Frontal Non-Invasive Neurostimulation Modulates Antisaccade Preparation in Non-Human Primates

A combination of oculometric measurements, invasive electrophysiological recordings and microstimulation have proven instrumental to study the role of the Frontal Eye Field (FEF) in saccadic activity. We hereby gauged the ability of a non-invasive neurostimulation technology, Transcranial Magnetic Stimulation (TMS), to causally interfere with frontal activity in two macaque rhesus monkeys trained to perform a saccadic antisaccade task. We show that online single pulse TMS significantly modulated antisaccade latencies. Such effects proved dependent on TMS site (effects on FEF but not on an actively stimulated control site), TMS modality (present under active but not sham TMS on the FEF area), TMS intensity (intensities of at least 40% of the TMS machine maximal output required), TMS timing (more robust for pulses delivered at 150 ms than at 100 post target onset) and visual hemifield (relative latency decreases mainly for ipsilateral AS). Our results demonstrate the feasibility of using TMS to causally modulate antisaccade-associated computations in the non-human primate brain and support the use of this approach in monkeys to study brain function and its non-invasive neuromodulation for exploratory and therapeutic purposes

Blood Pressure, Heart Rate Variability, and Adiposity in Caribbean Pre-pubertal Children

International audienceChildhood obesity prevalence has increased over the last 30 years. The Heart Rate Variability (HRV) studies performed in adults suggest a possible relation between abnormal autonomic regulation and hypertension in the situation of overweight or obesity. Objective: The aims of this study were to explore the early relationships between adiposity and blood pressure and HRV in pre-pubertal children. Methods: Data were collected during the medical examination of the follow-up at 7 years of the TIMOUN mother-child cohort in Guadeloupe. Body Mass Index z-score (zBMI), sum of tricipital and subscapular skinfold thickness, percentage of fat mass, and Waist-to-Height Ratio were measured. A global corpulence score was computed using a Principal Component Analysis (PCA). Systolic Blood Pressure (SBP) and HRV parameters (cardiac holter monitoring) were collected under 2 conditions (calm and tachycardial period). Relations between HRV, SBP, each adiposity indicator and the corpulence score were studied with restricted cubic splines models, and linear regression models. The age at adiposity rebound (AR) was estimated from the individual growth curves. Results: 575 children were included in the SBP study (mean age: 7.7 years, from 85 to 99 months). SBP was linearly correlated with the corpulence score and the zBMI. An increase of 1 in the zBMI was associated with an increase of 2.3 (±0.28) mmHg in SBP. The effect-size of zBMI on SBP was higher in children with early age at AR. Compared to children with normal BMI, children with a zBMI 2SD, we observed a global increase in all HRV parameters (under tachycardial conditions), particularly the LF [β = 0.43 (±0.18)]. Conclusion: In pre-pubertal period a positive correlation between adiposity excess and SBP was observed with significant changes of HRV in boys, arguing for an early abnormal autonomic regulation and for early preventive intervention in the infancy period, particularly in case of overweight or obesity. Thinness was associated with a reduction in almost all the HRV parameters studied, when compared to normal corpulence, suggesting a decrease in autonomic influence

Developmental exposure to chlordecone induces transgenerational effects in somatic prostate tissue which are associated with epigenetic histone trimethylation changes

International audienceBACKGROUND: Chlordecone (CD), also known as Kepone, is an organochlorine insecticide that has been used in banana crops in the French West Indies. Due to long-term contamination of soils and water, the population is still exposed to CD. Exposure to CD in adulthood is associated with an increased risk of prostate cancer (PCa). OBJECTIVES: We examined the transgenerational effects of CD on murine prostate tissue. METHODS: We exposed pregnant Swiss mice to CD. The prostates from directly exposed (F1) and non-exposed (F3) male progeny were analyzed. We used immunofluorescence, RNA-seq and ChIP-seq techniques for the comprehensive analyses of chromatin states in prostate. RESULTS: We observed an increased prostatic intraepithelial neoplasia phenotype (PIN) in both F1 and F3 generations. Transcriptomic analysis in CD-derived F1 and F3 prostate using RNA-seq revealed that 970 genes in F1 and 218 in F3 genes were differentially expressed. The differentially expressed genes in both datasets could be clustered accordingly to common biological processes, "cell differentiation", "developmental process", "regulating of signaling", suggesting that in both generations similar processes were perturbed. We detected that in both datasets several Hox genes were upregulated; in F1, the expression was detected mainly in Hoxb and Hoxd, and in F3, in Hoxa family genes. Using a larger number of biological replicates and RT-qPCR we showed that genes implicated in testosterone synthesis (Akr1b3, Cyp11a1, Cyp17a1, Srd5a1) were dramatically upregulated in PIN samples; Cyp19a1, converting testosterone to estradiol was elevated as well. We found a dramatic increase in Esr2 expression both in F1 and F3 prostates containing PIN. The PIN-containing samples have a strong increase in expression of self-renewal-related genes (Nanog, Tbx3, Sox2, Sox3, Rb1). We observed changes in liver, F1 CD-exposed males have an increased expression of genes related to DNA repair, matrix collagen and inflammation related pathways in F1 but not in F3 adult CD-derived liver. The changes in RNA transcription were associated with epigenetic changes. Specifically, we found a global increase in H3K4 trimethylation (H3K4me3) and a decrease in H3K27 trimethylation (H3K27me3) in prostate of F1 mice. ChIP-seq analysis showed that 129 regions in F1 and 240 in F3 acquired altered H3K4me3 occupancy in CD-derived prostate, including highest increase at several promoters of Hoxa family genes in both datasets. The alteration in H3K4me3 in both generations overlap 73 genes including genes involved in proliferation regulation, Tbx2, Stat3, Stat5a, Pou2f3 and homeobox genes Hoxa13, Hoxa9. CONCLUSIONS: Our data suggest that developmental exposure to CD leads to epigenetic changes in prostate tissue. The PIN containing samples showed evidence of implication in hormonal pathway and self-renewal gene expression that have the capacity to promote neoplasia in CD-exposed mice

Determinants of children's exposure to pyrethroid insecticides in western France

Pyrethroids are insecticides frequently used in agriculture and in the home; exposure occurs through dietary and non-dietary pathways, including indoor and outdoor environmental contamination. Our objective was to study the potential determinants of pyrethroid metabolite concentrations measured in children's urine samples and in the dust of their homes. Specifically, we measured urinary metabolites from morning spot samples of 245 six-year- old children living in Brittany (France) in 2009-2012 and from dust vacuumed from the floor of their homes. Mothers reported home insecticide use, dietary habits, sociodemographic data; residential and school proximity to agricultural crops was assessed with spatialized data. The metabolites cis-DBCA, trans-DCCA, cisDCCA, 3-PBA, and F-PBA were detected in 84, 95, 64, 63, and 16% of the urine samples, respectively. Permethrin, cypermethrin, cyfluthrin, deltamethrin, and tetramethrin pyrethroids were detected in 100, 56, 9, 15, and 26% of the dust samples, respectively. Multiple regression analysis suggested diet plays a role in children's exposure, in particular, the food groups "pasta, rice or semolina" (for cis-DCCA and F-PBA), fruit (3-PBA), "breakfast cereals and whole grain bread" (cis-DBCA), and the global proportion of organic food in diet (for cis-DBCA, trans-DCCA). Children with a parent occupationally exposed to pesticides were about 3-times more likely to have higher urinary concentrations of 3-PBA (OR = 2.8, 95% CI [1.2; 6.5]). Dust content was correlated mainly with household insecticide use: higher mean concentrations of permethrin (beta = 0.8 [0.3; 1.3], in mu g/g) and an increased risk of a detectable level of cyfluthrin (OR = 4.7 [1.7; 12.9]) were observed in home dust, for indoor use of at least twice a year. Outdoor insecticide use at least once a year was associated with detection in dust of cypermethrin (OR = 3.0 [1.3; 6.7]) and tetramethrin (OR = 3.7 [1.6; 8.3]). Three positive and one negative correlations (out of 11) between urinary metabolite concentrations and home dust contents of their possible corresponding parent compounds were observed. The strength of this study lies in its concurrent use of biomarkers, environmental measurements, and potential sources of exposure. Its limitations include the use of a single urine sample and imprecise data about pyrethroid use in local agriculture

TMS coil positioning.

<p>Schematic drawing of the smallest of the commercially available coils, which was used for this experiment (<i>Upper panel</i>), a custom-made ∼25 mm loop radius figure-of-eight TMS coil (exact dimensions of the coil used indicated in the figure) (Magstim Company, Carmathenshire, Wales). (<i>Bottom panel)</i> X-rays photography of monkey ‘C’. The red target represents the estimated stereotaxic coordinates of the monkey’s FEF area. The length of the white bar illustrates approximate differences in bone thickness between the human and the macaque skull at frontal locations.</p

Behavioral paradigm illustrating the experimental antisaccade paradigm.

<p>(<i>Upper panel)</i> Antisaccade paradigm practiced by the two monkeys under the online impact of sham (left panel) or active (right panel) TMS single pulses. After fixating on a central stimulus (red), monkeys were to initiate a fast saccade to a location in the opposite direction with respect to a peripheral target (green) appearing on the screen, simultaneously (no gap) to the disappearance of the central fixation. Animals performed within each block, no-TMS trials (white small rectangles) yielding no stimulation at all (Upper Left) and TMS trials (grey small rectangles) during which a single TMS pulse was delivered at a given postarget onset SOA prior to the AS initiation, to modulate the planning of visually guided oculomotor activity <i>(Bottom panel)</i> Example of an experimental session. Animals performed a total of 4 blocks of AS training per session. In one of the blocks they did not receive TMS (white long rectangle), whereas in the remaining 3, they received in half of the trials TMS pulses (see long grey-filled rectangles) at one of the 3 intensities used in the study (30%, 40% and 50%). The order of the four blocks (3 TMS blocks at 30%, 40% or 50% absolute TMS intensities and 1 noTMS block) was randomized within each session. Monkeys performed 100 trials per block (50 no-TMS and 50 TMS trials) for a total of 400 trials per session, and received 50 pulses per TMS block (i.e., only in 50% of the trials), amounting to 150 pulses per experimental session. Independent sessions comprising active TMS pulses delivered at 100 ms or 150 ms SOA post target onset on the FEF, sham TMS pulses and active TMS stimulation in a control location were carried over.</p

Estimate of discomfort induced by TMS.

<p>Estimation of discomfort based on the interaction between percent of initiated trials (used as an indirect correlate of the level of discomfort; the higher the discomfort the lower the number of initiated trials) and TMS intensity (% of machine maximal output). (<i>Upper panel)</i> Note that below 50% intensity, low discomfort is inferred from the high percent of initiated trials (grey and black lines) respectively for monkey ‘C’ and ‘Y’, for the TMS condition (dotted lines) as compared to the sham TMS condition (solid lines). (<i>Bottom panel</i>) Representative traces of eye movement with/without TMS (respectively grey, black lines) at 50% intensity for one of the monkeys. At least for the two SOAs used in the study (100 and 150 ms) eye movement metrics were not affected by TMS. Furthermore, no saccades were elicited by the stimulation.</p

Schematic of TMS sites.

<p>Modified picture showing a top view of each of the two monkey’s scalp profiles (animals ‘Y’ and ‘C’), while posted and under training. The dotted line corresponds to the stereotaxic zero bar; the grey dot signals the location and size of the head-post; the orange dot corresponds to the location where digit movements were evoked by TMS pulses; the red dot FEF region of stimulation; the double white/grey dots is an approximate schematic representation of the TMS figure-of-eight coil which was located on the FEF region.</p

Roman dogs from the Iberian Peninsula and the Maghreb - A glimpse into their morphology and genetics

International audienceIn this study, we integrate osteometric and palaeogenetic data to investigate dog variability in the Roman Empire in Iberia and North Africa. Osteometry was used to distinguish the status-domestic or wild, of approximately 2000 years old Canis remains and to understand to what extent teeth and long bones varied in dogs in the Roman provinces of Mauretania Tingitana, Lusitania and Tarraconensis. High-throughput 454-DNA sequencing technology was used to obtain mitochondrial DNA (mtDNA) sequences from 15 bone and teeth samples. We identified five dog haplotypes from partial sequences of the hypervariable D-loop region. MtDNA haplotypes were grouped into two of the four major clades found in present-day dogs. We detected three clade A haplotypes in 12 samples from Portugal, Spain and Morocco, and a single clade D haplotype in 3 samples from Spain. So far, this is the oldest evidence for the presence of dog clade D in Iberia. It is dated to the late Roman occupation in the 4th-5th cent. AD (ca. 1,600 years ago). Our results confirm the existence of distinct dog morphotypes in Roman times that also harboured distinct genetic lineages. According to our data, dogs from distinct mtDNA lineages (clades A and D) have been continuously bred in the Iberian Peninsula since at least 1600 years ago. Moreover, the sharing of matrilines between dogs from Spain and North Africa may indicate gene flow. Dogs could have been easily transported between these regions by humans along maritime and terrestrial trade routes. These results provide new insights into pre-Roman and Roman domestication practices, confirming selection practices were extensively applied to dogs during the first centuries of our era in the Iberian Peninsula. We show that the greater size variability of teeth length (and consequently cranium) and long bone breadths (and consequently phenotype) of Roman dogs in the Iberian Peninsula, is concomitant with the detection of diverse and rare maternal lineages. This would reflect an intensification of dog breeding and the use of non-local dogs for breeding. (C) 2017 Elsevier Ltd and INQUA. All rights reserved