HOMEONEWS - Farmacia Natural · Homeonews Edición N° 4 Œ Enero-Febrero de 2006 Director: Fernando Estevez Castillo 3 INDICE 1- Editorial, pÆg. 4. 2- Fitoterapia: -Efecto de los

HOMEONEWS

Edición N° 4

Enero/Febrero de 2006

Director: Farm. Fernando Estevez Castillo

PUBLICACION BIMESTRAL DISTRIBUCION GRATUITA

PARA PROFESIONALES DE LA SALUD

Homeonews Edición N° 4 � Enero-Febrero de 2006

Director: Fernando Estevez Castillo

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Edición 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) Dirección postal: Pte. Quintana 414 � Lanús Oeste � Pcia de Buenos Aires (B1824NVJ), Argentina Tel.: (54-11) 4241-4441 E.Mail: [email protected] ó [email protected]



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INDICE

1- Editorial, pág. 4.

2- Fitoterapia:

-Efecto de los brotes del rábano japonés (Raphanus sativus) sobre el metabolismo de los lípidos y carbohidratos en ratas normales o con diabetes inducida por estreptozotocina; pág. 5. -Actividad antifúngica y mecanismos de acción de compuestos extraidos de Tríbulus terrestris L.; pág. 6.

3- Alopatía:

Comparación del ácido azelaico y la antralina en el tratamiento de la alopecia areata circunscripta: estudio piloto; pág. 20.

4- Homeopatía:

Estimulación de la actividad mitocondrial del esperma bovino mediante diluciones homeopáticas de monensina; pág. 21.

5- Nutrición:

Los ácidos grasos Omega-3 disminuyen la irritabilidad de pacientes con desórdenes bipolares: estudio abierto; pág. 22.

6- Notas de interés: I Curso Internacional de Farmacia Homeopática (I Diplomatura en Terapias Alternativas, Universidad Nacional Mayor de San Marcos, Lima, Perú); pág. 32.

7- Novedades:

CHIACAPSR: nuevo suplemento dietario; pág. 34. Homeodinamizador HiemusR; pág. 38.

8- Otros temas:

Cursos; pág. 39. Libros; pág. 43. 10- Formulario de suscripción, pág. 44.

Foto de tapa: Salvia hispánica L. (Labiatae)

Las opiniones vertidas en los artículos firmados son responsabilidad de sus autores.



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EDITORIAL

Estimados colegas del equipo de salud: Nuevamente nos reencontramos con un nuevo número de Homeonews, con mucha información y novedades. Quiero aprovechar esta oportunidad para alertar a mis colegas sobre ciertos Cursos de Farmacia Homeopática que una Farmacia está difundiendo en Argentina, con una larga enumeración de antecedentes (Universidades Nacionales y Extranjeras) que toman como propios, omitiendo que los mismos corresponden al profesional (que ejercía la Dirección Técnica) que los ha dictado durante años por invitación directa a su persona y no a esa institución. Es por esto que a modo de consejo, y para no crear falsas expectativas, sugiero verificar siempre los antecedentes académicos de los docentes de los Cursos, así como también la validez de los certificados emitidos, fundamentalmente para la certificación profesional. En este número podrán encontrar dos trabajos muy interesantes de Fitoterapia realizados con el Raphanus sativus y el Tríbulus terrestris; la comparación de dos sustancias (ácido azelaico y antralina) para el tratamiento de la alopecía areata circunscripta; la estimulación de la actividad mitocondrial del esperma bovino mediante diluciones homeopáticas de monensina y un estudio que demuestra que los ácidos grasos Omega-3 disminuyen la irritabilidad de pacientes con desórdenes bipolares. En el capítulo NOTAS DE INTERES, las repercusiones del I Curso Internacional de Farmacia Homeopática dictado en la Universidad Nacional Mayor de San Marcos de Perú. También dos NOVEDADES muy interesantes: Laboratorios Dr. Madaus & Co. presentó CHIACAPSR, el primer suplemento dietario aprobado en Argentina con aceite de Chía y la elección del HOMEODINAMIZADOR HIEMUS para la realización de los trabajos prácticos del Curso de Farmacia Homeopática dictado en la UNMSM (Perú). En el capítulo de Cursos, dos actividades para recomendar, el Curso 2006 de Medicina Biológica, inmunomodulación, terapias complementarias y oculoanálisis 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 debería tener. Próximamente SAIDAH organizará un Taller y una Jornada que oportunamente les comunicaré por este medio. Llego el momento esperado para recorrer Homeonews en toda su extensión, deseando que sea de vuestro agrado, los saludo hasta el próximo número. Farm. Fernando Estévez Castillo



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FITOTERAPIA

EFECTO DE LOS BROTES DEL RÁBANO JAPONÉS (RAPHANUS SATIVUS) SOBRE EL METABOLISMO DE LOS LÍPIDOS 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 1Department of Nutritional Science and Culinary Arts, Toita Woman's College, 2-21-17 Shiba, Minato-ku, Tokyo 105-0014, Japan 2Department of Nutritional Sciences, Faculty of Applied Bio-Science, Tokyo University of Agriculture, 1-1-1 Sakuragaoka, Setagaya-ku, Tokyo 156-8502, Japan 3Department of Bioproductive Sciences, Faculty of Agriculture, Utsunomiya University, 350 Minemachi, Utsunomiya, Tochigi 321-8505, Japan 4Division of Microbiology, National Institute of Health Sciences, 1-18-1 Kamiyoga, Setagaya-ku, Tokyo 158-8501, Japan 5Department 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 información disponible sobre los efectos de los brotes del rábano japonés (JRS) sobre la diabetes. Para aclarar el tema, la influencia del JRS sobre el metabolismo de los lípidos y carbohidratos fue investigada en ratas normales y diabéticas inducidas por estreptozotocina. Estos animales fueron alimentados con una dieta conteniendo 0%, 2.5% o 5% de JRS ad libitum durante 21 días. Comparados con los correspondientes grupos controles, las ratas normales alimentadas con JRS mostraron niveles plasmáticos más bajos de colesterol total (TC), triglicéridos (TG), fosfolípidos (PL), fructosamina, glucosa e insulina y niveles más altos de LDL-Colesterol, mientras que las ratas diabéticas alimentadas con JRS mostraron niveles plasmáticos más bajos de fructosamina, glucosa e insulina sin modificaciones en los parámetros lipídicos del plasma. El JRS también disminuyó los niveles hepáticos de TC, TG and PL en las ratas normales y los TG en las ratas diabéticas. Estos resultados muestran que el JRS tuvo una actividad hipoglucemiante tanto en las ratas normales como diabéticas y parcialmente mejoraron el metabolismo lipídico en las ratas normales. El JRS tiene el potencial de mejorar a los individuos con hiperglucemia en los casos donde la diabetes está presente y en la prevención primaria de la diabetes mellitus.



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Palabras clave: Diabetes. Brotes de rábano japonés. Metabolismo de carbohidratos. Metabolismo lipídico. 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, aDepartment of Pharmacology, College of Pharmacy, Second Military Medical University, 325 Guo He Road, Shanghai 200433, PR China bDepartment 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 antifúngica de los productos naturales está siendo ampliamente estudiada. Las saponinas son conocidas por sus propiedades antifúngicas 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-β-D-glucopyranosyl (1 → 4)-β-D-galactopyranoside (TTS-8), tigogenin-3-O-β-D-glucopyranosyl (1 → 4)-β-D-galactopyranoside (TTS-9), hecogenin-3-O-β-D-glucopyranosyl (1 → 2)-β-D-glucopyranosyl (1 → 4)-β-D-galactopyranoside (TTS-10), hecogenin-3-O-β-D-xylopyranosyl (1 → 3)-β-D-glucopyranosyl (1 → 4)-β-D-galactopyranoside (TTS-11), tigogenin-3-O-β-D-xylopyranosyl (1 → 2)-[β-D-xylopyranosyl (1 → 3)]-β-D-glucopyranosyl (1 → 4)-[α-L-rhamnopyranosyl (1 → 2)]-β-D-galactopyranoside (TTS-12), 3-O-{β-D-xylopyranosyl (1 → 2)-[β-D-xylopyranosyl (1 → 3)]-β-D-glucopyranosyl (1 → 4)-[α-L-rhamnopyranosyl (1 → 2)]-β-D-galactopyranosyl}-26-O-β-D-glucopyranosyl-22-methoxy-(3β,5α,25R)-furostan-3,26-diol (TTS-13), hecogenin-3-O-β-D-glucopyranosyl (1 → 2)-[β-D-xylopyranosyl (1 → 3)]-β-D-glucopyranosyl (1 → 4)-β-D-galactopyranoside (TTS-14), tigogenin-3-O-β-D-glucopyranosyl (1 → 2)-[β-D-xylopyranosyl (1 → 3)]-β-D-glucopyranosyl (1 → 4)-β-D-galactopyranoside (TTS-15). La actividad antifúngica 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 dilución en microcaldo. Se estudió en particular, la actividad in vivo de TTS-12 en un modelo de infección vaginal con Candida albicans. Los resultados mostraron que TTS-12 y TTS-15 fueron muy efectivas in vitro contra muchas especies de cándidas patogénicas y Cryptococcus neoformans. Es digno de destacar que TTS-12 y TTS-15 fueron muy activas contra Candida albicans (MIC80 = 10 y 2.3 µg/mL) y Cryptococcus neoformans (MIC80 = 1.7 y 6.7 µg/mL). La microscopía de contraste de fases



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mostró que TTS-12 inhibe la formación de hifas, un factor de virulencia importante de Candida albicans, mientras que la microscopía de transmisión electrónica que TTS-12 destruye la membrana celular de Candida albicans. Como conclusión, TTS-12 tiene una actividad antifúngica in vitro e in vivo, debilitando la virulencia de Candida albicans y matando a los hongos a través de la destrucción de la membrana celular. Palabras clave: Tribulus terrestris L.; Saponinas; Antifúngicos; Candida albicans Abreviaturas: MICs, concentración inhibitoria mínima; TEM: microscopía de transferencia electrónica.

Artículo 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 3.6�7.2% 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



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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 n-butanol 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), TTS-14 (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 Candida albicans

Candida glabrata

Candida parapsilosis

Candida tropicalis

Cryptococcus neoformans

1 TTS-8 >128.0 >128.0 >128.0 >128.0 >128.0



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No. Compound Candida albicans

Candida glabrata

Candida parapsilosis

Candida tropicalis

Cryptococcus neoformans

2 TTS-9 >128.0 >128.0 >128.0 >128.0 >128.0

3 TTS-10 >128.0 >128.0 >128.0 >128.0 >128.0

4 TTS-11 >128.0 >128.0 >128.0 >128.0 >128.0

5 TTS-12 1.0 ± 0.6 8.8 ± 4.4 45.3 ± 21.3 21.3 ± 8.3 1.7 ± 0.6

6 TTS-13 >128.0 >128.0 >128.0 >128.0 >128.0

7 TTS-14 41.7 ± 20.5 57.6 ± 41.7 >128.0 >128.0 48.0 ± 27.7

8 TTS-15 2.3 ± 1.0 19.2 ± 12.1 74.7 ± 26.1 106.7 ± 33.0 6.7 ± 2.3

9 FLC 1.3 ± 0.7 1.4 ± 0.5 5 2 ± 3.3 1.5 ± 0.6 1.33 ± 0.58

10 AMB 0.29 ± 0.15 0.28 ± 0.21 0.75 ± 027 0.63 ± 0.25 0.21 ± 0.07

11 ICZ 0.12 ± 0.06 0.35 ± 0.14 0.38 ± 0.14 0.38 ± 0.14 0.17 ± 0.07

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 Sprague�Dawley (SD) rats weighing 100�120 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



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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 µg/mL (111.30�0.220 µmol/L), and the final concentrations of FLC, ICZ and AMB were 64.0�0.125 µg/mL (209.15�0.410 µmol/L), 2.0�0.004 µg/mL (2.83�0.006 µmol/L) and 2.0�0.004 µg/mL (2.16�0.004 µmol/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 µg/mL (0, 1.74, 3.48, 6.96, 13.91 µmol/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 µg/mL (0, 3.48 and 13.91 µmol/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 µg/mL.



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2.10. Ultrastructure analysis by transmission electron microscopy 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 µg/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 H-800, Japan) with 2 × 104 magnification. At the same time, the untreated cells were used as control, and FLC (8 µg/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-β-D-glucopyranosyl (1 → 4)-β-D-galactopyranoside (TTS-8), tigogenin-3-O-β-D-glucopyranosyl (1 → 4)-β-D-galactopyranoside (TTS-9), hecogenin-3-O-β-D-glucopyranosyl (1 → 2)-β-D-glucopyranosyl (1 → 4)-β-D-galactopyranoside (TTS-10), hecogenin-3-O-β-D-xylopyranosyl (1 → 3)-β-D-glucopyranosyl (1 → 4)-β-D-galactopyranoside (TTS-11), tigogenin-3-O-β-D-xylopyranosyl (1 → 2)-[β-D-xylopyranosyl (1 → 3)]-β-D-glucopyranosyl (1 → 4)-[α-L-rhamnopyranosyl (1 → 2)]-β-D-galactopyranoside (TTS-12), 3-O-{β-D-xylopyranosyl (1 → 2)-[β-D-xylopyranosyl (1 → 3)]-β-D-glucopyranosyl (1 → 4)-[α-L-rhamnopyranosyl (1 → 2)]-β-D-galactopyranosyl}-26-O-β-D-glucopyranosyl-22-methoxy-(3β,5α,25R)-furostan-3,26-diol (TTS-13), hecogenin-3-O-β-D-glucopyranosyl (1 → 2)-[β-D-xylopyranosyl (1 → 3)]-β-D-glucopyranosyl (1 → 4)-β-D-galactopyranoside (TTS-14), tigogenin-3-O-β-D-glucopyranosyl (1 → 2)-[β-D-xylopyranosyl (1 → 3)]-β-D-glucopyranosyl (1 → 4)-β-D-galactopyranoside (TTS-15), 2S,3S,4S,5S-hexitol (TTS-16), inorganic salt (a mixture of NaNO3, KNO3 and K2NO2F·HF 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 1�8.



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3.2. Antifungal susceptibility results 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 µg/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 µg/mL, respectively. They were also very effective against Cryptococcus neoformans at 1.7 and 6.7 µg/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 µg/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



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16 µg/mL) (A) or TTS-15 (0, 2, 4, 8 and 16 µg/mL) (B) in the subsequent for 48 h.

3.4. The vaginal infection model 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 TTS-12 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 µg/mL TTS-12, or 4 µg/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 µg/mL TTS-12 or FLC groups, hyphal formation of Candida albicans was inhibited markedly (Fig. 4).



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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 µg/mL; C, 16 µg/mL), and FLC (D, 4 µg/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 µm (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



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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 µg/mL); (C) ultrastructure of Candida albicans treated with FLC (8 µg/mL) as the positive control. The cell membrane of Candida albicans was seriously destroyed by TTS-12 or FLC (arrows indicate destroyed cell membrane). The white bar represents a length of 2 µm (A).

4. Discussion and conclusion

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 TTS-13 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 µg/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 µg/mL and that of individual saponins is between 4 and 20.8 µg/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 µg/mL (3.1 µmol/L) and 7.7 µg/mL (6.2 µmol/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).



16

Prosapogenin A of dioscin exhibited antifungal activity against human pathogenic yeasts Candida albicans (MIC 20.8 µg/mL), Candida glabrata (MIC 6.25 µg/mL) and Candida tropicalis (MIC 25 µg/mL) (Sautour et al., 2004). TTS-12 and TTS-15 were also very active on Cryptococcus neoformans (MIC80 = 1.7 and 6.7 µg/mL, respectively), similar to the antifungal activities of several other saponins reported with the MICs of 2�12.5 µg/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 4�7 (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 µg/mL, and MIC values against Cryptococcus neoformans were 2.00 and 3.12 µg/mL, respectively. However, our result failed to show that TTS-14 had strong activity against Candida albicans and



17

Cryptococcus neoformans, with MIC being 41.7 and 48.0 µg/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



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Basic Research Projects (02DJ14016) and Shanghai R&D Key Fund (02DZ19120).

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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. 628�631. 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. 221�222. 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. 9�35. 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. 325�326. 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. 504�512. 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. 75�81. 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. 27�33. 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. 90�92. 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. 1263�1268. 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. 203�211. 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.



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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. 363�369. 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. 163�169. 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. 1274�1277. 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. 1847�1849. 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. 539�543. 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. 1677�1681. 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. 545�550. 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. 223�228.

ALOPATIA

COMPARACION DEL ACIDO AZELAICO Y LA ANTRALINA EN

ELTRATAMIENTO DE LA ALOPECIA AREATA CIRCUNSCRIPTA: ESTUDIO PILOTO

Sasmaz, Sezai1; Arican, Ozer1



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1Department of Dermatology, School of Medicine, Kahramanmaras Sutcuimam University, Kahramanmaras, Turkey

[American Journal of Clinical Dermatology,Volume 6, Number 6, 2005, pp.403-406]

RESUMEN

Antecedentes: Aunque el ácido azelaico ha sido utilizado en forma tópica para el tratamiento de la alopecía, no existen hasta la fecha ensayos controlados con esta droga para esta patología.

Objetivo: El objetivo de este estudio fue determinar la eficacia, tolerabilidad y seguridad del tratamiento de la ALOPECIA AREATA CIRCUNSCRIPTA (AA) con ácido azelaico comparado con la antralina.

Métodos: Este estudio incluyó 31 sujetos con AA circunscripta quienes no habían recibido ningún tratamiento por lo menos un año antes del comienzo del estudio. Se registraron las características demográficas y clínicas iniciales de todos los individuos que participaron de la experiencia. Los sujetos fueron elegidos al azar para aplicarle ácido azelaico al 20% (15) o antralina al 0,5% (16) durante 12 semanas. Luego se continuó con un período de seguimiento de 8 semanas sin ningún tipo de crema aplicada. Dos investigadores independientes realizaron, a las 20 semanas, una evaluación de eficacia con examinación clínica 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 ácido 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 ácido azelaico (8 de 15) comparados con el 56.2% (9 de 16) del grupo de antralina (p > 0.05). Ningún efecto adverso serio fue observado en ambos grupos durante el estudio.

Conclusión: el presente estudio piloto mostró que el uso de ácido azelaico da resultados similares a la antralina con respecto al crecimiento capilar, y que puede ser una agente terapéutico tópico efectivo para el tratamiento de la Alopecía Areata circunscripta. Sin embargo, se necesitan más ensayos para poder llegar a una conclusión definitiva.

HOMEOPATIA

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



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DM Aziz1, and H Enbergs2 1Department of Surgery and Obstetrics, College of Veterinary Medicine, University of Mosul, Mosul, Iraq 2Institute 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 parámetro importante de la viabilidad de los espermatozoides y se relaciona con la motilidad espermática. La monensina es comúnmente utilizada en laboratorio como un inhibidor de la actividad mitocondrial espermática. Este estudio fue realizado para evaluar la influencia de algunas diluciones homeopáticas de monensina sobre la actividad mitocondrial espermática. Se utilizaron para el estudio, eyaculados frescos provenientes de seis toros adultos. Las muestras de semen fueron evaluadas usando un citómetro de flujo para determinar la actividad mitocondrial y la viabilidad espermática, utilizando Rhodamina 123 y SYBR-14, respectivamente. La monensina 9X produjo un resultado muy altamente significativo (P<0.001) en la estimulación de la actividad mitocondrial espermática, mientras que la 5X, 7X, 8X y 13X fue altamente significativo (P<0.01). Otras diluciones homeopáticas de monensina (6×, 10×, 11×, 12× and 14×) también tuvieron un efecto estimulante significativo (P<0.05). El uso de monensina no tuvo ningún efecto negativo sobre la viabilidad espermática. Podemos concluir que algunas diluciones homeopáticas de monensina incrementan la actividad mitocondrial de los espermatozoides bovinos sin efecto negativo con respecto a su viabilidad, siendo la 9X la dilución más efectiva. Es necesario realizar más estudios in vivo para estimar el efecto de las diluciones homeopáticas de monensina sobre la calidad del semen. Palabras clave: esperma; actividad mitocondrial, monensina, homeopatía.

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 Ayşegül Yõldõz6 1University of Missouri � Kansas City, Missouri, 8801 West 148th Terrace, Overland Park, KS 66221, USA.



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2Washington University, School of Medicine, Department of Psychiatry, Campus Box: 8134, 660 South Euclid Avenue, St. Louis, Missouri, 63110, USA. 3Department of Psychiatry, School of Medicine, University of Missouri-Kansas City, Resource Development Institute, 601 Walnut Street, Kansas City, MO 64106, USA. 4Mood Disorders Psychopharmacology Unit, University Health Network, Toronto Western Hospital, 399 Bathurst Street, ECW-3D-010, Toronto, Ontario M5T 2S8, Canada. 5Mood and Anxiety Disorders Clinic, Department of Psychiatry, University of Pennsylvania, 3535 Market Street, 2nd floor, Philadelphia, PA 19104, USA 6Dokuz Eylül Medical School, Department of Psychiatry, İzmir, Turkey. [Nutrition Journal 2005, 4:6]

RESUMEN Este es el reporte de la continuación de un estudio abierto con 37 pacientes sobre ácidos 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 tenían el diagnóstico de desórdenes bipolares y visitaban regularmente una Clínica para estas patologías a lo largo de toda la experiencia. En cada visita, el estado de cada paciente fue seguido utilizando la planilla de monitoreo clínico. Los sujetos reportaban la frecuencia y severidad de los episodios de irritabilidad experimentados en los últimos 10 días previos a la consulta; la frecuencia se midió por el porcentaje de días en los cuales experimentaban irritabilidad, mientras que para la severidad se utilizó la escala de Likert de 1 a 4 (si se presentaba). El componente de irritabilidad de acuerdo a la Escala de evaluación de Manía de Young (YMRS) fue registrado en 13 de los 39 pacientes. Los pacientes tuvieron una persistente irritabilidad a pesar de su tratamiento continuo farmacológico y psicoterapéutico.

La ingesta de ácidos Omega-3 ayudó con el componente de irritabilidad de pacientes sufriendo desórdenes bipolares con una presencia significativa de signos de irritabilidad. Bajas dosis de Omega- 3 (1 a 2 gramos por día), pueden ayudar también con el componente de irritabilidad de diferentes condiciones clínicas, tales como esquizofrenia, problemas de personalidad y otras condiciones psiquiátricas con un signo común presente de irritabilidad.

ARTICULO ORIGINAL INTRODUCTION



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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 O-3FA 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 (1.5�1.8 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



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influence both aggression and depression [12]. Although attention-deficit/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 O-3FA 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



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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 Last Recorded FinalT1

N 39 37 13 Mean 1833.33 2878.38 2615.38 Mode 1000T2, T3 1000 2000 Median 2000 2000 2000 SD 1071.91 2011.79 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 t-test (t = 4.36, 36 df, p < .001). The difference between the distributions was examined using the non-parametric sign test. The number of negative



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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 t-test (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



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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



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.



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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 Tesla-2 (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 1�2 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-expert-dependent 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



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for more aggressive funding for research into medicinal foods, such as omega-three 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 acid-containing food administration on symptoms of attention-deficit/hyperactivity disorder - a placebo-controlled double-blind study. Eur J Clin Nutr 2004, 58(3):467-473.



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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):133-140.

NOTAS DE INTERES

I CURSO INTERNACIONAL DE FARMACIA HOMEOPATICA (I DIPLOMATURA EN TERAPIAS ALTERNATIVAS

UNIVERSIDAD NACIONAL MAYOR DE SAN MARCOS LIMA - PERÚ

Del 4 al 8 de marzo del corriente año, tuvo lugar en la ciudad de Lima, Perú, el I Curso Internacional de Farmacia Homeopática (teórico-práctico), 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 único docente del mismo. El día previo (3 de marzo) se realizaron dos conferencias públicas, con el título Avances en Homeopatía I y II, en el Auditorio del Hospital �Rebagliatti� y de la Municipalidad de Lince, respectivamente.



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El Farm. Fernando Estevez Castillo exponiendo durante la parte teórica del Curso. Toda la organización estuvo a cargo de la Asociación Peruana de Salud Integral (ASPESIN), que a través 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 más mínimo detalle. Para el desarrollo de la parte teórica los alumnos contaron con material bibliográfico impreso y un CD con más información, y en la parte práctica cada uno en forma individual pudo elaborar los distintos productos intermedios y formas farmacéuticas homeopáticas, 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 prác- ticas realizadas en el laboratorio del Departamento de Química de la Facultad de Farmacia y Bioquímica de la UNMSM.



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El esfuerzo fue muy grande pero valió la pena, ya que el entusiasmo por aprender e intercambiar experiencias, superó ampliamente al cansancio lógico de actividades tan extensas en carga horaria. Pero todo no terminaba con la parte académica, ya que gracias a la excelente predisposición, hospitalidad y el don de buenas personas de todo el grupo de ASPESIN, en el poco tiempo que duró mi estadía en Perú, tuve la oportunidad de conocer numerosos lugares de Lima, costumbres de su pueblo e historia, y un montón de cosas más que dejaron una huella muy importante en mi persona para volver en un futuro no muy lejano y seguir conociendo a ese maravilloso país que es Perú.

El Farm. Fernando Estevez Castillo junto a los organizadores del

Curso y algunos alumnos del mismo, luego de la culminación de la parte teórica. ¡Felicitaciones a ASPESIN por la idoneidad y calidez de todos sus miembros, así como también 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 ChiacapsR, un nuevo suplemento dietario a base de Omega �3 obtenido a partir de la semilla de CHIA (Salvia hispánica), que es considerada la fuente natural más rica en éstos ácidos grasos, superando inclusive al lino, mostrando una ventaja muy



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

ChiacapsR se presenta en cápsulas blandas por 30 ó 60 unidades, conteniendo cada una 1000 mg de aceite de chía equivalente a 600 mg de Omega-3.

BENEFICIOS DE LOS OMEGA-3 presentes en la CHIA PARA EL CORAZON

• Disminución de la presión arterial. • Reducción de la arritmia y de la probabilidad de muerte súbita. • Disminución de triglicéridos y LDL.

EN LOS PROCESOS INFLAMATORIOS

• Inhiben los eventos pro-inflamatorios. • Protege al pulmón de procesos inflamatorios. • Reduce la sintomatología 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 neurológico del feto. • Previenen la hipertensión de la madre relacionada con el embarazo.

EN LA LACTANCIA

• Estimulan el crecimiento y desarrollo neurológico del niño. • Mejoran el índice de desarrollo mental de niños prematuros. • Refuerzan el sistema inmunológico del recién nacido. • Contribuyen al desarrollo psicomotor del niño. • Reducen la dermatitis seborreica del recién nacido.

ARTICULO PUBLICADO EN EL DIARIO CLARIN



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Domingo 9 de Abril de 2006 Diario Clarín – Suplemento Económico

FARMACIA NATURAL

Omega 3, de la planta a la cápsula

Cecilia de Castro. [email protected]

Los aztecas no sabían nada de Omega 3. Tampoco los incas ni los mayas. Pero es muy probable que el colesterol no taponara sus venas. Gracias a la chía, una planta que junto con el maíz y algunos porotos era su principal alimento. Con la llegada de los españoles la chía cayó en el olvido. Hoy los científicos desempolvaron la planta. En los Estados Unidos descubrieron que las tribus que consumían este alimento no sabían lo que era ir al médico �al brujo, más bien� por problemas de diabetes o hipertensión. Es que un buen plato de chía al ajillo, a la portuguesa o cualquiera que fuera la receta del chef proporcionaba a los indígenas una buena dosis de Omega 3. Para obtener los beneficios de la chía en el siglo 21 no será necesario aprender a cocinar la planta. Bastará con abrir un frasco y tomar unas pastillas. Muy pronto Farmacia Natural, el emprendimiento de Patricia Smolinski (47), empezará a vender las cápsulas con aceite de chía. "Todo empezó por casualidad. Una pariente mía muy viejita me comentó que el médico le había recetado un remedio. Yo le pedí a una farmacéutica que buscara algo sobre el tema pero sin darle demasiada importancia. 'Yo sé lo que es porque mi tío la está estudiando en los Estados Unidos', me dijo ella".

Patricia Smolinski, presenta el producto

CHIACAPSR Así Smolinski, que también es dueña de Laboratorios Dr. Madaus, se puso en contacto con la gente de la Universidad de Arizona. Y empezaron a trabajar



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 práctica, hicimos la investigación para desarrollar el suplemento dietario". Las cápsulas se venderán en la Argentina y también se exportarán a los Estados Unidos. La marca local será 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 únicos que lo encapsulamos y que tenemos el registro". Además Smolinski tiene contrato de exclusividad con la gente que tiene las mayores plantaciones de chía, una planta que crece en el norte argentino. La idea de crear una farmacia natural que ofreciera remedios y cosméticos hechos a partir de plantas se le ocurrió en 2002 porque "me pareció que no había un nicho en el mercado. La gente que busca productos naturales termina yendo a una dietética pero allí no hay medicamentos", explica. El target de los que recorren las góndolas en busca de remedios naturales "no es tan claro. Hay hombres y mujeres de 30 años en adelante pero es muy amplio". Algunos llegan con las recetas que les dieron los médicos homeópatas y los naturistas. Pero también va gente con recetas firmadas y selladas por médicos convencionales. "El Chofitol o el Bagóhepat, por caso, son remedios naturales hechos con alcachofa. Lo mismo que muchos laxantes elaborados con fibras". En la farmacia también hay cosméticos, con la marca Oms. El negocio marcha viento en popa, según su fundadora. La empresa, que factura 180.000 pesos por mes, inauguró el año 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". Según Smolinski, la característica principal de la farmacia es el asesoramiento. "Por eso no es un autoservicio". En el auditorio del local organizan charlas que dictan médicos. Pero a veces son los médicos los que se sientan a escuchar las conferencias que preparan para ellos.



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HOMEODINAMIZADOR HIEMUSR Una vez más el HOMEODINAMIZADOR HIEMUSR, equipo de fluxión continua diseñado y patentado por el Ing. José Angel Musmarra, fue elegido por sus características técnicas, para la realización de los Trabajos Prácticos del I Curso Internacional de Farmacia Homeopática, correspondiente a la I Diplomatura en Terapias Alternativas de la Universidad Nacional Mayor de San Marcos, Perú.

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

Muchos Colegas que participaron de la actividad, tanto del ámbito privado (Farmacias) o estatal (Instituciones de Investigación) de Perú, se mostraron muy interesados en la compra del Homeodinamizador HiemusR, ya que no existe hasta el momento ningún otro equipo de fluxión continua que reproduzca



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las condiciones de dilución y sucusión de acuerdo a los lineamientos dados por el Dr. C.S.F. Hahnemann, aunque con un sistema de frasco único, cumpliendo con las Buenas Prácticas de Preparación de Medicamentos Homeopáticos. Para más información, comunicarse a:

Farmacia+Natural

Av. Luis María Campos 575 (C1426BOD) Buenos Aires � Argentina

Tel.: 54-11-4774-5010 e.mail: [email protected] Web: www.farmaciamasnatural.com.ar

OTROS TEMAS

CURSOS

CURSO 2006 DE MEDICINA BIOLOGICA, INMUNOMODULACION Y

TERAPIAS COMPLEMENTARIAS. OCULOANALISIS El día 20 de Abril comenzará el CURSO 2006 DE MEDICINA BIOLOGICA, INMUNOMODULACION, TERAPIAS COMPLEMENTARIAS y OCULOANALISIS, organizado por Laboratorios Dr. Madaus & Co y dictado por la Dra. Ana María Soerensen y otros docentes colaboradores. Cada módulo se desarrollará los 2° jueves de cada mes, de 9 a 18 hs., desde el mes de Abril hasta Noviembre, inclusive y está dirigido a médicos, odontólogos y farmacéuticos. Se entregará en cada clase un CD con material complementario y se otorgará certificados de asistencia y aprobación a aquellos alumnos que reúnan los siguientes requisitos: 1. Asistencia del 75 % a clases 2. Trabajos prácticos aprobados. Lugar: Salón Auditorio de Laboratorios Dr. Madaus & Co (Luis María Campos 573 1° Piso, Ciudad Autónoma de Buenos Aires) Carga Horaria: 80 hs Arancel: $ 800 (8 cuotas de $ 100 c/u)



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PROGRAMA ACADEMICO

( 20/04/06) MODULO I: INMUNOMODULACION Y NUTRICION

9hs: El paradigma actual: la medicina basada en la evidencia. 10hs: El oculoanálisis en la práctica médica. 11hs: La enfermedad como necesidad psicobiológica. 12hs: Homeostasis de la inflamación. Leyes biológicas. 13hs: Receso. 14hs: Eicosanoides en la alimentación. Prostaglandinas y leucotrienos. 15hs: Práctica de oculoanálisis (OA): cartografia. Ojo normal y patológico. 16hs: Terapias complementarias. Medicamentos biológicos. 17hs: Conferencia a cargo del profesional invitado.

(11/05/06) MODULO II: DRENAJE BIOLOGICO. SATURACION MACROFAGICA

9hs: Implicancias clínicas de la toxemia intestinal. 10hs: Oculoanálisis: diverticulosis, colon irritable, disbacteriosis. Nefropatías. 11hs: Higiene intestinal y terapia biológica. Fitoterapia. 12hs: Principios generales de la nutriterapia. IFA. Acido butírico. 13hs: Receso. 14hs: Nefritis. Nefrosis. Litiasis renal. 15hs: Práctica de OA: órganos representados en la gola. 16hs: Eczemas, psoriasis, dermatitis alérgica. Enfoque holístico. 17hs: Conferencia a cargo del profesional invitado.

(8/06/06) MODULO III: CONSECUENCIAS CLINICAS DE LA DISFUNCION INMUNE

9hs: Inmunidad natural y adquirida. Interleuquinas, hormonas y neurotransmisores. 10hs: Cartografía iridológica del sistema psico- neuro-inmune. 11hs: Enfoque integrador y recursos complementarios en alergias respiratorias. 12hs: Esplenopatías, alergias y alteraciones endocrinas. 13hs: Receso. 14hs: Autoinmunidad, vasculitis, disfunción endotelial. 15hs: Práctica de OA: bazo, timo, ganglios linfáticos y glándula suprarrenal. 16hs: Inmunodeficiencias y terapia Kousmine. Oncograma. 17hs: Conferencia a cargo del profesional invitado.

(13/07/06) MODULO IV: PATOLOGIAS MÁS FRECUENTES DEL SISTEMA

DIGESTIVO. 9 hs: El �cerebro� gastrointestinal. Dispepsia y psicopatías. Oroanálisis. 10hs: Signos oculares de interés en las dispepsias. 11hs: Abordaje biológico en disfunción gastrointestinal, colitis ulcerosa, colon irritable. 12hs: Disfunción hepática grado I, II y III. 13hs: Receso



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14hs: Lisadoterapia, medicamentos biológicos y fitoterapia en hepatopatías. 15hs: Práctica de OA: signología de la esclerótica. 16hs: Dispepsia fermentativa y putrefactiva. Dieta amucosa. 17hs: Conferencia a cargo del profesional invitado.

(10/08/06) MODULO V: ESTRES, DEPRESIÓN Y DETERIORO NEUROCOGNITIVO

9hs: Concepto integrador de la enfermedad psíquica. 10hs: Oculoanálisis y psiconeuropatías. 11hs: Síndromes de hiperexcitación. 12hs: Alteración del ritmo circadiano en niños y adultos. Terapia naturista. 13hs: Receso. 14hs: Deterioro neurocognitivo, shear stress y procesos desmielinizantes. 15hs: Práctica de OA: anillos de contracción, signo de neurosis, distonía neurovegetativa. Representación cerebral según el Dr. Jensen. 16hs: Enfoque sistémico 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 córtico-hipotálamo-hipófiso-pineal. Sistema floral de Bach. 10hs: Oculoanálisis: representación neuroendocrina en el iris. 11hs: Eje hipotálamo-hipófiso-tiroideo. Función inmunoendócrina del tejido adiposo. 12hs: La diabetes como enfermedad vascular. Terapia inmunorreguladora. 13hs: Receso. 14hs: Ginecopatías: dismenorrea, esterilidad, síndrome menopáusico. 15hs: Práctica de OA: área 8 y sexualidad masculina y femenina. 16hs: Andropatías: H.P.V. Disfunción eréctil. 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 patología articular. Recursos locales y sistémicos. 10hs: Oculoanálisis: acidosis, desmineralización, colagenopatías. 11hs: Equilibrio ácido base y trastornos de la mineralización. 12hs: El paciente osteoporótico. Terapias alcalinizantes. 13hs: Receso. 14hs: Miopatías. Nutrición complementaria. Antiradicalares. 15hs: Práctica de OA: estructuras iridológicas y patología articular. 16hs: Termalismo. Terapias reflejas. Hidroterapia colónica. Apiterapia. 17hs: Conferencia a cargo del profesional invitado.

(9/11/06) MODULO VIII: NUTRICION, INTEGRACION Y DEPURACION ORGÁNICA

9hs: Fisiopatología del sistema básico de Pichinger.



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10hs: Oculoanálisis: la córnea y el sistema vascular. 11hs: Dislipemia y disfunción hepatoesplénica. Recursos naturales. 12hs: Patogénesis inflamatoria de la ateroesclerosis y su enfoque holístico. 13hs: Receso. 14hs: Síndrome gastrocardial. Quiropraxia y cardiopatías. 15hs: Práctica de OA: clase integradora. 16hs: Regulación de la presión arterial con medicación biológica. 17hs: Conferencia a cargo del profesional invitado. Para más información:

Laboratorios Dr. Madaus & Co. S.A. Av. Luis María Campos 585 � Buenos Aires � Argentina (C1426BOD)

Tel.: (54) (11) 4771-1734 / 4772-2428 Fax: (54) (11) 4775-4380

E.Mail: [email protected]

CURSO ANUAL DE FITOMEDICINA

El martes 4 de abril del corriente año dio inicio el XIº Curso Anual de Fitomedicina exclusivo para médicos, farmacéuticos, nutricionistas, biólogos, veterinarios y odontólogos. Tópicos más importantes a tratar: Legislación argentina y mundial sobre fitoterápicos - Antecedentes históricos - Descripción de principios activos - Formas Galénicas - Controles de Calidad - Repaso fisiopatológico de las principales enfermedades de cada sistema en la clínica diaria y en A.P.S. y su correspondiente abordaje fitoterapéutico avalado por farmacopeas mundiales - Interacciones medicamentosas - Contraindicaciones - Trabajos Prácticos. Días de Dictado: Martes de 20 a 21:45 horas. Evaluación: Al finalizar el curso el alumno deberá realizar una monografía sobre una especie medicinal o temática a determinar. Valor del Curso: $ 70 por mes + 1 matrícula de $ 80 que incluye membresía anual, código de acceso a nuestra base de datos en Internet + apuntes. Finaliza: 28 de noviembre de 2006. Director: Dr. Jorge R. ALONSO. Pre-inscripción - Informaciones: Personalmente en Av. Santa Fe 3553 2° �8� Capital Federal, de lunes a viernes de 10 a 19hs. Por teléfono al 4831-0378, 4832-4657. Por mail a: [email protected].



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LIBROS ! Plantas Medicinales Autóctonas de la Argentina. Bases científicas para

su aplicación en Atención Primaria de la Salud (2005). Dr. Jorge Alonso y Dr. Cristian Desmarchelier. Editorial Lola

Este libro será de mucha importancia para todos los profesionales de la salud de Argentina que se relacionen con la fitomedicina, encontrando las respuestas adecuadas para su utilización en la atención primaria de la salud de la población. Desde ya, aprovecho esta oportunidad, para felicitar al Dr. Jorge Alonso por la calidad de su obra y su cortesía en enviarme un ejemplar del mismo dedicado a mi persona.

! Tratado de Fitofármacos y Nutracéuticos (2004). Dr. Jorge Alonso. Editorial Corpus.

Este tratado contiene más de 300 monografías de las principales especies medicinales de todo el mundo, totalmente actualizadas e incluye un CD con fotos de las mismas.



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Si quiere que otro profesional reciba Homeonews por favor complete y envíe esta planilla al e.mail: [email protected]

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Documents

HOMEONEWS - Farmacia Natural · Homeonews Edición N° 4 Œ Enero-Febrero de 2006 Director: Fernando Estevez Castillo 3 INDICE 1- Editorial, pÆg. 4. 2- Fitoterapia: -Efecto de los