between. Severe changes in energy and behavior go along with these changes in mood. The periods of highs and lows are called episodes of mania and depression. Most people with bipolar disorder can achieve substantial stabilization of their mood swings and related symptoms over time with proper treatment. A strategy that combines medication and psychosocial treatment is optimal for managing the disorder over time. BACKGROUND Omega-3 fatty acids (0-3FA) may have a beneficial effect on irritable mood. Low O-3FA levels in red blood cell membranes of depressed patients hint that O3FA may be helpful in treating mood disorders [1]. A recent article has given an excellent review of O-3FA and studies showing their effectiveness in depression, bipolar disorder and aggression [2]. In this article, two published studies are discussed that have reported on similar therapeutic effects of O-3FA [2]. One placebo-controlled study of 20 patients revealed that ethyl ester of eicosapentaenoic acid (E-EPA) was effective in stabilizing the moods of depressed patients [3]. Another report, a double-blind, placebo controlled study (N = 22/19), measured the effect of O-3FA docosahexaenoic acid (DHA) on the aggressive tendencies of college students. The O-3FA DHA group (1.51.8 g O-FA DHA/day) did not display any increase in aggressive tendencies when external stressors peaked, while the placebo group displayed a significant increase in their aggressive tendencies under similar circumstances [4]. In a recent study, 25% of 111 patients with bipolar-I disorder who met criteria for a DSM-IV major depressive episode also experienced substantial irritability in the absence of associated symptoms of mania. These findings suggest that abnormal irritability is not limited to mania or mixed states [5]. However, recent studies give caution that at a 6 gram per day average daily dose, as a single agent, omega 3 fatty acids may not be as effective as an antidepressant [6-9]. O-3FA may also help with the irritability component of different clinical conditions, such as depression, mania, schizophrenia, borderline personality disorder and other psychiatric conditions with a common presenting sign of irritability. Numerous other conditions have an irritability component, including Borderline Personality Disorder, Alzheimer's disease, Premenstrual Dysphoric Disorder, to name a few [10-12]. There is one report suggesting beneficial effect of Omega-3 Fatty acid treatment for Borderline Personality Disorder. This double-blind, placebo-controlled pilot study specifically showed that EPA may Director: Fernando Estevez Castillo

Homeonews
Edicin N 4 Enero-Febrero de 2006

25

influence both aggression and depression [12]. Although attentiondeficit/hyperactivity disorder (ADHD) also has an irritability component, recent publications bring doubt to the O3FA connection in ADHD [13,14]. A recent, open ended, O-3FA add-on study has shown beneficial effect of O3FA on irritability in 19 patients with mood disorders [15]. These patients had already been receiving different combinations of pharmacotherapy and talk therapy. Despite their treatment, the irritability component of their illness was still causing social, occupational and other life disturbances. Hence, they were chosen for the O-3FA add-on component of the study. In the nineteen-patient phase of the study, bipolar patients of every subtype, ages 18 to 65 years, with significant irritability were studied. All patients received a systematic assessment battery at entry and were treated by a psychiatrist, trained to deliver care and measure outcomes in patients with bipolar disorder, consistent with expert recommendations. At every follow-up visit, the treating psychiatrist completed a standardized assessment and assigns a clinical status based on DSM-IV criteria. Patients had independent evaluations at regular intervals throughout the study and remain under the care of the same treating psychiatrist while receiving variable medications and talk therapy, depending on their need [15]. In the 19-patient study, a paired sample t-test revealed a large decrease in the percent of days irritable after O-3FA was administered. Before treatment, the mean irritability percentage was 81.05 (SD = 23.31) and after treatment the mean irritability percentage dropped to 30.00 (SD = 36.67). Despite the small number of patients in the study (n = 19), the difference between means was statistically significant (t (18) 4.512, p < .001). Using a paired sample t-test, a significant difference was also found between the highest irritability score (mean = 2.79; SD = 0.92) and the last recorded irritability (mean = 0.79; SD = 0.85) while taking O-3FA (t(18) = 8.270; p < .001) [15]. METHODS This is a report on a 37-patient continuation phase of the open ended, O-3FA add-on study. Subjects consisted of the original 19 patients, in addition to the 18 new patients recruited and followed in the same fashion as the first nineteen [15]. Subjects carried a DSM-IV-TR [16] diagnosis of Bipolar Disorder and were visiting a Mood Disorder Clinic regularly throughout the length of the study. At each visit, patients' clinical status was monitored using the Clinical Monitoring Form [17]. Subjects reported on the frequency and severity of irritability experienced during the preceding ten days; frequency was measured by way of percentage of days in which subjects experienced irritability, while severity of that irritability was rated on a Likert scale of 1 4 (if present). The irritability component of Young Mania Rating Scale [18] (YMRS) was also recorded quarterly on 13 of the 39 patients consistently. The patients were asked about general dietary omega-3 intake before the fish oil was added on, and basic

Director: Fernando Estevez Castillo

Homeonews
Edicin N 4 Enero-Febrero de 2006

26

nutritional guidance was given to subjects at the clinic. Patients in general were not heavy fish/product consumers. Dosage Starting dose and last maintenance dose were available for 37 subjects (Table 1). Subjects self-medicated, and therefore, the last maintenance dose of O-3FA was chosen by each subject. The mean starting dose was 1824.32 mg (SD 1075.07), and the mean for the last maintenance dose was considerably higher at 2878.38 mg (SD 2011.79). The increase was statistically significant using a paired sample t-test (t = -3.44, 36df, p = .001). Table 1 Initial, Last and Final Omega 3 Dosages (mg). summarizes dosage under three conditions. Figures for the Initial Dose include two subjects (n = 39) for whom no corresponding follow-up data were available. Initial, Last Recorded and Final Omega 3 Dosages (mg) Inicial N Mean Mode Median SD 39 1833.33 1000 2000 1071.91
T2, T3

Last Recorded 37 2878.38 1000 2000 2011.79

FinalT1 13 2615.38 2000 2000 1894.66

T1 = Final group results (n = 13) are discussed below. T2 = Multiple modes exist. The smallest value is shown. T3 = One gram (1,000 miligram) of fish oil; of which about 180 milligrams is (eicosapentaenoic acid) EPA and 120 milligrams is DHA (docosahexaenoic acid), (for a total of 300 milligrams of omega 3's) in each clear capsule.

STATISTICAL RESULTS Percentage of Irritable (Days) The initial mean was 63.51 (SD 34.17), indicating that on average, subjects were irritable for about six of the previous ten days. The mean for the last recorded percentage was less than half of the initial score: 30.27 (SD 34.03). The decrease was found to be statistically significant using a paired sample ttest (t = 4.36, 36 df, p < .001). The difference between the distributions was examined using the non-parametric sign test. The number of negative Director: Fernando Estevez Castillo

Homeonews
Edicin N 4 Enero-Febrero de 2006

27

differences (25) significantly exceeded positive differences (7); there were five ties, and the pre/post distributions were significantly different (p < .003). YMRS Irritability Sub-score Thirty four subjects had initial and last recorded YMRS irritability sub-scores. As with the above means there was a sizable decrease. The initial mean score was 3.18 (SD 1.09). The mean for the last recorded percentage was 1.68 (SD 1.89). The decrease was found to be statistically significant using a paired sample ttest (t = 4.21, 33 df, p < .001). YMRS Total Score Starting and last recorded YMRS scores were available for 34 subjects. The mean starting score 10.71 (SD 6.77), and the mean for the last recorded score was 4.85 (SD 5.63). The decrease found to be statistically significant using a paired sample t-test (t = 4.14, 33 df, p < .001). Severity Thirty six subjects had initial and last recorded severity scores on the ADE. Again, a decrease was found. The initial mean score was 2.14 (SD 1.22). The mean for the last recorded score was 0.94 (SD 0.92). This decrease was found to be statistically significant using a paired sample t-test (t = 5.23, 35 df, p < .001). Composite: Severity and Irritability As an exploratory measure, a composite score was created by multiplying the ADE severity score, which has a maximum of 4 points, by the percentage of the ten days prior to measurement which the patient was rated as irritable. The initial mean on this composite was 159.72. As with other measures, there was wide variation: SD = 122.92. The mean for this measure on the last recorded scores was percentage was about one-fourth of the initial score: 43.89 (SD 64.38). The decrease was found to be statistically significant using a paired sample t-test (t = 5.00, 35 df, p < .001). Last Recorded Maintenance Dose and Percentage of Irritability After Because of apparent wide variation on these two measures and a concern that outliers may have affected some results, the last recorded irritability scores were plotted against the maintenance dose. This revealed a rather bimodal pattern, in which relatively lower irritability measures ( 50%) clustered in the quadrant with lower dosage levels ( 4,000 mg). Duration and YMRS Total In response to a similar observation regarding wide variation in the last recorded values (84 days to 5.5 years) the values were also plotted. A clearly

Director: Fernando Estevez Castillo

Homeonews
Edicin N 4 Enero-Febrero de 2006

28

bimodal pattern appeared in which 11 subjects (about one-third of study participants) clustered in the quadrant representing short duration (<500 days) and higher YMRS totals (>7). The remaining two-thirds of subjects clustered in the quadrant representing short duration and lower YMRS totals (<6). Subject Weight The mean start weight was 176.97 lbs (SD 43.13), and the mean for the last weight recorded was slightly higher at 178.59 lbs (SD 43.24). The increase was not statistically significant. FOLLOW-UP SUBJECTS Follow-up information, recorded after the collection of the "last" scores for most of the above variables, was available for 13 of the 37 subjects. Final YMRS total or scale scores were not available for this sub-group. Omega 3 Duration The final date recorded for the duration of O3 was derived based from an O3 start date and a "final" date recorded for O3. The time period ranged 84 days to 1995 days (5.46 years). The mean duration of O3 for this group was 439.62 days (SD = 487.46). Dosage For these subjects, the mean starting dose was 1807.69 mg (SD 990.34), and the mean for the last maintenance dose was higher at 2615.38 mg (SD 1894.66). The increase was not significant. Percentage Irritable (Days) The initial mean was 82.31 (SD 20.88). The mean for the last recorded percentage was dramatically lower: 25.38 (SD 32.04). The decrease was found to be statistically significant using a paired sample t-test (t = 6.52 12 df, p < .001). The difference between the distributions was examined using a sign test. The number of negative differences (12) significantly exceeded positive differences (0); there was one tie, and the pre/post distributions were significantly different (p < .001). Severity The initial mean score for the 13 subjects with final scores was 2.69 (SD 0.95). The mean for the final score was 0.77 (SD 0.83). This decrease was found to be statistically significant using a paired sample t-test (t = 6.22, 12 df, p < .001). Composite: Severity and Irritability An exploratory composite score, described above, was also created for the subjects with final scores. For these subjects, the initial mean was higher than

Director: Fernando Estevez Castillo

Homeonews
Edicin N 4 Enero-Febrero de 2006

29

that of the total group, 223.08. Again, there was wide variation: SD = 104.19. The mean for this measure on the last recorded scores was percentage was much lower that the initial score: 33.08 (SD 39.87). The decrease was found to be statistically significant using a paired sample t-test (t = 6.70, 12 df, p < .001). Weight For these 13 subjects, the mean start weight was 166.23 lbs (SD 35.68), and the mean for the final weight recorded was also slightly higher at 168.23 lbs (33.62). As with the previous finding regarding weight, the increase was not statistically significant. RESULTS Omega-3 Fatty Acids added onto the existing treatment helped with the irritability component of a significant percentage of patients suffering from bipolar disorder with a persistent sign of irritability. DISCUSSION As seen from the standard deviations of several of the variables discussed here, measures ranged widely. This creates difficulty in using descriptive data, such as means, to adequately portray subject attributes and performance. Using data reduction techniques or grouping subjects according to high and low scores on various attributes may be one way to increase the descriptiveness, which would be possible and more reasonable with a larger pool of subjects. A potential limitation or interpretive consideration merits discussion. For many of the variables discussed above, noticeable differences in measures were observed between the "starting" versus "last recorded" group (n = 37) and the "starting" versus "final" measures group (n = 13). Given these differences and the smaller number of subjects in the second set of comparisons, "starting" versus "final" comparisons should be interpreted with caution until differences inherent in this "final" subgroup (n = 13) are more clearly understood. This is clearly seen in the results of sign tests, in which the apparent magnitude of the "final" effects is pronounced. Statistically significant within-subjects differences were found in several independent variables. This is especially notable given the small number of subjects. The preliminary findings suggest that a rigorously designed study tailored especially to the examination of the effects of O-3FA is warranted. The majority of data were collected within an ongoing "best-practice, outpatient bipolar disorder study" that involved medications and talk therapy which we have not reported or discussed herein. Results must, therefore, be interpreted with caution.

Director: Fernando Estevez Castillo

Homeonews
Edicin N 4 Enero-Febrero de 2006

30

There are several mechanisms through which O-3FA are theorized to help with mood, irritability, aggression etc. Suggested theories of mechanism converge on the theory of nerve cell membrane stabilization. A recent study has come closest to showing physical proof of effectiveness of O-3FA through indirect demonstration of greater membrane fluidity, as detected by reductions in Tesla2 (T2) values in MRI scans [19]. The overlapping beneficial effects of antipsychotics, antidepressants, anticonvulsants, O-3FA, and nonpsychoactive cannabinoids, as they relate to pain, stroke, schizophrenia, psychoneuroimmunology, Alzheimer's disease, and stress, may be because of their common effects at protein kinases, thus affecting the structure and function of the cell membrane and the cell [20]. These changes should help the cell operate within an optimal level of excitation, which may be related to emerging evidence that these therapeutic agents have neuroprotective value [20]. A recent randomized placebo controlled double blind intervention study suggests an adaptogenic role for O-3FA in stress [21]. We would like to discuss briefly the issue of daily dosing of O-3FA for nutrition and medicinal purpose: Recent studies give caution that at a 6 gram per day average daily dose, as a single agent, omega 3 fatty acids may not be as effective as an antidepressant [6-9]. However, these studies may have given too high of a dose of O-3FA, above 6 grams daily, with possibly beyond a therapeutic window of effectiveness for O-3FA. Our scatter plots indicate that the optimum effective dose for irritability is at 12 gram of EPA plus DHA per day, which would be the dosing we suggest. A recent exploratory dose study of O-3FA for schizophrenic patients showed that 2 g/day EPA-treated patients had lower symptom scores, and needed less medication greatest. In this study, there was a positive relationship between improvement on rating scales and rise in red blood cell arachidonic acid concentration as well [22]. The United States (US) accounts for more than 51% of the 430.3 billion dollar expended on pharmaceutical products worldwide each year [23]. World healthcare society first needs access to low-cost, nontoxic, non-expertdependent interventions to ensure basic health outcomes. Food may represent the most cost-effective means of promoting public health [23]. The American Heart Association recommends consumption of two servings of fish per week for persons with no history of coronary heart disease and at least one serving of fish daily for those with known coronary heart disease [24]. Approximately 1 g per day of EPA acid plus DHA acid is recommended for cardioprotection [24]. Higher dosages of omega-3 fatty acids are required to reduce elevated triglyceride levels (2 to 4 g per day) and to reduce morning stiffness and the number of tender joints in patients with rheumatoid arthritis (at least 3 g per day) [24]. We conclude that it is beneficial in many ways to establish a regular intake of 1 2 g per day EPA acid plus DHA, similar to daily intake of vitamins with minerals. Dietary interventions to remedy omega-3 deficiency is necessary [23]. It is time Director: Fernando Estevez Castillo

Homeonews
Edicin N 4 Enero-Febrero de 2006

31

for more aggressive funding for research into medicinal foods, such as omegathree fatty acids [23]. REFERENCES 1.Peet M, Murphy B, Shay J, Horrobin D: Depletion of omega-3 fatty acid levels in red blood cell membranes of depressive patients. Biol Psychiatry 1998, 43(5):315-319. 2.Lake J: Omega-3 Fatty Acids: Theory, Clinical Trials and Safety Issues. Psychiatric Times 2002, 19(10):28-34. 3.Nemets B, Stahl Z, Belmaker RH: Addition of omega-3 fatty acid to maintenance medication treatment for recurrent unipolar depressive disorder. Am J Psychiatry 2002, 159(3):477-479. 4.Hamazaki T, Sawazaki S, Itomura M, Asaoka E, Nagao Y, Nishimura N, Yazawa K, Kuwamori T, Kobayashi M: The effect of docosahexaenoic acid on aggression in young adults. A placebo-controlled double-blind study. J Clin Invest 1996, 97(4):1129-1133. 5.Deckersbach T, Perlis RH, Frankle WG, Gray SM, Grandin L, Dougherty DD, Nierenberg AA, Sachs GS: Presence of irritability during depressive episodes in bipolar disorder. CNS Spectr 2004, 9(3):227-231. 6.Marangell LB, Martinez JM, Zboyan HA, Kertz B, Kim HF, Puryear LJ: A double-blind, placebo-controlled study of the omega-3 fatty acid docosahexaenoic acid in the treatment of major depression. Am J Psychiatry 2003, 160(5):996-998. 7.Keck PE, McElroy SL, Freeman MP: Randomized, placebo-controlled trial of eicosapentaenoic acid in bipolar depression. Bipolar Disord 2003, 5(suppl 1):58. 8.Keck PE, McElroy SL, Freeman MP: Randomized, placebo-controlled trial of eicosapentaenoic acid in rapid cycling bipolar disorder. Bipolar Disord 2003, 5(suppl 1):58. 9.Stoll AL, Severus WE, Freeman MP, Rueter S, Zboyan HA, Diamond E, Cress KK, Marangell LB: Omega 3 fatty acids in bipolar disorder: a preliminary double-blind, placebo-controlled trial. Arch Gen Psychiatry 1999, 56(5):407-412. 10.Mirakhur A, Craig D, Hart DJ, McLlroy SP, Passmore AP: Behavioural and psychological syndromes in Alzheimer's disease. Int J Geriatr Psychiatry 2004, 19(11):1035-1039. 11.McHichi alami K, Tahiri SM, Moussaoui D, Kadri N: [Assessment of premenstrual dysphoric disorder symptoms: population of women in Casablanca]. Encephale 2002, 28(6 Pt 1):525-530. 12.Zanarini MC, Frankenburg FR: omega-3 Fatty acid treatment of women with borderline personality disorder: a double-blind, placebo-controlled pilot study. Am J Psychiatry 2003, 160(1):167-169. 13.Young GS, Maharaj NJ, Conquer JA: Blood phospholipid fatty acid analysis of adults with and without attention deficit/hyperactivity disorder. Lipids 2004, 39(2):117-123. 14.Hirayama S, Hamazaki T, Terasawa K: Effect of docosahexaenoic acidcontaining food administration on symptoms of attention-deficit/hyperactivity disorder - a placebo-controlled double-blind study. Eur J Clin Nutr 2004, 58(3):467-473. Director: Fernando Estevez Castillo

Homeonews
Edicin N 4 Enero-Febrero de 2006

32

15.Sagduyu K: Omega-3 Fatty Acids for Irritable Mood?. Psychiatric Times 2003, 20(3):9. 16.Diagnostic and Statistical Manual of Mental Disorders, Text Revision 4th edition. Washington, D.C., American Psychiatric Association; 2000. 17.Sachs GS, Guille C, McMurrich SL: A clinical monitoring form for mood disorders. Bipolar Disord 2002, 4(5):323-327. 18.Young RC, Biggs JT, Ziegler VE, Meyer DA: A rating scale for mania: reliability, validity and sensitivity. Br J Psychiatry 1978, 133:429-435. 19.Hirashima F, Parow AM, Stoll AL, Demopulos CM, Damico KE, Rohan ML, Eskesen JG, Zuo CS, Cohen BM, Renshaw PF: Omega-3 fatty acid treatment and T(2) whole brain relaxation times in bipolar disorder. Am J Psychiatry 2004, 161(10):1922-1924. 20.Ryback R: Bioelectrical modulators and the cell membrane in psychiatric medicine. Psychopharmacol Bull 2001, 35(4):5-44. 21.Bradbury J, Myers SP, Oliver C: An adaptogenic role for omega-3 fatty acids in stress; a randomised placebo controlled double blind intervention study (pilot)ISRCTN22569553. Nutr J 2004, 3(1):20. 22.Peet M, Horrobin DF: A dose-ranging exploratory study of the effects of ethyl-eicosapentaenoate in patients with persistent schizophrenic symptoms. J Psychiatr Res 2002, 36(1):7-18. 23.Plotnikoff GA: Food as medicine--cost-effective health care? The example of omega-3 fatty acids. Minn Med 2003, 86(11):41-45. 24.Covington MB: Omega-3 fatty acids. Am Fam Physician 2004, 70(1):133140.

NOTAS DE INTERES
I CURSO INTERNACIONAL DE FARMACIA HOMEOPATICA (I DIPLOMATURA EN TERAPIAS ALTERNATIVAS UNIVERSIDAD NACIONAL MAYOR DE SAN MARCOS LIMA - PER Del 4 al 8 de marzo del corriente ao, tuvo lugar en la ciudad de Lima, Per, el I Curso Internacional de Farmacia Homeoptica (terico-prctico), correspondiente a la I Diplomatura en Terapias Alternativas de la Unidad de Posgrado de Medicina Humana de la Universidad Nacional Mayor de San Marcos, al cual tuve el honor de ser invitado para participar como nico docente del mismo. El da previo (3 de marzo) se realizaron dos conferencias pblicas, con el ttulo Avances en Homeopata I y II, en el Auditorio del Hospital Rebagliatti y de la Municipalidad de Lince, respectivamente.

Director: Fernando Estevez Castillo

Homeonews
Edicin N 4 Enero-Febrero de 2006

33

El Farm. Fernando Estevez Castillo exponiendo durante la parte terica del Curso.

Toda la organizacin estuvo a cargo de la Asociacin Peruana de Salud Integral (ASPESIN), que a travs del esfuerzo de todos los colegas que la integran, lograron que todas las actividades se desarrollen con total normalidad y prolijidad, estando atentos en todo momento para cumplir hasta con el ms mnimo detalle. Para el desarrollo de la parte terica los alumnos contaron con material bibliogrfico impreso y un CD con ms informacin, y en la parte prctica cada uno en forma individual pudo elaborar los distintos productos intermedios y formas farmacuticas homeopticas, contando para ello con todos los materiales necesarios, inclusive con el HOMEODINAMIZADOR HIEMUSR, que fue adquirido por ASPESIN, para poder cumplir con los objetivos previstos.

Los alumnos del Curso durante el desarrollo de las clases prcticas realizadas en el laboratorio del Departamento de Qumica de la Facultad de Farmacia y Bioqumica de la UNMSM.

Director: Fernando Estevez Castillo

Homeonews
Edicin N 4 Enero-Febrero de 2006

34

El esfuerzo fue muy grande pero vali la pena, ya que el entusiasmo por aprender e intercambiar experiencias, super ampliamente al cansancio lgico de actividades tan extensas en carga horaria. Pero todo no terminaba con la parte acadmica, ya que gracias a la excelente predisposicin, hospitalidad y el don de buenas personas de todo el grupo de ASPESIN, en el poco tiempo que dur mi estada en Per, tuve la oportunidad de conocer numerosos lugares de Lima, costumbres de su pueblo e historia, y un montn de cosas ms que dejaron una huella muy importante en mi persona para volver en un futuro no muy lejano y seguir conociendo a ese maravilloso pas que es Per.

El Farm. Fernando Estevez Castillo junto a los organizadores del Curso y algunos alumnos del mismo, luego de la culminacin de la parte terica.

Felicitaciones a ASPESIN por la idoneidad y calidez de todos sus miembros, as como tambin por el logro alcanzado con la I Diplomatura en Terapias Alternativas de la UNMSM!

NOVEDADES CHIACAPS: NUEVO SUPLEMENTO DIETARIO


(OMEGA 3 PROVENIENTE DEL ACEITE DE CHIA) Laboratorios Dr. Madaus & Co. ha presentado Chiacaps , un nuevo suplemento dietario a base de Omega 3 obtenido a partir de la semilla de CHIA (Salvia hispnica), que es considerada la fuente natural ms rica en stos cidos grasos, superando inclusive al lino, mostrando una ventaja muy
R

Director: Fernando Estevez Castillo

Homeonews
Edicin N 4 Enero-Febrero de 2006

35

importante sobre las otras fuentes de omega-3 (aceites de pescado), ya que no contiene colesterol y su contenido en cidos grasos saturados es mucho menor. Adems contiene una cantidad de compuestos con potente actividad antioxidante, que evitan la necesidad de utilizar antioxidantes artificiales para estabilizar el aceite y disminuir la prdida de sus propiedades naturales.

ChiacapsR

se presenta en cpsulas blandas por 30 60 unidades, conteniendo cada una 1000 mg de aceite de cha equivalente a 600 mg de Omega-3. BENEFICIOS DE LOS OMEGA-3 presentes en la CHIA PARA EL CORAZON Disminucin de la presin arterial. Reduccin de la arritmia y de la probabilidad de muerte sbita. Disminucin de triglicridos y LDL. EN LOS PROCESOS INFLAMATORIOS Inhiben los eventos pro-inflamatorios. Protege al pulmn de procesos inflamatorios. Reduce la sintomatologa de enfermedades inflamatorias: enfermedad inflamatoria intestinal, asma, artritis reumatoidea, colitis ulcerosa, eczema y psoriasis. EN LA GESTACION Indispensables en el desarrollo visual del feto. Esenciales en el desarrollo neurolgico del feto. Previenen la hipertensin de la madre relacionada con el embarazo. EN LA LACTANCIA Estimulan el crecimiento y desarrollo neurolgico del nio. Mejoran el ndice de desarrollo mental de nios prematuros. Refuerzan el sistema inmunolgico del recin nacido. Contribuyen al desarrollo psicomotor del nio. Reducen la dermatitis seborreica del recin nacido. ARTICULO PUBLICADO EN EL DIARIO CLARIN Director: Fernando Estevez Castillo

Homeonews
Edicin N 4 Enero-Febrero de 2006

36

Domingo 9 de Abril de 2006

Diario Clarn Suplemento Econmico


FARMACIA NATURAL

Omega 3, de la planta a la cpsula


Cecilia de Castro. cdecastro@clarin.com Los aztecas no saban nada de Omega 3. Tampoco los incas ni los mayas. Pero es muy probable que el colesterol no taponara sus venas. Gracias a la cha, una planta que junto con el maz y algunos porotos era su principal alimento. Con la llegada de los espaoles la cha cay en el olvido. Hoy los cientficos desempolvaron la planta. En los Estados Unidos descubrieron que las tribus que consuman este alimento no saban lo que era ir al mdico al brujo, ms bien por problemas de diabetes o hipertensin. Es que un buen plato de cha al ajillo, a la portuguesa o cualquiera que fuera la receta del chef proporcionaba a los indgenas una buena dosis de Omega 3. Para obtener los beneficios de la cha en el siglo 21 no ser necesario aprender a cocinar la planta. Bastar con abrir un frasco y tomar unas pastillas. Muy pronto Farmacia Natural, el emprendimiento de Patricia Smolinski (47), empezar a vender las cpsulas con aceite de cha. "Todo empez por casualidad. Una pariente ma muy viejita me coment que el mdico le haba recetado un remedio. Yo le ped a una farmacutica que buscara algo sobre el tema pero sin darle demasiada importancia. 'Yo s lo que es porque mi to la est estudiando en los Estados Unidos', me dijo ella".

Patricia Smolinski, presenta el producto CHIACAPSR

As Smolinski, que tambin es duea de Laboratorios Dr. Madaus, se puso en contacto con la gente de la Universidad de Arizona. Y empezaron a trabajar Director: Fernando Estevez Castillo

Homeonews
Edicin N 4 Enero-Febrero de 2006

37

juntos. "El estudio de la planta lo hicieron ellos y nosotros nos encargamos de los estudios en bioterio, con las ratas. Lo llevamos a la prctica, hicimos la investigacin para desarrollar el suplemento dietario". Las cpsulas se vendern en la Argentina y tambin se exportarn a los Estados Unidos. La marca local ser ChiacapsR y el precio, 25 pesos para 30 unidades y 47 pesos para 60. "Lo tenemos registrado como suplemento dietario pero al ser un aceite no hay un proceso que se pueda patentar. Por ahora somos los nicos que lo encapsulamos y que tenemos el registro". Adems Smolinski tiene contrato de exclusividad con la gente que tiene las mayores plantaciones de cha, una planta que crece en el norte argentino. La idea de crear una farmacia natural que ofreciera remedios y cosmticos hechos a partir de plantas se le ocurri en 2002 porque "me pareci que no haba un nicho en el mercado. La gente que busca productos naturales termina yendo a una diettica pero all no hay medicamentos", explica. El target de los que recorren las gndolas en busca de remedios naturales "no es tan claro. Hay hombres y mujeres de 30 aos en adelante pero es muy amplio". Algunos llegan con las recetas que les dieron los mdicos homepatas y los naturistas. Pero tambin va gente con recetas firmadas y selladas por mdicos convencionales. "El Chofitol o el Baghepat, por caso, son remedios naturales hechos con alcachofa. Lo mismo que muchos laxantes elaborados con fibras". En la farmacia tambin hay cosmticos, con la marca Oms. El negocio marcha viento en popa, segn su fundadora. La empresa, que factura 180.000 pesos por mes, inaugur el ao pasado la primera franquicia, en el shopping de Misiones. "Vamos a lanzar las franquicias oficialmente en agosto pero en este caso la otorgamos igual porque eran clientes nuestros y nos pidieron con insistencia". Segn Smolinski, la caracterstica principal de la farmacia es el asesoramiento. "Por eso no es un autoservicio". En el auditorio del local organizan charlas que dictan mdicos. Pero a veces son los mdicos los que se sientan a escuchar las conferencias que preparan para ellos.

Director: Fernando Estevez Castillo

Homeonews
Edicin N 4 Enero-Febrero de 2006

38

HOMEODINAMIZADOR HIEMUSR
Una vez ms el HOMEODINAMIZADOR HIEMUSR, equipo de fluxin continua diseado y patentado por el Ing. Jos Angel Musmarra, fue elegido por sus caractersticas tcnicas, para la realizacin de los Trabajos Prcticos del I Curso Internacional de Farmacia Homeoptica, correspondiente a la I Diplomatura en Terapias Alternativas de la Universidad Nacional Mayor de San Marcos, Per.

Todos los alumnos escucharon con detalle las explicaciones del Farm. Fernando Estevez Castillo referida al uso del equipo y su mantenimiento, as como tambin lo utilizaron para preparar las altas dinamizaciones durante los trabajos prcticos.

Muchos Colegas que participaron de la actividad, tanto del mbito privado (Farmacias) o estatal (Instituciones de Investigacin) de Per, se mostraron muy interesados en la compra del Homeodinamizador HiemusR, ya que no existe hasta el momento ningn otro equipo de fluxin continua que reproduzca Director: Fernando Estevez Castillo

Homeonews
Edicin N 4 Enero-Febrero de 2006

39

las condiciones de dilucin y sucusin de acuerdo a los lineamientos dados por el Dr. C.S.F. Hahnemann, aunque con un sistema de frasco nico, cumpliendo con las Buenas Prcticas de Preparacin de Medicamentos Homeopticos. Para ms informacin, comunicarse a: Farmacia+Natural Av. Luis Mara Campos 575 (C1426BOD) Buenos Aires Argentina Tel.: 54-11-4774-5010 e.mail: info@farmaciamasnatural.com.ar Web: www.farmaciamasnatural.com.ar

OTROS TEMAS CURSOS


CURSO 2006 DE MEDICINA BIOLOGICA, INMUNOMODULACION Y TERAPIAS COMPLEMENTARIAS. OCULOANALISIS El da 20 de Abril comenzar el CURSO 2006 DE MEDICINA BIOLOGICA, INMUNOMODULACION, TERAPIAS COMPLEMENTARIAS y OCULOANALISIS, organizado por Laboratorios Dr. Madaus & Co y dictado por la Dra. Ana Mara Soerensen y otros docentes colaboradores. Cada mdulo se desarrollar los 2 jueves de cada mes, de 9 a 18 hs., desde el mes de Abril hasta Noviembre, inclusive y est dirigido a mdicos, odontlogos y farmacuticos. Se entregar en cada clase un CD con material complementario y se otorgar certificados de asistencia y aprobacin a aquellos alumnos que renan los siguientes requisitos: 1. Asistencia del 75 % a clases 2. Trabajos prcticos aprobados. Lugar: Saln Auditorio de Laboratorios Dr. Madaus & Co (Luis Mara Campos 573 1 Piso, Ciudad Autnoma de Buenos Aires) Carga Horaria: 80 hs Arancel: $ 800 (8 cuotas de $ 100 c/u)

Director: Fernando Estevez Castillo

Homeonews
Edicin N 4 Enero-Febrero de 2006 PROGRAMA ACADEMICO ( 20/04/06) MODULO I: INMUNOMODULACION Y NUTRICION 9hs: El paradigma actual: la medicina basada en la evidencia. 10hs: El oculoanlisis en la prctica mdica. 11hs: La enfermedad como necesidad psicobiolgica. 12hs: Homeostasis de la inflamacin. Leyes biolgicas. 13hs: Receso. 14hs: Eicosanoides en la alimentacin. Prostaglandinas y leucotrienos. 15hs: Prctica de oculoanlisis (OA): cartografia. Ojo normal y patolgico. 16hs: Terapias complementarias. Medicamentos biolgicos. 17hs: Conferencia a cargo del profesional invitado. (11/05/06) MODULO II: DRENAJE BIOLOGICO. SATURACION MACROFAGICA 9hs: Implicancias clnicas de la toxemia intestinal. 10hs: Oculoanlisis: diverticulosis, colon irritable, disbacteriosis. Nefropatas. 11hs: Higiene intestinal y terapia biolgica. Fitoterapia. 12hs: Principios generales de la nutriterapia. IFA. Acido butrico. 13hs: Receso. 14hs: Nefritis. Nefrosis. Litiasis renal. 15hs: Prctica de OA: rganos representados en la gola. 16hs: Eczemas, psoriasis, dermatitis alrgica. Enfoque holstico. 17hs: Conferencia a cargo del profesional invitado.

40

(8/06/06) MODULO III: CONSECUENCIAS CLINICAS DE LA DISFUNCION INMUNE 9hs: Inmunidad natural y adquirida. Interleuquinas, hormonas y neurotransmisores. 10hs: Cartografa iridolgica del sistema psico- neuro-inmune. 11hs: Enfoque integrador y recursos complementarios en alergias respiratorias. 12hs: Esplenopatas, alergias y alteraciones endocrinas. 13hs: Receso. 14hs: Autoinmunidad, vasculitis, disfuncin endotelial. 15hs: Prctica de OA: bazo, timo, ganglios linfticos y glndula suprarrenal. 16hs: Inmunodeficiencias y terapia Kousmine. Oncograma. 17hs: Conferencia a cargo del profesional invitado. (13/07/06) MODULO IV: PATOLOGIAS MS FRECUENTES DEL SISTEMA DIGESTIVO. 9 hs: El cerebro gastrointestinal. Dispepsia y psicopatas. Oroanlisis. 10hs: Signos oculares de inters en las dispepsias. 11hs: Abordaje biolgico en disfuncin gastrointestinal, colitis ulcerosa, colon irritable. 12hs: Disfuncin heptica grado I, II y III. 13hs: Receso Director: Fernando Estevez Castillo

Homeonews
Edicin N 4 Enero-Febrero de 2006 14hs: Lisadoterapia, medicamentos biolgicos y fitoterapia en hepatopatas. 15hs: Prctica de OA: signologa de la esclertica. 16hs: Dispepsia fermentativa y putrefactiva. Dieta amucosa. 17hs: Conferencia a cargo del profesional invitado.

41

(10/08/06) MODULO V: ESTRES, DEPRESIN Y DETERIORO NEUROCOGNITIVO 9hs: Concepto integrador de la enfermedad psquica. 10hs: Oculoanlisis y psiconeuropatas. 11hs: Sndromes de hiperexcitacin. 12hs: Alteracin del ritmo circadiano en nios y adultos. Terapia naturista. 13hs: Receso. 14hs: Deterioro neurocognitivo, shear stress y procesos desmielinizantes. 15hs: Prctica de OA: anillos de contraccin, signo de neurosis, distona neurovegetativa. Representacin cerebral segn el Dr. Jensen. 16hs: Enfoque sistmico del paciente depresivo. Terapia ortomolecular. 17hs: Conferencia a cargo del profesional invitado. (14/09/06) MODULO VI: EJES NEUROENDOCRINOS. LABORATORIO NORMAL Y PATOLOGICO. TERAPIA COMPLEMENTARIA ORAL E INYECTABLE 9hs: Eje crtico-hipotlamo-hipfiso-pineal. Sistema floral de Bach. 10hs: Oculoanlisis: representacin neuroendocrina en el iris. 11hs: Eje hipotlamo-hipfiso-tiroideo. Funcin inmunoendcrina del tejido adiposo. 12hs: La diabetes como enfermedad vascular. Terapia inmunorreguladora. 13hs: Receso. 14hs: Ginecopatas: dismenorrea, esterilidad, sndrome menopusico. 15hs: Prctica de OA: rea 8 y sexualidad masculina y femenina. 16hs: Andropatas: H.P.V. Disfuncin erctil. Esterilidad. 17hs: Conferencia a cargo del profesional invitado. (12/10/06) MODULO VII: EQUILIBRIO ACIDO BASE Y OSTEOPATIAS. TRATAMIENTO BIOLOGICO DE LA PATOLOGIA ARTICULAR 9hs: Metabolismo de las purinas y patologa articular. Recursos locales y sistmicos. 10hs: Oculoanlisis: acidosis, desmineralizacin, colagenopatas. 11hs: Equilibrio cido base y trastornos de la mineralizacin. 12hs: El paciente osteoportico. Terapias alcalinizantes. 13hs: Receso. 14hs: Miopatas. Nutricin complementaria. Antiradicalares. 15hs: Prctica de OA: estructuras iridolgicas y patologa articular. 16hs: Termalismo. Terapias reflejas. Hidroterapia colnica. Apiterapia. 17hs: Conferencia a cargo del profesional invitado. (9/11/06) MODULO VIII: NUTRICION, INTEGRACION Y DEPURACION ORGNICA 9hs: Fisiopatologa del sistema bsico de Pichinger. Director: Fernando Estevez Castillo

Homeonews
Edicin N 4 Enero-Febrero de 2006 10hs: Oculoanlisis: la crnea y el sistema vascular. 11hs: Dislipemia y disfuncin hepatoesplnica. Recursos naturales. 12hs: Patognesis inflamatoria de la ateroesclerosis y su enfoque holstico. 13hs: Receso. 14hs: Sndrome gastrocardial. Quiropraxia y cardiopatas. 15hs: Prctica de OA: clase integradora. 16hs: Regulacin de la presin arterial con medicacin biolgica. 17hs: Conferencia a cargo del profesional invitado. Para ms informacin: Laboratorios Dr. Madaus & Co. S.A. Av. Luis Mara Campos 585 Buenos Aires Argentina (C1426BOD) Tel.: (54) (11) 4771-1734 / 4772-2428 Fax: (54) (11) 4775-4380 E.Mail: info@drmadaus.com.ar

42

CURSO ANUAL DE FITOMEDICINA El martes 4 de abril del corriente ao dio inicio el XI Curso Anual de Fitomedicina exclusivo para mdicos, farmacuticos, nutricionistas, bilogos, veterinarios y odontlogos. Tpicos ms importantes a tratar: Legislacin argentina y mundial sobre fitoterpicos - Antecedentes histricos - Descripcin de principios activos Formas Galnicas - Controles de Calidad - Repaso fisiopatolgico de las principales enfermedades de cada sistema en la clnica diaria y en A.P.S. y su correspondiente abordaje fitoteraputico avalado por farmacopeas mundiales Interacciones medicamentosas - Contraindicaciones - Trabajos Prcticos. Das de Dictado: Martes de 20 a 21:45 horas. Evaluacin: Al finalizar el curso el alumno deber realizar una monografa sobre una especie medicinal o temtica a determinar. Valor del Curso: $ 70 por mes + 1 matrcula de $ 80 que incluye membresa anual, cdigo de acceso a nuestra base de datos en Internet + apuntes. Finaliza: 28 de noviembre de 2006. Director: Dr. Jorge R. ALONSO. Pre-inscripcin - Informaciones: Personalmente en Av. Santa Fe 3553 2 8 Capital Federal, de lunes a viernes de 10 a 19hs. Por telfono al 4831-0378, 4832-4657. Por mail a: fitomedicina@uolsinectis.com.ar.

Director: Fernando Estevez Castillo

Homeonews
Edicin N 4 Enero-Febrero de 2006

43

LIBROS
! Plantas Medicinales Autctonas de la Argentina. Bases cientficas para su aplicacin en Atencin Primaria de la Salud (2005). Dr. Jorge Alonso y Dr. Cristian Desmarchelier. Editorial Lola Este libro ser de mucha importancia para todos los profesionales de la salud de Argentina que se relacionen con la fitomedicina, encontrando las respuestas adecuadas para su utilizacin en la atencin primaria de la salud de la poblacin. Desde ya, aprovecho esta oportunidad, para felicitar al Dr. Jorge Alonso por la calidad de su obra y su cortesa en enviarme un ejemplar del mismo dedicado a mi persona.

! Tratado de Fitofrmacos y Nutracuticos (2004). Dr. Jorge Alonso. Editorial Corpus. Este tratado contiene ms de 300 monografas de las principales especies medicinales de todo el mundo, totalmente actualizadas e incluye un CD con fotos de las mismas.

Director: Fernando Estevez Castillo

Homeonews
Edicin N 4 Enero-Febrero de 2006

44

Si quiere que otro profesional reciba Homeonews por favor complete y enve esta planilla al e.mail: festevezcastillo@fullzero.com.ar

SUSCRIPCIN GRATUITA A HOMEONEWS


Nombre y Apellido:........................................................................................................... Profesin:............................................................................................................................ Domicilio particular: Calle:........................................................... N:................................................................. Localidad:....................................................Prov.:............................................................

C.P.:..Tel.:................................Fax:........................................... E.Mail: Domicilio laboral: Calle:........................................................... N:................................................................. Localidad:....................................................Prov.:............................................................ C.P.:..Tel.:................................Fax:........................................... E.Mail: Temas de inters: ............................................................................................................................................. ............................................................................................................................................. .............................................................................................................................................

Director: Fernando Estevez Castillo