Consideraciones de los estudios toxicológicos preclínicos para vacunas / Preclinical toxicology considerations for vaccines

Con respecto al artículo publicado en la revista Vaccimonitor, volumen 28 número 1 del año 2019, donde se presentan los resultados del estudio toxicológico realizado a la vacuna antimeningocóccica VA-MENGOC-BC® después de 24 y 36 meses de almacenada de 4 a 8°C, quisiera emitir un grupo de consideraciones sobre algunos de los parámetros evaluados en los estudios toxicológicos para vacunas, con el objetivo de optimizar los resultados. En este sentido, podríamos contribuir a un mejor diseño de los estudios toxicológicos preclínicos, para alcanzar un valor predictivo más cercano a lo esperado en los ensayos clínicos. Por otra parte, estas consideraciones pudieran tenerse en cuenta para el análisis de los aspectos éticos, bienestar animal y determinación del punto final humanitario. Los estudios toxicológicos preclínicos de las vacunas se convirtieron en obligatorios desde la última década del pasado siglo XX, donde se han propuesto y diseñado diferentes estudios para cumplir con estos requisitos. Podemos mencionar, por ejemplo, los estudios de seguridad farmacológica, dosis única, tolerancia local y dosis repetida, entre otros.1 En cada uno de ellos existen parámetros a evaluar que se repiten, como son la observación clínica de los animales, peso corporal, consumo de agua y alimentos, así como la respuesta local y sistémica. Sin embargo, la información que pudiéramos obtener de estos parámetros, desde las pruebas de concepto hasta los propios estudios toxicológicos, no es explotada al máximo. Hoy en día, las guías emitidas por la Organización Mundial de la Salud (OMS), en mi opinión, son una de las más completas, porque recogen gran parte de las regulaciones y guías emitidas por otras entidades como la FDA (del inglés: Food and Drug Administration), EMA (del inglés: European Medicine Agency), OECD (del inglés: Organisation for Economic Co-operation and Development), ICH (del inglés: International Committee of Harmonization), entre otras. Contemplan un amplio número de parámetros a investigar en los estudios toxicológicos, que abarcan desde ensayos in vitro hasta in vivo.1 Sobre estos últimos, cada vez más, y para bien, son tenidos en cuenta los aspectos del bienestar animal y el principio de las 3Rs (Reemplazo, Reducción, Refinamiento).1,2 En este sentido, consideramos que se pueden introducir elementos que pudieran potenciar las respuestas de algunos de los parámetros investigados. En nuestro caso los realizamos para brindar nuevos elementos a la seguridad de un producto con años de aplicación, tanto en niños...(AU)

Comparative safety assessments of the biosimilar APZ001 and Erbitux in pre-clinical animal models

Abstract Purpose: To evaluate the toxicity of Erbitux as well as its biosimilar APZ001 antibody (APZ001) in pre-clinical animal models including mice, rabbits and cynomolgus monkeys. Methods: We performed analysis of normal behavior activity, autonomic and non-autonomic nervous functions, nervous-muscle functions, nervous excitability and sensorimotor functions on CD-1 mice. Subsequently, we studied that effects of APZ001 and Erbitux on respiratory system, cardiovascular system and kidney in Cynomolgus monkey models and performed local tolerance experiments on New Zealand rabbits. Results: The comparisons between APZ001 and Erbitux showed no significant differences in mice autonomic nervous system, nervous muscle functions, non-autonomic nervous functions, nervous excitability and sensorimotor functions between treated and untreated group (p>0.05). APZ001 and Erbitux showed negative effect on CD-1 mice in the present of pentobarbital sodium anesthesia (p>0.05). Single administrations of high, medium or low doses of APZ001 did not lead to monkey urine volume alterations (p>0.05). In human tissues, APZ001 and Erbitux showed positive signals in endocardium, lung type II alveolar epithelial cell and surrounding vessels, but showed negative results in kidney and liver tissues. No hemolysis phenomenon and serious side-effects in vessels and muscles were observed in rabbits when administrated with APZ001 and Erbitux respectively. Conclusion: The safety comparisons between APZ001 antibody and Erbitux showed that these two antibodies showed highly similarities in mice, rabbits and cynomolgus monkey animal models in consideration of pharmaceutical effects, indicating APZ001 might be a suitable substitute for Erbitux.

Achievement of constitutive fluorescent pLEXSY-egfp Leishmania braziliensis and its application as an alternative method for drug screening in vitro

BACKGROUND Gene reporter-fluorescent cells have emerged as alternative method for drug screening. OBJECTIVE Achievement of constitutive expression of fluorescent protein GFP by Leishmania braziliensis as alternative method for drug screening. METHODS L. braziliensis-GFP was generated using Leishmania tarentolae pLEXSY-egfp for constitutive expression of GFP. Fluorescent cells were selected and subjected to standardisation tests of anti-promastigote and anti-intracellular amastigote assays. FINDINGS Our results showed that L. braziliensis-GFP method is faster and more sensitive than Allamar Blue-resazurin. MAIN CONCLUSION Transfected parasites maintained stable fluorescence after successive in vitro passages and pLEXSY system can be used to achieve non-L. tarentolae fluorescent cells.

Non-clinical studies required for new drug development - Part I: early in silico and in vitro studies, new target discovery and validation, proof of principles and robustness of animal studies

This review presents a historical overview of drug discovery and the non-clinical stages of the drug development process, from initial target identification and validation, through in silico assays and high throughput screening (HTS), identification of leader molecules and their optimization, the selection of a candidate substance for clinical development, and the use of animal models during the early studies of proof-of-concept (or principle). This report also discusses the relevance of validated and predictive animal models selection, as well as the correct use of animal tests concerning the experimental design, execution and interpretation, which affect the reproducibility, quality and reliability of non-clinical studies necessary to translate to and support clinical studies. Collectively, improving these aspects will certainly contribute to the robustness of both scientific publications and the translation of new substances to clinical development.

Sudden Cardiac Death in Brazil: A Community-Based Autopsy Series (2006-2010) / Morte Súbita Cardíaca no Brasil: Análise dos Casos de Ribeirão Preto (2006-2010)

Background: Sudden cardiac death (SCD) is a sudden unexpected event, from a cardiac cause, that occurs in less than one hour after the symptoms onset, in a person without any previous condition that would seem fatal or who was seen without any symptoms 24 hours before found dead. Although it is a relatively frequent event, there are only few reliable data in underdeveloped countries. Objective: We aimed to describe the features of SCD in Ribeirão Preto, Brazil (600,000 residents) according to Coroners’ Office autopsy reports. Methods: We retrospectively reviewed 4501 autopsy reports between 2006 and 2010, to identify cases of SCD. Specific cause of death as well as demographic information, date, location and time of the event, comorbidities and whether cardiopulmonary resuscitation (CPR) was attempted were collected. Results: We identified 899 cases of SCD (20%); the rate was 30/100000 residents per year. The vast majority of cases of SCD involved a coronary artery disease (CAD) (64%) and occurred in men (67%), between the 6th and the 7th decades of life. Most events occurred during the morning in the home setting (53.3%) and CPR was attempted in almost half of victims (49.7%). The most prevalent comorbidity was systemic hypertension (57.3%). Chagas’ disease was present in 49 cases (5.5%). Conclusion: The majority of victims of SCD were men, in their sixties and seventies and the main cause of death was CAD. Chagas’ disease, an important public health problem in Latin America, was found in about 5.5% of the cases. .

Fundamento: Morte súbita cardíaca (MSC) é um evento súbito e inesperado, de causa cardiovascular, que ocorre em menos de uma hora após o início dos sintomas, em indivíduo sem qualquer condição clínica prévia potencialmente fatal ou assintomático nas últimas 24 horas antes do óbito, em caso de morte não testemunhada. Apesar de ser um evento relativamente frequente, há poucos dados confiáveis na literatura sobre países em desenvolvimento. Objetivo: Descrever as características da MSC em Ribeirão Preto (SP 600.000 habitantes) baseando-se nos relatórios de autopsias do Serviço de Verificação de Óbitos do Interior. Métodos: Foram revisados retrospectivamente 4.501 relatórios de autopsias entre 2006 e 2010, para identificar casos de MSC. Foram coletados dados como causa específica do óbito, características demográficas e comorbidades das vítimas, data, local e hora do evento, e se foram realizadas manobras de ressuscitação cardiopulmonar (RCP). Resultados: Foram identificados 899 casos de MSC (20%; razão 30/100.000 habitantes por ano). A principal causa de MSC foi doença arterial coronariana (DAC - 64%), acometendo homens (67%) entre a sexta e a sétima década de vida. A maior parte dos eventos ocorreu durante a manhã, no domicílio (53,3%), e a RCP foi realizada em quase metade das vítimas (49,7%). A comorbidade mais prevalente foi hipertensão arterial sistêmica (57,3%). Doença de Chagas foi detectada em 49 casos (5,5%). Conclusão: A maioria dos casos de MSC ocorreu por DAC em homens entre a sexta e a sétima década de vida. Doença de Chagas, um importante problema de saúde pública na América Latina, foi detectada em 5,5% dos casos. .

In vitro assessment of commercial sunscreens available in Latin America / Valoración in vitro de protectores solares comerciales existentes en Latinoamérica

In Latin America, people have largely abandoned the practice of wearing hats and traditional clothing that provided skin protection. Sunscreen application has therefore become essential to protect against the increased sun exposure. The physician-prescribed medical-grade sunscreens provide sufficient sun protection but the requirement for regular use puts a financial burden on the patient that is often not sustainable. An appropriate sunscreen should provide a high and broad ultraviolet (UV) protection against UVB and UVA. Several over-the-counter (OTC) sunscreens have been developed for sale at affordable prices and are available for purchase in convenient locations, such as local grocery stores. The aim of this study was to assess the in vitro UV protection of 34 popular OTC sunscreens found in the Latin American market. UV absorbance/transmittance was quantified by diffusion transmission spectroscopy using coarse silica plaques. Photostability was tested by irradiating them with simulated solar light and calculating the sun protection factor (SPF), critical length of absorption (C λ ), UVA/UVB ratio, and the spectral uniformity index (SUI). The results indicated that the in vitro SPFs were significantly lower than the value declared on the labels, particularly for those claiming high SPF values; however, the majority of these sunscreens offered high levels of UV protection. Considering the advantages of low cost and ample accessibility, we concluded that this sample of OTC sunscreens can be beneficial to the general public by providing some level of skin protection from solar radiation, and may be promoted to improve compliance with recommended photoprotection behavior.

En Latinoamérica, la población ha abandonado la costumbre de usar sombrero y ropa tradicional para protegerse del sol. En consecuencia, es básico el uso de protectores solares si se realizan actividades bajo sol. Los protectores solares que se usan en la práctica médica son adecuados, pero su uso frecuente condiciona una carga económica que muchos pacientes no pueden solventar debido a sus costos considerables. Un protector apropiado contiene una amplia y elevada protección ultravioleta (UV) A y B. En las tiendas de conveniencia, existen numerosos protectores solares a precios más accesibles. El objetivo del estudio fue determinar la protección UV in vitro de 34 protectores solares con amplia presencia comercial (de venta sin prescripción médica) en el mercado latinoamericano. La absorbancia/transmitancia de la radiación UV se cuantificó mediante espectroscopía de transmisión difusa. Placas de sílice esmerilado fueron recubiertas con el producto y expuestas a radiación solar simulada para conocer su fotoestabilidad. Se calcularon índices como el factor de protección solar (SPF), longitud crítica de absorción (C λ), relación UVA/UVB y el índice de uniformidad espectral (SUI). Se encontró que el SPF in vitro fue inferior al establecido en las etiquetas, especialmente en aquellos con valores altos. No obstante, la mayoría de los protectores incluidos ofrecen niveles de protección UV elevados. Considerando su amplia accesibilidad y menor costo, concluimos que esta muestra comercial de protectores solares podría utilizarse en el entorno clínico para favorecer su apego junto a las otras medidas de fotoprotección sugeridas.

Granulocyte-colony stimulating factor decreases the extent of ovarian damage caused by cisplatin in an experimental rat model / 부인종양

OBJECTIVE: To investigate whether granulocyte-colony stimulating factor (G-CSF) can decrease the extent of ovarian follicle loss caused by cisplatin treatment. METHODS: Twenty-one adult female Sprague-Dawley rats were used. Fourteen rats were administered 2 mg/kg/day cisplatin by intraperitoneal injection twice per week for five weeks (total of 20 mg/kg). Half of the rats (n=7) were treated with 1 mL/kg/day physiological saline, and the other half (n=7) were treated with 100 microg/kg/day G-CSF. The remaining rats (n=7, control group) received no therapy. The animals were then euthanized, and both ovaries were obtained from all animals, fixed in 10% formalin, and stored at 4degrees C for paraffin sectioning. Blood samples were collected by cardiac puncture and stored at -30degrees C for hormone assays. RESULTS: All follicle counts (primordial, primary, secondary, and tertiary) and serum anti-Mullerian hormone levels were significantly increased in the cisplatin+G-CSF group compared to the cisplatin+physiological saline group. CONCLUSION: G-CSF was beneficial in decreasing the severity of follicle loss in an experimental rat model of cisplatin chemotherapy.

Establishment of an Allo-Transplantable Hamster Cholangiocarcinoma Cell Line and Its Application for In Vivo Screening of Anti-Cancer Drugs

Opisthorchis viverrini (O. viverrini) is a well-known causative agent of cholangiocarcinoma (CCA) in humans. CCA is very resistant to chemotherapy and is frequently fatal. To understand the pathogenesis of CCA in humans, a rodent model was developed. However, the development of CCA in rodents is time-consuming and the xenograft-transplantation model of human CCA in immunodeficient mice is costly. Therefore, the establishment of an in vivo screening model for O. viverrini-associated CCA treatment was of interest. We developed a hamster CCA cell line, Ham-1, derived from the CCA tissue of O. viverrini-infected and N-nitrosodimethylamine-treated Syrian golden hamsters. Ham-1 has been maintained in Dulbecco's Modified Essential Medium supplemented with 10% fetal bovine serum for more than 30 subcultures. These cells are mostly diploid (2n=44) with some being polyploid. Tumorigenic properties of Ham-1 were demonstrated by allograft transplantation in hamsters. The transplanted tissues were highly proliferative and exhibited a glandular-like structure retaining a bile duct marker, cytokeratin 19. The usefulness of this for in vivo model was demonstrated by berberine treatment, a traditional medicine that is active against various cancers. Growth inhibitory effects of berberine, mainly by an induction of G1 cell cycle arrest, were observed in vitro and in vivo. In summary, we developed the allo-transplantable hamster CCA cell line, which can be used for chemotherapeutic drug testing in vitro and in vivo.

Validación de un método in vivo para evaluar la actividad diurética / Validation of an in vivo method to assess the diuretic activity

Con el objetivo de proporcionar un modelo farmacológico in vivo para determinar la actividad diurética de plantas medicinales, se prepararon extractos acuosos a partir de la droga seca de 8 plantas con actividad diurética atribuida por la medicina tradicional cubana, pero que carecían de validación experimental. Se distribuyeron al azar 88 ratas machos Sprague-Dawley a razón de 8 animales por grupo: controles positivos (furosemida 20 mg/kg e hidroclorotiazida 10 mg/kg); control negativo (NaCl 0,9 por ciento) y 8 grupos tratados con extractos acuosos de plantas que se administraron por vía oral a dosis de 400 mg/kg, sobre la base de la determinación de los sólidos totales. La dosis fue completada con solución salina fisiológica para lograr una sobrecarga hidrosalina con un volumen total de administración constante de 40 mL/kg de peso vivo. Las ratas se colocaron en jaulas metabólicas y se midieron los volúmenes de orina excretados a las ¢, 1, 2, 3, 4, 5 y 6 h posadministración y las concentraciones de electrolitos (Na+ y K+) en la orina total colectada a las 24 h. Se observó que todos los grupos tratados incrementaron el volumen de orina en relación con el grupo control negativo. La excreción urinaria, acción y actividad diurética fueron mayores en los grupos experimentales: Persea americana Miller (similar a la furosemida) y Cassia alata L, Zanthoxylum fagara L. (similar a las tiazidas)

To supply a in vivo pharmacological model to determine the diuretic activity of medicinal plants, aqueous extracts were prepared from dry drug of 8 medicinal plants with diuretic activity attributed by the Cuban traditional medicine, but there was an experimental validation. Eighty Sprague-Dawley eight male rats were random distributed at a rate of eight rats by group: positive controls ( 20 mg/kg furosemide and 10 mg/kg hydrochlorothiazide); negative control (NaCI 0.9 percent) and 8 groups treated with aqueous extracts from plants supplied by oral route at doses of 400 mg/kg, on the base of determination of total solids. Dose was completed with physiological saline solution to achieve a hydrosaline overload with a total volume of constant administration of 40 mL/kg of weight lives. Rats were placed in metabolic cages measuring the volumes of urine extracted at ¢, 1, 2, 3, 4, 5 and 6 h post-administration and the electrolytes concentrations (Na+ and K+) in total urine collected at 24 h. It was noted that all the treated groups increased the urine volume in relation to negative control group. Urinary excretion, diuretic action and activity were higher in experimental groups: Persea Americana Miller (similar to Furosemide) and Cassia alata L, Zanthoxylum fagara L. (similar to thiazides)

Visceral leishmaniasis: Experimental models for drug discovery.

Visceral leishmaniasis (VL) or kala-azar is a chronic protozoan infection in humans associated with significant global morbidity and mortality. The causative agent is a haemoflagellate protozoan Leishmania donovani, an obligate intracellular parasite that resides and multiplies within macrophages of the reticulo-endothelial system. Most of the existing anti-leishmanial drugs have serious side effects that limit their clinical application. As an alternate strategy, vaccination is also under experimental and clinical trials. The in vitro evaluation designed to facilitate rapid testing of a large number of drugs has been focussed on the promastigotes milt little attention on the clinically relevant parasite stage, amastigotes. Screening designed to closely reflect the situation in vivo is currently time consuming, laborious, and expensive, since it requires intracellular amastigotes and animal model. The ability to select transgenic Leishmania expressing reporter proteins, such as the green fluorescent proteins (GFP) or the luciferase opened up new possibilities for the development of drug screening models. Many experimental animal models like rodents, dogs and monkeys have been developed, each with specific features, but none accurately reproduces what happens in humans. Available in vitro and in vivo methodologies for antileishmanial drug screening and their respective advantages and disadvantages are reviewed.

Cytotoxic effects of commercial wheatgrass and fiber towards human acute promyelocytic leukemia cells [Hl60]

Cytotoxicity, the possible selective activity upon HL60 as well as the anti-proliferation effect of local health supplement wheatgrass and mixture of fibers were investigated in vitro using various cancerous cell line and normal blood cell culture. The IC[50] of wheatgrass-treated HL60 [17.5 +/- 1.1, 12.5 +/- 0.3, and 16 +/- 0.5 microgram/ml for 24, 48 and 72 h, respectively] and fibers-treated HL60 [86.0 +/- 5.5, 35.0 +/- 2.5, and 52.5 +/- 4.5 microgram/ml for 24, 48 and 72 h, respectively] showed that both extracts possessed optimum effect after 48 hours of treatment. No significant cytotoxic effect was observed on other type of cells. For trypan blue dye exclusion method, wheatgrass reduced the number of viable cells by 13.5% [ +/- 1.5], 47.1% [ +/- 3.6], and 64.9% [ +/- 2.7] after 24, 48 and 72 h exposure, respectively. Mixture of fibers reduced the number of viable cells by 36.4% [ +/- 2.3], 57.1% [ +/- 3.1], and 89.0% [ +/- 3.4] after 24, 48 and 72 h exposure, respectively, indicated that necrosis is also an alternative to the apoptotic mechanism of cell death. Annexin-V/propidium iodide staining revealed that both extracts induced apoptosis where early apoptosis had been detected concurrently with the reduction of percentage of cell viability. Cell cycle analysis revealed that in HL60, the percentage of apoptosis increased with time [wheatgrass: 16.0% +/- 2.4, 45.3% +/- 3.4 and 39.6% +/- 4.1; mixture of fibers: 14.6% +/- 1.8, 45.4% +/- 2.3 and 45.9% +/- 1.2] after exposure for 24, 48 and 72 h, respectively at the concentration of 100 microgram/ml and showed optimum effect at 48 hours. Thus, these health products can be a potential alternative supplement for leukaemia patients

Pharmacological profile of salvadora persica

This work was conducted to investigate the various pharmacological activities of Salvadora. persica family Salvadoracea and that includes anti inflammatory, analgesic, CNS, bleeding and clotting time activity by oral administration at the dose of 300 and 500mg/kg of body weight in animal models. Acute oral toxicity results showed that crude extract of S. persica is safe up to the dose of 5g/kg body weight of animals. Carraganeen induced hind paw edema method for anti inflammatory activity, tail immersion test method for analgesic activity, Rota rod and grip strength test for CNS activity were carried out in animal models. The analgesic activity was compared with aspirin, 300mg/kg body weight, anti inflammatory activity was compared with indomethacine, lOmg/kg body weight, Transamin 250mg/kg and Vitamin K lOmg were used for bleeding and clotting time activity respectively while diazepam 5mg/kg were used as standard for behavior and CNS activities. In all activities S. persica showed prolonged and dose dependent effects. Phytochemical analysis was also carried out which showed the presence of certain phytoconstituents which possesses these properties. Therefore the results justified the traditional use of the plant

Phytochemical analysis, antioxidant and antibacterial effects of sea buckthorn berries

Sea buckthorn berries are therapeutically used as folk medicine for a variety of diseases, however, the scientific evidence is hardly available to support their role. This study explored their chemical constituents and their role as antioxidant and antibacterial agents. Three common solvents such as petroleum ether [40° - 60°C], chloroform and methanol were successively used for the extraction of active principles from sea buckthorn berries. Five major fractions [F1-F5] were isolated from the active methanol extract by column and thin layer chromatography. An attempt was made to identify the chemical nature of pooled fractions by available spectral means. Antioxidant potential of methanol extract and its fractions was measured by DPPH, formation of phosphomolybdenum complex and TEA methods. The hole-plate diffussion method was used to find out the antibacterial activity. A very brief structure-activity relationship of the potent antioxidant and antimicrobial compounds is discussed. Methanolic extract and its fractions contain numerous phenolic compounds such as flavonoids, which may be responsible for antioxidant and antibacterial effects

Flavonoids of enhydra fluctuans exhibits analgesic and anti-inflammatory activity in different animal models

Enhydra fluctuans [Compositae], an edible semi aquatic herbaceous vegetable plant, widely used in traditional system of Indian medicine. Total flavonoids of E. fluctuans [TFEF] were screened for analgesic and anti-inflammatory activity. Analgesic activity was studied in acetic acid induced writhing response and by hot plate method in Swiss albino mice. Anti-inflammatory activity was estimated by carrageenan and histamine induced acute inflammation and Freund's complete adjuvant [FCA] induced chronic inflammation in rats. Two flavonoids, baicalein 7-O-glucoside and baicalein 7-O-diglucoside, were isolated from the ethyl acetate fraction. Oral administration of TFEF at the doses of 200 and 400 mg/kg provide 27.05 and 55.49% protection respectively in acetic acid induced writhing method. It also increased the pain threshold in mice evidenced by hot plate method. TFEF showed more potent anti-inflammatory activity. The results of this study may be attributed to high free radical scavenging and antioxidant potential of the flavonoids present in ethyl acetate fraction of Enhydra fluctuans

Hepatoprotective studies on haloxylon salicornicum: a plant from cholistan desert

The objective was to study the in-vivo hepatoprotective effect of aerial parts vi Haloxylon salicornicum [Moq.] Bunge [Family: Chenopodiaceae] in order to validate its traditional use in hepatobiliary disorders, by native people of Cholistan desert, Pakistan. Aerial parts [ethanolic extract] ofHaloxylon salicornicum [HS], [500 and 750 mg/kg/day, p.o. for 7 days] were evaluated on CC1[4] intoxicated rabbits [0.75 ml/kg., s/c.] by serum biochemical parameters and liver histopathological observations. Silymarin [100 mg/kg/day, p.o. for 7 days] was used as a standard hepatoprotective drug. CC1[4] intoxicated group had elevated levels of SCOT, SGPT and ALP significantly but TB level was normal as compared to control group. HS extract [both doses of 500 and 750 mg/kg] showed hepatoprotective effect by significant restoration of SCOT, SGPT, ALP and TB levels as compared to CC1[4] control. 500 mg/kg doses of HS extract produced more significant results as compared to 750 mg/kg doses and Silymarin. Histopathological examination of liver tissues further substantiated these findings. Therefore, outcome of the present study validate the traditional claims on hepatoprotective effects ofHaloxylon salicornicum [aerial parts]

Formulation and evaluation of ficus glomerata mucilage sustained release matrix tablets of gliclazide

The main aim of present investigation was to develop sustained release matrix tablets of Gliclazide using fruit mucilage from the plant Ficus glomerata. Varying ratios of drug and polymer viz. 1:0.25, 1:0.5, 1:0.75, 1:1.0 and 1:1.25 were selected for the study. The flow properties of powdered mucilage and physical properties of matrix tablets were performed. The swelling behavior and release rate characteristics were studied. The in vitro drug release data was analyzed by zero order, first order, Higuchi plot, Peppas plot and Hixon-Crowell Models. It was observed that as the proportion of mucilage increased the release of drug from the matrix tablets was retarded. Stability studies were conducted at 40 +/- 2°C and RH 75 +/- 5% for 3 months indicates that Gliclazide was stable in the matrix tablets. The Differential Scanning Calorimetric [DSC] and Fourier Transform Infrared [FTIR] study revealed that there was no negative chemical interaction between drug and the mucilage used. From the dissolution study, it was concluded that dried Ficus glomerata mucilage can be used as an excipient for making sustained release matrix tablets

Anti-inflammatory and analgesic activities of ethanolic extract of sphaeranthusindicus linn

The aim of the present study was to evaluate the intensity of the anti-inflammatory and analgesic activities of Sphaeranthus indicus on albino mice and rat of either sex. The flowers of S. indicus are an important herb used in folk eastern medicines. In this study, the ethanolic extract of S. indicus in doses of 300 and 500 mg/kg was used. Anti-inflammatory activity was evaluated by measuring the mean decrease in hind paw volume after the sub planter injection of carrageenan. The analgesic activity was tested against acetic acid induced writhing response using albino rats. Result of the study shows that at the end of one hour the inhibition of paw edema was 42.66 and 50.5% respectively and the percentage of protection from writhing was 62.79 and 68.21 respectively. S. indicus possesses several important pharmaceutical and pharmacological properties. The current study describes that flower of S. indicus has significant anti-inflammatory and analgesic properties. Conclusion of the study is that this herbal medicine can be used as an alternative therapy for the treatment of minor to moderate types of inflammation and as a painkiller

Evaluation of anti-Wnt/β-catenin signaling agents by pGL4-TOP transfected stable cells with a luciferase reporter system

Refractory and relapsed leukemia is a major problem during cancer therapy, which is due to the aberrant activation of Wnt/β-catenin signaling pathway. Activation of this pathway is promoted by wingless (Wnt) proteins and induces co-activator β-catenin binding to lymphoid enhancer factor (LEF)/T-cell factor protein (TCF). To provide a convenient system for the screening of anti-Wnt/β-catenin agents, we designed a bi-functional pGL4-TOP reporter plasmid that contained 3X β-catenin/LEF/TCF binding sites and a selectable marker. After transfection and hygromycin B selection, HEK 293-TOP and Jurkat-TOP stable clones were established. The luciferase activity in the stable clone was enhanced by the recombinant Wnt-3A (rWnt-3A; 100-400 ng/mL) and GSK3β inhibitor (2’Z,3’E)-6-bromoindirubin-3’-oxime (BIO; 5 µM) but was inhibited by aspirin (5 mM). Using this reporter model, we found that norcantharidin (NCTD; 100 µM) reduced 80 percent of rWnt-3A-induced luciferase activity. Furthermore, 50 µM NCTD inhibited 38 percent of BIO-induced luciferase activity in Jurkat-TOP stable cells. Employing ³H-thymidine uptake assay and Western blot analysis, we confirmed that NCTD (50 µM) significantly inhibited proliferation of Jurkat cells by 64 percent, which are the dominant β-catenin signaling cells and decreased β-catenin protein in a concentration-dependent manner. Thus, we established a stable HEK 293-TOP clone and successfully used it to identify the Wnt/β-catenin signaling inhibitor NCTD.

Adulticidal activity of essential oil of Lantana camara leaves against mosquitoes.

Background & objectives: Development of insect resistance to synthetic pesticides, high operational cost and environmental pollution have created the need for developing alternative approaches to control vector-borne diseases. In the present study we have investigated the insecticidal activity of essential oil isolated from the leaves of Lantana camara against mosquito vectors. Methods: Essential oil was isolated from the leaves of L. camara using hydro-distillation method. Bioassay test was carried out by WHO method for determination of adulticidal activity against mosquitoes. Different compounds were identified by gas chromatography-mass spectrometry analysis. Results: LD50 values of the oil were 0.06, 0.05, 0.05, 0.05 and 0.06 mg/cm2 while LD90 values were 0.10, 0.10, 0.09, 0.09 and 0.10 mg/cm2 against Ae. aegypti, Cx. quinquefasciatus, An. culicifacies, An. fluvialitis and An. stephensi respectively. KDT50 of the oil were 20, 18, 15, 12, and 14 min and KDT90 values were 35, 28 25, 18, 23 min against Ae. aegypti, Cx. quinquefasciatus, An. culicifacies, An. fluviatilis and An. stephensi, respectively on 0.208 mg/cm2 impregnated paper. Studies on persistence of essential oil of L. camara on impregnated paper revealed that it has more adulticidal activity for longer period at low storage temperature. Gas chromatographic-mass spectrometric analysis of essential oil showed 45 peaks. Caryophyllene (16.37%), eucalyptol (10.75%), α-humelene (8.22%) and germacrene (7.41%) were present in major amounts and contributed 42.75 per cent of the total constituents. Interpretation &conclusion: Essential oil from the leaves of L. camara possesses adulticidal activity against different mosquito species that could be utilized for development of oil-based insecticide as supplementary to synthetic insecticides.

In vitro and in vivo experimental models for drug screening and development for Chagas disease

Chagas disease, a neglected illness, affects nearly 12-14 million people in endemic areas of Latin America. Although the occurrence of acute cases sharply has declined due to Southern Cone Initiative efforts to control vector transmission, there still remain serious challenges, including the maintenance of sustainable public policies for Chagas disease control and the urgent need for better drugs to treat chagasic patients. Since the introduction of benznidazole and nifurtimox approximately 40 years ago, many natural and synthetic compounds have been assayed against Trypanosoma cruzi, yet only a few compounds have advanced to clinical trials. This reflects, at least in part, the lack of consensus regarding appropriate in vitro and in vivo screening protocols as well as the lack of biomarkers for treating parasitaemia. The development of more effective drugs requires (i) the identification and validation of parasite targets, (ii) compounds to be screened against the targets or the whole parasite and (iii) a panel of minimum standardised procedures to advance leading compounds to clinical trials. This third aim was the topic of the workshop entitled Experimental Models in Drug Screening and Development for Chagas Disease, held in Rio de Janeiro, Brazil, on the 25th and 26th of November 2008 by the Fiocruz Program for Research and Technological Development on Chagas Disease and Drugs for Neglected Diseases Initiative. During the meeting, the minimum steps, requirements and decision gates for the determination of the efficacy of novel drugs for T. cruzi control were evaluated by interdisciplinary experts and an in vitro and in vivo flowchart was designed to serve as a general and standardised protocol for screening potential drugs for the treatment of Chagas disease.