Inflammatory markers and bone mass in children with overweight/obesity: the role of muscular fitness

Objectives To examine which inflammatory markers are associated with bone mass and whether this association varies according to muscular fitness in children with overweight/obesity. Methods Plasma interleukin-1β (IL-1β), IL-6, tumor necrosis factor-α (TNF-α), epidermal growth factor, vascular endothelial growth factor A (VEGF), and C-reactive protein were analyzed in 55 children aged 8–11 years. A muscular fitness score was computed. Bone mineral content (BMC) of the total body-less head (TBLH) and lumbar spine (LS) were assessed using dual-energy x-ray absorptiometry. Results IL-6 (β = −0.136) and VEGF (β = −0.099) were associated with TBLH BMC, while TNF-α (β = −0.345) and IL-1β (β = 0.212) were associated with LS BMC (P < 0.05). The interaction effect of muscular fitness showed a trend in the association of VEGF with TBLH BMC (P = 0.122) and TNF-α with LS BMC (P = 0.057). Stratified analyses by muscular fitness levels showed an inverse association of VEGF with TBLH BMC (β = −0.152) and TNF-α with LS BMC (β = −0.491) in the low-fitness group, while no association was found in the high-fitness group. Conclusion IL-6, VEGF, TNF-α, and IL-1β are significantly associated with bone mass. Higher muscular fitness may attenuate the adverse effect of high VEGF and TNF-α on bone mass

Analysis of local and peripheral immune response developed in sheep experimentally infected with Toxoplasma gondii at different times of gestation

Trabajo presentado al: 4th International Meeting on Apicomplexa in Farm Animals. (11-14 October 2017 - Madrid, Spain).Toxoplasmosis is one the main infectious causes of reproductive failure in sheep where the time of gestation when sheep are infected affects the clinical and lesional outcome of the disease. In order to investigate the variations of local and peripheral immune responses during gestation, pregnant sheep were infected at early, mid and late gestation and subsequently culled at 2, 3 and 4 weeks post infection. In those sheep inoculated during the second term, serological antibodies were detected earlier and the increase in serological γ-IFN was higher than in the other infected animals. Regarding the local immune response at the placenta, infiltration of inflammatory cells was mainly found in the maternal septa, although it also invaded foetal mesenchyme adjacent to the lesions. The increase in the number of T lymphocytes was observed only in ewes infected during the second and last terms of gestation while the increase of B cells occurred in sheep infected at the first and second terms. The expression of iba-1 antigen by macrophages was more frequent after infection during the first term whereas macrophages expressing lysozyme, CD163 or calprotectin were more frequent in infections at mid-gestation. A significant increase in the transcription of γ-IFN, when compared to control animals, occurred after infections in the first and second term, while TNF-α and IL-10 transcription increased only in the second and last term, respectively. There were no differences when comparing transcription of cytokines between animals infected at different terms of gestation. This study shows that the time of gestation when infection occurs has a clear influence over the pathogenesis of ovine toxoplasmosis as a greater inflammatory response was found after inoculating sheep at the second term of gestation. This finding may explain the later invasion of the placenta by the parasite at early and mid gestation described in previous studies.This study was founded by AGL2016-75935-C2-2-R and LE080U16 research grants.Peer Reviewe

In Vitro versus in Mice: Efficacy and Safety of Decoquinate and Quinoline-O-Carbamate Derivatives against Experimental Infection with Neospora caninum Tachyzoites.

The effects of decoquinate (DCQ) and three O-quinoline-carbamate-derivatives were investigated using human foreskin fibroblasts (HFF) infected with Neospora caninum tachyzoites. These compounds exhibited half-maximal proliferation inhibition (IC50s) from 1.7 (RMB060) to 60 nM (RMB055). Conversely, when applied at 5 (DCQ, RMB054) or 10µM (RMB055, RMB060), HFF viability was not affected. Treatments of infected cell cultures at 0.5µM altered the ultrastructure of the parasite mitochondrion and cytoplasm within 24 h, most pronounced for RMB060, and DCQ, RMB054 and RMB060 did not impair the viability of splenocytes from naïve mice. Long-term treatments of N. caninum-infected HFF monolayers with 0.5µM of each compound showed that only exposure to RMB060 over a period of six consecutive days had a parasiticidal effect, while the other compounds were not able to kill all tachyzoites in vitro. Thus, DCQ and RMB060 were comparatively assessed in the pregnant neosporosis mouse model. The oral application of these compounds suspended in corn oil at 10 mg/kg/day for 5 d resulted in a decreased fertility rate and litter size in the DCQ group, whereas reproductive parameters were not altered by RMB060 treatment. However, both compounds failed to protect mice from cerebral infection and did not prevent vertical transmission/pup mortality. Thus, despite the promising in vitro efficacy and safety characteristics of DCQ and DCQ-derivatives, proof of concept for activity against neosporosis could not be demonstrated in the murine model

A Serosurvey of Selected Cystogenic Coccidia in Spanish Equids: First Detection of Anti-\u3cem\u3eBesnoitia\u3c/em\u3e spp. Specific Antibodies in Europe

Background: Equine besnoitiosis, caused by Besnoitia bennetti, and equine protozoal myeloencephalitis (EPM), caused by Sarcocystis neurona and Neospora hughesi are relevant equine diseases in the Americas that have been scarcely studied in Europe. Thus, a serosurvey of these cystogenic coccidia was carried out in Southern Spain. A cross-sectional study was performed and serum samples from horses (n = 553), donkeys (n = 85) and mules (n = 83) were included. An in-house enzyme-linked immunosorbent assay (ELISA) was employed to identify a Besnoitia spp. infection and positive results were confirmed by an a posteriori western blot. For Neospora spp. and Sarcocystis spp., infections were detected using in-house ELISAs based on the parasite surface antigens N. hughesi rNhSAG1 and S. neurona rSnSAG2/3/4. Risk factors associated with these protozoan infections were also investigated. Results: Antibodies against Besnoitia spp., Neospora spp. and Sarcocystis spp. infections were detected in 51 (7.1%), 46 (6.4%) and 20 (2.8%) of 721 equids, respectively. The principal risk factors associated with a higher seroprevalence of Besnoitia spp. were the host species (mule or donkey), the absence of shelter and the absence of a rodent control programme. The presence of rodents was the only risk factor for Neospora spp. infection. Conclusions: This study was the first extensive serosurvey of Besnoitia spp. infection in European equids accomplished by two complementary tests and gives evidence of the presence of specific antibodies in these populations. However, the origin of the infection is still unclear. Further parasite detection and molecular genotyping are needed to identify the causative Besnoitia and Neospora species. Finally, cross-reactions with antibodies directed against other species of Sarcocystis might explain the positive reactions against the S. neurona antigens

Isolation and genetic characterization of Toxoplasma gondii in Spanish sheep flocks

© The Author(s) 2020.[Background]: Toxoplasma gondii is a major cause of abortion in small ruminants and presents a zoonotic risk when undercooked meat containing cysts is consumed. The aim of the present study was to investigate the genetic diversity among the T. gondii strains circulating in ovine livestock in Spain.[Methods]: Selected samples collected from abortion outbreaks due to toxoplasmosis (n = 31) and from chronically infected adult sheep at slaughterhouses (n = 50) in different Spanish regions were bioassayed in mice, aiming at parasite isolation. In addition, all original clinical samples and the resulting isolates were genotyped by multi-nested PCR-RFLP analysis of 11 molecular markers and by PCR-DNA sequencing of portions of the SAG3, GRA6 and GRA7 genes.[Results]: As a result, 30 isolates were obtained from 9 Spanish regions: 10 isolates from abortion-derived samples and 20 isolates from adult myocardial tissues. Overall, 3 genotypes were found: ToxoDB#3 (type II PRU variant) in 90% (27/30) of isolates, ToxoDB#2 (clonal type III) in 6.7% (2/30), and ToxoDB#1 (clonal type II) in 3.3% (1/30). When T. gondii-positive tissue samples (n = 151) were directly subjected to RFLP genotyping, complete restriction profiles were obtained for 33% of samples, and up to 98% of the specimens belonged to the type II PRU variant. A foetal brain showed a clonal type II pattern, and four specimens showed unexpected type I alleles at the SAG3 marker, including two foetal brains that showed I + II alleles as co-infection events. Amplicons of SAG3, GRA6 and GRA7 obtained from isolates and clinical samples were subjected to sequencing, allowing us to confirm RFLP results and to detect different single-nucleotide polymorphisms.[Conclusions]: The present study informed the existence of a predominant type II PRU variant genotype (ToxoDB#3) infecting domestic sheep in Spain, in both abortion cases and chronic infections in adults, coexisting with other clonal (ToxoDB#1 and ToxoDB#2), much less frequent genotypes, as well as polymorphic strains as revealed by clinical sample genotyping. The use of multilocus sequence typing aided in accurately estimating T. gondii intragenotype diversity.This research was supported by projects funded by the Spanish Ministry of Science and Innovation (AGL2016-75935-C2-R) and the Community of Madrid (PLATESA2-CM-P2018/BAA-4370). MF and RC were funded by UCM-Santander/2017 pre-doctoral grants, and PLATESA2 post-doctoral grants, respectively. CG was funded by DGAPA, National Autonomous University of Mexico (UNAM). RC, EC and LO are part of the TOXOSOURCES consortium, supported by funding from the European Union’s Horizon 2020 Research and Innovation programme under grant agreement No. 773830: One Health European Joint Programme.Peer reviewe

Infección experimental por Toxoplasma gondii en el primer, segundo y último tercio de gestación en ovejas. Respuesta lesional y distribución del parásito

6 páginas.--Trabajo presentado al: XL Congreso Nacional y el XVI Congreso Internacional de la Sociedad Española de Ovinotecnia y Caprinotecnia. (Castellón de la Plana, España, 16-18 septiembre 2015).Peer Reviewe

Graphene-Assisted Integrated Nonlinear Optics

‎The unique linear and massless band structure of graphene in a purely two-dimensional Dirac fermionic structure has ignited intense research since the first monolayer graphene was isolated in the laboratory‎. ‎Not only does it offer new inroads into low-dimensional physics; graphene exhibits several peculiar properties that promise to widen the realm of opportunities for integrated optics and photonics‎. ‎This thesis is an attempt to shed light on the exceptional nonlinear optical properties of graphene and their potential applications in integrated photonics‎. ‎Following a theoretical exploration of light-graphene interaction‎, ‎disruptive new insight into the nonlinear optics of graphene was generated‎. ‎It now appears that graphene can efficiently enable photon-photon interaction in a fully integrated fashion‎. ‎This property‎, ‎taken together with ultrawideband tunability and ultrafast carrier dynamics could be fully exploited within integrated photonics for a variety of applications including harmonic generation and all-optical signal processing‎. ‎The multidisciplinary work described herein combines theoretical modeling and experimentation to proceed one step further toward this goal‎. ‎This thesis begins by presenting a semiclassical theory of light-graphene interaction‎. ‎The emphasis is placed on the nonlinear optical response of graphene from the standpoint of its underlying chiral symmetry‎. ‎The peculiar energy‎- ‎momentum dispersion of the quasiparticles in graphene entails a diverging field-induced interband coupling‎. ‎Following a many-body study of the carrier relaxations dynamics in graphene‎, ‎it will be shown that the charged carriers in the vicinity of the Dirac point undergo an unconventional saturation effect that can be induced by an arbitrarily weak electromagnetic field‎. ‎The perturbative treatment of the optical response of graphene is revisited and a theoretical model is developed to estimate the nonlinear optical coefficients including the Kerr coefficient of graphene‎. ‎The theoretical models are complimented by the experimental results‎. ‎The peculiar nonlinear optical properties of graphene together with its ablity to being integrated with optical platforms would render it possible to perform nonlinear optics in graphene integrated nanophotonic structures‎. ‎Here‎, ‎the suitability of graphene for nonlinear optical applications is investigated both theoretically and experimentally‎. ‎The emphasis is placed on an on-chip platform for ultrafast all-optical amplitude modulation‎. ‎The experimental results indicate strong all-optical modulation in a graphene-cladded planar photonic crystal nanocavity‎. ‎This development relies heavily on the unique properties of graphene‎, ‎including its fast carrier dynamics and the special phonon induced relaxation mechanism‎. ‎Finally‎, ‎the potential application of graphene based all-optical modulation in time resolved nonlinear spectroscopy is also discussed‎

Influencia de la variabilidad en la virulencia de diferentes aislados de Toxoplasma gondii sobre las lesiones de encéfalos fetales ovinos

Trabajo presentado a la: XXXII Reunión de la Sociedad Española de Anatomía Patológica Veterinaria (SEAPV). 1 octubre. Congreso virtual.Peer reviewe

Endochin-like quinolones (ELQs) and bumped kinase inhibitors (BKIs): Synergistic and additive effects of combined treatments against Neospora caninum infection in vitro and in vivo.

The apicomplexan parasite Neospora caninum is an important causative agent of congenital neosporosis, resulting in abortion, birth of weak offspring and neuromuscular disorders in cattle, sheep, and many other species. Among several compound classes that are currently being developed, two have been reported to limit the effects of congenital neosporosis: (i) bumped kinase inhibitors (BKIs) target calcium dependent protein kinase 1 (CDPK1), an enzyme that is encoded by an apicoplast-derived gene and found only in apicomplexans and plants. CDPK1 is essential for host cell invasion and egress; (ii) endochin-like quinolones (ELQs) are inhibitors of the cytochrome bc1 complex of the mitochondrial electron transport chain and thus inhibit oxidative phosphorylation. We here report on the in vitro and in vivo activities of BKI-1748, and of ELQ-316 and its respective prodrugs ELQ-334 and ELQ-422, applied either as single-compounds or ELQ-BKI-combinations. In vitro, BKI-1748 and ELQ-316, as well as BKI-1748 and ELQ-334, acted synergistically, while this was not observed for the BKI-1748/ELQ-422 combination treatment. In a N. caninum-infected pregnant BALB/c mouse model, the synergistic effects observed in vitro were not entirely reproduced, but 100% postnatal survival and 100% inhibition of vertical transmission was noted in the group treated with the BKI-1748/ELQ-334 combination. In addition, the combined drug applications resulted in lower neonatal mortality compared to treatments with single drugs