Stream-bank shade and larval distribution of the Philippine malaria vector Anopheles flavirostris

The principal malaria vector in the Philippines, Anopheles flavirostris (Ludlow) (Diptera: Culicidae), is regarded as 'shade-loving' for its breeding sites, i.e. larval habitats. This long-standing belief, based on circumstantial observations rather than ecological analysis, has guided larval control methods such as 'stream-clearing' or the removal of riparian vegetation, to reduce the local abundance of An. flavirostris . We measured the distribution and abundance of An. flavirostris larvae in relation to canopy vegetation cover along a stream in Quezon Province, the Philippines. Estimates of canopy openness and light measurements were obtained by an approximation method that used simplified assumptions about the sun, and by hemispherical photographs analysed using the program hemiphot(C) . The location of larvae, shade and other landscape features was incorporated into a geographical information system (GIS) analysis. Early larval instars of An. flavirostris were found to be clustered and more often present in shadier sites, whereas abundance was higher in sunnier sites. For later instars, distribution was more evenly dispersed and only weakly related to shade. The best predictor of late-instar larvae was the density of early instars. Distribution and abundance of larvae were related over time (24 days). This pattern indicates favoured areas for oviposition and adult emergence, and may be predictable. Canopy measurements by the approximation method correlated better with larval abundance than hemispherical photography, being economical and practical for field use. Whereas shade or shade-related factors apparently have effects on larval distribution of An. flavirostris , they do not explain it completely. Until more is known about the bionomics of this vector and the efficacy and environmental effects of stream-clearing, we recommend caution in the use of this larval control method.

Inhibition of rat platelet aggregation by the diazeniumdiolate nitric oxide donor MAHMA NONOate

1 Inhibition of rat platelet aggregation by the nitric oxide (NO) donor MAHMA NONOate (Z-1-{N-methyl-N-[6-(N-methylammoniohexyl)amino]}diazen-l-ium-1,2-diolate) was investigated. The aims were to compare its anti-aggregatory effect with vasorelaxation, to determine the effects of the soluble guanylate cyclase inhibitor, ODQ (1H-[1,2,4]oxadiazolo[4,3-ajquinoxalin-1-one), and to investigate the possible role of activation of sarco-encloplasmic reticulum calcium-ATPase (SERCA), independent of soluble guanylate cyclase, using thapsigargin. 2 MAHMA NONOate concentration-dependently inhibited sub-maximal aggregation responses to collagen (2 - 10 mug ml(-1)) and adenosine diphosphate (ADP; 2 mum) in platelet rich plasma. It was (i) more effective at inhibiting aggregation induced by collagen than by ADP, and (ii) less potent at inhibiting platelet aggregation than relaxing rat pulmonary artery. 3 ODQ (10 mum) caused only a small shift (approximately half a log unit) in the concentration-response curve to MAHMA NONOate irrespective of the aggregating agent. 4 The NO-independent activator of soluble guanylate cyclase, YC-1 (3-(5'-hydroxymethyl-2'-furyl)-1-benzy] indazole; 1 - 100 mum), did not inhibit aggregation. The cGMP analogue, 8-pCPT-cGMP (8-(4-chlorophenylthio)guanosine 3'5' cyclic monophosphate; 0.1 - 1 mm), caused minimal inhibition. 5 On collagen-aggregated platelets responses to MAHMA NONOate (ODQ 10 PM present) were abolished by thapsigargin (200 nm). On ADP-aggregated platelets thapsigargin caused partial inhibition. 6 Results with S-nitrosoglutathione (GSNO) resembled those with MAHMA NONOate. Glyceryl trinitrate and sodium nitroprusside were poor inhibitors of aggregation. 7 Thus inhibition of rat platelet aggregation by MAHMA NONOate (like GSNO) is largely ODQ-resistant and, by implication, independent of soluble guanylate cyclase. A likely mechanism of inhibition is activation of SERCA.

Prolonged retention after aggregation into secretory granules of human R183H-growth hormone (GH), a mutant that causes autosomal dominant GH deficiency type II

Human R183H-GH causes autosomal dominant GH deficiency type II. Because we show here that the mutant hormone is fully bioactive, we have sought to locate an impairment in its progress through the secretory pathway as assessed by pulse chase experiments. Newly synthesized wild-type and R183H-GH were stable when expressed transiently in AtT20 cells, and both formed equivalent amounts of Lubrol-insoluble aggregates within 40 min after synthesis. There was no evidence for intermolecular disulfide bond formation in aggregates of wild-type hormone or the R183H mutant. Both wildtype and R183H-GH were packaged into secretory granules, assessed by the ability of 1 mm BaCl2 to stimulate release and by immunocytochemistry. The mutant differed from wildtype hormone in its retention in the cells after packaging into secretory granules; 50% more R183H-GH than wild-type aggregates were retained in AtT20 cells 120 min after synthesis, and stimulated release of R183H-GH or a mixture of R183H-GH and wild-type that had been retained in the cell was reduced. The longer retention of R183H-GH aggregates indicates that a single point mutation in a protein contained in secretory granules affects the rate of secretory granule release.

Expression of anterior Hox genes during larval development of the gastropod Haliotis asinina

We report the spatial expression patterns of five anterior Hox genes during larval development of the gastropod mollusc Haliotis asinina, an unsegmented spiralian lophotrochozoan. Molecular alignments and phylogenetic analysis indicate that these genes are homologues of Drosophila HOM-C genes labial, proboscipedia, zen, Deformed, and Sex combs reduced, the abalone genes are named Has-Hox1, -Hox2, -Hox3, -Hox4, and -Hox5. Has-Hox transcripts are first detected in the free-swimming trochophore larval stage- and restricted to the posttrochal ectoderm. Has-Hox2, -Hox3, and -Hox4 are expressed in bilaterally symmetrical and overlapping patterns in presumptive neuroectodermal cells on the ventral side of the trochophore. Has-Hox1 expression is restricted to a ring of cells on the dorsoposterior surface, corresponding to the outer mantle edge where new larval shell is being synthesized. There appears to be little change in the expression domains of these Has-Hox genes in pre- and posttorsional veliger larvae, with expression maintained in ectodermal and neuroectodermal tissues. Has-Hox2, -Hox3, -Hox4, and-Hox5 appear to be expressed in a colinear manner in the ganglia and connectives in the twisted nervous system. This pattern is not evident in older larvae. Has-Hox1 and-Hox4 are expressed in the margin of the mantle in the posttorsional veliger, suggesting that Hox genes play a role in gastropod shell formation.

5-Hydroxytryptamine and platelets: uptake and aggregation in hypoxic pulmonary hypertensive rats

Pulmonary hypertension is associated with various alterations in 5-hydroxytryptamine (5-HT) physiology. In this study in platelets from hypoxic pulmonary hypertensive rats (10% O-2; 1 week) and normoxic rats (room air), (i) initial rates of specific [H-3]5-HT uptake were measured and (ii) potentiation of collagen- and ADP-induced aggregation by 5-HT was quantified. The platelet count was almost halved in hypoxic rats. In uptake experiments, there was a decrease in 5-HT uptake in platelets from hypoxic compared with normoxic rats, due to a 36% reduction in the maximal initial rate of uptake. The aggregation experiments showed that 5-HT (1-100 muM) increased the magnitude of responses to collagen and the duration of responses to ADP, but there was no difference between hypoxic and normoxic rats. Abnormalities in platelet function may conceivably lead to increases in plasma 5-HT levels in hypoxic pulmonary hypertension, but are unlikely to aggravate pulmonary thromboembolism. (C) 2002 Elsevier Science B.V. All rights reserved.

Effect of cylindrical filler aggregation on the electrical conductivity of composites

This work reports on the effect of carbon nanotube aggregation on the electrical conductivity and other network properties of polymer/carbon nanotube composites by modeling the carbon nanotubes as hard-core cylinders. It is shown that the conductivity decreases for increasing filler aggregation, and that this effect is more significant for higher cylinder volume fractions. It is also demonstrated, for volume fractions at which the giant component is present, that increasing the fraction of cylinders within clusters leads to a break of the giant component and the formation of a set of finite clusters. The decrease of the giant component with the increase of the fraction of cylinders within the cluster can be related to a decrease of the spanning probability due to a decrease of the number of cylinders between the clusters. Finally, it is demonstrated that the effect of aggregation can be understood by employing the network theory.

Biologia de Anopheles (Kerteszia) neivai H., D. & K., 1913 (Diptera: Culicidae) en la costa Pacífica de Colombia: I. Fluctuación de la población larval y características de sus criaderos

En Charambirá, localidad del município de Istmina (Chocó) en la costa Pacífica de Colombia, donde existe un problema de malaria endémica, se estudió la fluctuación de la población larval y las características de los criaderos de Anopheles (Kerteszia) neivai, especie considerada como posible vectora de malaria en esa región del país. La vegetación circundante fue dividida en tres estratos de acuerdo al grado de cobertura foliar. Se demarcaron cuadrantes de 100 m² en cada estrato para el muestreo de plantas epífitas de la família Bromeliaceae, en las cuales se acumula agua que sirve como criadero para esta especie de anofelino. Se tomaron datos de temperatura, pH y volumen del agua contenida en cada bromelia. El mayor número de larvas se detectó en el estrato 1 (manglar) a una altitud inferior a 4 m, pero no se encontró evidencia significativa de estratificación vertical de la población larval de A. neivai hasta los 8 m. Se evidenció una correlación lineal positiva entre el número de larvas y el volumen de agua contenida en cada bromelia; por otra parte se observó también una correlación directa entre la precipitación mensual acumulada y la fluctuación poblacional de esta especie. Los índices larvales mas altos se observaron entre los meses de marzo a abril y de julio a agosto. La mortalidad larval fue alta en el primer estadío (43,5%) y solo un 23,7% sobrevivió hasta el cuarto.

Avaliação do desenvolvimento larval de Anopheles darlingi criado em laboratório sob diferentes dietas

São testadas três dietas para larvas de A. darlingi, buscando-se os seguintes parâmetros indicativos de desenvolvimento nesta fase: tempo de evolução total e de cada estádio, sobrevivência ao estádio, diária e total. A metodologia utilizada na determinação desses parâmetros constitui-se da combinação de dois métodos de análise da estatística vital, adaptados ao estudo de populações mantidas em laboratório. Os tempos de evolução total e de cada estádio foram determinados gráficamente a partir das curvas de tendência dos estádios medianos da colônia, em inspeções sucessivas. Os valores de sobrevivência diária, ao estádio e total foram calculados a partir de tábuas de sobrevivência. Os resultados permitiram eleger a dieta composta de uma parte de farinha de peixe para duas partes de farinha de pão e duas partes de germe de trigo como a mais adequada ao desenvolvimento larval, com duração de 12,9 dias entre o primerio estádio e a emersão do adulto e sobrevivência total de 95%.

Duration of larval and pupal development stages of Aedes albopictus in natural and artificial containers

Aedes albopictus were reared in different containers: a tree hole, a bamboo stump and an auto tire. The total times from egg hatching to adult emergence were of 19.6,27.3 and 37.5 days, respectively, according to the container. The first, second and third-instar larvae presented growth periods with highly similar durations. The fourth-instar larvae was longer than the others stages. The pupation time was longer than the fourth-instar larvae growth period. The temperature of the breeding sites studied, which was of 18° C to 22° C on average, was also taken into consideration. The mortality of the immature stages was analysed and compared as between the experimental groups; it was lower in the natural containers than in the discarded tire. The average wing length of adult females emerging from tree hole was significantly larger (p < 0.05) than that of those emerging from the tire.

Human case of gastric infection by a fourth larval stage of Pseudoterranova decipiens (Nematoda, Anisakidae)

Only three cases of human infection by anisakid nematodes have been reported in Chile since 1976. In the present case, an anisakid worm, identified as a fourth-stage Pseudoterranova decipiens larva, was removed with a gastroendoscopic biopsy clipper from the stomach of a 45 year-old man from southern Chile. The patient, who presented acute epigastric pain and a continuous sensation of having an empty stomach, reported having eaten smoked fish. The worm was fixed in 70% ethanol and cleaned in lactophenol for morphological study. The morphometric characteristics of the worm are described and drawn. Anisakid larvae in fish flesh can be killed by freezing or cooking.

Susceptibilidade larval de duas populações de Aedes egypti a inseticidas químicos

OBJETIVO: A susceptibilidade dos insetos tem sido um dos mais importantes aspectos a ser monitorados em programas de saúde pública que tratam do controle de vetores. O estudo objetiva avaliar a susceptibilidade de larvas de Aedes aegypti a inseticidas químicos em áreas sujeitas ou não a controle. MÉTODOS: Bioensaios foram realizados com concentração de diagnóstico e concentração múltipla, segundo padrão da Organização Mundial de Saúde para as coletas de larvas de Aedes aegypti, em uma área não sujeita -- Campinas, SP -- e em uma outra área sujeita -- Campo Grande, MS -- a tratamentos químicos de controle. RESULTADOS: Larvas de Aedes aegypti coletadas em Campinas indicaram resistência potencial à concentração-diagnóstico (CD) de 0,04 ppm do organofosforado temephos. O teste de concentração múltipla registrou sobrevivência de 24,5% à concentração de 0,0125 ppm. A susceptibilidade dessa mesma linhagem foi avaliada para o organofosforado fenitrothion (CD=0,08 ppm) e o piretróide cipermetrina (CD=0,01 ppm), resultando em valores normais para essas concentrações. Larvas de Ae. aegypti coletadas em Campo Grande mostraram susceptibilidade normal ao temephos (CD=0,04 ppm) e à cipermetrina (CD=0,01 ppm). Também foram estabelecidas as CL50 e as CL95 de cipermetrina 25 CE, cyfluthrin 5 CE, betacyfluthrin 1,25 SC e propoxur 20 CE para Ae. aegypti. Com base nos dados da linhagem-padrão Rockefeller, foram estimadas as razões de resistência de 2,9, 2,2, 2,4 e 1,3, respectivamente, pela CL50, e de 3,5, 2,6, 3,9 e 1,3 pela CL95. CONCLUSÃO: Os resultados reforçam a necessidade de avaliações prévias e monitoramento da efetividade dos inseticidas que devem ser usados em programas de controle de mosquitos.

Susceptibilidade larval de populações de Aedes aegypti e Culex quinquefasciatus a inseticidas químicos

OBJETIVO: Avaliar a susceptibilidade a inseticidas químicos de larvas de Culex quinquefasciatus e Aedes aegypti, provenientes de áreas sujeitas ou não a tratamentos de controle. MÉTODOS: Foram coletadas larvas de mosquitos em uma área não sujeita a tratamentos com inseticidas (Campinas, SP) e em áreas sujeitas a esses tratamentos (Campo Grande, MS e Cuiabá, MT). Foram usados bioensaios com concentrações diagnóstico e concentrações múltipla de inseticidas organofosforados e piretróides, segundo padrão da Organização Mundial de Saúde, para avaliar a susceptibilidade dessas larvas. RESULTADOS: Ensaios com larvas de Culex quinquefasciatus de Campinas, SP, permitiram a suspeita de resistência à cipermetrina e evidenciaram resistência à ciflutrina. Larvas dessa espécie coletadas em Campo Grande, MS, e Campinas, SP, apresentaram resistência ao temephos. Para a colônia campineira desta espécie, foram estabelecidas as razões de resistência: RR50=6,36 e RR95=4,94, com base em linhagem susceptível padrão. Adicionalmente, os testes com Aedes aegypti mostraram susceptibilidade similar ao temephos em uma população de campo (Cuiabá, MT) e uma de laboratório. CONCLUSÕES: Os resultados indicam resistência a organofosforado e piretróides em Culex quinquefasciatus, evidenciando a necessidade de avaliações e monitoramento da efetividade dos inseticidas a serem usados nos programas de controle de mosquitos.

Pseudomonas aeruginosa amidase: Aggregation in recombinant Escherichia coli

The effect of cultivation parameters such as temperature incubation, IPTG induction and ethanol shock on the production of Pseudomonasaeruginosa amidase (E.C.3.5.1.4) in a recombinant Escherichia coli strain in LB ampicillin culture medium was investigated. The highest yield of solubleamidase, relatively to other proteins, was obtained in the condition at 37 degrees C using 0.40 mM IPTG to induce growth, with ethanol. Our results demonstrate the formation of insoluble aggregates containing amidase, which was biologically active, in all tested growth conditions. Addition of ethanol at 25 degrees C in the culture medium improved amidase yield, which quantitatively aggregated in a biologically active form and exhibited in all conditions an increased specific activity relatively to the soluble form of the enzyme. Non-denaturing solubilization of the aggregated amidase was successfully achieved using L-arginine. The aggregates obtained from conditions at 37 degrees C by Furier transform infrared spectroscopy (FTIR) analysis demonstrated a lower content of intermolecular interactions, which facilitated the solubilization step applying non-denaturing conditions. The higher interactions exhibited in aggregates obtained at suboptimal conditions compromised the solubilization yield. This work provides an approach for the characterization and solubilization of novel reported biologically active aggregates of this amidase.

Living bacteria rheology: Population growth, aggregation patterns, and collective behaviour under diferente shear flows

The activity of growing living bacteria was investigated using real-time and in situ rheology-in stationary and oscillatory shear. Two different strains of the human pathogen Staphylococcus aureus-strain COL and its isogenic cell wall autolysis mutant, RUSAL9-were considered in this work. For low bacteria density, strain COL forms small clusters, while the mutant, presenting deficient cell separation, forms irregular larger aggregates. In the early stages of growth, when subjected to a stationary shear, the viscosity of the cultures of both strains increases with the population of cells. As the bacteria reach the exponential phase of growth, the viscosity of the cultures of the two strains follows different and rich behaviors, with no counterpart in the optical density or in the population's colony-forming units measurements. While the viscosity of strain COL culture keeps increasing during the exponential phase and returns close to its initial value for the late phase of growth, where the population stabilizes, the viscosity of the mutant strain culture decreases steeply, still in the exponential phase, remains constant for some time, and increases again, reaching a constant plateau at a maximum value for the late phase of growth. These complex viscoelastic behaviors, which were observed to be shear-stress-dependent, are a consequence of two coupled effects: the cell density continuous increase and its changing interacting properties. The viscous and elastic moduli of strain COL culture, obtained with oscillatory shear, exhibit power-law behaviors whose exponents are dependent on the bacteria growth stage. The viscous and elastic moduli of the mutant culture have complex behaviors, emerging from the different relaxation times that are associated with the large molecules of the medium and the self-organized structures of bacteria. Nevertheless, these behaviors reflect the bacteria growth stage.