Added value of the measles-rubella supplementary immunization activity in reaching unvaccinated and under-vaccinated children, a cross-sectional study in five Indian districts, 2018-20.

INTRODUCTION: Supplementary immunization activities (SIAs) aim to interrupt measles transmission by reaching susceptible children, including children who have not received the recommended two routine doses of MCV before the SIA. However, both strategies may miss the same children if vaccine doses are highly correlated. How well SIAs reach children missed by routine immunization is a key metric in assessing the added value of SIAs. METHODS: Children aged 9 months to younger than 5 years were enrolled in cross-sectional household serosurveys conducted in five districts in India following the 2017-2019 measles-rubella (MR) SIA. History of measles containing vaccine (MCV) through routine services or SIA was obtained from documents and verbal recall. Receipt of a first or second MCV dose during the SIA was categorized as "added value" of the SIA in reaching un- and under-vaccinated children. RESULTS: A total of 1,675 children were enrolled in these post-SIA surveys. The percentage of children receiving a 1st or 2nd dose through the SIA ranged from 12.8% in Thiruvananthapuram District to 48.6% in Dibrugarh District. Although the number of zero-dose children prior to the SIA was small in most sites, the proportion reached by the SIA ranged from 45.8% in Thiruvananthapuram District to 94.9% in Dibrugarh District. Fewer than 7% of children remained measles zero-dose after the MR SIA (range: 1.1-6.4%) compared to up to 28% before the SIA (range: 7.3-28.1%). DISCUSSION: We demonstrated the MR SIA provided considerable added value in terms of measles vaccination coverage, although there was variability across districts due to differences in routine and SIA coverage, and which children were reached by the SIA. Metrics evaluating the added value of an SIA can help to inform the design of vaccination strategies to better reach zero-dose or undervaccinated children.

Effects of prostaglandin administration 10 days apart on reproductive parameters of cyclic dairy nulliparous goats / Efeitos da aplicação de prostaglandinas intervaladas de 10 dias sobre características reprodutivas de cabras leiteiras nulíparas cíclicas

This study reported the effects of prostaglandin (PGF2a) administration 10 days apart on reproductive parameters of cyclic artificial inseminated (AI) nulliparous Alpine (n=9) and Saanen (n=9) goats. Animals received two doses of 22.5mg PGF2a 10 days apart. After 1st and 2nd PGF2a administrations, estrus was monitored at 12 h intervals, with a buck teaser. Plasma progesterone concentration (ng/mL) was determined from blood sampled on day 0 (1st PGF2a) and the following 5, 10 (2nd PGF2a), 15, 20, 25 and 30 days. After the onset of the second estrus, females were transrectally (5 MHz probe) scanned at 4 hour intervals until at least 8h after ovulation. Pregnancy was checked through transrectal ultrasound on days 20, 25, 30, 35 and 90 after insemination. All parameters studied did not differ between breeds (P>0.05). Estrous response and interval to estrus, respectively, after 1st (78.9% and 50.6±17.2h) and 2nd PGF2a (88.9% and 50.0±14.8h) administration did not differ (P>0.05). Overall animals ovulating (100.0%), interval to ovulation after 2nd PGF2a (64.5±19.5h) and after estrous onset (18.0±9.1h), ovulation rate (1.3±0.5), diameter of ovulatory follicle (8.1±1.1mm) were recorded. Embryo loss occurred before day 30 of pregnancy. Estrus can be efficiently synchronized in nulliparous Alpine and Saanen goats with two doses of prostaglandin 10 days apart.

Relataram-se os efeitos da aplicação de prostaglandina sobre características reprodutivas de cabras leiteiras nulíparas cíclicas. Cabras Alpinas (n=9) e Saanen (n=9) receberam duas doses de 22,5mg PGF2a com 10 dias de intervalo. A progesterona plasmática (ng/mL) foi determinada a partir de amostras de sangue coletadas nos dias 0 (primeira dose), 5, 10 (segunda dose), 15, 20, 25 e 30. Após início do segundo estro, as fêmeas foram monitoradas por ultrassonografia transretal a cada quatro horas até oito horas após a ovulação. A gestação foi verificada por ultrassonografia transretal nos dias 20, 25, 30, 35 e 90 após a segunda dose. As características estudadas foram semelhantes entre as raças (P>0,05). Animais em estro e o intervalo parto-estro de, respectivamente, 78,9% e 50,6±17,2h e 88,9% e 50,0±14,8h após a primeira e segunda administrações de prostaglandina, não diferiram (P>0,05). Todas as cabras ovularam e registraram-se valores do intervalo parto-ovulação após a segunda aplicação de prostaglandina de 64,5±19,5h e após início do estro de 18,0±9,1h, a taxa de ovulação de 1,3±0,5 e diâmetro do folículo ovulatório de 8,1±1,1mm. Perda embrionária ocorreu antes de 30 dias de gestação. O estro pode ser eficientemente sincronizado em cabras leiteiras núliparas com duas doses de prostaglandina intervaladas de 10 dias.

Cianobacterial bloom and animal mass mortality in a reservoir from Central Argentina.

Piedras Moras reservoir (32° 10'27" S and 64° 16' 29" W; 832 ha), integrates a series of artificial lakes belonging to the Rio Tercero basin (Córdoba, Argentina). During March 2009 an algal bloom occurred, coinciding with several animal species mortality, mainly wild birds. The goal of this work was to establish the trophic status of the reservoir in relation to that mortality. Variables were evaluated in situ (temperature and water transparency) and samples were taken in order to identify algal species, Chl-a concentration (spectrophotometry) and toxins - total microcystines- (inmuno-enzymatic assay, ELISA). Histopathology studies were made on Fulica sp. A strong heterogenity in water transparency was observed, and "patches" of Potamogeton berteroanus distributed all along the lake, with Secchi disk minimal and medium values of 0.15 and 0.94 m. Chl-a concentration oscillated from 35.7 to 320.9 mg.m-3. Predominant phytoplankton species were Anabaena spiroides and Microcystis aeruginosa (Cyanophyceae). Water temperature was 27.8 °C (±0.88). Maximal value of total microcystine concentration was 0.23 µg.L-1. Chl-a concentration at the moment when mass mortality occurred (2.022 mg.m-3), and histopathological observations, strongly suggest that the animals' death was due to cianotoxins.

Cianobacterial bloom and animal mass mortality in a reservoir from Central Argentina

Piedras Moras reservoir (32° 10'27" S and 64° 16' 29" W; 832 ha), integrates a series of artificial lakes belonging to the Rio Tercero basin (Córdoba, Argentina). During March 2009 an algal bloom occurred, coinciding with several animal species mortality, mainly wild birds. The goal of this work was to establish the trophic status of the reservoir in relation to that mortality. Variables were evaluated in situ (temperature and water transparency) and samples were taken in order to identify algal species, Chl-a concentration (spectrophotometry) and toxins - total microcystines- (inmuno-enzymatic assay, ELISA). Histopathology studies were made on Fulica sp. A strong heterogenity in water transparency was observed, and "patches" of Potamogeton berteroanus distributed all along the lake, with Secchi disk minimal and medium values of 0.15 and 0.94 m. Chl-a concentration oscillated from 35.7 to 320.9 mg.m-3. Predominant phytoplankton species were Anabaena spiroides and Microcystis aeruginosa (Cyanophyceae). Water temperature was 27.8 °C (±0.88). Maximal value of total microcystine concentration was 0.23 g.L-1. Chl-a concentration at the moment when mass mortality occurred (2.022 mg.m-3), and histopathological observations, strongly suggest that the animals' death was due to cianotoxins.

O reservatório de Piedras Moras (32° 10'27" S e 64° 16' 29" W; 832 ha) faz parte de uma série de lagos artificiais que estão localizados na bacia do rio Tercero (Córdoba, Argentina). Durante março de 2009, ocorreu um florescimento maciço de cianobactérias que resultou na morte de muitas espécies de animais, especialmente aves aquáticas silvestres. O objetivo deste trabalho foi o de estabelecer o estado trófico do reservatório em função desta mortalidade. As variáveis foram avaliadas in situ (temperatura e transparência da água). Foram coletadas amostras para analisar as espécies de algas encontradas nos florescimentos. Foram determinados chl-a (espectrofotometria) e toxinas (microcistinas totais - ensaio imunoenzimático ELISA). Estudos histopalógicos foram realizados em Fulica sp. Uma grande heterogeneidade em transparência de água foi observada e agrupamentos de Potamogeton berteroanus estavam distribuídos em todo o reservatório com valores mínimos e médios do disco de Secchi de 0,15 a 0,54 m. Concentrações de chl-a oscilaram de 35,7 a 320,9 mg.m-3. As espécies fitoplanctônicas dominantes identificadas foram Anabaena spiroides e Microcystis aeruginosa (Cyanophyceae). A temperatura da água foi de 27,8 °C (±0.88). O valor máximo de concentração de microcistinas foi de 0,23 µg.L-1. A concentração de chl-a, quando ocorreu a mortalidade em massa (2,022 mg.m-3), e as lesões histopatológicas indicam que a mortalidade de animais foi devida a cianotoxinas.

Main diseases of pejerrey (Odontesthes bonariensis) in central Argentina

Argentina's central region includes an important area covered by shallow pampean lakes and dams. In these environments, fishing of pejerrey Odontesthes bonariensis Valenciennes, 1835 (Pisces, Atherinopsidae), the most important fresh-water fish of the country, is a relevant social activity and also a considerable economic resource. The main diseases found in this species were studied from 1992 to 2003 in the provinces of Córdoba, La Rioja and Santa Fe (30º and 35º S, 61º and 67º W). Most cases were registered in high temperature months. Lernaea sp and Aeromonas hydrophila were the etiological agents most frequently found. The trophic characteristics of the aquatic environments enhanced disease processes and caused massive death of O. bonariensis, due to complex hydrochemical interactions.

A região central de Argentina possui uma importante superfície coberta por represas e lagunas pampianas. Nestes ambientes, a pesca do peixe-rei Odontesthes bonariensis Valenciennes, 1835 (Pisces, Atherinopsidae), peixe de água doce mais importante do país, é uma atividade social relevante e de significado valor econômico. Estudaram-se as principais enfermidades que afetaram esta espécie de peixe no período de 1992-2003, nas províncias de Córdoba, La Rioja e Santa Fe (30 e 35ºS, 61 e 67ºW). A maior quantidade de casos foi registrada nos meses de temperaturas elevadas. Lernaea sp e Aeromonas hydrophila foram os agentes etiológicos mais importantes. As características tróficas particulares dos ambientes aquáticos estudados foram importantes por potenciarem alguns casos, mas em outros a causa da mortandade dos peixes esteve relacionada a complexas interações hidroquímicas.

Taxa de gestação em cabras Alpinas e Saanen tratadas com hCG no terceiro dia após o estro / Pregnancy rate and progesterone concentration of Alpine and Saanen goats treated with hCG the third day after estrus

Avaliou-se o efeito da aplicação da gonadotrofina coriônica humana (hCG) sobre a taxa e duração da gestação e sobre a concentração plasmática de progesterona (P4) em cabras lactantes das raças Alpina (83) e Saanen (60), na estação de acasalamento induzida pelo programa de fotoperíodo artificial. Os animais foram distribuídos em dois tratamentos após a cobrição; as cabras do tratamento 1 (T1 controle) receberam solução salina via intramuscular (1ml), e as do tratamento 2 (T2) receberam 250UI do hCG, no terceiro dia no pós-estro. As taxas de gestação, detectadas por exame ultra-sonográfico, realizado no 35° após a cobrição, para as cabras das raças Alpina e Saanen controle e tratadas, foram de 80,4; 70,3; 72,7; 81,5 por cento, respectivamente, não diferindo entre os tratamentos e entre as raças (P>0,05). A taxa média de gestação foi 77,2 e 75,0 por cento para os animais controle e tratados, respectivamente. A duração da gestação não foi influenciada pelos tratamentos (P>0,05). As amostras de sangue para análise da concentração plasmática de progesterona (P4) foram coletadas de cinco cabras Alpina por tratamento, no dia do estro (dia 0) e aos 3, 8, 15, 21, 42 e 60 dias após o estro. Não houve diferença na concentração plasmática média de P4 entre os tratamentos (P>0,05), sendo de 5,84ng/ml para T1 e 5,76ng/ml para T2. Observou-se diferença em relação aos dias de coleta (P<0,05). O hCG aplicado no terceiro dia pós-estro não alterou a taxa de gestação e nem a concentração de P4.

Utilização de gonadotrofina coriônica humana e cipionato de estradiol associado ao dispositivo de liberação controlada de drogas para sincronização de ovulação em cabras da raça Saanen / Use of human chorionic gonadodrophin and estradiol cipionate plus controlled internal drug release for synchronization of ovulation in Saanen goats

Dezesseis cabras nulíparas da raça Saanen foram distribuídas em dois grupos de tratamentos (T1 e T2) para sincronização da ovulação. Inicialmente, ambos os tratamento consistiram na aplicação concomitante do dispositivo de liberação controlada de drogas (CIDR-G®), de 5mg de dinoprost e de 1mg de cipionato de estradiol (CE) (dia 0). No quarto dia aplicaram-se 250UI de eCG e no quinto dia retirou-se o CIDR-G®. As cabras do T1 (n=8) receberam 1mg de CE 24 horas depois da retirada do CIDR-G® e as do T2 (n=8) receberam 250UI de hCG 30 horas após. Sete cabras do T1 e oito do T2 entraram em estro depois da retirada do CIDR-G®. Cabras que receberam hCG permaneceram em estro por 42,0± 6,9 horas e as que receberam CE por 45,0± 5,5 horas (P>0,05). As características ovulatórias não foram influenciadas pelos tratamentos. O intervalo da retirada do CIDR-G® à ovulação para ambos os protocolos de sincronização da ovulação não diferiu (P>0,05) entre tratamentos. As ovulações promovidas pelo CE ocorreram em menor intervalo de tempo.

Comparison between flurbiprofen and its nitric oxide-releasing derivatives HCT-1026 and NCX-2216 on Abeta(1-42)-induced brain inflammation and neuronal damage in the rat.

Brain inflammation is an underlying factor in the pathogenesis of Alzheimers disease (AD). We investigated, in vivo, whether differences exist in the anti-inflammatory and neuroprotective actions of flurbiprofen and its two nitric oxide-donor derivatives, HCT-1026 and NCX-2216, and the ability of these two derivatives to release nitric oxide in the brain. In adult rats injected into the nucleus basalis with preaggregated Abeta(1-42) we investigated glia reaction, the induction of inducible nitric oxide synthase (iNOS), the activation of p38 mitogen-activated protein kinase (p38MAPK) pathway and the number of choline acetyltransferase (ChAT)-positive neurons and, in naive rats we investigated, by microdialysis, cortical extracellular levels of nitrite. Injection of Abeta(1-42) induced iNOS and activation of p38MAPK 7 days after injection and an intense microglia and astrocyte reaction along with a marked reduction in the number ChAT-positive neurons, persisting up to at least 21 days. Flurbiprofen, HCT-1026 and NCX-2216 (15 mg/kg) significantly attenuated the Abeta(1-42)-induced glia reaction, iNOS induction and p38MAPK activation 7 days after treatment and astrocytes reaction 21 days after treatment. On an equimolar basis, HCT-1026 resulted the most active agent in reducing the Abeta(1-42)-induced microglia reaction. The cholinergic cell loss was also significantly reduced by 21 days of HCT-1026 treatment. No differences in body weight were found between the animals treated for 21 days with 15 mg/kg of either HCT-1026 or NCX-2216 and the controls. Oral administration of HCT-1026 (15 mg/kg) or NCX-2216 (100 mg/kg) to naive rats was followed by significant and long lasting increases in cortical nitrite levels. These findings indicate that the addition of a nitric oxide donor potentiates the anti-inflammatory activity of flurbiprofen in a model of brain inflammation.

Experimental brain inflammation and neurodegeneration as model of Alzheimer's disease: protective effects of selective COX-2 inhibitors.

Epidemiological studies indicate that long-term treatment with non-steroidal anti-inflammatory drugs reduces the risk of Alzheimer Disease and may delay its onset or slow its progression. Neuroinflammation occurs in vulnerable regions of the Alzheimer's disease (AD) brain where highly insoluble beta-amyloid (Abeta) peptide deposits and neurofibrillary tangles, as well as damaged neurons and neurites, provide stimuli for inflammation. To elucidate the complex role of inflammation in neurodegenerative processes and the efficacy of selective COX-2 inhibitors in AD, we examined whether the attenuation of brain inflammatory reaction by selective COX-2 inhibitors may protect neurons against neurodegeneration. The data reported in this review show that in in vivo models of brain inflammation and neurodegeneration, the administration of selective COX-2 inhibitors prevent not only the inflammatory reaction, but also the cholinergic hypofunction. Our data may help elucidate the epidemiological findings indicating that anti-inflammatory agents, in particular NSAIDs, reduce the risk of developing AD and may slow its progression.

Monitoreo de reservorios del centro de Córdoba (Argentina) como base para una adecuada gestión ambiental

La actividad agroganadera produce impacto sobre la calidad del agua superficial y subterránea, y es una causa importante de eutroficación en los reservorios de agua. Los objetivos del presente trabajo fueron evaluar las fluctuaciones bacteriológicas y fitoplanctónicas temporales de la calidad de agua, para uso recreacional y protección de la vida acuática, en los embalses Piedras Moras (32º 12S, 64º 19O) y La Viña (31º 47 S, 65º 01 W) en la provincia de Córdoba, Argentina. Se investigaron bacterias patógenas y de riesgo para la salud pública y animal. Se determinó el NMP/100 ml de coliformes totales y termotolerantes, Escherichia coli, Streptococcus faecalis, Salmonella spp., Pseudomonas spp. y Aeromonas spp. y se realizaron estudios taxonómicos sobre las especies de algas fitoplanctónicas, especialmente las bioindicadoras de eutroficación.Se evaluaron variables físico-químicas y biológicas según metodología estandarizada. El aislamiento de especies del grupo Aeromonas móviles, A. hydrophila, A. caviae, A. media, P. aeruginosa y E. coli, requiere monitoreos periódicos de los embalses que conduzcan a profundizar estudios en virtud de su interrelación con el ambiente, la salud pública y animal. En el embalse de Piedras Moras pudo observarse que la dispersión del nitrógeno total fue diferente según la estación del año. El fósforo total en primavera presentó sus mayores niveles en las estaciones del río Soconcho y el Balneario. Los riesgos de florecimientos de algas disminuirían en otoño, encuadrándose a los embalses como eutrófico-hipereutrófico. Hubo también una dominancia alternada entre Cianofíceas (representadas principalmente por los géneros Microcystis y Anabaena) y Dinofíceas (representadas por Ceratium hirundinella), condicionada por las variaciones en el estado trófico de los lagos

Hidrobiología del Embalse La Quebrada (Córdoba), Argentina : bacteriología y fitoplancton

El Embalse La Quebrada, localizado en Río Ceballos, a 32 Km al Noroeste de la ciudad de Córdoba, es el único reservorio para provisión de agua potable de aproximadamente 30.000 personas que viven en los alrededores, en las localidades de Río Ceballos, Unquillo y Mendiolaza. Desde 1997 se ha detectado un incremento en el proceso de eutroficación del lago, manifestándose principalmente en florecimientos de algas. En la actualidad el lago tiene periódicamente importantes blooms de Ceratium hirundinella (Dinofíceae), que se ven en la superficie del agua como manchones rojizos. También ha habido en los últimos años una interesante variación en dominancia entre Cianobacterias indicadoras de eutroficación, tales como las especies pertenecientes a los géneros Anabaena y Microcystis. En esta contribución se presentan datos varios correspondientes a muestreos efectuados no solamente en el Embalse sino además en sus dos tributarios.Los parámetros estudiados incluyen las más importantes características físico-químicas, microbiológicas y fitoplanctónicas que sirven para estimar calidad de aguaLos resultados indican que el Embalse podría ubicarse en un estadio mesotrófico, con marcada tendencia a eutrófico. Sus aguas se caracterizan por abundante productividad primaria, baja transparencia, procesos anaeróbicos en el hipolimnion que generan gases odoríferos, y recirculación de nutrientes con una alta carga de materia orgánica.Todas estas condiciones son indeseables en un ambiente utilizado como recurso turístico y para provisión de agua potable

Hidrobiología del Embalse La Quebrada (Córdoba), Argentina: bacteriología y fitoplancton

El Embalse La Quebrada, localizado en Río Ceballos, a 32 Km al Noroeste de la ciudad de Córdoba, es el único reservorio para provisión de agua potable de aproximadamente 30.000 personas que viven en los alrededores, en las localidades de Río Ceballos, Unquillo y Mendiolaza. Desde 1997 se ha detectado un incremento en el proceso de eutroficación del lago, manifestándose principalmente en florecimientos de algas. En la actualidad el lago tiene periódicamente importantes blooms de Ceratium hirundinella (Dinofíceae), que se ven en la superficie del agua como manchones rojizos. También ha habido en los últimos años una interesante variación en dominancia entre Cianobacterias indicadoras de eutroficación, tales como las especies pertenecientes a los géneros Anabaena y Microcystis. En esta contribución se presentan datos varios correspondientes a muestreos efectuados no solamente en el Embalse sino además en sus dos tributarios.Los parámetros estudiados incluyen las más importantes características físico-químicas, microbiológicas y fitoplanctónicas que sirven para estimar calidad de aguaLos resultados indican que el Embalse podría ubicarse en un estadio mesotrófico, con marcada tendencia a eutrófico. Sus aguas se caracterizan por abundante productividad primaria, baja transparencia, procesos anaeróbicos en el hipolimnion que generan gases odoríferos, y recirculación de nutrientes con una alta carga de materia orgánica.Todas estas condiciones son indeseables en un ambiente utilizado como recurso turístico y para provisión de agua potable

Monitoreo de reservorios del centro de Córdoba (Argentina) como base para una adecuada gestión ambiental

La actividad agroganadera produce impacto sobre la calidad del agua superficial y subterránea, y es una causa importante de eutroficación en los reservorios de agua. Los objetivos del presente trabajo fueron evaluar las fluctuaciones bacteriológicas y fitoplanctónicas temporales de la calidad de agua, para uso recreacional y protección de la vida acuática, en los embalses Piedras Moras (32º 12S, 64º 19O) y La Viña (31º 47 S, 65º 01 W) en la provincia de Córdoba, Argentina. Se investigaron bacterias patógenas y de riesgo para la salud pública y animal. Se determinó el NMP/100 ml de coliformes totales y termotolerantes, Escherichia coli, Streptococcus faecalis, Salmonella spp., Pseudomonas spp. y Aeromonas spp. y se realizaron estudios taxonómicos sobre las especies de algas fitoplanctónicas, especialmente las bioindicadoras de eutroficación.Se evaluaron variables físico-químicas y biológicas según metodología estandarizada. El aislamiento de especies del grupo Aeromonas móviles, A. hydrophila, A. caviae, A. media, P. aeruginosa y E. coli, requiere monitoreos periódicos de los embalses que conduzcan a profundizar estudios en virtud de su interrelación con el ambiente, la salud pública y animal. En el embalse de Piedras Moras pudo observarse que la dispersión del nitrógeno total fue diferente según la estación del año. El fósforo total en primavera presentó sus mayores niveles en las estaciones del río Soconcho y el Balneario. Los riesgos de florecimientos de algas disminuirían en otoño, encuadrándose a los embalses como eutrófico-hipereutrófico. Hubo también una dominancia alternada entre Cianofíceas (representadas principalmente por los géneros Microcystis y Anabaena) y Dinofíceas (representadas por Ceratium hirundinella), condicionada por las variaciones en el estado trófico de los lagos

The selective cyclooxygenase-2 inhibitor rofecoxib suppresses brain inflammation and protects cholinergic neurons from excitotoxic degeneration in vivo.

Brain inflammatory processes underlie the pathogenesis of Alzheimer's disease, and non-steroidal anti-inflammatory drugs have a protective effect in the disease. The aim of this work was to study in vivo whether attenuation of brain inflammatory response to excitotoxic insult by the selective cyclooxygenase-2 inhibitor, rofecoxib, may prevent neurodegeneration, as a contribution to a better understanding of the role inflammation plays in the pathology of Alzheimer's disease. We investigated, by immunohistochemical methods, glia reaction, the activation of p38 mitogen-activated protein kinase (p38MAPK) pathway with an antibody selective for the phosphorylated form of the enzyme and the number of choline acetyltransferase-positive neurons and, by in vivo microdialysis, cortical extracellular levels of acetylcholine following the injection of quisqualic acid into the right nucleus basalis of adult rats. Seven days after injection, a marked reduction in the number of choline acetyltransferase-positive neurons was found, along with an intense glia reaction, selective activation of p38MAPK at the injection site and a significant decrease in the extracellular levels of acetylcholine in the cortex ipsilateral to the injection site. The loss of cholinergic neurons persisted for at least up to 28 days. Rofecoxib (3 mg/kg/day, starting 1 h prior to injection of quisqualic acid) treatment for 7 days significantly attenuated glia activation and prevented the loss of choline acetyltransferase-positive cells and a decrease in cortical acetylcholine release. The prevention of cholinergic cell loss by rofecoxib occurred concomitantly with the inhibition of p38MAPK phosphorylation. Our findings suggest an important role of brain inflammatory reaction in cholinergic degeneration and demonstrate a neuroprotective effect of rofecoxib, presumably mediated through the inhibition of p38MAPK phosphorylation.

De novo backbone and sequence design of an idealized alpha/beta-barrel protein: evidence of stable tertiary structure.

We have designed, synthesized, and characterized a 216 amino acid residue sequence encoding a putative idealized alpha/beta-barrel protein. The design was elaborated in two steps. First, the idealized backbone was defined with geometric parameters representing our target fold: a central eight parallel-stranded beta-sheet surrounded by eight parallel alpha-helices, connected together with short structural turns on both sides of the barrel. An automated sequence selection algorithm, based on the dead-end elimination theorem, was used to find the optimal amino acid sequence fitting the target structure. A synthetic gene coding for the designed sequence was constructed and the recombinant artificial protein was expressed in bacteria, purified and characterized. Far-UV CD spectra with prominent bands at 222nm and 208nm revealed the presence of alpha-helix secondary structures (50%) in fairly good agreement with the model. A pronounced absorption band in the near-UV CD region, arising from immobilized aromatic side-chains, showed that the artificial protein is folded in solution. Chemical unfolding monitored by tryptophan fluorescence revealed a conformational stability (DeltaG(H2O)) of 35kJ/mol. Thermal unfolding monitored by near-UV CD revealed a cooperative transition with an apparent T(m) of 65 degrees C. Moreover, the artificial protein did not exhibit any affinity for the hydrophobic fluorescent probe 1-anilinonaphthalene-8-sulfonic acid (ANS), providing additional evidence that the artificial barrel is not in the molten globule state, contrary to previously designed artificial alpha/beta-barrels. Finally, 1H NMR spectra of the folded and unfolded proteins provided evidence for specific interactions in the folded protein. Taken together, the results indicate that the de novo designed alpha/beta-barrel protein adopts a stable three-dimensional structure in solution. These encouraging results show that de novo design of an idealized protein structure of more than 200 amino acid residues is now possible, from construction of a particular backbone conformation to determination of an amino acid sequence with an automated sequence selection algorithm.

NO-flurbiprofen attenuates excitotoxin-induced brain inflammation, and releases nitric oxide in the brain.

Brain inflammation underlies the pathogenesis of Alzheimer's disease (AD) and nonsteroidal anti-inflammatory drug therapy may delay the onset of AD. We investigated, in vivo, the effects of NO-flurbiprofen on brain inflammation in rats injected with quisqualic acid into the nucleus basalis and on the release of nitric oxide from the drug in naive rat brains. We showed that the excitotoxin-induced microglia reaction, the expression of inducible nitric oxide synthase-positive cells and the production of interleukin-1beta and prostaglandin-E2 in the injected area were attenuated by the NO-flurbiprofen (15 mg/kg, p.o.) treatment. An oral administration of NO-flurbiprofen (25, 50 and 100 mg/kg) to naive rats was followed by significant increases in cortical nitrite levels. This drug may have important therapeutic implications for the treatment of AD.

Strychnogucines A and B, two new antiplasmodial bisindole alkaloids from Strychnos icaja.

A reinvestigation of Strychnos icaja roots has resulted in the isolation of two tertiary quasi-symmetric bisindole alkaloids named strychnogucines A (1) and B (2). Their structures were identified by means of spectroscopic data interpretation. Compound 2 was highly active in vitro and compound 1 moderately active against four strains of Plasmodium falciparum. Strychnogucine B (2) was more active against a chloroquine-resistant strain than against a chloroquine-sensitive one (best CI(50), 80 nM against the W2 strain). In addition, this compound showed a selective antiplasmodial activity with 25-180 times greater toxicity toward P. falciparum, relative to cultured human cancer cells (KB) or human fibroblasts (WI38).

Brain inflammatory reaction in an animal model of neuronal degeneration and its modulation by an anti-inflammatory drug: implication in Alzheimer's disease.

Brain inflammatory processes underlie the pathogenesis of Alzheimer's disease, and nonsteroidal anti-inflammatory drugs have a protective effect in the disease. The aim of this study was to characterize in vivo in the rat brain the inflammatory reaction in response to excitotoxic insult and to investigate the efficacy of nimesulide treatment. Quisqualic acid was injected into the right nucleus basalis of rats. The excitotoxin induced cholinergic degeneration, an intense glial reaction and the production of inflammatory mediators. Three hours after injection, a five-fold elevation in the concentration of interleukin-1beta in the injected area was observed. This elevation was reduced by 50% by nimesulide (10 mg/kg, i.m.) pretreatment. Electron microscope examination and immunocytochemical staining revealed an intense activation of microglia and astrocytes at both 24 h and 7 days after injection. Cyclooxygenase-2-immunoreactivity was induced in the blood vessels of the injected hemisphere in perivascular microglial and endothelial cells 24 h after injection. Seven days postinjection, a cyclooxygenase-2-positive signal was induced in the parenchymal microglia and large amounts of prostaglandin-E2 were measured in the injected area. Twenty-four hours and 7 days after injection, many inducible nitric oxide synthase-positive cells and a high level of nitrite were detected at the injection site. Seven days of nimesulide (10 mg/kg/day, i.m.) treatment strongly attenuated the microglial reaction, reduced the number of inducible nitric oxide synthase-positive cells and completely abolished the increase in prostaglandin-E2 formation. These data provide valuable support in vivo for the potential efficacy of cyclooxygenase-2 inhibitors in Alzheimer's disease therapy.

Interleukin-1beta activates forebrain glial cells and increases nitric oxide production and cortical glutamate and GABA release in vivo: implications for Alzheimer's disease.

Interleukin-1beta (10 U) was injected into the nucleus basalis of adult male Wistar rats. The inflammation-induced changes in glial cell morphology and expression of inducible nitric oxide synthase in the injected area, the release of acetylcholine, GABA and glutamate from the ipsilateral cortex, the production of nitrite levels in the injected area and ipsilateral cortex, and changes in motor activity were investigated. Saline-injected rats were used as control. Interleukin-1beta induced an activation of both microglia and astrocytes which was already evident 24 h after injection. Seven days after injection, many reactive microglial cells and astrocytes were seen in the injected area and in other brain regions of the same hemisphere. Microglia reaction, but not astrocyte activation, disappeared 30 days post-injection. Seven days after interleukin-1beta injection, many cells immunopositive for inducible nitric oxide synthase were found surrounding the injection site. Inducible nitric oxide synthase-positive cells were identified, by double staining immunohistochemistry, in the reactive microglial cells and, by electron microscope examination, in the perineuronal subpopulation of resident activated microglia. Microdialysis investigations revealed a transient increase in reactive nitrogen intermediates (at seven days post-injection), a delayed (at 30 days post-injection) increase in GABA and glutamate release, and no changes in acetylcholine release in the ipsilateral cortex in interleukin-1beta, but not saline, injected rats. Inhibition of inducible nitric oxide synthase expression by N(G)-nitro-L-arginine methyl ester administration prevented the increase in nitrogen intermediates and GABA release, but not in glutamate release. Our findings suggest that an inflammatory reaction of the basal forebrain facilitates GABA release through the production of nitric oxide.

Beta(1-40) amyloid peptide injection into the nucleus basalis of rats induces microglia reaction and enhances cortical gamma-aminobutyric acid release in vivo.

The nucleus basalis of adult rats was injected with beta(1-40) amyloid peptide. A marked increase in basal and K(+)-evoked GABA release in the ipsilateral cortex and a significant decrease in GAD activity in the injected NB were found 30 days after injection. An intense activation of microglial cells that surrounded and infiltrated the deposit was observed. These data demonstrate that a local injection of beta(1-40) peptide into the NB induces glia activation and affects GABAergic neurons.