Larvicidal activity of silver nanoparticles synthesized using <Emphasis Type="Italic">Plumeria rubra</Emphasis> plant latex against <Emphasis Type="Italic">Aedes aegypti</Emphasis> and <Emphasis Type="Italic">Anopheles stephensi</Emphasis>

In the present study activity of silver nanoparticles (AgNPs) synthesized using Plumeria rubra plant latex against second and fourth larval instar of Aedes aegypti and Anopheles stephensi was determined. Range of concentrations of synthesized AgNps (10, 5, 2.5, 1.25, 0.625, 0.3125 ppm) and aqueous crude latex (1,000, 500, 250, 125, 62.50, 31.25 ppm) were tested against larvae of A. aegypti and A. Stephensi. The synthesized AgNps from P. rubra latex were highly toxic than crude latex extract in both mosquito species. The LC₅₀ values for second and fourth larval instars after 24 h of crude latex exposure were 1.49, 1.82 ppm against A. aegypti and 1.10, 1.74 ppm against A. stephensi respectively. These figures were 181.67, 287.49 ppm against A. aegypti and 143.69, 170.58 ppm against A. stephensi respectively for crude latex extract. The mortality rates were positively correlated with the concentration of AgNPs. The characterization studies of synthesized AgNPs by UV–Vis spectrophotometry, transmission electron microscopy (TEM), Particle size analysis (PSA) and zeta potential confirmed the spherical shape and size (32–200 nm) of silver nanoparticles alongwith stability. Toxicity studies carried out against non-target fish species Poecilia reticulata, the most common organism in the habitats of A. aegypti and A. stephensi showed no toxicity at LC₅₀ and LC₉₀ doses of the AgNPs. This is the first report on mosquito larvicidal activity of latex synthesized nanoparticles.     

Effect of bioactive fractions of <Emphasis Type="Italic">Citrullus vulgaris</Emphasis> Schrad. leaf extract against <Emphasis Type="Italic">Anopheles stephensi</Emphasis> and <Emphasis Type="Italic">Aedes aegypti</Emphasis>

The benzene extract of Citrullus vulgaris was tested against Anopheles stephensi and Aedes aegypti for the larvicidal activity and ovicidal properties. The crude benzene extract was found to be more effective against A. stephensi than A. aegypti. The LC₅₀ values were 18.56 and 42.76 ppm respectively. The LC₅₀ values for silica gel fractions (bioactive fractions I, II, III and IV) were 11.32, 14.12, 14.53 and 16.02 ppm respectively. The mean per cent hatchability of the egg rafts were observed after 48 h post treatment. The crude extract of benzene exerted 100% mortality at 250 ppm against A. stephensi and at 300 ppm against A. aegypti. The silica gel fractions I and II afforded 100% mortality at 100 ppm and III and IV exerted the hatchability rate of 4.9 and 5.3% at the same concentration against A. stephensi.     

Efficacy of fungus mediated silver and gold nanoparticles against <Emphasis Type="Italic">Aedes aegypti</Emphasis> larvae

Chrysosporium tropicum is a pathogenic fungus. It is known to be an effective mosquito control agent. In the present study, we have synthesized the silver and gold nanoparticles using C. tropicum. These nanoparticles have been characterized through Microscan reader, X-ray diffractometer, transmission electron microscopy, and further confirmed by scanning electron microscopy. The characterization study confirmed the spherical shape and size (2–15 and 20–50 nm) of gold and silver nanoparticles. These silver and gold nanoparticles have been tested as a larvicide against the Aedes aegypti larvae. The larvicidal efficacy was noted when performed against all instars of A. aegypti at six different log concentrations, and significant results could be observed. The gold nanoparticles used as an efficacy enhancer have shown mortality at three times higher concentration than the silver nanoparticles. The larval mortality was observed after different time of exposures. The mortality values were obtained using the probit analysis. The larvae of A. aegypti were found to be highly susceptible for the silver nanoparticles. The second instar larvae have shown 100% mortality against the silver nanoparticles after 1 h, whereas the first, third, and fourth instars have shown efficacy (LC₅₀ = 3.47, 4, and 2; LC₉₀ = 12.30, 8.91, and 4; LC₉₉ = 13.18, 13.18, and 7.58, respectively) after 1 h. The results could suggest that the use of fungus C. tropicum, silver, and gold nanoparticles is a rapid, environmentally safer, and greener approach for mosquito control. This could lead us to a new possibility in vector control strategy.     

Adulticidal and repellent properties of indigenous plant extracts against <Emphasis Type="Italic">Culex quinquefasciatus</Emphasis> and <Emphasis Type="Italic">Aedes aegypti</Emphasis> (Diptera: Culicidae)

Several diseases are associated to the mosquito–human interaction. Mosquitoes are the carriers of severe and well-known illnesses such as malaria, arboviral encephalitis, dengue fever, chikunguniya fever, West Nile virus, and yellow fever. These diseases produce significant morbidity and mortality in humans and livestock around the world. The adulticidal and repellent activities of crude hexane, ethyl acetate, benzene, chloroform, and methanol extracts of leaf of Eclipta alba and Andrographis paniculata were assayed for their toxicity against two important vector mosquitoes, viz., Culex quinquefasciatus and Aedes aegypti (Diptera: Culicidae). The adult mortality was observed after 24 h of exposure. All extracts showed moderate adulticide effects; however, the highest adult mortality was found in methanol extract of A. paniculata against the adults of C. quinquefasciatus and A. aegypti with the LC₅₀ and LC₉₀ values were 149.81, 172.37 ppm and 288.12, 321.01 ppm, respectively. The results of the repellent activity of hexane, ethyl acetate, benzene, chloroform, and methanol extract of E. alba and A. paniculata plants at three different concentrations of 1.0, 2.5, and 5.0 mg/cm² were applied on skin of forearm in man and exposed against adult female mosquitoes. In this observation, these two plant crude extracts gave protection against mosquito bites without any allergic reaction to the test person, and also, the repellent activity is dependent on the strength of the plant extracts. These results suggest that the leaf solvent plant extracts have the potential to be used as an ideal ecofriendly approach for the control of mosquitoes. This is the first report on the mosquito adulticidal and repellent activities of the reported E. alba and A. paniculata plants.     

Effect of <Emphasis Type="Italic">Myracrodruon urundeuva</Emphasis> leaf lectin on survival and digestive enzymes of <Emphasis Type="Italic">Aedes aegypti</Emphasis> larvae

Aedes aegypti transmits the viruses that cause yellow and dengue fevers. Vector control is essential, since a vaccine for dengue has not as yet been made available. This work reports on the larvicidal activity of Myracrodruon urundeuva leaf lectin (MuLL) against A. aegypti fourth-stage larvae (L₄). Also, the resistance of MuLL to digestion by L₄ gut proteases and the effects of MuLL on protease, trypsin-like and α-amylase activities from L₄ gut were evaluated to determine if lectin remains active in A. aegypti gut and if insect enzyme activities can be modulated by MuLL. MuLL promoted mortality of L₄ with LC₅₀ of 0.202 mg/ml. Haemagglutinating activity of MuLL was detected even after incubation for 96 h with L₄ gut preparation containing protease activity. MuLL affected the activity of gut enzymes, inhibiting protease and trypsin activities and stimulating α-amylase activity. The results suggest that MuLL may become a new biodegradable larvicidal agent for dengue control. Larvicidal activity of MuLL may be linked to its resistance to proteolysis by larval enzymes and interference in the activity of digestive larval enzymes.     

Bioactivity of flavonoid compounds from <Emphasis Type="Italic">Poncirus trifoliata</Emphasis> L. (Family: Rutaceae) against the dengue vector, <Emphasis Type="Italic">Aedes aegypti</Emphasis> L. (Diptera: Culicidae)

The bioactivity of four flavonoid compounds, namely poncirin, rhoifolin, naringin and marmesin, from Poncirus trifoliata was studied against the Aedes aegypti. Larvicidal assays were conducted to evaluate the 24 h LC(50) and LC(90) values of the flavonoid compounds. The lethal concentration (LC(50) and LC(90)) values ranged from 0.082 to 0.122 mg/l and 0.152 to 0.223 mg/l, respectively. The result of ovicidal test suggests that the ovicidal activity of the flavonoid compounds was influenced by the concentration of flavonoid compounds and age of the eggs. The result of oviposition test showed that the four flavonoid compounds exhibited oviposition-deterrent activity against gravid female mosquitoes. Oviposition decreased with an increase in concentration of flavonoid compounds. A laboratory test was carried out to evaluate protection period and percentage of repellency of four compounds diluted in ethanol (10 mg/l). The compound rhoifolin provided maximum 365.0 +/- 12.0 min protection and also 100.0% +/- 0.0 repellency against mosquito bite followed by poncirin, marmesin and naringin. None of the 25 volunteers of either sex exposed to 10% (w/v) flavonoid compounds (4-h patch test) showed a positive skin irritant reaction. All of the tested compounds proved to have various activities against different life stages of A. aegypti. Therefore, flavonoid compounds from P. trifoliata can be a potential candidate for use in the development of commercial mosquitocidal products that may be an alternative to conventional synthetic chemicals, particularly in integrated vector control applications.     

Electroantennogram,flight orientation,and oviposition responses of <Emphasis Type="Italic">Aedes aegypti</Emphasis> to the oviposition pheromone <Emphasis Type="Italic">n</Emphasis>-heneicosane

Oviposition pheromones specifically influence the females of many insects to lay eggs in the sites resulting in more egg deposition. A previous report describes the principal role of n-heneicosane (C₂₁) identified and characterized from the larval cuticle of Aedes aegypti (L.) in attracting the gravid mosquitoes to oviposit in treated substrates among other chemical components. However, the means by which this compound is perceived by the females for oviposition has not been reported. In this study, we have recorded the peripheral olfactory responses from the antenna of Ae. aegypti from 10⁻⁷ g to 10⁻³ g doses of n-heneicosane. The EAG response of female mosquitoes increased in a dose-dependent manner with increasing stimulus strength. In the orientation assay using Y-maze olfactometer, female mosquitoes were attracted to the odor plume of 10⁻⁶ g and 10⁻⁵ g dose, while the higher dose of 10⁻³ g plume enforced repellency to gravid mosquitoes. The response to oviposition substrates by gravid Ae. aegypti females differed across the range of concentrations of n-heneicosane under multiple choice conditions, larger number of eggs were deposited in 10 ppm (10 mg/l) solutions compared to lower and higher concentrations indicating 10 ppm was most attractive. Application of n-heneicosane at 10 ppm in breeding habitats will be a useful method to attract the gravid mosquitoes using ovitraps for surveillance and monitoring. The possible use of this compound in monitoring of mosquito population in endemic areas in relevance to integrated vector management strategies is discussed in detail.     

Exsheathment and midgut invasion of nocturnally subperiodic <Emphasis Type="Italic">Brugia malayi</Emphasis> microfilariae in a refractory vector, <Emphasis Type="Italic">Aedes aegypti</Emphasis> (Thailand strain)

Exsheathment and midgut invasion of nocturnally subperiodic Brugia malayi microfilariae were analyzed using light and scanning electron microscopy in a refractory vector, Aedes aegypti (Thailand strain). Results showed that exsheathed microfilariae represented only approximately 1 % of the total microfilaria midguts dissected at 5-min post-infected blood meal (PIBM). The percentage of exsheathed microfilariae found in midguts progressively increased to about 20, 60, 80, 90, and 100 % at 1-, 2–5-, 6–12-, 18–36-, and 48-h PIBM, respectively. Importantly, all the microfilariae penetrating the mosquito midguts were exsheathed. Midgut invasion by the exsheathed microfilariae was observed between 2- and 48-h PIBM. SEM analysis revealed sheathed microfilariae surrounded by small particles and maceration of the microfilarial sheath in the midguts, suggesting that the midguts of the refractory mosquitoes might have protein(s) and/or enzyme(s) and/or factor(s) that induce and/or accelerate exsheathment. The microfilariae penetrated the internal face of the peritrophic matrix (PM) by their anterior part and then the midgut epithelium, before entering the hemocoel suggesting that PM was not a barrier against the microfilariae migrating towards the midgut. Melanized microfilariae were discovered in the hemocoel examined at 96-h PIBM suggesting that the refractory mosquitoes used melanization reactions against this parasite. This study provided evidence that A. aegypti (Thailand strain) has refractory mechanisms against B. malayi in both midgut and hemocoel.     

Larvicidal activity of micronized aqueous suspension of calcium hydroxide against <Emphasis Type="Italic">Aedes aegypti</Emphasis> and <Emphasis Type="Italic">Culex quinquefasciatus</Emphasis> (Diptera: Culicidae)

In the search of alternatives for the control of mosquitoes of medical importance, we evaluated the larvicidal activity of micronized suspensions of calcium hydroxide [Ca(OH)₂] against Aedes aegypti and Culex quinquefasciatus. Tests conducted under laboratory conditions determined a LC50 = 0.027% (LC90 = 0.096%) for A. aegypti and a LC50 = 0.092% (LC90 = 0.2%) for C. quinquefasciatus, at 24 h post-treatment. Considering that the LC50 for the less susceptible species killed 100% of larvae for both species at 48 h post-treatment, we decided to use the diagnostic concentration of 0.1% which eliminated 100% of larvae at 48 h under a simulated field trial. There was a residual effect for up to 84 and 70 days for A. aegypti and C. quinquefasciatus, respectively. Evaluation of Ca(OH)₂ on breeding sites showed a larvicidal activity of 100% for up to 56 days. When the micronized Ca(OH)₂ was incorporated at concentrations from 0.02% (w/v), a superficial film was formed which killed 100% of the larvae of both species after 24 h of contact, and the activity remains until the film broke apart due to stirring. The fact that Ca(OH)₂ is cheap and the people in rural areas of Mexico and other countries know the handling procedures for this compound led us to consider that 0.1% suspensions of Ca(OH)₂ could be used for mosquito control in deposits of water placed in human premises both in urban and rural areas.     

Susceptibility of immature stages of <Emphasis Type="Italic">Aedes (Stegomyia) aegypti</Emphasis>; vector of dengue and chikungunya to insecticides from India

Susceptibility of Aedes aegypti to some insecticides in different geographic areas was conducted during dengue and chikungunya outbreak. At present, the only method of preventing dengue and chikungunya is to control the vector, which is the weakest link in vector-borne diseases. In our study, the susceptibility of A. aegypti collected from urban areas of Delhi, Mumbai, Jodhpur, Chennai and Coimbatore was evaluated against temephos, fenthion, malathion and DDT. The A. aegypti from different locations exhibited 0.33–7.11, 0.36–3.00, 0.65–2.84 and 2.16–20.8 fold more lethal concentration value of 50% (LC50) to temephos, fenthion, malathion and DDT, respectively, compared to susceptible reference strain. The result reveals that A. aegypti from various locations studied are still susceptible to temephos, fenthion and malathion, whereas low level of DDT resistance was noticed in field-collected A. aegypti. Amongst the insecticides tested, temephos was found to be relatively more effective in controlling A. aegypti, followed by fenthion, malathion and DDT.     

Cloning,expression, and characterization of salivary apyrase from <Emphasis Type="Italic">Aedes albopictus</Emphasis>

Apyrases (ATP diphosphohydrolase) hydrolyze the phosphodiester bonds of nucleoside tri- and diphosphates to orthophosphate and mononucleodides. They can inhibit platelet activation by depletion of adenosine diphosphate. In the current study, the Escherichia coli expression vector pET-19b equipped with an N-terminal histidine tag was applied to express the apyrase of Aedes albopictus. The gene-coding mature apyrase protein was amplified by RT-PCR and cloned into pET-19b. Soluble apyrase protein with high purity was successfully obtained by utilization of the suitable renaturation approach and Ni-NTA purification column. Four monoclonal antibodies to apyrase from A. albopictus were produced in male BALB/c mice immunized with the renatured apyrase. Using immunofluorescence assay and immunoblotting analysis, recombinant apyrase showed fine consistency with native apyrase. From kinetic analysis, it had a K _m of 11.6 μM and V _max of 1.02 nM/S/μg protein for adenosine triphosphate. Adenosine diphosphate-induced platelet aggregation was inhibited by approximately 6% when 0.4 μM recombinant apyrase was added and by about 9.5% when the concentration of recombinant apyrase was 0.8 μM. The effect on platelet aggregation was dose dependent. In conclusion, the apyrase of A. albopictus was cloned and expressed highly in the E. coli expression system. Recombinant apyrase protein showed biological activity, and anti-apyrase monoclonal antibody was also prepared.     

Oviposition-altering and ovicidal potentials of five essential oils against female adults of the dengue vector, <Emphasis Type="BoldItalic">Aedes aegypti</Emphasis> L.

The oviposition deterrence and ovicidal potential of five different essential oils, peppermint oil (Mentha piperita), basil oil (Ocimum basilicum), rosemary oil (Rosemarinus officinalis), citronella oil (Cymbopogon nardus), and celery seed oil (Apium graveolens), were assessed against female adults of the dengue vector, Aedes aegypti L. Multiple concentration tests were carried out where cups containing 1 mL of different concentrations (100%, 10%, 1%, 0.1%) of the oils and 199 mL of water were used for oviposition. The number of eggs laid and the larvae hatched in each cup were scored to evaluate the oviposition deterrent and ovicidal potentials of the oils. Our investigations revealed that the addition of 100% oil (pure oil) caused complete oviposition deterrence except in A. graveolens which resulted in 75% effective repellency. The use of 10% oil resulted in the maximum deterrence of 97.5% as shown by the M. piperita oil while other oils caused 36–97% oviposition deterrence as against the control. The oviposition medium with 1% oil showed decreased deterrent potential with 30–64% effective repellency, the M. piperita oil being exceptional. However, as the concentrations of the oil were reduced further to 0.1%, the least effective oil observed was A. graveolens (25% ER). Also, the M. piperita oil showed much reduced activity (40%) as compared to the control, while the other oils exhibited 51–58% repellency to oviposition. The studies on the ovicidal effects of these oils revealed that the eggs laid in the water with 100% essential oils did not hatch at all, whereas when 10% oils were used, only the R. officinalis oil resulted in 28% egg hatch. At lower concentrations (1%), the oils of M. piperita, O. basilicum, and C. nardus showed complete egg mortality while those of A. graveolens and R. officinalis resulted in 71% and 34% egg hatches, respectively. When used at 0.1%, the O. basilicum oil was found to be the only effective oil with 100% egg mortality, whereas other oils resulted in 16–76% egg mortality, the least mortality caused by the A. graveolens oil. These results suggest that these essential oils can be employed in a resistance-management program against A. aegypti. Further detailed research is needed to identify the active ingredient in the extracts and implement the effective mosquito management program.     

First report on susceptibility of wild <Emphasis Type="Italic">Aedes aegypti</Emphasis> (Diptera: Culicidae) using <Emphasis Type="Italic">Carapa guianensis</Emphasis> (Meliaceae) and <Emphasis Type="Italic">Copaifera</Emphasis> sp. (Leguminosae)

Oils of Carapa guianensis and Copaifera spp. are well known in the Amazonian region as natural insect repellent, and studies have reported their efficiency as larvicide against some laboratory mosquito species. However, in wild populations of mosquitoes, these oils have not yet been evaluated. Thus, the objective of this study was to investigate their efficiency as larvicide in wild populations of Aedes aegypti with a history of exposure to organophosphate. The susceptibility of larvae was determined under three different temperatures, 15°C, 20°C, and 30°C. For each test, 1,000 larvae were used (late third instar and early fourth instar—four replicates of 25 larvae per concentration). Statistical tests were used to identify significant differences. The results demonstrated that as the laboratory A. aegypti, the wild populations of A. aegypti were also susceptible to C. guianensis and Copaifera sp. oils. The lethal concentrations for Copaifera sp. ranged from LC₅₀ 47 to LC₉₀ 91 (milligrams per liter), and for C. guianensis, they were LC₅₀ 136 to LC₉₀ 551 (milligrams per liter). In relation to different temperature, the effectiveness of the oils on larvae mortality was directly related to the increase of temperature, and better results were observed for temperature at 25°C. The results presented here indicate the potential larvicidal activity of C. guianensis and species of Copaifera, in populations of A. aegypti from the wild. Therefore, the results presented here are very important since such populations are primarily responsible for transmitting the dengue virus in the environment.     

Efficacy of the <Emphasis Type="Italic">Trichophyton ajelloi</Emphasis> and <Emphasis Type="Italic">Lagenidium giganteum</Emphasis> metabolites against mosquitoes after flash chromatography

Entomopathogens are significant natural enemies for mosquitoes. We have investigated the adulticidal efficacies of metabolites of Trichophyton ajelloi and Lagenidium giganteum against Culex quinquefasciatus and Aedes aegypti simultaneously. The T. ajelloi was grown on Sabouraud’s dextrose broth medium at 25 ± 2°C and relative humidity at 75 ± 5% for 15 days. L. giganteum was grown in peptone yeast extract glucose broth at 25 ± 2°C and relative humidity 75 ± 5% for 15 days. The filtrations of metabolites have been made by using Whatman-1 filter paper then with the flash chromatograph. The bioassays were conducted as per the World Health Organization’s methods and protocols (2006). In this significant investigations, the metabolites of T. ajelloi have been found highly susceptible against A. aegypti with LC₉₉-7.24 ml after an exposure time of 24 h with a comparison, the LC₉₉-66 ml was observed against C. quinquefasciatus after exposure of 24 h. Moreover, the L. giganteum metabolites have shown higher pathogenicity against C. quinquefasciatus with LC₉₉-11.3 ml and A. aegypti with LC₉₉-15.49 ml. Although, the efficacy in adults could be achieved with higher concentration can be significant also. Their adulticidal activities in different climatic zones are plausible with metabolites which have better LT values of T. ajelloi.     

<Emphasis Type="Italic">Leishmania</Emphasis> (<Emphasis Type="Italic">Viannia</Emphasis>) <Emphasis Type="Italic">shawi</Emphasis> purified antigens confer protection against murine cutaneous leishmaniasis

Objective  

Leishmania (Viannia) shawi was characterized only recently, and few studies concerning the immunogenic and protective properties of its antigens have been performed. The present study aimed to evaluate the protective potential of the five antigenic fractions isolated from L. (V.) shawi promastigotes in experimental cutaneous leishmaniasis.     

Repellent and adulticide efficacy of a combination containing 10% imidacloprid and 50% permethrin against <Emphasis Type="Italic">Aedes aegypti</Emphasis> mosquitoes on dogs

This study was conducted to assess the repellent and adulticide efficacy of the combination containing 10% imidacloprid and 50% permethrin against Aedes aegypti mosquitoes on dogs. Blood-feeding success rates of the mosquitoes that were exposed to the treated dogs were 4.9 and 4.4% on days 3 and 7 post the combination application (PCA), respectively, and blood-feeding success rates increased to 6.3, 12.8, and 24.5% on days 14, 21, and 28 PCA, respectively. Blood-feeding success rates between the mosquitoes that were exposed to the treated and untreated control dogs on days 3, 7, 14, and 21 PCA were significantly different. All mosquitoes that were exposed to the treated dogs on day 3 PCA died, and mortality rates decreased to 97.1, 77.8, 40.4, and 2.1% on days 7, 14, 21, and 28 PCA, respectively. Mortality rates between the mosquitoes that were exposed to the treated and untreated control dogs on days 3, 7, 14, and 21 PCA were significantly different. This study suggested that this combination can be used to repel and kill mosquitoes on dogs; however, the application of this insecticide combination on dogs needs to be repeated every 3–4 weeks.     

Cercarial shedding from <Emphasis Type="Italic">Galba</Emphasis><Emphasis Type="Italic">truncatula</Emphasis> infected with <Emphasis Type="Italic">Fasciola</Emphasis><Emphasis Type="Italic">gigantica</Emphasis> of distinct geographic origins

Galba truncatula snails were experimentally infected with either of two different isolates of Fasciola gigantica, originating from Egypt or China, to determine the influence of these isolates on the characteristics of snail infections. The survival rates of G. truncatula on day 30 post-exposure were 90.0% and 60.2% in the Egyptian and Chinese groups, respectively. The frequency of cercaria-shedding snails within the Egyptian group was 79.8%, whereas in the Chinese group it was 22.4%. The parasite origin had a significant effect on the durations of the prepatent and patent periods. The mean number of cercariae shed from the Egyptian group was significantly greater than that shed from the Chinese group (a mean of 275.5 per cercaria-shedding snail compared with 29.0). These results could be explained by the fact that G. truncatula might be a natural intermediate host for F. gigantica in Egypt, and the greater adaptability of the Egyptian miracidia of F. gigantica to unusual snail hosts. These results demonstrate the influence of the geographic origin of the parasite on the success of trematodes infecting snails.     

Cysteine proteinases from promastigotes of <Emphasis Type="Italic">Leishmania</Emphasis> (<Emphasis Type="Italic">Viannia</Emphasis>) <Emphasis Type="Italic">braziliensis</Emphasis>

Leishmania (Viannia) braziliensis is the major causative agent of American tegumentary leishmaniasis, a disease that has a wide geographical distribution and is a severe public health problem. The cysteine proteinase B (CPB) from Leishmania spp. represents an important virulence factor. In this study, we characterized and localized cysteine proteinases in L. (V.) braziliensis promastigotes. By a combination of triton X-114 extraction, concanavalin A-affinity, and ion exchange chromatographies, we obtained an enriched fraction of hydrophobic proteins rich in mannose residues. This fraction contained two proteinases of 63 and 43 kDa, which were recognized by a CPB antiserum, and were partially sensitive to E-64 in enzymatic assays with the peptide Glu-Phe-Leu. In confocal microscopy, the CPB homologues localized in the peripheral region of the parasite. This data together with direct agglutination and flow cytometry assays suggest a surface localization of the CPB homologues. The incubation of intact promastigotes with phospholipase C reduced the number of CPB-positive cells, while anti-cross-reacting determinant and anti-CPB antisera recognized two polypeptides (63 and 43 kDa) derived from phospholipase C treatment, suggesting that some CPB isoforms may be glycosylphosphatidylinositol-anchored. Collectively, our results suggest the presence of CPB homologues in L. braziliensis surface and highlight the need for further studies on L. braziliensis cysteine proteinases, which require enrichment methods for enzymatic detection.     

<Emphasis Type="Italic">In vivo</Emphasis> activity of <Emphasis Type="Italic">Sapindus saponaria</Emphasis> against azole-susceptible and -resistant human vaginal <Emphasis Type="Italic">Candida</Emphasis> species

Background  

Study of in vivo antifungal activity of the hydroalcoholic extract (HE) and n-BuOH extract (BUTE) of Sapindus saponaria against azole-susceptible and -resistant human vaginal Candida spp.     

Functional polymorphisms in <Emphasis Type="Italic">XRCC</Emphasis>-<Emphasis Type="Italic">1</Emphasis> and <Emphasis Type="Italic">APE</Emphasis>-<Emphasis Type="Italic">1</Emphasis> contribute to increased apoptosis and risk of ulcerative colitis

Objective  

The present study was designed to investigate the role of X-ray cross-complementing group 1 (XRCC1) and apurinic/apyrimidinic endonuclease 1 (APE1) polymorphisms in apoptosis and the risk of ulcerative colitis (UC).