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Brain injuries cause hemodynamic changes in several distant, spared areas from the lesion. Our objective was to better understand the neuronal correlates of this reorganization in awake, behaving female monkeys. We used reversible inactivation techniques to “injure” the primary motor cortex, while continuously recording neuronal activity of the ventral premotor cortex in the two hemispheres, before and after the onset of behavioral impairments. Inactivation rapidly induced profound alterations of neuronal discharges that were heterogeneous within each and across the two hemispheres, occurred during movements of either the affected or nonaffected arm, and varied during different phases of grasping. Our results support that extensive, and much more complex than expected, neuronal reorganization takes place in spared areas of the bihemispheric cortical network involved in the control of hand movements. This broad pattern of reorganization offers potential targets that should be considered for the development of neuromodulation protocols applied early after brain injury.SIGNIFICANCE STATEMENTIt is well known that brain injuries cause changes in several distant, spared areas of the network, often in the premotor cortex. This reorganization is greater early after the injury and the magnitude of early changes correlates with impairments. However, studies to date have used noninvasive brain imaging approaches or have been conducted in sedated animals. Therefore, we do not know how brain injuries specifically affect the activity of neurons during the generation of movements. Our study clearly shows how a lesion rapidly impacts neurons in the premotor cortex of both hemispheres. A better understanding of these complex changes can help formulate hypotheses for the development of new treatments that specifically target neuronal reorganization induced by lesions in the brain.

The integration of the circular economy in agriculture has promoted sustainable innovation in food production systems such as horticulture. The present paper illustrates how horticulture is transitioning to the circular economy. This research field’s performance approaches and trends were assessed through a bibliometric and text-mining analysis of the literature. Our findings revealed that circular horticulture is a recent research field that is constantly growing. Its approach has been neither systemic nor integrative but fragmented. Bioeconomy, urban agriculture, recycled nutrients, biochar, fertigation, and desalination have been positioned as research hotspots. Vegetables and fruits are the most studied crops. Resource circulation has focused primarily on biowaste recovery to provide benefits such as biofertilizers and linear-substrate substitutes, and on water reuse for the establishment of hydroponic systems. The One Health approach is scarcely explored and, therefore, weakly articulated, wherein the absence of assessment methodologies encompassing the health of ecosystems, animals, and people is a notable limitation. Science-policy interfaces between One Health and food systems need to be improved. Lastly, greenhouse technologies are aligned with bioenergy, sustainable materials, and sensing technologies. Challenges and directions for future research have been raised to promote the redesign of horticultural production systems, integrating long-term circularity.

The development of neurostimulation techniques to evoke motor patterns is an active area of research. It serves as a crucial experimental tool to probe computation in neural circuits, and has applications in neuroprostheses used to aid recovery of motor function after stroke or injury to the nervous system. There are two important challenges when designing algorithms to unveil and control neurostimulation-to-motor correspondences, thereby linking spatiotemporal patterns of neural stimulation to muscle activation: (1) the exploration of motor maps needs to be fast and efficient (exhaustive search is to be avoided for clinical and experimental reasons) (2) online learning needs to be flexible enough to deal with noise and occasional spurious responses. We propose a stimulation search algorithm to address these issues, and demonstrate its efficacy with experiments in the motor cortex (M1) of a non-human primate model. Our solution is a novel iterative process using Bayesian Optimization via Gaussian Processes on a hierarchy of increasingly complex signal spaces. We show that our algorithm can successfully and rapidly learn correspondences between complex stimulation patterns and evoked muscle activation patterns, where standard approaches fail. Importantly, we uncover nonlinear circuit-level computations in M1 that would have been difficult to identify using conventional mapping techniques.

In rats, forelimb movements are evoked from two cortical regions, the caudal and rostral forelimb areas (CFA and RFA, respectively). These areas are densely interconnected and RFA induces complex and powerful modulations of CFA outputs. CFA and RFA also have interhemispheric connections, and these areas from both hemispheres send projections to common targets along the motor axis, providing multiple potential sites of interactions for movement production. Our objective was to characterize how CFA and RFA in one hemisphere can modulate motor outputs of the opposite hemisphere. To do so, we used paired-pulse protocols with intracortical microstimulation techniques (ICMS), while recording electromyographic (EMG) activity of forelimb muscles in sedated rats. A subthreshold conditioning stimulation was applied in either CFA or RFA in one hemisphere simultaneously or before a suprathreshold test stimulation in either CFA or RFA in the opposite hemisphere. Both CFA and RFA tended to facilitate motor outputs with short (0–2.5 ms) or long (20–35 ms) delays between the conditioning and test stimuli. In contrast, they tended to inhibit motor outputs with intermediate delays, in particular 10 ms. When comparing the two areas, we found that facilitatory effects from RFA were more frequent and powerful than the ones from CFA. In contrast, inhibitory effects from CFA on its homolog were more frequent and powerful than the ones from RFA. Our results demonstrate that interhemispheric modulations from CFA and RFA share some similarities but also have clear differences that could sustain specific functions these cortical areas carry for the generation of forelimb movements. NEW & NOTEWORTHY We show that caudal and rostral forelimb areas (CFA and RFA) have distinct effects on motor outputs from the opposite hemisphere, supporting that they are distinct nodes in the motor network of rats. However, the pattern of interhemispheric modulations from RFA has no clear equivalent among premotor areas in nonhuman primates, suggesting they contribute differently to the generation of ipsilateral hand movements. Understanding these interspecies differences is important given the common use of rodent models in motor control and recovery studies.

Nonhuman primate (NHP) movement kinematics have been decoded from spikes and local field potentials (LFPs) recorded during motor tasks. However, the potential of LFPs to provide network-like characterizations of neural dynamics during planning and execution of sequential movements requires further exploration. Is the aggregate nature of LFPs suitable to construct informative brain state descriptors of movement preparation and execution? To investigate this, we developed a framework to process LFPs based on machine-learning classifiers and analyzed LFP from a primate, implanted with several microelectrode arrays covering the premotor cortex in both hemispheres and the primary motor cortex on one side. The monkey performed a reach-to-grasp task, consisting of five consecutive states, starting from rest until a rewarding target (food) was attained. We use this five-state task to characterize neural activity within eight frequency bands, using spectral amplitude and pairwise correlations across electrodes as features. Our results show that we could best distinguish all five movement-related states using the highest frequency band (200-500 Hz), yielding an 87% accuracy with spectral amplitude, and 60% with pairwise electrode correlation. Further analyses characterized each movement-related state, showing differential neuronal population activity at above-γ frequencies during the various stages of movement. Furthermore, the topological distribution for the high-frequency LFPs allowed for a highly significant set of pairwise correlations, strongly suggesting a concerted distribution of movement planning and execution function is distributed across premotor and primary motor cortices in a specific fashion, and is most significant in the low ripple (100-150 Hz), high ripple (150-200 Hz), and multiunit frequency bands. In summary, our results show that the concerted use of novel machine-learning techniques with coarse grained queue broad signals such as LFPs may be successfully used to track and decode fine movement aspects involving preparation, reach, grasp, and reward retrieval across several brain regions.

Background Hypophosphatasia (HPP) is an inherited disorder characterized by defective mineralization of the bone and teeth that is also associated with a deficiency of serum alkaline phosphatase (ALP). Patients with HPP exhibit a broad range of symptoms including stillbirth with an unmineralized skeleton, premature exfoliation and dental caries in childhood, and pseudo-fractures in adulthood. The broad clinical spectrum of HPP is attributed to various mutations in the ALPL gene, which encodes tissue-nonspecific alkaline phosphatase (TNSALP). Nevertheless, the molecular mechanisms underlying the genotypic and phenotypic relationship of HPP remain unclear. Highlight The expression of HPP-related TNSALP mutants in mammalian cells allows us to determine for the effects of mutations on the properties of TNSALP, which could contribute to a better understanding of the relationship between structure and function of TNSALP. Conclusion Molecular characterization of TNSALP mutants helps establish the etiology and onset of HPP.

The Myrtaceae is one of the most common plant families in Brazil, including >1000 species of native fruit spread from North to South, where around 50% of all species are endemic to the Atlantic Rain Forest Biome. Most Brazilian species belong to Eugenia, Campomanesia, Psidium and Myrciaria genera. In general, they are characterized by the presence of leaves opposite, simple, entire, with pellucid glands containing ethereal oils and produces berry-like, fleshy fruits with a wide diversity of beneficial characteristics. Several parts of these plants are widely used in regional folk medicine as an astringent, anti-inflammatory, antihypertensive, to treat gastrointestinal disorders, and so on. Some species have already been studied and most of their metabolic effects are attributed to the presence of polyphenols, carotenoids as well as sesquiterpenes and monoterpenes. In addition, the Myrtaceae family has great economic potential, because of the sensory characteristics which encourages their commercial exploitation plus the presence of phytochemicals which play an important role in managing several degenerative chronic diseases besides representing a hotspot of technological innovation for food, cosmetic and pharmaceutical purposes. In this context, this review discusses about the importance of some Eugenia genera representatives such as Eugenia uniflora, Eugenia stipitata, Eugenia brasiliensis, Eugenia pyriformis and Eugenia dysenterica, emphasizing its profile of bioactive compounds as well as the knowledge of the nutritional and functional fruit potential, providing information for further studies aiming to stimulate the cultivation of Brazilian wild berries, for consumption and new food ingredients.

With few reported exceptions, G protein–coupled receptors (GPCRs) are modified by Cys palmitoylation (S-palmitoylation). In multiple GPCRs, S-palmitoylation targets a canonical site within the C-terminal cytoplasmic tail adjacent to the C terminus of the seventh transmembrane domain, but modification of additional sites is exemplified by the β-adrenergic receptors (βARs). The β(1)AR is S-palmitoylated at a second, more distal site within the C-terminal tail, and the β(2)AR is modified at a second site within the third intracellular loop, neither of which is conserved in other βAR isoforms. The functional roles of S-palmitoylation of disparate sites are incompletely characterized for any GPCR family. Here, we describe S-palmitoylation of the β(3)AR. We compared mouse and human β(3)ARs and found that both were S-palmitoylated at the canonical site within the C-terminal tail, Cys-358 and Cys-361/363 in mouse and human β(3)ARs, respectively. Surprisingly, the human β(3)AR was S-palmitoylated at two additional sites, Cys-153 and Cys-292 within the second and third intracellular loops, respectively. Cys-153 is apparently unique to the human β(3)AR, and Cys-292 is conserved primarily in primates. Mutational substitution of C-tail Cys in human but not mouse β(3)ARs resulted in diminished ligand-induced cAMP production. Substitution of Cys-153, Cys-292, or Cys-361/363 within the human β(3)AR diminished membrane-receptor abundance, but only Cys-361/363 substitution diminished membrane-receptor half-life. Thus, S-palmitoylation of different sites differentially regulates the human β(3)AR, and differential S-palmitoylation distinguishes human and rodent β(3)ARs, potentially contributing to species-specific differences in the clinical efficacy of β(3)AR-directed pharmacological approaches to disease.

The identification of key biodiversity areas (KBA) was initiated by the International Union for Conservation of Nature in 2004 to overcome taxonomic biases in the selection of important areas for conservation, including freshwater ecosystems. Since then, several KBAs have been identified mainly based on the presence of trigger species (i.e., species that trigger either the vulnerability and or the irreplaceability criterion and thus identify a site as a KBA). However, to our knowledge, many of these KBAs have not been validated. Therefore, classical surveys of the taxa used to identify freshwater KBAs (fishes, molluscs, odonates, and aquatic plants) were conducted in Douro (Iberian Peninsula) and Sebou (Morocco) River basins in the Mediterranean Biodiversity Hotspot. Environmental DNA analyses were undertaken in the Moroccan KBAs. There was a mismatch between the supposed and actual presence of trigger species. None of the trigger species were found in 43% and 50% of all KBAs surveyed in the Douro and Sebou basins, respectively. Shortcomings of freshwater KBA identification relate to flawed or lack of distribution data for trigger species. This situation results from a misleading initial identification of KBAs based on poor (or even inaccurate) ecological information or due to increased human disturbance between initial KBA identification and the present. To improve identification of future freshwater KBAs, we suggest selecting trigger species with a more conservative approach; use of local expert knowledge and digital data (to assess habitat quality, species distribution, and potential threats); consideration of the subcatchment when delineating KBAs boundaries; thoughtful consideration of terrestrial special areas for conservation limits; and periodic field validation., La identificación de las áreas clave de biodiversidad (ACB) fue iniciada por la Unión Internacional para la Conservación de la Naturaleza en 2004 con el objetivo de sobreponerse a los sesgos taxonómicos en la selección de áreas importantes para la conservación, incluyendo los ecosistemas de agua dulce. Desde entonces, varias ACB han sido identificadas principalmente con base en la presencia de especies desencadenantes (es decir, especies que desencadenan el criterio de vulnerabilidad o de carácter irremplazable y por lo tanto identifican a un sitio como una ACB). Sin embargo, a nuestro conocimiento, muchas de estas ACB no han sido validadas. Por lo tanto, los censos clásicos de taxones utilizados para identificar las ACB de agua dulce (peces, moluscos, odonatos y plantas acuáticas) fueron realizados en las cuencas de los ríos Duero (Península Ibérica) y Sebou (Marruecos) en el Punto Caliente de Biodiversidad del Mediterráneo. Realizamos análisis de ADN ambiental en las ACB de Marruecos. Hubo una discrepancia entre la supuesta presencia y la actual presencia de especies desencadenantes. Ninguna de las especies desencadenantes se encontró en 43% y 50% de las ACB censadas en las cuencas del Duero y del Sebou, respectivamente. Las deficiencias en la identificación de las ACB de agua dulce están relacionadas con la carencia de datos o datos erróneos sobre la distribución de las especies desencadenantes. Esta situación resulta en una identificación inicial engañosa de las ACB con base en información ecológica deficiente (o incluso incorrecta) o también puede deberse al incremento en las perturbaciones humanas ocurridas entre la identificación de la ACB y el presente. Para mejorar la identificación de ACB de agua dulce en el futuro, sugerimos que la selección de especies desencadenantes se realice con un enfoque más conservador; que se usen el conocimiento local de los expertos y los datos digitales (para evaluar la calidad del hábitat, la distribución de las especies y las amenazas potenciales); que se consideren las subcuencas cuando se delimiten las fronteras de las ACB; que se consideren cuidadosamente las áreas de especies terrestres para los límites de conservación; y que se realicen validaciones periódicas de campo., Financial support was provided by the Portuguese Foun dation for Science and Technology (FCT) Grant to J.N. (2020.04637.BD). We thank the editor and 3 anonymous ref erees for the valuable suggestions made, which increased the clarity of our manuscript. This study was partially funded by the MAVA Foundation through the action plan Ensuring Integrated Resource Management in River Basins.

Adult mammalian hearts are not regenerative. However, recent studies have evidenced that hypoxia enhances their regeneration. Islet1 (isl1) is known as a cardiac progenitor marker, which is quiescent in adult mammal hearts. In Xenopus hearts, transcriptional activation of isl1 was shown during cardiac regeneration of froglets at 3 months after metamorphosis. In this study, we examined transcriptional regulation of isl1 focusing on hypoxia-inducible factor 1α (hif1α) in Xenopus heart. We found that hif1α expression was increased in response to cardiac injury and overexpression of hif1α upregulated mRNA expression of isl1. Multiple conservation analysis including 9 species revealed that 8 multiple conserved regions (MCRs) were present upstream of isl1. DNA sequence analysis using JASPAR showed hif1α binding motifs in MCRs. By luciferase reporter assay and chromatin immunoprecipitation analysis, we found that hif1α directly bound to hif1α motifs in the most distant MCR8 and showed a specific transcriptional activity on the MCR8. In the luciferase assay using constructs carrying MCR8 without a responsive motif of hif1α, the reporter activity was lost. Pharmacologically inhibition of hif1α affected isl1 transcription and downstream events including cardiac phenotypes, suggesting functional defects of islet1. Contrarily in murine hearts, transcription of isl1 was unresponsive even after cryoinjury to adult hearts while hif1α mRNA was induced. In comparative analysis of multiple alignment, hif1α elements present in MCR8 of Xenopus or zebrafish were found to be disrupted as species are evolutionarily distant from Xenopus and zebrafish. Our results suggested an altered switch of isl1 transcription between mammals and Xenopus laevis.

Prompt responses to invasive Latrodectus spiders introduced unintentionally are needed worldwide due to their medical and ecological importance. Latrodectus species are chemically controlled using pyrethroid insecticides despite concerns about the ecological impacts of these compounds on biodiversity/ecosystems. Here, the relative sensitivities (acute toxicity: 48-h LC50) of Latrodectus hasseltii Thorell and Latrodectus geometricus C.L. Koch from Japan to the conventional neurotoxic insecticide bifenthrin (pyrethroid) and a new candidate insecticide, fipronil (phenylpyrazole), were examined. Acute residual toxicity tests of these compounds in two nontarget spiders (Parasteatoda tepidariorum C.L. Koch (Araneae: Theridiidae), Badumna insignis L. Koch (Araneae: Desidae)) were conducted for comparison. To test whether bifenthrin and fipronil toxicities differed among the four spiders, corresponding species sensitivity distributions (SSDs) were compared, and hazardous concentrations were determined. Sensitivity (especially in the nontarget species) was two to four orders of magnitude higher for bifenthrin than for fipronil. The SSD patterns of the two insecticides differed significantly, with the spider communities being more sensitive to bifenthrin than to fipronil. The lethal bifenthrin concentration for Latrodectus may reduce spider populations by over 70–90%. If L. hasseltii (established throughout Japan) is targeted for effective population suppression rather than L. geometricus (with a limited distribution range) using the specified insecticide concentration (LC50 value) for fipronil, less than 20% of spider communities will be impacted. Chemical operations aimed at the effective population management and subsequent eradication of invasive Latrodectus spiders while supporting local biodiversity conservation would benefit from considerations of fipronil dosages and target species sensitivities.

Serotonin (5-HT) is a common neurotransmitter in mammals, playing a central role in the regulation of various processes such as sleep, perception, cognitive and autonomic functions in the nervous system. Previous studies have demonstrated that 5-HT type 3 (5-HT

Ivermectin is the most common endectocide used to control parasites affecting livestock. Short-term physiological and behavioural effects of ivermectin on dung beetles may have long-term consequences for beetle populations and ecosystem functioning. Long-term effects of the use of ivermectin can be estimated by comparing dung assemblages and ecosystem functions in areas with conventional ivermectin-treated livestock and environmentally similar areas in which livestock are not treated with veterinary medical products (organic farming). In this study, we investigated both short-term and long-term effects of the administration of ivermectin on the characteristics of dung beetle assemblages and the services they provided in a protected area (Doñana National Park, SW Spain). We examined short-term dung colonization, dwelling, relocation, and disaggregation rates and the associations between these processes and the key assemblage parameters of species richness, abundance, biomass and functional diversity. Furthermore, we analysed changes in soil physical-chemical properties and processes. Short-term differences were observed in the total amount of dung relocated by dung beetles at different colonization vs. emigration stages, suggesting that dung beetles in this area were affected by the recent treatments of livestock with ivermectin. Moreover, short-term effects could also be responsible for the significant differences in dung spreading rates between sites. Conventional use of ivermectin disrupted ecosystem functioning by affecting species richness, abundance and biomass. The decrease in diversity parameters was related to a reduction in the functional efficiency, which resulted in the long-term accumulation of dung on the ground and considerable changes in soil functionality., Financial support was provided by the projects CGL2015-68207-R of the Secretaría de Estado de Investigación– Ministerio de Economía y Competitividad, and OAPN 762/2012 of the Organismo Autónomo de Parques Nacionales-Ministerio de Agricultura, Alimentación y Medio Ambiente.

This review examines the nutritional and functional aspects of some representatives of the Cactaceae family, as well as its technological potential in the most diverse industrial fields. The studied species are good sources of nutrients and phytochemicals of biological interest, such as phenolic compounds, carotenoids, betalains, phytosterols, tocopherols, etc. They also have shown great potential in preventing some diseases, including diabetes, obesity, cancer, and others. As to technological applications, the Cactaceae family can be explored in the production of food (e.g., cakes, yogurts, bread, ice cream, and juices), as natural dyes, sources of pectins, water treatment and in animal feed. In addition, they have great potential for many technological domains, including food chemistry, pharmacy, biotechnology, and many others.

A novel antifungal strategy targeting the inhibition of calcineurin is described. To develop a calcineurin based inhibitor of pathogenic fungi, analogs of FK506 were synthesized that were able to permeate mammalian but not fungal cells. Antagonists in combination with FK506 were not immunosuppressive and retained antifungal activity in A. fumigatus. To reduce the dosage burden of the antagonist, murine oral PK was improved an order of magnitude relative to previous FK506 antagonists.

The dorsal and ventral premotor cortices (PMd and PMv, respectively) each take part in unique aspects for the planning and execution of hand movements. These premotor areas are components of complex anatomical networks that include the primary motor cortex (M1) of both hemispheres. One way that PMd and PMv could play distinct roles in hand movements is by modulating the outputs of M1 differently. However, patterns of effects from PMd and PMv on the outputs of M1 have not been compared systematically. Our goals were to study how PMd within the same (i.e., ipsilateral or iPMd) and in the opposite hemisphere (i.e., contralateral or cPMd) can shape M1 outputs and then compare these effects with those induced by PMv. We used paired-pulse protocols with intracortical microstimulation techniques in sedated female cebus monkeys while recording EMG signals from intrinsic hand and forearm muscles. A conditioning stimulus was delivered in iPMd or cPMd concurrently or before a test stimulus in M1. The patterns of modulatory effects from PMd were compared with those from PMv collected in the same animals. Striking differences were revealed. Conditioning stimulation in iPMd induced more frequent and powerful inhibitory effects on M1 outputs compared with iPMv. In the opposite hemisphere, cPMd conditioning induced more frequent and powerful facilitatory effects than cPMv. These contrasting patterns of modulatory effects could allow PMd and PMv to play distinct functions for the control of hand movements and predispose them to undertake different, perhaps somewhat opposite, roles in motor recovery after brain injury. SIGNIFICANCE STATEMENT The dorsal and ventral premotor cortices (PMd and PMv, respectively) are two specialized areas involved in the control of hand movements in primates. One way that PMd and PMv could participate in hand movements is by modulating or shaping the primary motor cortex (M1) outputs to hand muscles. Here, we studied the patterns of modulation from PMd within the same and in the opposite hemisphere on the outputs of M1 and compared them with those from PMv. We found that PMd and PMv have strikingly different effects on M1 outputs. These contrasting patterns of modulation provide a substrate that may allow PMd and PMv to carry distinct functions for the preparation and execution of hand movements and for recovery after brain injury.

Lesion size and location affect the magnitude of impairment and recovery following stroke, but the precise relationship between these variables and functional outcome is unknown. Herein, we systematically varied the size of strokes in motor cortex and surrounding regions to assess effects on impairment and recovery of function. Female Sprague Dawley rats (N = 64) were evaluated for skilled reaching, spontaneous limb use, and limb placement over a 7 week period after stroke. Exploration and reaching were also tested in a free ranging, more naturalistic, environment. MRI voxel-based analysis of injury volume and its likelihood of including the caudal forelimb area (CFA), rostral forelimb area (RFA), hindlimb (HL) cortex (based on intracranial microstimulation), or their bordering regions were related to both impairment and recovery. Severity of impairment on each task was best predicted by injury in unique regions: impaired reaching, by damage in voxels encompassing CFA/RFA; hindlimb placement, by damage in HL; and spontaneous forelimb use, by damage in CFA. An entirely different set of voxels predicted recovery of function: damage lateral to RFA reduced recovery of reaching, damage medial to HL reduced recovery of hindlimb placing, and damage lateral to CFA reduced recovery of spontaneous limb use. Precise lesion location is an important, but heretofore relatively neglected, prognostic factor in both preclinical and clinical stroke studies, especially those using region-specific therapies, such as transcranial magnetic stimulation. SIGNIFICANCE STATEMENT By estimating lesion location relative to cortical motor representations, we established the relationship between individualized lesion location, and functional impairment and recovery in reaching/grasping, spontaneous limb use, and hindlimb placement during walking. We confirmed that stroke results in impairments to specific motor domains linked to the damaged cortical subregion and that damage encroaching on adjacent regions reduces the ability to recover from initial lesion-induced impairments. Each motor domain encompasses unique brain regions that are most associated with recovery and likely represent targets where beneficial reorganization is taking place. Future clinical trials should use individualized therapies (e.g., transcranial magnetic stimulation, intracerebral stem/progenitor cells) that consider precise lesion location and the specific functional impairments of each subject since these variables can markedly affect therapeutic efficacy.

Premotor areas of primates are specialized cortical regions that can contribute to hand movements by modulating the outputs of the primary motor cortex (M1). The goal of the present work was to study how the supplementary motor area (SMA) located within the same hemisphere [i.e., ipsilateral SMA (iSMA)] or the opposite hemisphere [i.e., contralateral (cSMA)] modulate the outputs of M1. We used paired-pulse protocols with intracortical stimulations in sedated capuchin monkeys. A conditioning stimulus in iSMA or cSMA was delivered simultaneously or before a test stimulus in M1 with different interstimulus intervals (ISIs) while electromyographic activity was recorded in hand and forearm muscles. The pattern of modulation from iSMA and cSMA shared some clear similarities. In particular, both areas predominantly induced facilitatory effects on M1 outputs with shorter ISIs and inhibitory effects with longer ISIs. However, the incidence and strength of facilitatory effects were greater for iSMA than cSMA. We then compared the pattern of modulatory effects from SMA to the ones from the dorsal and ventral premotor cortexes (PMd and PMv) collected in the same series of experiments. Among premotor areas, the impact of SMA on M1 outputs was always weaker than the one of either PMd or PMv, and this was regardless of the hemisphere, or the ISI, tested. These results show that SMA exerts a unique set of modulations on M1 outputs, which could support its specific function for the production of hand movements. NEW & NOTEWORTHY We unequivocally isolated stimulation to either the ipsilateral or contralateral supplementary motor area (SMA) using invasive techniques and compared their modulatory effects on the outputs of primary motor cortex (M1). Modulations from both SMAs shared many similarities. However, facilitatory effects evoked from ipsilateral SMA were more common and more powerful. This pattern differs from the ones of other premotor areas, which suggests that each premotor area makes unique contributions to the production of motor outputs.

The ventral premotor cortex (PMv) is a key node in the neural network involved in grasping. One way PMv can carry out this function is by modulating the outputs of the primary motor cortex (M1) to intrinsic hand and forearm muscles. As many PMv neurons discharge when grasping with either arm, both PMv within the same hemisphere (ipsilateral; iPMv) and in the opposite hemisphere (contralateral; cPMv) could modulate M1 outputs. Our objective was to compare modulatory effects of iPMv and cPMv on M1 outputs to intrinsic hand and forearm muscles. We used paired-pulse protocols with intracortical microstimulations in capuchin monkeys. A conditioning stimulus was applied in either iPMv or cPMv simultaneously or prior to a test stimulus in M1 and the effects quantified in electromyographic signals. Modulatory effects from iPMv were predominantly facilitatory, and facilitation was much more common and powerful on intrinsic hand than forearm muscles. In contrast, while the conditioning of cPMv could elicit facilitatory effects, in particular to intrinsic hand muscles, it was much more likely to inhibit M1 outputs. These data show that iPMv and cPMv have very different modulatory effects on the outputs of M1 to intrinsic hand and forearm muscles.

In primates, the refinement of motor behaviors, in particular hand use, is associated with the establishment of more direct projections from primary motor cortex (M1) onto cervical motoneurons [1, 2] and the appearance of additional premotor and sensory cortical areas [3]. All of these areas have reciprocal connections with M1 [4-7]. Thus, during the evolution of the sensorimotor network, the number of interlocutors with which M1 interacts has tremendously increased. It is not clear how these additional interconnections are organized in relation to one another within the hand representation of M1. This is important because the organization of connections between M1 and phylogenetically newer and specialized cortical areas is likely to be key to the increased repertoire of hand movements in primates. In cebus monkeys, we used injections of retrograde tracers into the hand representation of different cortical areas of the sensorimotor network (ventral and dorsal premotor areas [PMv and PMd], supplementary motor area [SMA], and posterior parietal cortex [area 5]), and we analyzed the pattern of labeled neurons within the hand representation of M1. Instead of being uniformly dispersed across M1, neurons sending projections to each distant cortical area were largely segregated in different subregions of M1. These data support the view that primates split the cortical real estate of M1 into modules, each preferentially interconnected with a particular cortical area within the sensorimotor network. This modular organization could sustain parallel processing of interactions with multiple specialized cortical areas to increase the behavioral repertoire of the hand.

Background: Microstimulation of brain tissue plays a key role in a variety of sensory prosthetics, clinical therapies and research applications. However, the effects of stimulation parameters on the responses they evoke remain widely unknown. Objective: We aimed to investigate the contribution of each stimulation parameter to the response and identify interactions existing between parameters. Methods: Parameters of the constant-current, biphasic square waveform were examined in acute terminal experiments under ketamine anaesthesia. The motor cortex of 7 Sprague-Dawley rats was stimulated while recording motor evoked potentials (MEP) from the forelimb. Intracortical microstimulation (ICMS) parameters were systematically tested in a pair-wise fashion to observe the influence of each parameter on the amplitude and latency of the MEP. Results: The amplitude of the MEP increased continually with stimulus amplitude (p

Tissue-nonspecific alkaline phosphatase (TNSALP) is a membrane glycoprotein with a proposed role in bone mineralization. Indeed, mutations in TNSALP have been identified in patients with hypophosphatasia (HPP), a genetic disease characterized by hypomineralization of bone and teeth and a deficiency in serum ALP activity. TNSALP has five potential N-glycosylation sites at N140, N230, N271, N303 and N430 by standard nomenclature. A mutation at one of these sites, N430, was recently detected in a patient with infantile HPP. Using site-directed mutagenesis, we demonstrated that TNSALP has five N-glycans in transfected COS-1 cells and that individual single N-glycan deletion mutants of TNSALP retain the dimeric structure required for ALP activity, excluding the possibility that any single N-glycan plays a vital role in the structure and function of TNSALP. However, we found that TNSALP (N430Q) and TNSALP (N430E) mutants, but not a TNSALP (N430D) mutant, failed to form dimers. The TNSALP (N430S) mutant linked to infantile HPP was glycosylation-defective and unable to dimerise, similar to TNSALP (N430Q) and TNSALP (N430E) mutants; therefore, TNSALP (N430S) was established as a severe allele without strong ALP activity. By contrast to individual single N-glycan deletion mutants, TNSALP devoid of all five N-glycans was present to a much lesser extent than wild-type TNSALP in transfected cells, possibly reflecting its instability. A comprehensive analysis of a series of multiple N-glycan depletion mutants in TNSALP revealed that three N-glycans on N230, N271 and N303 were the minimal requirement for the structure and function of TNSALP and a prerequisite for its stable expression in a cell.

Class switch recombination (CSR) is the process by which B cells switch production from IgM/IgD to other immunoglobulin isotypes, enabling them to mount an effective immune response against pathogens. Timely resolution of CSR prevents damage due to an uncontrolled and prolonged immune response. While many positive regulators of CSR have been described, negative regulators of CSR are relatively unknown. Using an shRNA library screen targeting more than 28,000 genes in a mouse B cell line, we have identified a novel, uncharacterized protein of 82kD (KIAA1841, NM_027860), which we have named SANBR (SANT and BTB domain regulator of CSR), as a negative regulator of CSR. The purified, recombinant BTB domain of SANBR exhibited characteristic properties such as homodimerization and interaction with corepressor proteins, including HDAC and SMRT. Overexpression of SANBR inhibited CSR in primary mouse splenic B cells, and inhibition of CSR is dependent on the BTB domain while the SANT domain is largely dispensable. Thus, we have identified a new member of the BTB family that serves as a negative regulator of CSR. Future investigations to identify transcriptional targets of SANBR in B cells will reveal further insights into the specific mechanisms by which SANBR regulates CSR as well as fundamental gene regulatory activities of this protein.

Aims Animal studies using various stress models have shown that excessive environmental stress induces depression? and anxiety?like behaviors through inflammatory responses in the brain and periphery. Although the leptomeningeal cells have multiple functions related to inflammatory responses in the brain, whether environmental stress influences the leptomeninges remains unknown. In this study, we aimed to examine phosphorylation of the extracellular signal‐regulated kinase (ERK) in the leptomeninges. Methods We subjected C57BL/6 male mice to a single episode of social defeat stress and analyzed the expression of phosphorylated ERK in the leptomeninges by immunohistochemistry. Results Social defeat stress in mice induced phosphorylation of ERK in the leptomeninges, adjacent to vascular endothelial cells and the glia limitans. This ERK phosphorylation was maintained for at least one hour after the stress. Conclusions This study shows the effect of environmental stress on the leptomeninges for the first time and paves the way for elucidating its functional role in stress‐induced changes in neural functions., This study shows for the first time that social defeat stress induces phosphorylation of extracellular signal‐regulated kinase in the leptomeninges in mice. This finding paves the way for elucidating a potential role of the leptomeninges in mediating stress‐induced inflammatory signals from the periphery to the brain.

The aim of this study was to determine the heavy metals content of milk from cows fed with forage irrigated with waste water from industrial sources and the health risk to children. Milk samples were taken from milk collection tanks of eight localities in the state of Puebla. On average, the heavy metals in the milk had the following order Zn As Pb Cr Cu Ni. Pb (0.03 mg kg

An iridescent yellow pigmented bacterium isolated from the Antarctic continent, named Cellulophaga fucicola strain 416, was found to be able to tolerate UV-B radiation. Its crude pigment extract was tested for antioxidant capacity, UV light stability and phototoxicity profile against murine fibroblast lines. The pigments were further isolated and chemically identified by ultra-high-performance liquid chromatography with photodiode array and mass spectrometry detectors. The results showed that the pigment extract presented weak stability under exposure to UV light, a phototoxic profile in the 3t3 Neutral Red Uptake test and a very high antioxidant activity, suggesting that it could be used as food and feed colourants. Zeaxanthin and two isomers of zeaxanthin, β-cryptoxanthin and β-carotene, were identified using a C18 column. These five carotenoids were the major pigments isolated from C. fucicola 416. In conclusion, the identification of pigments produced by the bacterial strain under study may help us understand how bacteria thrive in high UV and cold environments, and opens avenues for further biotechnological application towards a more sustainable and environmentally friendly way of pigment exploitation.

The primary motor cortex (M1) plays an essential role in the control of hand movements in primates and is part of a complex cortical sensorimotor network involving multiple premotor and parietal areas. In a previous study in squirrel monkeys, we found that the ventral premotor cortex (PMv) projected mainly to 3 regions within the M1 forearm representation [rostro-medial (RM), rostro-lateral (RL), and caudo-lateral (CL)] with very few caudo-medial (CM) projections. These results suggest that projections from premotor areas to M1 are not uniform, but rather segregated into subregions. The goal of the present work was to study how inputs from diverse areas of the ipsilateral cortical network are organized within the M1 hand representation. In Cebus apella, different retrograde neuroanatomical tracers were injected in 4 subregions of the hand area of M1 (RM, RL, CM, and CL). We found a different pattern of input to each subregion of M1. RM receives inputs predominantly from dorsal premotor cortex, RL from PMv, CM from area 5, and CL from area 2. These results support that the M1 hand representation is composed of several subregions, each part of a unique cortical network.

Hypophosphatasia (HPP) is a genetic disease characterized by defective calcification of hard tissues such as bone and teeth accompanying deficiency of serum alkaline phosphatase (ALP) activity. Its development results from various mutations in the ALPL gene encoding tissue-nonspecific ALP (TNSALP). HPP is known to be transmitted in an autosomal recessive or autosomal dominant manner. A point mutation (c.323C>T) in the ALPL gene leading to a proline to leucine substitution at position 108 of TNSALP was first reported in a patient diagnosed with odonto-HPP (M Herasse et al., J Med Genet 2003;40:605–609), although the effects of this mutation on the TNSALP molecule have not been elucidated. To understand the molecular basis of this dominantly transmitted HPP, we first characterized TNSALP (P108L) by expressing it in COS-1 cells transiently. In contrast to wild-type TNSALP (WT), TNSALP (P108L) showed virtually no ALP activity. When coexpressed with TNSALP (WT), TNSALP (P108L) significantly inhibited the enzyme activity of TNSALP (WT), confirming that this mutant TNSALP exerts a dominant negative effect on TNSALP (WT). Using immunofluorescence and digestion with phosphatidylinositol-specific phospholipase C, we demonstrated that TNSALP (P108L) was anchored to the cell surface via glycosylphosphatidylinositol-like TNSALP (WT) in a Tet-On CHO cell expression system. Consistent with this, TNSALP (P108L) acquired endo- β - N -acetylglucosaminidase H resistance and sialic acids, as evidenced by glycosidase treatments. Importantly, TNSALP (WT) largely formed a functional dimeric structure, while TNSALP (P108L) was found to be present as a monomer in the cell. This indicates that the molecular structure of TNSALP is affected by a missense mutation at position 108, which is in contact with the active site, such that it no longer assembles into the functional dimeric form. Collectively, these results may explain why TNSALP (P108L) loses its ALP activity, even though it is able to gain access to the cell surface.

In rats, forelimb movements can be evoked from two distinct cortical regions, the rostral (RFA) and the caudal (CFA) forelimb areas. RFA and CFA have numerous reciprocal connections, and their projections reach several common targets, which allows them to interact at multiple levels of the motor axis. Lesions affecting these areas result in profound and persistent deficits, supporting their essential role for the production of arm and hand movements. Whereas rats are widely used to study motor control and recovery following lesions, little is known as to how cortical motor areas in this model interact to generate movements. To study interactions between RFA and CFA, we used paired-pulse protocols with intracortical microstimulation techniques (ICMS). A conditioning stimulus (C) in RFA was applied simultaneously, or before a test stimulus (T) in CFA. The impact of RFA conditioning on CFA outputs was quantified by recording electromyographic signals (EMG) signals from the contralateral arm muscles. We found that stimulation of RFA substantially modulates the intensity of CFA outputs while only mildly affecting the latency. In general, the effect of RFA conditioning changed from predominantly facilitatory to inhibitory with increasing delays between the C and the T stimulus. However, inspection of individual cortical sites revealed that RFA has a wide range of influence on CFA outputs with each interstimulation delay we used. Our results show that RFA has powerful and complex modulatory effects on CFA outputs that can allow it to play a major role in the cortical control of forelimb movements.

© The Author(s) 2018., Among macrocyclic lactones (ML), ivermectin (IVM) and moxidectin (MOX) potentially affect all Ecdysozoan species, with dung beetles being particularly sensitive. The comparative effects of IVM and MOX on adult dung beetles were assessed for the first time to determine both the physiological sub-lethal symptoms and pre-lethal consequences. Inhibition of antennal response and ataxia were tested as two intuitive and ecologically relevant parameters by obtaining the lowest observed effect concentration (LOEC) values and interpolating other relevant toxicity thresholds derived from concentration-response curves (IC50, as the concentration of each ML where the antennal response is inhibited by half; and pLC50, as the quantity of ingested ML where partial paralysis was observed by half of treated individuals) from concentration-response curves. Both sub-lethal and pre-lethal symptoms obtained in this study coincided in that IVM was six times more toxic than MOX for adult dung beetles. Values of LOEC, IC50 and pLC50 obtained for IVM and MOX evaluated in an environmental context indicate that MOX, despite needing more time for its elimination in the faeces, would be half as harmful to dung beetles as IVM. This approach will be valuable to clarify the real impact of MLs on dung beetle health and to avoid the subsequent environmental consequences., Financial support was provided by the project CGL2015-68207-R of the Secretaría de Estado de Investigación–Ministerio de Economía y Competitividad.

The aim of this work was to determine Ni, Cr, Cu, Zn, Pb, and As levels in raw milk and Oaxaca and ranchero type cheeses, produced in areas irrigated with waste water from Puebla in Mexico. Milk results showed a mean Pb level of 0.03 mg kg−1, which is above the maximum limit as set by Codex Alimentarius and the European Commission standards. For As a mean value of 0.12 mg kg−1 in milk was obtained. Mean As and Pb levels in milk were below the Mexican standard. Milk whey and ranchero cheese had mean Pb levels of 0.07 and 0.11 mg kg−1, respectively. As was higher in Oaxaca and ranchero cheese at 0.17 and 0.16 mg kg−1, respectively. It was concluded that cheeses made from cow’s milk from areas irrigated with waste water are contaminated with Pb and As, which may represent a health risk.

In previous studies, we reported that pretreatment with the antioxidant Tempol attenuated the development and expression of cocaine-induced psychomotor sensitization in rats and diminished cocaine-induced oxidative stress (OS) in the prefrontal cortex (PFC) and nucleus accumbens (NAc), suggesting a potential role for Tempol in interfering with cocaine-related psychomotor sensitization. The aim of the current study was to examine the role of Tempol in reward and reinforcement using the conditioned place preference (CPP) paradigm. We found that administration of Tempol during the conditioning session abolished the expression of cocaine-induced CPP. We also found that OS was significantly elevated following the establishment of CPP, and that cocaine-induced OS was significantly diminished by pretreatment with Tempol during conditioning. Furthermore, we found that repeated, but not single, administration of Tempol for seven days during withdrawal from CPP resulted in significant attenuation in the expression of CPP. Moreover, Tempol did not affect the expression of food reward. Taken together, these findings provide evidence for the involvement of Tempol in regulating cocaine rewarding properties without affecting natural rewards. Since Tempol was found to be effective in reducing OS and expression of CPP following withdrawal, it may be a potential treatment for cocaine addiction.

Stroke recovery research involves distinct biological and clinical targets compared to the study of acute stroke. Guidelines are proposed for the pre-clinical modeling of stroke recovery and for the alignment of pre-clinical studies to clinical trials in stroke recovery.

This study aimed to determine the heavy metal content in cow’s milk produced in areas irrigated with waste water and to evaluate the health risk with daily consumption of milk for children. The sample consisted of four zones in which small farmers were selected and the milk of 160 cows in two seasons of the year. On average, the metals in the milk in decreasing order were 0.36; 0.046; 0.035; 0.029; 0.015; 0.012, and 0.002 mg kg−1 for Zn, Pb, As, Cu, Cr, Ni, and Cd, respectively. The Pb exceeded the limits allowed by Codex. The values shown in the hazard quotient for the As of more than 1 and HI were higher. On the other hand, the individual risk of cancer showed a descending order As> Cd> Cr> Pb, while the total risk indicated that the combined effect of metals put girls and boys at serious risk.

Growing awareness in favor of innovative and healthier alternatives is creating a noticeable shift from synthetic colorants to natural additives. And, such a swing in the consumer market is growing slowly but noticeably. In this context, genipap (Genipa americana L.) fruit represents an emerging source of blue colorants in Latin America with extensive application possibilities. This is despite the fact that there are few studies concerning its toxicity predictive factors. In this early-stage study we propose to investigate safety issues around genipap extract (IBBP); we also attempt to identify fingerprint profiling of both IBBP extract and solid lipid microparticles containing IBBP extract (SLM-IBBP) using in vitro assays. The main compounds identified were genipin, and genipin 1-β-gentiobioside. Results indicated that IBBP extract, at 25 µg/mL, was able to promote DNA damage in CHO-K1 cells, suggesting a genotoxic effect. On the other hand, the SLM-IBBP inhibited almost all cancer cell lines with GI50 ranging from 0.25 μg/mL to 43.5 μg/mL. Also, IBBP-SLM seems to exert a desirable apoptosis induction (at 25 µg/mL dosage). The next steps for our work, therefore, will focus on other nanoparticle formulation approaches, in particular with the use of natural Brazilian starch. An evaluation of the metabolism and distribution of microparticles, and their safety for food and pharmaceutical purposes, are also required.

We recently reported that dopamine D1 receptor in the medial prefrontal cortex (mPFC) is activated by subthreshold social defeat stress and suppresses the induction of depressive-like behavior in mice. However, which mPFC projection(s) mediates this antidepressant-like effect remains poorly understood. Here we show that social defeat stress specifically increased c-Fos expression, a marker for neuronal activity, in distinct brain regions involved in emotional regulation, relative to novelty-induced exploration. Among these brain areas, D1 knockdown in the mPFC decreased social defeat stress-induced c-Fos expression in the interstitial nucleus of the posterior limb of the anterior commissure (IPAC), a subregion of the extended amygdala. Using retrograde adeno-associated virus vectors and transgenic mice expressing Cre recombinase under the D1 promoter, we also found that D1-expressing deep-layer pyramidal neurons in the mPFC send direct projections to the IPAC. These findings indicate that social defeat stress specifically activates neurons in distinct brain areas, among which the IPAC is regulated by dopamine D1 receptor in the mPFC perhaps through direct projections. Thus, this study provides hints toward identifying neural circuits that underlie antidepressant-like effects of stress-induced dopamine D1 receptor signaling in the mPFC.

A variety of mechanisms has been suggested for cocaine toxicity, including the possibility that cocaine induces an increase in oxidative stress (OS) due to excessive oxidation of dopamine (e.g. dopamine quinine), or by redox cycling of cocaine oxidized metabolites. However, the association between oxidative status in the brain and cocaine induced-behavior is poorly understood. Therefore, we examined the ability of the unique antioxidant tempol to attenuate cocaine-induced oxidative damage and behavioral response. Acute cocaine treatment significantly elevated OS markers in prefrontal cortex (PFC) and nucleus accumbens (NAc) in rats, both in slices and following a single cocaine injection, which corresponded with a decrease in total antioxidant capacity (TAC). Tempol, at the optimal concentration we determined that was needed to observe an antioxidant non-toxic effect in vitro (1 mM) and in vivo (200 mg/kg), completely abolished the elevation of OS markers and prevented the reduction in TAC in these areas. Importantly, tempol injections, at a dose that does not affect the basal levels of locomotor activity, attenuated both the development and expression of cocaine-induced locomotor sensitization. Finally, in cocaine-sensitized animals, tempol prevented the elevation of OS markers in both PFC and NAc. Our findings suggest that oxidation of specific sites in the brain reward system by cocaine is accompanied with behavioral changes. Tempol has a neuro-protective effect against cocaine toxicity in these regions, and it may be beneficial in the treatment of cocaine addiction.

Our earlier efforts to document the cortical connections of the ventral premotor cortex (PMv) revealed dense connections with a field rostral and lateral to PMv, an area we called the frontal rostral field (FR). Here, we present data collected in FR using electrophysiological and anatomical methods. Results show that FR contains an isolated motor representation of the forelimb that can be differentiated from PMv based on current thresholds and latencies to evoke electromyographic activity using intracortical microstimulation techniques. In addition, FR has a different pattern of cortical connections compared with PMv. Together, these data support that FR is an additional, previously undescribed motor-related area in squirrel monkeys.

Microstimulation of brain tissue plays a key role in a variety of sensory prosthetics, clinical therapies and research applications. At present, tailoring the parameters of a stimulation signal to a specific goal relies heavily on parameters from literature. Optimization methods seek to improve tried and tested waveforms developed for specific purposes, however the fundamental understanding of how stimulation parameters interact and the effects these interactions have on brain tissue remains widely unknown. This study explores the interactions between parameters of the constant-current, biphasic square waveform with the intention of developing a stimulation efficient strategy. We find that, the traditional premise of a waveform's effectiveness being dominated by its amplitude does apply, however exceptions are noted which may be of essential importance to the development of electrical stimuli in restrictive paradigms.

Microstimulation of brain tissue plays a key role in a variety of sensory prosthetics, clinical therapies and research applications, however the effects of stimulation parameters on the responses they evoke remain widely unknown. In particular, the effects of parameters when delivered in the form of a stimulus train as opposed to a single pulse are not well understood despite the prevalence of stimulus train use. We aimed to investigate the contribution of each parameter of a stimulus train to the duration of the motor responses they evoke in forelimb muscles. We used constant-current, biphasic, square wave pulse trains in acute terminal experiments under ketamine anaesthesia. Stimulation parameters were systematically tested in a pair-wise fashion in the caudal forelimb region of the motor cortex in 7 Sprague-Dawley rats while motor evoked potential (MEP) recordings from the forelimb were used to quantify the influence of each parameter in the train. Stimulus amplitude and train duration were shown to be the dominant parameters responsible for increasing the total duration of the MEP, while interphase interval had no effect. Increasing stimulus frequency from 100-200 Hz or pulse duration from 0.18-0.34 ms were also effective methods of extending response durations. Response duration was strongly correlated with peak time and amplitude. Our findings suggest that motor cortex intracortical microstimulations are often conducted at a higher frequency rate and longer train duration than necessary to evoke maximal response duration. We demonstrated that the temporal properties of the evoked response can be both predicted by certain response metrics and modulated via alterations to the stimulation signal parameters.

Clinical and experimental data support a role for the intact cortex in recovery of function after stroke, particularly ipsilesional areas interconnected to the infarct. There is, however, little understanding of molecular events in the intact cortex, as most studies focus on the infarct and peri-infarct regions. This study investigated neuronal immunoreactivity for hypoxia-inducible factor-1alpha (HIF-1alpha) and vascular endothelial growth factor (VEGF) receptor-2 (VEGFR-2) in remote cortical areas 3 days after a focal ischemic infarct, as both HIF-1alpha and VEGFR-2 have been implicated in peri-infarct neuroprotection. For this study, intracortical microstimulation techniques defined primary motor (M1) and premotor areas in squirrel monkeys (genus Saimiri). An infarct was induced in the M1 hand representation, and immunohistochemical techniques identified neurons, HIF-1alpha and VEGFR-2. Stereologic techniques quantified the total neuronal populations and the neurons immunoreactive for HIF-1alpha or VEGFR-2. The results indicate that HIF-1alpha upregulation is confined to the infarct and peri-infarct regions. Increases in VEGFR-2 immunoreactivity occurred; however, in two remote regions: the ventral premotor hand representation and the M1 hindlimb representation. Neurons in these representations were previously shown to undergo significant increases in VEGF protein immunoreactivity, and comparison of the two data sets showed a significant correlation between levels of VEGF and VEGFR-2 immunoreactivity. Thus, while remote areas undergo a molecular response to the infarct, we hypothesize that there is a delay in the initiation of the response, which ultimately may increase the 'window of opportunity' for neuroprotective interventions in the intact cortex.

Background. Cortical stimulation (CS) combined with rehabilitative training (RT) has proven effective for enhancing poststroke functional recovery in rats, but human clinical trials have had mixed outcomes. Objective. To assess the efficacy of CS/RT versus RT in a nonhuman primate model of cortical ischemic stroke. Methods. Squirrel monkeys learned a pellet retrieval task, then received an infarct to the distal forelimb (DFL) representation of primary motor cortex. A subdural monopolar electrode was implanted over the spared DFL representation in dorsal premotor cortex (PMD). Seven weeks postinfarct, monkeys underwent 4 to 6 weeks of RT (n = 8) or CS/RT (n = 7; 100 Hz, cathodal current) therapy. Behavioral performance was assessed before and after infarct, prior to therapy, and 1 and 12 weeks posttherapy (follow-up). The primary outcome measure was motor performance at 1 week posttherapy. Secondary outcomes included follow-up performance at 12 weeks and treatment-related changes in neurophysiological maps of spared DFL representations. Results. While postinfarct performance deficits were found in all monkeys, both groups demonstrated similar recovery profiles, with no difference in motor recovery between the RT and CS/RT groups. Posttherapy, PMD DFL area was significantly expanded in the RT group but not the CS/RT group. A significant relationship was found between motor recovery and DFL expansion in premotor cortex. Conclusions. Results suggest that the specific parameters utilized here were not optimal for promoting behavioral recovery in nonhuman primates. Though CS/RT has consistently shown efficacy in rat stroke models, the present finding has cautionary implications for translation of CS/RT therapy to clinical populations.

Background: Cortical injections of the vasoconstrictor endothelin-1 (ET1) have widely been used to induce focal circumscribed ischemic lesions in the motor cortex of rodents in the context of stroke recovery studies. In order to apply this model correctly, it is essential to understand the time course of regional flow changes and of the development of penumbra and infarction. Methods: Multitracer micro-PET of ET1 focal ischemia in rats was performed using [11C]-flumazenil ([11C]FMZ) as a flow- and viability tracer and [18F]-fluoromisonidazole ([18F]FMISO) as hypoxia marker in order to characterize the physiological time-course of this model. Nine adult Sprague-Dawley rats received stereotaxic injections of ET1 into the right primary motor cortex, 3 served as controls. PET imaging was started 2, 3 and 20 h after the last ET1 injection. Histology was obtained at the end of the scans. Standardized uptake value ratios reflecting cerebral blood flow (CBF), [11C]FMZ-binding and [18F]FMISO-retention were calculated for the region of hypoperfusion and the normoperfused cortex. Results: CBF in the hypoperfused cortex was significantly reduced (p < 0.01) at 5 h (0.58 ± 0.025), 6 h (0.54 ± 0.043) and 23 h (0.66 ± 0.024) compared to controls (1.00 ± 0.011) and moderately reduced (p < 0.05) in the remainder of the affected hemisphere at 5 h (0.93 ± 0.036). [11C]FMZ-binding was within the control range at all time points. Significant [18F]FMISO-retention (1.16 ± 0.091, p < 0.05) was observed only after 6 h in the ischemic core that later turned into infarct. Conclusion: ET1 injections yield reproducible, slowly developing ischemic lesions with constant levels of hypoperfusion. This multitracer micro-PET study suggests that the ET1 model is appropriate for inducing chronic circumscribed ischemic lesions but seems to be less suited for studying acute stroke pathophysiology.

The last decade of neuroscience research has revealed that the adult brain can undergo substantial reorganization following injury. Plasticity after stroke has traditionally been perceived as adaptive and supporting recovery, but recent studies have suggested that some plasticity may also be detrimental. In particular, increased activity in the unaffected (contralesional) hemisphere has been proposed to contribute to motor deficits of the paretic hand in some patients. Longitudinal imaging studies in humans have reported a progressive behavioral improvement associated with a decrease of contralesional activity and have correlated the intensity of contralesional hemisphere activity with the degree of motor impairment. Consequently, inhibitory neuromodulatory protocols have been applied to the contralesional hemisphere of stroke patients. Such protocols can facilitate the activation of the ipsilesional motor cortex and improve the function of the paretic limb. Although the use of noninvasive techniques after brain injury shows promise, much work needs to be done to understand better how these approaches affect postlesion plasticity and motor recovery. Ultimately, this knowledge will allow for the design of more effective treatments and will potentially lead to protocols adapted to the specific condition of each patient. In this chapter, we review the literature on the basic pathways that can support the effects of contralesional inhibition, interhemispheric interactions, and some of the changes that can occur in the sensorimotor network after stroke. Finally, we show work in rats that demonstrates how parameters of contralesional inactivation can affect postlesion recovery.

Background. There is growing interest in the use of d-amphetamine (d-AMPH) as a pharmacological treatment to supplement rehabilitative therapy following stroke. Based on the success of earlier animal models, several clinical studies have demonstrated beneficial effects of applying physical rehabilitation while stroke patients are under the influence of d-AMPH. To begin to understand the neural mechanisms underlying this promising adjuvant therapy, the authors examined the effects of a single pairing of d-AMPH and rehabilitative training on motor performance after cortical infarct in squirrel monkeys. Methods. Microelectrode stimulation techniques were used to delineate hand movement areas in the primary motor cortex prior to delivering a unilateral infarct to the complete hand representation. Postinfarct recovery was assessed for 3 groups of monkeys: d-AMPH + training, saline + training, and spontaneous recovery (SR). Postinfarct training groups received 14 consecutive days of motor skill training on a reach and retrieval task. A single injection of d-AMPH (0.25 mg/kg) or saline was given only on the 1st day of training (postinfarct day 10). Monkeys in the SR group had only minimal exposure to the training task once per week to monitor recovery. Results. The results show that a single coupling of d-AMPH + training initiated 10 days after cortical infarct facilitated the rate of recovery and improved performance (68% improvement from 1st day of training) beyond the level achieved by the monkeys in the saline + training group (27% improved from 1st day of training). Conclusions. D-AMPH is a potent modulator of behavioral recovery following an ischemic infarct in nonhuman primates.

We explore the physiological constraints of body temperature as related to body mass and ambient temperature during flight in endothermic dung beetles showing a mass-related breakpoint where species show strong vs. weak endothermy. We found two different strategies in the dung beetles prior to flight; larger beetles (>1.9 g) elevate and maintain their body temperature (T(b)) at levels well above ambient temperature (T(a)) whereas smaller beetles' (

Vascular endothelial growth factor (VEGF) is thought to contribute to both neuroprotection and angiogenesis after stroke. While increased expression of VEGF has been demonstrated in animal models after experimental ischemia, these studies have focused almost exclusively on the infarct and peri-infarct regions. The present study investigated the association of VEGF to neurons in remote cortical areas at three days after an infarct in primary motor cortex (M1). Although these remote areas are outside of the direct influence of the ischemic injury, remote plasticity has been implicated in recovery of function. For this study, intracortical microstimulation techniques identified primary and premotor cortical areas in a non-human primate. A focal ischemic infarct was induced in the M1 hand representation, and neurons and VEGF protein were identified using immunohistochemical procedures. Stereological techniques quantitatively assessed neuronal-VEGF association in the infarct and peri-infarct regions, M1 hindlimb, M1 orofacial, and ventral premotor hand representations, as well as non-motor control regions. The results indicate that VEGF protein significantly increased association to neurons in specific remote cortical areas outside of the infarct and peri-infarct regions. The increased association of VEGF to neurons was restricted to cortical areas that are functionally and/or behaviorally related to the area of infarct. There was no significant increase in M1 orofacial region or in non-motor control regions. We hypothesize that enhancement of neuronal VEGF in these functionally related remote cortical areas may be involved in recovery of function after stroke, through either neuroprotection or the induction of remote angiogenesis.

The present study describes the pattern of connections of the ventral premotor cortex (PMv) with various cortical regions of the ipsilateral hemisphere in adult squirrel monkeys. Particularly, we 1) quantified the proportion of inputs and outputs that the PMv distal forelimb representation shares with other areas in the ipsilateral cortex and 2) defined the pattern of PMv connections with respect to the location of the distal forelimb representation in primary motor cortex (M1), primary somatosensory cortex (S1) and the supplementary motor area (SMA). Intracortical microstimulation techniques (ICMS) were used in four experimentally naïve monkeys to identify M1, PMv and SMA forelimb movement representations. Multi-unit recording techniques and myelin staining were used to identify the S1 hand representation. Then, biotinylated dextran amine (BDA; 10000MW) was injected in the center of the PMv distal forelimb representation. Following tangential sectioning, the distribution of BDA-labeled cell bodies and terminal boutons was documented. In M1, labeling followed a rostro-lateral pattern, largely leaving the caudo-medial M1 unlabeled. Quantification of somata and terminals showed that two areas share major connections with PMv: M1 and frontal areas immediately rostral to PMv, designated as frontal rostral area (FR). Connections with this latter region have not been described previously. Moderate connections were found with PMd, SMA, anterior operculum and posterior operculum/inferior parietal area. Minor connections were found with diverse areas of the precentral and parietal cortex, including S1. No statistical difference between the proportion of inputs and outputs for any location was observed, supporting the reciprocity of PMv intracortical connections.

A focal injury within the cerebral cortex results in functional reorganization within the spared cortex through time-dependent metabolic and physiological reactions. Physiological changes are also associated with specific post-injury behavioral experiences. Knowing how these factors interact can be beneficial in planning rehabilitative intervention after a stroke. The purpose of this study was to assess the functional impact of delaying the rehabilitative behavioral experience upon movement representations within the primary motor cortex (M1) in an established nonhuman primate, ischemic infarct model. Five adult squirrel monkeys were trained on a motor-skill task prior to and 1 month after an experimental ischemic infarct was induced in M1. Movement representations of the hand were derived within M1 using standard electrophysiological procedures prior to the infarct and again one and two months after the infarct. The results of this study show that even though recovery of motor skills was similar to that of a previous study in squirrel monkeys after early training, unlike early training, delayed training did not result in maintenance of the spared hand representation within the M1 peri-infarct hand area. Instead, delaying training resulted in a large decrease in spared hand representation during the spontaneous recovery period that persisted following the delayed training. In addition, delayed training resulted in an increase of simultaneously evoked movements that are typically independent. These results indicate that post-injury behavioral experience, such as motor skill training, may modulate peri-infarct cortical plasticity in different ways in the acute versus chronic stages following stroke.

Previously, we showed that the ventral premotor cortex (PMv) underwent neurophysiological remodeling after injury to the primary motor cortex (M1). In the present study, we examined cortical connections of PMv after such lesions. The neuroanatomical tract tracer biotinylated dextran amine was injected into the PMv hand area at least 5 months after ischemic injury to the M1 hand area. Comparison of labeling patterns between experimental and control animals demonstrated extensive proliferation of novel PMv terminal fields and the appearance of retrogradely labeled cell bodies within area 1/2 of the primary somatosensory cortex after M1 injury. Furthermore, evidence was found for alterations in the trajectory of PMv intracortical axons near the site of the lesion. The results suggest that M1 injury results in axonal sprouting near the ischemic injury and the establishment of novel connections within a distant target. These results support the hypothesis that, after a cortical injury, such as occurs after stroke, cortical areas distant from the injury undergo major neuroanatomical reorganization. Our results reveal an extraordinary anatomical rewiring capacity in the adult CNS after injury that may potentially play a role in recovery.