Effects of miltefosine treatment in fibroblast cell cultures and in mice experimentally infected with Neospora caninum tachyzoites

Miltefosine was investigated for its activity against Neospora caninum tachyzoites in vitro, and was shown to inhibit the proliferation of N. caninum tachyzoites cultured in human foreskin fibroblasts (HFF) with an IC50 of 5·2 μM. Treatment of infected cells with 25 μM miltefosine for a period of 10 h had only a parasitostatic effect, while after 20 h of treatment parasiticidal effects were observed. This was confirmed by transmission electron microscopy of N. caninum-infected and miltefosine-treated HFF. Administration of miltefosine to N. caninum-infected Balb/c female mice at 40 mg/kg/day for 14 days resulted in 6 out of 10 mice exhibiting weight loss, ruffled coat and apathy between days 7 and 13 post-infection. In the group that received placebo, only 2 out of 8 mice succumbed to infection, but the cerebral burden was significantly higher compared to the miltefosine treatment group. In a second experiment, the time-span of treatment was reduced to 5 days, and mice were maintained without further treatment for 4 weeks. Only 2 out of 9 mice in the miltefosine treatment group exhibited signs of disease, while 8 out of 10 mice succumbed to infection in the placebo group. These results showed that miltefosine hampered the dissemination of parasites into the CNS during experimental N. caninum infection in mic

Molecular cloning and characterization of NcROP2Fam-1, a member of the ROP2 family of rhoptry proteins in Neospora caninum that is targeted by antibodies neutralizing host cell invasion in vitro

Recent publications demonstrated that a fragment of a Neospora caninum ROP2 family member antigen represents a promising vaccine candidate. We here report on the cloning of the cDNA encoding this protein, N. caninum ROP2 family member 1 (NcROP2Fam-1), its molecular characterization and localization. The protein possesses the hallmarks of ROP2 family members and is apparently devoid of catalytic activity. NcROP2Fam-1 is synthesized as a pre-pro-protein that is matured to 2 proteins of 49 and 55 kDa that localize to rhoptry bulbs. Upon invasion the protein is associated with the nascent parasitophorous vacuole membrane (PVM), evacuoles surrounding the host cell nucleus and, in some instances, the surface of intracellular parasites. Staining was also observed within the cyst wall of ‘cysts' produced in vitro. Interestingly, NcROP2Fam-1 was also detected on the surface of extracellular parasites entering the host cells and antibodies directed against NcROP2Fam-1-specific peptides partially neutralized invasion in vitro. We conclude that, in spite of the general belief that ROP2 family proteins are intracellular antigens, NcROP2Fam-1 can also be considered as an extracellular antigen, a property that should be taken into account in further experiments employing ROP2 family proteins as vaccine

Intraperitoneal and intra-nasal vaccination of mice with three distinct recombinant Neospora caninum antigens results in differential effects with regard to protection against experimental challenge with Neospora caninum tachyzoites

Recombinant NcPDI(recNcPDI), NcROP2(recNcROP2), and NcMAG1(recNcMAG1) were expressed in Escherichia coli and purified, and evaluated as potential vaccine candidates by employing the C57Bl/6 mouse cerebral infection model. Intraperitoneal application of these proteins suspended in saponin adjuvants lead to protection against disease in 50% and 70% of mice vaccinated with recNcMAG1 and recNcROP2, respectively, while only 20% of mice vaccinated with recNcPDI remained without clinical signs. In contrast, a 90% protection rate was achieved following intra-nasal vaccination with recNcPDI emulsified in cholera toxin. Only 1 mouse vaccinated intra-nasally with recNcMAG1 survived the challenge infection, and protection achieved with intra-nasally applied recNcROP2 was at 60%. Determination of cerebral parasite burdens by real-time PCR showed that these were significantly reduced only in recNcROP2-vaccinated animals (following intraperitoneal and intra-nasal application) and in recNcPDI-vaccinated mice (intra-nasal application only). Quantification of viable tachyzoites in brain tissue of intra-nasally vaccinated mice showed that immunization with recNcPDI resulted in significantly decreased numbers of live parasites. These data show that, besides the nature of the antigen, the protective effect of vaccination also depends largely on the route of antigen delivery. In the case of recNcPDI, the intra-nasal route provides a platform to generate a highly protective immune respons

Synthesis of heteroarenes dyads from heteroarenes and heteroarylsulfonyl chlorides via Pd-catalyzed desulfitative C-H bond heteroarylations

International audienceWe report herein on the palladium-catalyzed direct heteroarylation of heteroarenes (e.g., pyrroles, furans, and thiophenes) in which heteroarylsulfonyl chlorides are used as coupling partners through a desulfitative cross-coupling. These C-H bond functionalizations occurred at the α-position in the case of pyrrole and furan derivs., while in the case of thiophenes the C-H bonds at the β-position have been heteroarylated. This methodol. represents a very simple route to heteroaryl dyads. Moreover, some examples of heteroaryl triads have been synthesized via iterative C-H bond arylations

N-[(2-Chloro-8-methyl­quinolin-3-yl)meth­yl]-4-meth­oxy­aniline

In the title compound, C18H17ClN2O, the quinoline ring system is essentially planar; the r.m.s. deviation for the non-H atoms is 0.04 Å with a maximum deviation from the mean plane of 0.026 (4) Å for the C atom bonded to the –CH2– group. The meth­oxy-substituted benzene ring forms a dihedral angle of 70.22 (4)° with this ring system. The crystal structure can be described as zigzag layers in which the quinoline ring systems are parallel to (011) and molecules are connected via inter­molecular N—H⋯N hydrogen bonds, forming chains along [100]. The crystal studied was an inversion twin with a 0.86 (5):0.14 (5) domain ratio

A DMAP-catalyzed mild and efficient synthesis of 1,2-dihydroquinazolines via a one-pot three-component protocol

International audienceAn efficient and simple method for the synthesis of 1,2-dihydroquinazolines catalyzed by 4-(N,N-dimethylamino) pyridine (DMAP) from readily available aromatic or heteroaromatic aldehydes, 2-aminobenzophenone, and ammonium acetate under mild conditions is described. The scope and limitations of the method are discussed

2-Hy­droxy-N-(4-meth­oxy­benz­yl)-4-nitro­anilinium chloride

The crystal structure of the title compound, C14H15N2O4 +·Cl−, can be described as being composed of layers containing both cations and anions that are staggered along [010]. Two types of the hydrogen bonds are observed, viz. cation–anion and cation–cation. The chloride anions are acceptors of the strong hydrogen bonds donated by the secondary amine and the hy­droxy groups. The packing is also stabilized by weak C—H⋯O inter­molecular hydrogen bonds. An intra­molecular N—H⋯O inter­action also occurs

Integration of the social dimension in urban wastelands reconversion projects : case of the Guelma railway station, Algeria

The railway station of Guelma (city located in the Northeast of Algeria) is an imposing building, built by the French colonization, which is closed now after so many years of prosperity. The development of the city and the urban sprawl have meant that after having been located for a long time on the periphery, this station now occupies a central position, where the city is now made. However, the future of this station is so far unclear since it will no longer be operational, given that the new Guelma’s railway station will be located on the outskirts of the city. What would be the fate of this architectural good left to itself? By preserving the memory of the station by offering it a new permanent use, this architectural jewel with its fallow site is a real opportunity for urban regeneration for the city. Therefore, the purpose of this paper is to try to give new life to Guelma station, through a new global approach that would channel the useful and sustainable reuse of the site from its immediate environment and the current executive context. The questionnaire was used as an investigative tool intended for residents living near the station, in total, 40 respondents have been surveyed, to find out their opinions and expectations relating to the future of this lost part of the city. The data from the study were descriptively analyzed, using percentages. Results revealed that the inhabitants are aware of the importance of the reconquest of the station, which will constitute for them a real opportunity for urban and economic development for the region. A reconquest that wants to be sustainable by adopting a socio-spatial approach, which will integrate socio-economic and environmental aspects

Synthesis of Some New 3-Pyrrolidinylquinoline Derivatives via 1,3-Dipolar Cycloaddition of Stabilized Azomethine Ylides to Quinolinyl α,β- Unsaturated Ketones

International audienceN-Metallated azomethine ylide generated from methyl (E)-N-benzylideneglycinate, LiBr and triethylamine underwent cycloaddition to quinolyl α,β- unsaturated ketones with excellent diastereoselectivity to afford new functionalised 3-pyrrolidinylquinoline derivatives

Diethyl 2,6-dimethyl­pyridine-3,5-dicarboxyl­ate at 100 K

In the structure of the title compound, C13H17NO4, the packing is stabilized by weak C—H⋯O and C—H⋯π inter­actions, resulting in the formation of a three-dimensional network