First levantine fossil murines shed new light on the earliest intercontinental dispersal of mice.

Recent extensive field prospecting conducted in the Upper Miocene of Lebanon resulted in the discovery of several new fossiliferous localities. One of these, situated in the Zahleh area (Bekaa Valley, central Lebanon) has yielded a particularly diverse vertebrate fauna. Micromammals constitute an important part of this assemblage because not only do they represent the first Neogene rodents and insectivores from Lebanon, but they are also the only ones from the early Late Miocene of the Arabian Peninsula and circumambient areas. Analyses of the murines from Zahleh reveal that they belong to a small-sized early Progonomys, which cannot be assigned to any of the species of the genus hitherto described. They are, thereby, shown to represent a new species: Progonomys manolo. Morphometric analyses of the outline of the first upper molars of this species suggest a generalist and omnivorous diet. This record sheds new light onto a major phenomenon in the evolutionary history of rodents, which is the earliest dispersal of mice. It suggests that the arrival of murines in Africa got under way through the Levant rather than via southern Europe and was monitored by the ecological requirements of Progonomys.

The in vivo antinociceptive and µ-opioid receptor activating effects of the combination of N-phenyl-2',4'-dimethyl-4,5'-bi-1,3-thiazol-2-amines and naloxone.

Morphine is widely used for the treatment of severe pain. This analgesic effect is mediated principally by the activation of µ-opioid receptors (MOR). However, prolonged activation of MOR also results in tolerance, dependence, addiction, constipation, nausea, sedation, and respiratory depression. To address this problem, we sought alternative ways to activate MOR - either by use of novel ligands, or via a novel activation mechanism. To this end, a series of compounds were screened using a sensitive CHO-K1/MOR/Gα15 cell-based FLIPR® calcium high-throughput screening (HTS) assay, and the bithiazole compound 5a was identified as being able activate MOR in combination with naloxone. Structural modifications of 5a resulted in the discovery of lead compound 5j, which could effectively activate MOR in combination with the MOR antagonist naloxone or naltrexone. In vivo, naloxone in combination with 100 mg/kg of compound 5j elicited antinociception in a mouse tail-flick model with an ED50 of 17.5 ±â€¯4 mg/kg. These results strongly suggest that the mechanism by which the 5j/naloxone combination activates MOR is worthy of further study, as its discovery has the potential to yield an entirely novel class of analgesics.

Evaluation of Qualea grandiflora Mart effect on HIF-1alpha and MMP-14 modulation in cell lines of pre-osteoblasts and fibroblasts / Avaliação do efeito da Qualea grandiflora Mart na modulação de HIF-1alpha e MMP-14 em linhagens celulares de fibroblastos e pré-osteoblastos

A espécie vegetal Qualea grandiflora (QG), popularmente conhecida como "pauferro", "pau-terra-da-folha-grande", "pau-terra" ou "pau-de-tucano", muito comum no Cerrado brasileiro, é bem conhecida devido às suas variadas propriedades terapêuticas. Suas indicações incluem ações preventivas no aparecimento de lesões de mucosa gástrica, efeitos analgésicos, antibacterianos, anti-inflamatórios e antifúngicos. Assim, os componentes da QG poderiam ter alguma ação sobre moléculas amplamente envolvidas em processos angiogênicos e de desenvolvimento/reparo, como a Metaloproteinase de matriz 14 (MMP-14) e o Fator Induzido por hipóxia 1α (HIF-1alfa). Dessa maneira, o objetivo deste estudo foi investigar os efeitos do extrato hidroalcoólico das folhas de QG na viabilidade celular e expressão de MMP-14 e HIF-1alpha em culturas de fibroblastos da linhagem NIH/3T3 e pré-osteoblastos da linhagem MC3T3-E1. Para o teste de viabilidade celular e expressão das moléculas, concentrações de 0.1; 1.0 e 10 µg/mL do extrato hidroalcoólico das folhas de QG foram administrados por períodos de 24, 48, 72 e 96h. Após cada período, a viabilidade celular foi avaliada pelo método de redução de MTT e a análise da expressão das moléculas foi feita por meio da técnica de imunofluorescência. Os resultados mostram que o extrato de QG não promove redução da viabilidade celular de fibroblastos e pré-osteoblastos em concentrações até 10 µg/mL, nos períodos iniciais (24 e 48h). Porém, uma redução significativa da viabilidade pode ser verificada nos períodos de 72h e 96h para os fibroblastos e 96h para os pré-osteoblastos, expostos a mais alta concentração do extrato (10 µg/mL). O ensaio de imunofluorescência indica que o extrato, nas concentrações de 0.1; 1.0 e 10 µg/mL foi capaz de aumentar a expressão de MMP-14 e HIF-1alpha, em ambos os tipos celulares. Em conclusão, nossos resultados indicam que o extrato de QG exerce um efeito capaz de aumentar a expressão das duas moléculas em estudo (MMP-14 e HIF-1alpha), tanto para os fibroblastos da linhagem NIH/3T3 como para os pré- osteoblastos da linhagem MC3T3-E1. Assim, os compostos de QG podem apresentar potencial para serem utilizados como agentes terapêuticos moduladores da angiogênese, por meio do aumento da expressão de MMP-14 e HIF-1alpha.(AU)

The vegetable specie Qualea grandiflora (QG), popularly known as "pau-ferro", "pauterra-da-folha-grande", "pau-terra" or "pau-de-tucano", very common in the Brazilian Cerrado, is well known due to its varied therapeutic properties. Its indications include preventive actions in the appearance of lesions of gastric mucosa, analgesic, antibacterial, anti-inflammatory and antifungal effects. Thus, QG components could have some action on molecules widely involved in angiogenic and developmental / repair processes, such as Matrix metalloproteinase 14 (MMP-14) and HypoxiaInducible Factor-1α (HIF-1alpha). Thus, the objective of our study was to investigate the effects of QG hydroalcoholic extract on cell viability and expression of MMP-14 and HIF-1alpha in NIH/3T3 fibroblasts and MC3T3-E1 pre-osteoblasts cell lines. For the cell viability assay and expression of the molecules, concentrations of 0.1; 1.0 and 10 µg / mL of the hydroalcoholic extract of leaves of QG, were administered for periods of 24, 48, 72 and 96h. After each period, the cell viability was evaluated by MTT assay and the expression of the molecules was analyzed using the immunofluorescence technique. The results show that the QG extract does not promote reduction of the cellular viability of fibroblasts and pre-osteoblasts in concentrations up to 10 µg/mL in the initial periods (24 and 48h). However, a significant reduction in viability can be observed in 72h and 96h for fibroblasts and 96h for pre-osteoblasts exposed to the highest extract concentration (10 µg/mL). The immunofluorescence assay indicates that the extract, at concentrations of 0.1; 1.0 and 10 µg/mL was able to increase the expression of MMP-14 and HIF-1alpha in both cell types. In conclusion, our results indicate that the QG extract exerts an effect capable of increasing the expression of the two molecules under study (MMP-14 and HIF-1alpha) both for the NIH/3T3 fibroblasts as well as for the MC3T3-E1 pre-osteoblasts cells. Thus, the QG compounds could have potential to be used as angiogenesis modulating therapeutic agents, by increasing the expression of MMP-14 and HIF-1alpha.(AU)

Fraccionamiento guiado por bioensayo e identificación de blancos moleculares de potencial acción antipsicótica de cuatro plantas medicinales peruanas / Fractionation guided by bioassay and identification of molecular targets of potential antipsychotic action of four Peruvian medicinal plants

El principal síntoma de la esquizofrenia es la psicosis, caracterizada principalmente por la aparición de alucinaciones y delusiones. En esta investigación, se llevó a cabo el fraccionamiento biodirigido de extractos etanólicos (EE) de 4 plantas (6078, 6518, 7158 y 6521) usadas en la medicina tradicional Peruana para el tratamiento de psicosis. Los extractos y fracciones fueron probados en el modelo animal murino de hiperactividad inducida por Dizocilpina o MK-801(antagonista del receptor de glutamato tipo N-metil D-aspartato). Se utilizó la prueba de campo abierto (OFT) para medir la hiperactividad y la prueba de inhibición del sobresalto por prepulso (PPI) para medir la respuesta de sobresalto. En la prueba de campo abierto, la fracción apolar A del EE 7158 (p<0.05), la fracción D del EE 6078 (p <0.001) y la fracción A (p<0.05) y D (p<0.001) del EE 6518 (p<0.001) suprimieron los efectos del MK-801. En la prueba de PPI fueron activas las fracciones A, C y D (7158); A, B y C (6078); y todas las fracciones del extracto 6518. Adicionalmente, los ensayos de unión a radioligando y ensayos funcionales indican que los cuatro EEs tienen un efecto sobre receptores involucrados en esquizofrenia.

Control de calidad y evaluación del efecto inflamatorio de los extractos de Xanthium spinosum L. y Urtica urens L. en modelo murino / Quality control and evaluation of the inflammatory effect of extracts of Xanthium spinosum L. and Urtica urens L. in murine model

La tesis se enmarca dentro de la problemática de la elevada incidencia de los efectos adversos provocados por el uso crónico de antiinflamatorios y la posibilidad de contar con una terapia alternativa o coadyuvante para el tratamiento de cuadros inflamatorios basada en la utilización de plantas medicinales empleadas tradicionalmente en Bolivia. En este sentido, la investigación se realizó con las especies vegetales Xanthium spinosum L. y Urtica urens L. procedentes de la Provincia Ingavi del Departamento de La Paz. El objetivo principal de esta investigación fue el de profundizar en el conocimiento del efecto antiinflamatorio de los extractos de Xanthium spinosum L. y Urtica urens L. a través de la evaluación de procesos inflamatorios inducidos, en modelo murino. Para alcanzar este objetivo, se desarrolló un plan de trabajo que se dividió en cuatro partes fundamentales: la autentificación de ambas especies vegetales estudiadas, el estudio de sus parámetros de calidad, el estudio fitoquímico cualitativo y el estudio farmacológico in vivo, de los diferentes extractos obtenidos. La autentificación fue realizada sobre muestras representativas de las plantas recolectadas, en el Herbario Nacional de Bolivia, identificándose posteriormente como X. spinosum L. y U. urens L. El grado de su calidad fue determinado empleando las directrices propuestas por la Real Farmacopea Española y por comparación con muestras auténticas, lo que reveló la presencia de tricomas en X. spinosum L. y pelos urticantes unicelulares y pluricelulares en U. urens L. y valores de humedad, cenizas totales, cenizas ácidas, cenizas solubles, elementos extraños, índice de hinchamiento, dentro de los límites aceptables de calidad. Por otro lado, el protocolo desarrollado para la obtención de extractos por el método de polaridad creciente permitió la obtención de extractos etéreo, diclorometánico, etanólico y acuoso, los que mediante un screening fitoquímico cualitativo reveló la presencia mayoritaria de esteroles y flavonoides en X. spinosum L. y taninos, alcaloides, compuestos reductores, esteroles, flavonoides y cumarinas en U. urens L. Adicionalmente, el perfil cromatográfico de los distintos extractos reveló la presencia de diferentes manchas en fases móviles asociadas principalmente con flavonoides. El estudio del efecto antiinflamatorio de los extractos se realizó en ratones albinos Swiss, siguiendo el protocolo de edema plantar inducido por carragenina y el estudio del efecto analgésico siguiendo el protocolo de inducción de contorciones abdominales por ácido acético. Los mayores valores de inhibición de la inflamación fue registrado para los extractos acuoso y etanólico de X. spinosum L. y de U. urens L., en tanto que la inhibición de la algesia fue más importante cuando se empleó extractos diclorometánico y etanólico de ambas especies vegetales. Finalmente, la combinación de extractos acuosos de ambas especies estudiadas mostró un efecto sinérgico antiinflamatorio y analgésico, en tanto que la combinación de los extractos etanólicos de estas especies vegetales mostró sólo un efecto antiinflamatorio, no así analgésico. (AU)

Protective effect of Hypericum calabricum Sprengel on oxidative damage and its inhibition of nitric oxide in lipopolysaccharide-stimulated RAW 264.7 macrophages.

The present study shows for the first time the phenolic composition and the in vitro properties (antioxidant and inhibition of nitric oxide production) of Hypericum calabricum Sprengel collected in Italy. The content of hypericins (hypericin and pseudohypericin), hyperforin, flavonoids (rutin, hyperoside, isoquercetrin, quercitrin, quercetin and biapigenin) and chlorogenic acid of H. calabricum, have been determined. The ethyl acetate fraction from the aerial parts of H. calabricum exhibited activity against the radical 1,1-diphenyl-2-picrylhydrazyl (DPPH) with IC50 value of 1.6 jig/ml. The test for inhibition of nitric oxide (NO) production was performed using the murine monocytic macrophage cell line RAW 264.7. The ethyl acetate fraction had significant activity with an IC50 value of 102 jig/ml and this might indicate that it would have an anti-inflammatory effect in vivo.

Protective effect of Hypericum calabricum Sprengel on oxidative damage and its inhibition of nitric oxide in lipopolysaccharide-stimulated RAW 264.7 macrophages

The present study shows for the first time the phenolic composition and the in vitro properties (antioxidant and inhibition of nitric oxide production) of Hypericum calabricum Sprengel collected in Italy. The content of hypericins (hypericin and pseudohypericin), hyperforin, flavonoids (rutin, hyperoside, isoquercetrin, quercitrin, quercetin and biapigenin) and chlorogenic acid of H. calabricum, have been determined. The ethyl acetate fraction from the aerial parts of H. calabricum exhibited activity against the radical 1,1-diphenyl-2-picrylhydrazyl (DPPH) with IC50 value of 1.6 jig/ml. The test for inhibition of nitric oxide (NO) production was performed using the murine monocytic macrophage cell line RAW 264.7. The ethyl acetate fraction had significant activity with an IC50 value of 102 jig/ml and this might indicate that it would have an anti-inflammatory effect in vivo.

Actividad tripanocida in vitro e in vivo de extractos etanólicos de algunas plantas medicinales bolivianas / In vitro and in vivo trypanocidal activity, of ethanolic extracts from some bolivian medicinal plants

A 100 años del descubrimiento de la enfermedad de Chagas, aun no existen drogas que satisfagan completamente. Los extractos de plantas medicinales son una posibilidad de obtener nuevos compuestos que sean activos contra Trypanosoma cruzi. Se evaluó la actividad tripanocida in vitro sobre las formas tripomastigotes de ocho extractos etanólicos provenientes de 4 plantas medicinales bolivianas. El extracto etanólico de la corteza de Anacardium occidentale fue el más activo in vitro (CI50= 200 µg/ml), seguido de las hojas de Bowdichia virgilioides (350 µg/ml). A. occidentale, B. virgilioides y Senna reticulata, no son eficaces en la fase aguda de la enfermedad de Chagas en el modelo murino. (AU)

Determinación de la actividad antiinflamatoria e interacción de extractos de la planta Kiswara (Buddleja coriácea Rémy) con dexametasona, mediante los ensayos de edema plantar y auricular en modelo murino / Determination of the anti-inflammatory activity and interaction of extracts of the Kiswara plant (Buddleja coriácea Rémy) with dexamethasone, by tests of plantar and auricular edema in murine model

El propósito del presente trabajo fue el evaluar la actividad antiinflamatoria de extractos de la especie Buddleja coriácea R. (Kiswara), además de su combinación con el fármaco antiinflamatorio dexametasona, mediante modelos preclínicos. Porque esta especie es empleada en la medicina tradicional para patologías inflamatorias. Inicialmente se determino la presencia de compuestos fenólicos en los extractos: Hidroalcohólico y Diclorometano/Hidroalcohólico de Buddleja coriácea R., mediante pruebas cualitativas, donde se identificaron terpenos, alcaloides, saponinas, taninos y flavonoides presentes en las hojas de la especie. Posteriormente se evaluó el efecto de inhibición de inflamación en el modelo de edema auricular a dosis de 3g/Kg de los extractos Hidroalcohólico y Diclorometano/Hidroalcohólico, además de su interacción con dexametasona donde se obtuvieron 21.39 por ciento y 31.02 por ciento de inhibición respectivamente, mientras que la dexametasona presento un 63.35 por ciento de inhibición pero cuando interacciona con los extractos presenta 45.64 por ciento y 68.08 por ciento de inhibición respectivamente, siendo estos estadísticamente significativos con respecto al control (p <0.05). Los resultados de inhibición de inflamación en el modelo de edema plantar al cabo de las 7 horas de los extractos Hidroalcohólico y Diclorometano/Hidroalcohólico de Buddleja coriácea R. fueron 12.03 por ciento y 14.19 por ciento, datos que resultaron ser significativos respecto al grupo control, en el caso de las interacciones de los extractos con dexametasona se obtuvieron 21.95 por ciento y 28.04 por ciento de inhibición respectivamente, ambos resultaron ser significativos, la dexametasona revelo un 55.9 por ciento de inhibición de inflamación siendo este significativo con respecto al grupo control (p<0.05). (AU)

Análise do perfil de expressão gênica em melanomas cutâneos malignos murinos após tratamento sistêmico com o fator de necrose tumoral (TNF) associado ou não a melfalano (MEL) / Gene expression profile of murine malignat melanoma after systemic treatment with with tumor necrosis factor (TNF) associated or not with melphalan (MEL)

ILP (Perfusão Isolada de Membro) com melphalan (MEL) é uma obordagem alternativa no tratamento de melanomas cutâneos avançados, não-ressecáveis, de extremidades, poupando o membro da amputação. ... O objetivo do nosso estudo foi determinar o perfil de expressão gênica do melanoma e sua modificação após tratamento com TNF, MEL ou TMF+MEL. .... Os perfis de expressão gênica das células e dos tumores foram determinados através da estratégia de “microarray”, empregando um “biochip” contendo 16.128 oligonucleotídeos. O tratamento das células de melanoma com MEL durante 48h esteve associado com uma maior taxa de morte celular. O TNF não exerceu nenhuma atividade citotóxica sobre as células de melanoma in vitro. A administração de melfalano promoveu uma menor velocidade de crescimento tumoral (p<0,001), efeito este que não foi modificado pela co-administração de TNF. Entretanto, tumores tratados com TNF apenas ou em combinação com MEL apresentaram mais necrose (p=0,017 e p=0,014, respectivamente) e menos mitoses (p=0,001). Um número menor de mitoses também foi observado em resposta a MEL. O tratamento com MEL ou MEL+TNF esteve associado com aumento da sobrevida livre de progressão. ... Um grupo de genes grande teve sua expressão modulada pelo tratamento in vitro. Entre estes, nós identificamos Aff1, um regulador do ciclo celular, positivamente regulado por MEL. O mesmo efeito foi observado in vivo para este gene em especial. Da comparação entre os perfis de expressão gênica dos tumores por ANOVA, nós identificamos 118 genes diferencialmente expressos com p<0,05. Entre os elementos diferencialmente expressos estão genes que codificam proteínas envolvidas em adesão celular (Pecam1, Pard3, Dlg5), apoptose (Bcl11l), vias de sinalização (Arhgef6, Flt-1), regulação de transcrição (Tcfe3, Ifi202b, Irak1bp1, Fabp4,Trp73, Trim24), ciclo celular (Cdc7, Aff1), metabolismo de drogas (Akr1b7), potencial metastático (Trpm1, Mib2) e várias outras funções biológicas...

ILP (Isolated Limb Perfusion) with melphalan (MEL) is an alternative approach in the treatment of non-resectable locally advanced cutaneous melanomas of extremity, sparing the limb from amputation. ILP is a locoregional procedure to deliver high doses of drugs to a limb with minimal systemic and mild local toxicity. The association with the Tumor Necrosis Factor (TNF) improves total complete response rates from 40-50% to 70-90%. The molecular mechanisms underlining this synergistic effect are poorly understood. The aim of our study was to determine the gene expression profile of melanoma and its alterations after the treatment with TNF, MEL and TNF+MEL. To reach this objective B16F1 and B16F10 cells were exposed to MEL (36,9µM), TNF (10ng/ml) or both during 3 hours for microarray assays or for 24h, 48h and 72h for citoxicity assays. At same time, 6-8 weeks-old male C57BL6 mice were injected subcutaneously, on both flanks, with 5,0x105 of B16F10 and B16F1 melanoma cells. Animals bearing tumors of 1,0±0,3cm2 (n=9-10 mice/group) received an intravenous injection of TNF (0,05µg/Kg), MEL (12mg/Kg), TNF+MEL or saline (control). One of the tumors was surgically removed 3h after for RNA extraction and histology and the other one was left for growth measurements. Gene expression profiles from cells and tumors were determined by microarray strategy using a biochip containing 16.128 oligonucleotides. Treatment of melanoma cells with MEL for 48h was associated with a significantly higher cell death rate. TNF exerted no cytotoxic effects on melanoma cells in vitro. The administration of melphalan led to slower tumor growth rate (p<0,001), effect that was not modified by the TNF co-administration. Nevertheless, tumors treated with TNF alone or in combination with MEL presented more necrosis (p=0,017 and p=0,014, respectively) and less mitosis (p=0,001). Less mitosis was also observed in response to MEL alone. Treatment with MEL or MEL+TNF was associated with a longer progression free survival. Three-hourtreatment with MEL or TNF did not affect vascular density. A whole set of genes had its expression modulated by treatment in vitro. Amongst them, we identified Aff1, a regulator of cell proliferation, as positively regulated by MEL. The same effect was observed in vivo for this specific gene. From the comparison between the gene expression profile by ANOVA, we identified 118 genes differentially expressed with p<0,05. Among the differentially expressed elements are genes coding for proteins involved in cell adhesion (Pecam1, Pard3, Dlg5), apoptosis (Bcl11l), signaling pathways (Arhgef6, Flt-1), regulation of transcription (Tcfe3, Ifi202b, Irak1bp1, Fabp4,Trp73, Trim24), cell cycle (Cdc7, Aff1), drug metabolism (Akr1b7), metastatic potential (Trpm1, Mib2) and many other diverse biological functions. Supported by: FAPESP (AU)

Antihyperlipidemic and antioxidant effects of extracts from Pleurotus citrinopileatus.

Pleurotus citrinopileatus is a popular edible mushroom which is physiologically active in both humans and animals. In the study we investigate the effects of this mushroom on hyperlipidemic hamster rats. Four dietary forms of the mushroom were created as follows. The powdered dry fruiting body, hot-water extract, and two kinds of elutes were obtained, from ethyl acetate extract and methanol extract, respectively, in different mixed proportion solvents over silica gel column chromatography (referred to as EAE and MOE, respectively). They were tested at different dosages as a supplement to a high-fat diet in hyperlipidemic rats. Serum triglycerides and total cholesterol levels were significantly lower in groups supplemented with the highest dosages of EAE and MOE (0.5 g/kg, body weight daily) as compared with the control groups that received no mushroom additive. High-density lipoprotein levels in these same two experimental groups were also significantly higher than those in the negative control group. The tested rats that were fed with EAE had the highest serum glutathione peroxidase and superoxide dismutase activity, and those with the MOE and EAE had the highest DPPH free radical scavenging activities and ferric-reducing abilities, tested in vitro. The major constituents of MOE and EAE were identified as ergosterol and nicotinic acid, respectively. P. citrinopileatus extracts may have a significant antihyperlipidemia effect. Furthermore, antioxidant activities and antihyperlipidemic effects of MOE and EAE seemed to display similar tendencies.

Protective activity of picroliv on hepatic amoebiasis associated with carbon tetrachloride toxicity.

BACKGROUND AND OBJECTIVE: Picroliv, isolated from the root and rhizome of Picrorhiza kurroa, is known to have significant hepatoprotective activity. Its effects against Entamoeba histolytica induced liver damage are not studied. This study aims to evaluate the hepatoprotective action of picroliv against the hepatotoxic changes induced by carbon tetrachloride (CCl(4)) and E. histolytica infection in three animal models. METHODS: Mastomys, gerbils and albino Druckray rats were used in this study. A total of 30 animals were used for each model and divided into five groups of six animals each. Group I consisted of normal animals. The rest received six doses of CCl(4) intraperitoneally. Group II served as hepatotoxic control. The remaining animals were infected intraperitoneally with E. histolytica trophozoites, of which group III was the hepatotoxic plus amoeba infected control. The remaining animals were divided into two groups, one received hepatoprotective agent picroliv and the other silymarin. All animals were sacrificed seven days post amoeba infection. RESULTS: Increase in the enzyme levels induced by CCl(4) was further elevated after E. histolytica infection. Pinpoint abscesses were found to develop only in gerbils after E. histolytica infection. Picroliv was found to possess hepatoprotective activity against amoebic liver abscess. INTERPRETATION AND CONCLUSION: Significant recovery obtained in serum enzyme levels in all animal models and against amoebic liver abscess in gerbils on treatment with picroliv indicated that picroliv possesses therapeutic activity against E. histolytica induced hepatic damage.

Toxicity of Nerium indicum Miller seed extract on bandicoot rat, Bandicota bengalensis Gray.

Crude extract (12.5 ml/kg) of N. indicum seed gave 100% mortality of B. bengalensis. Humanness assessment study revealed that this plant orgin chemical caused low pain and sufferings to the target pests.

Immunomodulator tuftsin augments antifilarial activity of diethylcarbamazine against experimental brugian filariasis.

In the present study, we evaluated the potential of an immunomodulator tuftsin in increasing the efficacy of liposomised diethylcarbamazine (DEC) against experimental filarial infection of Brugia malayi. The liposomised form of DEC, when used at sub-optimal dose of 25 mg/kg body weight, successfully eliminated filarial parasite from systemic circulation in animals inflicted with B. malayi infection. However, the formulation was effective upto 60 days post infection only, followed by recurrence of the infection. In contrast, the co-administration of liposomal formulation of DEC along with an immunomodulator tuftsin was found to be competent enough to suppress microfilarial stage of parasite till 90 days post treatment. Interestingly, tuftsin bearing DEC liposomes were found to be effective against adult parasite as well.

Effects of Acacia condensed tannins on urinary parameters, body mass, and diet choice of an Acacia specialist rodent, Thallomys nigricauda.

The aim of this study was to investigate the dietary and physiological effects of condensed tannin ingestion on foregut fermenters, using Thallomys nigricauda, a folivorous rodent, as a model. We initially investigated the variability in physiological parameters, such as daily body mass (DMb), daily feed intake, daily fecal energy loss (FE), daily energy intake (DEI), daily urine pH, and daily urinary ammonia and urea concentrations, in response to different diets with low condensed tannin levels. This experiment was conducted to identify which physiological variables showed the least variation in the absence of tannin. In a second experiment, we investigated the response of the same dietary and physiological parameters to the effects of high dietary condensed tannin ingestion in T. nigricauda. We hypothesized that DMb, daily feed intake, FE, and DEI of T. nigricauda would be adversely affected by high dietary tannin content. We predicted that detoxification activity by T. nigricauda would increase at higher tannin levels. Ingestion of tannins affected the nutritional status of T. nigricauda, as shown by a decrease in body mass at high tannin levels. We also found that fewer ammonium ions were excreted in the urine by T. nigricauda, as would be expected if this were a means of regulating metabolic acidosis. The urine produced was more alkaline. This result indicates that T. nigricauda is not metabolizing these allelochemicals. Urea production was initially reduced, indicating conservation of bicarbonate ions that will neutralize blood acidity if there is detoxification. A diet choice experiment showed that tree rats avoid high tannin diets, even to the extent that they lose body mass on an alternative diet. This last-mentioned result is noteworthy because previous studies of the effects of tannins on herbivorous mammals have shown that there is physiological control rather than behavioral avoidance of the negative effects of tannin ingestion.

Bioavailability and trophic transfer of sediment-bound Ni and U in a southeastern wetland system.

Elemental composition of soil, herbaceous and woody plant species, and the muscle and liver tissue of two common small mammal species were determined in a wetland ecosystem contaminated with Ni and U from nuclear target processing activities at the Savannah River Site, Aiken, SC. Species studied were black willow ( Salix nigra L.), rushes ( Juncus effusus L.), marsh rice rat ( Oryzomys palustris), and cotton rat ( Sigmodon hispidus). Two mature trees were sampled around the perimeter of the former de facto settling basin, and transect lines sampling rushes and trapping small mammals were laid across the wetland area, close to a wooden spillway that previously enclosed the pond. Ni and U concentrations were elevated to contaminant levels; with a total concentration of 1,065 (+/- 54) mg kg(-1) U and 526.7 (+/-18.3) mg kg(-1) Ni within the soil. Transfer of contaminants into woody and herbaceous plant tissues was higher for Ni than for U, which appeared to remain bound to the outside of root tissues, with very little (0.03 +/- 0.001 mg kg(-1)) U detectable within the leaf tissues. This indicated a lower bioavailability of U than the cocontaminant Ni. Trees sampled from the drier margins of the pond area contained more Ni within their leaf tissues than the rushes sampled from the wetter floodplain area, with leaf tissues concentrations of Ni of approximately 75.5 (+/- 3.6) mg kg(-1) Ni. Ni concentrations were also elevated in small mammal tissues. Transfer factors of contaminants indicated that U bioavailability is negligable in this wetland ecosystem.

Comparative genetics and evolution of annexin A13 as the founder gene of vertebrate annexins.

Annexin A13 (ANXA13) is believed to be the original founder gene of the 12-member vertebrate annexin A family, and it has acquired an intestine-specific expression associated with a highly differentiated intracellular transport function. Molecular characterization of this subfamily in a range of vertebrate species was undertaken to assess coding region conservation, gene organization, chromosomal linkage, and phylogenetic relationships relevant to its progenitor role in the structure-function evolution of the annexin gene superfamily. Protein diagnostic features peculiar to this subfamily include an alternate isoform containing a KGD motif, an elevated basic amino acid content with polyhistidine expansion in the 5'-translated region, and the conservation of 15% core tetrad residues specific to annexin A13 members. The 12 coding exons comprising the 58-kb human ANXA13 gene were deduced from BAC clone sequencing, whereas internal repetitive elements and neighboring genes in chromosome 8q24.12 were identified by contig analysis of the draft sequence from the human genome project. A unique exon splicing pattern in the annexin A13 gene was corroborated by coanalysis of mouse, rat, zebrafish, and pufferfish genomic DNA and determined to be the most distinct of all vertebrate annexins. The putative promoter region was identified by phylogenetic footprinting of potential binding sites for intestine-specific transcription factors. Mouse annexin A13 cDNA was used to map the gene to an orthologous linkage group in mouse chromosome 15 (between Sdc2 and Myc by backcross analysis), and the zebrafish cDNA permitted its localization to linkage group 24. Comparative analysis of annexin A13 from nine species traced this gene's speciation history and assessed coding region variation, whereas phylogenetic analysis showed it to be the deepest-branching vertebrate annexin, and computational analysis estimated the gene age and divergence rate. The unique, conserved aspects of annexin A13 primary structure, gene organization, and genetic maps identify it as the probable common ancestor of all vertebrate annexins, beginning with the sequential duplication to annexins A7 and A11 approximately 700 MYA, before the emergence of chordates.

Evolutionary history of the most speciose mammals: molecular phylogeny of muroid rodents.

Phylogenetic relationships between 32 species of rodents representing 14 subfamilies of Muridae and four subfamilies of Dipodidae were studied using sequences of the nuclear protein-coding genes Lecithin Cholesterol Acyl Transferase (LCAT) and von Willebrand Factor (vWF). An examination of some evolutionary properties of each data matrix indicates that the two genes are rather complementary, with lower rates of nonsynonymous substitutions for LCAT. Both markers exhibit a wide range of GC3 percentages (55%-89%), with several taxa above 70% GC3 for vWF, which indicates that those exonic regions might belong to the richest class of isochores. The primary sequence data apparently harbor few saturations, except for transitions on third codon positions for vWF, as indicated by comparisons of observed and expected pairwise values of substitutions. Phylogenetic trees based on 1,962 nucleotidic sites from the two genes indicate that the 14 Muridae subfamilies are organized into five major lineages. An early isolation leads to the clade uniting the fossorial Spalacinae and semifossorial Rhizomyinae with a strong robustness. The second lineage includes a series of African taxa representing nesomyines, dendromurines, cricetomyines, and the sole living member of mystromyines. The third one comprises only the mouselike hamster CALOMYSCUS: The fourth clade represents the cricetines, myospalacines, sigmodontines, and arvicolines, whereas the fifth one comprises four "traditional" subfamilies (Gerbillinae, Murinae, Otomyinae, and Acomyinae). Within these groups, we confirm the monophyly of almost all studied subfamilies, namely, Spalacinae, Rhizomyinae, Nesomyinae, Cricetomyinae, Arvicolinae, Sigmodontinae, Cricetinae, Gerbillinae, Acomyinae, and Murinae. Finally, we present evidence that the sister group of Acomyinae is Gerbillinae, and we confirm a nested position of Myospalacinae within Cricetinae and Otomyinae within Murinae. From a biogeographical point of view, the five main lineages spread and radiated from Asia with different degrees of success: the first three groups are now represented by a limited number of species and genera localized in some regions, whereas the last two groups radiated in a large variety of species and genera dispersed all over the world.

Structure, chromosomal localization, and expression of the gene for mouse ecto-mono(ADP-ribosyl)transferase ART5.

Mono(ADP-ribosyl)transferases regulate the function of target proteins by attaching ADP-ribose to specific amino acid residues in their target proteins. The purpose of this study was to determine the structure, chromosomal localization, and expression profile of the gene for mouse ecto-ADP-ribosyltransferase ART5. Southern blot analyses indicate that Art5 is a single copy gene which maps to mouse chromosome 7 at offset 49.6 cM in close proximity to the Art1, Art2a and Art2b genes. Northern blot and RT-PCR analyses demonstrate prominent expression of Art5 in testis, and lower levels in cardiac and skeletal muscle. Sequence analyses reveal that the Art5 gene encompasses six exons spanning 8 kb of genomic DNA. The 5' end of the Art5 gene overlaps with that of the Art1 gene. A single long exon encodes the predicted ART5 catalytic domain. Separate exons encode the N-terminal leader peptide and a hydrophilic C-terminal extension. Sequencing of RT-PCR products and ESTs identified six splice variants. The deduced amino acid sequence of ART5 shows 87% sequence identity to its orthologue from the human, and 37 and 32% identity to its murine paralogues ART1 and ART2. Unlike ART1 and ART2, ART5 lacks a glycosylphosphatidylinositol-anchor signal sequence and is predicted to be a secretory enzyme. This prediction was confirmed by transfecting an Art5 cDNA expression construct into Sf9 insect cells. The secreted epitope-tagged ART5 protein resembled rat ART2 in exhibiting potent NAD-glycohydrolase activity. This study provides important experimental tools to further elucidate the function of ART5.

Effects of Bay 44-4400, a new cyclodepsipeptide, on developing stages of filariae (Acanthocheilonema viteae, Brugia malayi, Litomosoides sigmodontis) in the rodent Mastomys coucha.

Bay 44-4400 was used as a spot on formulation and administered in single doses of 25 and 100 mg/kg to Acanthocheilonema viteae, Brugia malayi, and Litomosoides sigmodontis infected Mastomys coucha on various dates during prepatency, aiming to affect third stage larvae, fourth stage larvae or preadult worms. Microfilaraemia levels were controlled in comparison to untreated controls until necropsies were performed 100 days p.i. (A. viteae, L. sigmodontis) and 150 days p.i. (B. malayi) to determine the numbers of surviving worms and the condition of intrauterine developing stages. A significant proportion (86-100%) of larval and preadult stages of A. viteae were killed by Bay 44-4400 at a dose of 100 mg/kg. A dose of 25 mg/kg had only insignificant effects on the developing parasites, however, it strongly reduced microfilaraemia levels caused by surviving worms in the early phase of patency. Larval and preadult B. malayi and L. sigmodontis were not killed by Bay 44-4400 to a significant degree. Microfilaraemia developing by surviving parasites was generally and significantly reduced throughout the observation period when treatment was performed to affect the preadult parasites. In the other cases variable results were obtained. Intrauterine early embryonic stages were found to be pathologically altered in worms which had been treated at a preadult stage.