"Cyclophosphamide and analogues; a matter of dose and schedule for dual anticancer activities".

Cyclophosphamide and ifosfamide are major alkylating agents but their therapeutics uses are limiting by the toxicity due to several toxicities. Indeed conventional chemotherapies are generally used with the maximum tolerated dose. In contrast, metronomic schedule aims to get a minimum dose for efficacy with a good safety. Depending on the dose, their mechanisms of action are different and offer a dual activity: at high dose, cyclophosphamide is mainly used in graft conditioning for its immunosuppressive properties, while at metronomic dose it is used as an immunoactive agent. Currently, at metronomic dose, cyclophosphamide is studied in clinic against various types of cancer, alone or in combination with others anticancer drugs (anti-angiogenic, immune-modulating agents, immune checkpoints blockers, vaccines, radiotherapy, others conventional anticancer agents), as a nth-line or first-line treatment. More than three quarters of clinical studies show promising results, mostly in breast, ovarian and prostate cancers. Taking advantage of the immune system, use dual antitumor action's chemotherapy is clearly a therapeutic strategy that deserves to be confirmed in order to improve the efficacy/toxicity balance of anticancer treatments, and to use CPM or analogs as a standard of care.

Implementing Gait Kinematic Trajectory Forecasting Models on an Embedded System.

Smart algorithms for gait kinematic motion prediction in wearable assistive devices including prostheses, bionics, and exoskeletons can ensure safer and more effective device functionality. Although embedded systems can support the use of smart algorithms, there are important limitations associated with computational load. This poses a tangible barrier for models with increased complexity that demand substantial computational resources for superior performance. Forecasting through Recurrent Topology (FReT) represents a computationally lightweight time-series data forecasting algorithm with the ability to update and adapt to the input data structure that can predict complex dynamics. Here, we deployed FReT on an embedded system and evaluated its accuracy, computational time, and precision to forecast gait kinematics from lower-limb motion sensor data from fifteen subjects. FReT was compared to pretrained hyperparameter-optimized NNET and deep-NNET (D-NNET) model architectures, both with static model weight parameters and iteratively updated model weight parameters to enable adaptability to evolving data structures. We found that FReT was not only more accurate than all the network models, reducing the normalized root-mean-square error by almost half on average, but that it also provided the best balance between accuracy, computational time, and precision when considering the combination of these performance variables. The proposed FReT framework on an embedded system, with its improved performance, represents an important step towards the development of new sensor-aided technologies for assistive ambulatory devices.

Hypothalamic CRH neurons represent physiological memory of positive and negative experience.

Recalling a salient experience provokes specific behaviors and changes in the physiology or internal state. Relatively little is known about how physiological memories are encoded. We examined the neural substrates of physiological memory by probing CRHPVN neurons of mice, which control the endocrine response to stress. Here we show these cells exhibit contextual memory following exposure to a stimulus with negative or positive valence. Specifically, a negative stimulus invokes a two-factor learning rule that favors an increase in the activity of weak cells during recall. In contrast, the contextual memory of positive valence relies on a one-factor rule to decrease activity of CRHPVN neurons. Finally, the aversive memory in CRHPVN neurons outlasts the behavioral response. These observations provide information about how specific physiological memories of aversive and appetitive experience are represented and demonstrate that behavioral readouts may not accurately reflect physiological changes invoked by the memory of salient experiences.

High-throughput gait acquisition system for freely moving mice.

Normal and pathological locomotion can be discriminated by analyzing an animal's gait on a linear walkway. This step is labor intensive and introduces experimental bias due to the handling involved while placing and removing the animal between trials. We designed a system consisting of a runway embedded within a larger arena, which can be traversed ad libitum by unsupervised, freely moving mice, triggering the recording of short clips of locomotor activity. Multiple body parts were tracked using DeepLabCut and fed to an analysis pipeline (GaitGrapher) to extract gait metrics. We compared the results from unsupervised against the standard experimenter-supervised approach and found that gait parameters analyzed via the new approach were similar to a previously validated approach (Visual Gait Lab). These data show the utility of incorporating an unsupervised, automated, approach for collecting kinematic data for gait analysis.NEW & NOTEWORTHY The acquisition and analysis of walkway data is a time-consuming task. Here, we provide an unmonitored approach for collecting gait metrics that reduces the handling and stress of mice and saves time. A detailed pipeline is outlined that provides for the collection and analysis of data using an integrated suite of tools.

Astrocytes amplify neurovascular coupling to sustained activation of neocortex in awake mice.

Functional hyperemia occurs when enhanced neuronal activity signals to increase local cerebral blood flow (CBF) to satisfy regional energy demand. Ca2+ elevation in astrocytes can drive arteriole dilation to increase CBF, yet affirmative evidence for the necessity of astrocytes in functional hyperemia in vivo is lacking. In awake mice, we discovered that functional hyperemia is bimodal with a distinct early and late component whereby arteriole dilation progresses as sensory stimulation is sustained. Clamping astrocyte Ca2+ signaling in vivo by expressing a plasma membrane Ca2+ ATPase (CalEx) reduces sustained but not brief sensory-evoked arteriole dilation. Elevating astrocyte free Ca2+ using chemogenetics selectively augments sustained hyperemia. Antagonizing NMDA-receptors or epoxyeicosatrienoic acid production reduces only the late component of functional hyperemia, leaving brief increases in CBF to sensory stimulation intact. We propose that a fundamental role of astrocyte Ca2+ is to amplify functional hyperemia when neuronal activation is prolonged.

A versatile computational algorithm for time-series data analysis and machine-learning models.

Here we introduce Local Topological Recurrence Analysis (LoTRA), a simple computational approach for analyzing time-series data. Its versatility is elucidated using simulated data, Parkinsonian gait, and in vivo brain dynamics. We also show that this algorithm can be used to build a remarkably simple machine-learning model capable of outperforming deep-learning models in detecting Parkinson's disease from a single digital handwriting test.

Visual Gait Lab: A user-friendly approach to gait analysis.

BACKGROUND: Gait analysis forms a critical part of many lab workflows, ranging from those interested in preclinical neurological models to others who use locomotion as part of a standard battery of tests. Unfortunately, while paw detection can be semi-automated, it becomes generally a time-consuming process with error corrections. Improvement in paw tracking would aid in better gait analysis performance and experience. NEW METHOD: Here we show the use of Visual Gait Lab (VGL), a high-level software with an intuitive, easy to use interface, that is built on DeepLabCut™. VGL is optimized to generate gait metrics and allows for quick manual error corrections. VGL comes with a single executable, streamlining setup on Windows systems. We demonstrate the use of VGL to analyze gait. RESULTS: Training and evaluation of VGL were conducted using 200 frames (80/20 train-test split) of video from mice walking on a treadmill. The trained network was then used to visually track paw placements to compute gait metrics. These are processed and presented on the screen where the user can rapidly identify and correct errors. COMPARISON WITH EXISTING METHODS: Gait analysis remains cumbersome, even with commercial software due to paw detection errors. DeepLabCut™ is an alternative that can improve visual tracking but is not optimized for gait analysis functionality. CONCLUSIONS: VGL allows for gait analysis to be performed in a rapid, unbiased manner, with a set-up that can be easily implemented and executed by those without a background in computer programming.

Impaired Hippocampal-Cortical Interactions during Sleep in a Mouse Model of Alzheimer's Disease.

Spatial learning is impaired in humans with preclinical Alzheimer's disease (AD). We reported similar impairments in 3xTg-AD mice learning a spatial reorientation task. Memory reactivation during sleep is critical for learning-related plasticity, and memory consolidation is correlated with hippocampal sharp wave ripple (SWR) density, cortical delta waves (DWs), cortical spindles, and the temporal coupling of these events-postulated as physiological substrates for memory consolidation. Further, hippocampal-cortical discoordination is prevalent in individuals with AD. Thus, we hypothesized that impaired memory consolidation mechanisms in hippocampal-cortical networks could account for spatial memory deficits. We assessed sleep architecture, SWR-DW dynamics, and memory reactivation in a mouse model of tauopathy and amyloidosis implanted with a recording array targeting isocortex and hippocampus. Mice underwent daily recording sessions of rest-task-rest while learning the spatial reorientation task. We assessed memory reactivation by matching activity patterns from the approach to the unmarked reward zone to patterns during slow-wave sleep (SWS). AD mice had more SWS, but reduced SWR density. The increased SWS compensated for reduced SWR density so there was no reduction in SWR number. In control mice, spindles were phase-coupled with DWs, and hippocampal SWR-cortical DW coupling was strengthened in post-task sleep and was correlated with performance on the spatial reorientation task the following day. However, in AD mice, SWR-DW and spindle-DW coupling were impaired. Thus, reduced SWR-DW coupling may cause impaired learning in AD, and spindle-DW coupling during short rest-task-rest sessions may serve as a biomarker for early AD-related changes in these brain dynamics.

Pharmacological challenges examining the underlying mechanism of altered response inhibition and attention due to circadian disruption in adult Long-Evans rats.

Endogenous circadian rhythms govern behavior and physiology, while circadian disruption is an environmental factor that impacts cognition by altering the circadian clock at a molecular level. We modeled the effects of 2 sources of circadian disruption - activity occurring during typical rest periods and untimely light exposure - to evaluate the effects of circadian disruption on behavior and underlying neurochemistry. Firstly, adult Long-Evans rats of both sexes were maintained on a 12 h:12 h light:dark cycle and tested using a 5-choice serial reaction time task (5-CSRTT) under 3 conditions: 4 h into the dark phase with no exposure to ambient light during testing (control), 4 h into the dark phase with exposure to ambient light during testing, and 4 h into the light phase. Both models resulted in impulsive behavior and reduced attention compared to control. We established that changes in the diurnal expression pattern occur in the clock gene Period 2 (Per2) in the light phase-tested model. Choline acetyltransferase (Chat) and Dopamine receptor 1 (Drd1) showed rhythmic expression with peak expression during the dark phase regardless of light-testing condition. Next, we performed drug challenges in a new rat cohort to examine the interaction between the cholinergic and dopaminergic neurotransmitter systems in regulating the behavioral changes caused by circadian disruption. We administered the cholinergic agonist nicotine and either the dopamine-1 receptor (DR1) antagonist SCH23390 or the DR2 antagonist eticlopride under the 3 circadian conditions to identify differential drug responses between treatment groups. Rats in both models demonstrated increased sensitivity to nicotine as compared to control, while SCH23390 and eticlopride ameliorated the effect of nicotine on 5-CSRTT performance in both models. Our study is the first to identify detrimental effects of both models of circadian disruption on impulsive behavior, and that the effects of circadian disruption are mediated by an interaction between cholinergic and dopaminergic systems.

Vision and Locomotion Combine to Drive Path Integration Sequences in Mouse Retrosplenial Cortex.

The retrosplenial cortex (RSC) is involved in a broad range of cognitive functions, integrating rich sensory, motor, and spatial signals from multiple brain areas, including the hippocampal system. RSC neurons show hippocampus-dependent activity reminiscent of place cell sequences. Using cellular calcium imaging in a virtual reality (VR)-based locomotion task, we investigate how the integration of visual and locomotor inputs may give rise to such activity in RSC. A substantial population shows neural sequences that track position in the VR environment. This activity is driven by the conjunction of visual stimuli sequences and active movement, which is suggestive of path integration. The activity is anchored to a reference point and predominantly follows the VR upon manipulations of optic flow against locomotion. Thus, locomotion-gated optic flow, combined with the presence of contextual cues at the start of each trial, is sufficient to drive the sequential activity. A subpopulation shows landmark-related visual responses that are modulated by animal's position in the VR. Thus, rather than fragmenting the spatial representation into equivalent locomotion-based ensemble versus optic-flow-based ensemble, in RSC, optic flow appears to override locomotion signals coherently in the population, when the gain between the two signals is altered.

An economical solution to record and control wheel-running for group-housed mice.

BACKGROUND: The effects of exercise on brain function are widely known; however, there is a need for inexpensive, practical solutions for monitoring and metering the activity of multiple mice. NEW METHOD: A contoured running wheel that has a built-in radio-frequency identification (RFID) receiver to monitor the activity of several mice in a single cage is presented. This system is scalable , the interface is easy to use, and the wheel can be dynamically locked so that each group-housed mouse receives a set exercise regimen. RESULTS: We were able to reliably monitor three mice that were group-housed. We were able to reliably meter the amount of exercise performed by the mice using the servo-controlled lock. COMPARISON WITH EXISTING METHODS: Current methods allow a wheel to be locked when a set distance is reached. However, an issue with this method is that the set distance includes the cumulative activity of all mice in the cage so one mouse could contribute a disproportionate amount to the total distance. Our solution ensures that the wheel is locked when an individual mouse reaches the target distance, but remains unlocked for individuals that have not reached the programmed distance. CONCLUSIONS: The dynamic locking wheel (DynaLok) is designed to allow a researcher to provide individually designed exercise plans for multi-housed mice; therefore, users are able to house mice conventionally rather than in individual cages. DynaLok reduces animal housing costs, allows for new experimental exercise regimens to be developed, and is scalable and cost-effective.

Using Neuron Spiking Activity to Trigger Closed-Loop Stimuli in Neurophysiological Experiments.

Closed-loop neurophysiological systems use patterns of neuronal activity to trigger stimuli, which in turn affect brain activity. Such closed-loop systems are already found in clinical applications, and are important tools for basic brain research. A particularly interesting recent development is the integration of closed-loop approaches with optogenetics, such that specific patterns of neuronal activity can trigger optical stimulation of selected neuronal groups. However, setting up an electrophysiological system for closed-loop experiments can be difficult. Here, a ready-to-apply Matlab code is provided for triggering stimuli based on the activity of single or multiple neurons. This sample code can be easily modified based on individual needs. For instance, it shows how to trigger sound stimuli and how to change it to trigger an external device connected to a PC serial port. The presented protocol is designed to work with a popular neuronal recording system for animal studies (Neuralynx). The implementation of closed-loop stimulation is demonstrated in an awake rat.

Pilot field trial of the EG95 vaccine against ovine cystic echinococcosis in Rio Negro, Argentina: 8 years of work.

Cystic echinococcosis (CE) is endemic in the Rio Negro province of Argentina. After 30 years of control using praziquantel in dogs the transmission rate to humans and sheep has decreased significantly, however transmission persists. The objective of the study was to assess the inclusion of the EG95 for sheep in the control program and to determine the vaccine's operative feasibility in field conditions. An intervention study was defined in Rio Negro Province in Argentina comprising, in total, an area of 5820 Km2. Lambs received two vaccinations with the EG95 vaccine followed by a single booster injection when the animals were 1-1.5 years of age. Vaccination of lambs born into one trial site was introduced and continued for 8 years. Evidence for Echinococcus granulosus transmission was monitored before and after vaccination by coproantigen ELISA in faecal samples of dog, purgation of dogs to detect E. granulosus worms, necropsy on adult sheep and by ultrasound screening in children of 6-14 years old. 29,323 doses of vaccine were applied between 2009 and 2017, which a vaccination coverage of 80.1%/85.7% (57.3% average for fully vaccinated). Before the introduction of the vaccine 56.3% of the 6-year-old sheep were infected with E. granulosus at necropsy and 84.2% of the farms had infected sheep; 4.3% of the dogs were positive for E. granulosus infection using the arecoline test, and with coproELISA 9.6% of dog fecal samples were positive and 20.3% of the farms had infected dog.After the vaccine was introduced, 21.6% of sheep older than 6 years were found to be infected at necropsy and 20.2% of the farms were found to be infected; in dogs, 4.5% were found positive for E. granulosus using arecoline purgation and with coproELISA 3.7% of samples were positive, with 8.9% of farms having a positive dog. In 2016 only one case of E. granulosus infection was diagnosed by US screening in a 6-14 years old child. Included in the analysis are discussions of difficulties experienced in the field which affected correct vaccine administration as well as social features and practices that may impact on echinococcosis control and the EG95 vaccination program in Rio Negro. Vaccination of sheep with the EG95 vaccine provides a valuable new tool which improves the effectiveness of CE control activities. Vaccination was effective even in a difficult, remote environment where only approximately half the lambs born into the communities were fully vaccinated.

Time series analysis of bovine venereal diseases in La Pampa, Argentina.

The venereal diseases bovine trichomoniasis (BT) and bovine genital campylobacteriosis (BGC) cause economic losses in endemic areas like La Pampa province in Argentina where beef cattle are usually extensively managed. This study used data compiled between 2007 and 2014 by a Provincial Program for the Control and Eradication of venereal diseases in order to develop and analyze retrospective models of time series for BT and BGC. Seasonality and long-term trend were explored with decomposition and simple regression methods. Autoregressive Integrated Moving Average models (ARIMA) were used to fit univariate models for the prevalence and persistence of BT and BGC. Autoregressive Integrated Moving Average with Explanatory Variable models (ARIMAX) were used to analyze the association between different time series, replacement entries and herd samplings. The prevalence and persistence of BT and BGC have decreased from 2007 to 2014. All the BT and BGC time series are seasonal and their long-term trend is decreasing. Seasonality of BT and BGC is similar, with higher rates of detection in autumn-winter than is spring-summer. Prevalence and persistence time series are correlated, indicating their changes are synchronic and follow a similar time pattern. Prevalence of BT and BGC showed the best fitting with the ARIMA (0,0,1)(0,1,1)12 model. While for persistence of BT and BGC, the best adjustment was with the same model with no seasonal difference where the current number of cases depends on the moving averages of the month and the previous season. Including covariates improve the fitting of univariate models, in addition, estimations using ARIMAX models are more precise than using ARIMA models. The time distribution of the samplings could be increasing the false negative ratio. According to the obtained results, the ARIMA and ARIMAX models can be considered an option to predict the BT and BGC prevalence and persistence in La Pampa (Argentina).

Lesions of dorsal striatum eliminate lose-switch responding but not mixed-response strategies in rats.

We used focal brain lesions in rats to examine how dorsomedial (DMS) and dorsolateral (DLS) regions of the striatum differently contribute to response adaptation driven by the delivery or omission of rewards. Rats performed a binary choice task under two modes: one in which responses were rewarded on half of the trials regardless of choice; and another 'competitive' one in which only unpredictable choices were rewarded. In both modes, control animals were more likely to use a predictable lose-switch strategy than animals with lesions of either DMS or DLS. Animals with lesions of DMS presumably relied more on DLS for behavioural control, and generated repetitive responses in the first mode. These animals then shifted to a random response strategy in the competitive mode, thereby performing better than controls or animals with DLS lesions. Analysis using computational models of reinforcement learning indicated that animals with striatal lesions, particularly of the DLS, had blunted reward sensitivity and less stochasticity in the choice mechanism. These results provide further evidence that the rodent DLS is involved in rapid response adaptation that is more sophisticated than that embodied by the classic notion of habit formation driven by gradual stimulus-response learning.

Acute NMDA receptor antagonism disrupts synchronization of action potential firing in rat prefrontal cortex.

Antagonists of N-methyl-D-aspartate receptors (NMDAR) have psychotomimetic effects in humans and are used to model schizophrenia in animals. We used high-density electrophysiological recordings to assess the effects of acute systemic injection of an NMDAR antagonist (MK-801) on ensemble neural processing in the medial prefrontal cortex of freely moving rats. Although MK-801 increased neuron firing rates and the amplitude of gamma-frequency oscillations in field potentials, the synchronization of action potential firing decreased and spike trains became more Poisson-like. This disorganization of action potential firing following MK-801 administration is consistent with changes in simulated cortical networks as the functional connections among pyramidal neurons become less clustered. Such loss of functional heterogeneity of the cortical microcircuit may disrupt information processing dependent on spike timing or the activation of discrete cortical neural ensembles, and thereby contribute to hallucinations and other features of psychosis induced by NMDAR antagonists.

Estructura factrrial y consistencia interna de a escala messy (versión docentee) en una muestra de niños argentinos / Factor strucure and internal consistency ofmessy scale (teacher version) iin a sample of argentinean children

La escala Matson para la Evaluación de Habilidades Sociales con Jóvenes, evalúa habilidades sociales específicas implicadas en comportamientos adaptativos y comportamientos no adaptativos. El objetivo de este estudio es evaluar las propiedades psicométricas del instrumento a partir del informe de docentes, en particular la estructura factorial y la consistencia interna. Participaron 26 docentes quienes evaluaron a niños cordobeses (n = 311) de 7 a 13 años (X = 9.16, DE = 1.48), de ambos sexos (52.7% niños). Los docentes participaron de manera voluntaria y completaron la escala del 60 al 80% de los niños de su aula autorizados por los padres a participar. Se realizaron estudios exploratorios y luego se examinó la estructura factorial, obteniéndose tres dimensiones: Agresividad/Conducta Antisocial, Habilidades Sociales Apropiadas y Sobreconfianza/Celos/Soberbia. La escala total y cada una de las dimensiones mostraron índices adecuados de consistencia interna. Además, las diferencias entre niñas y niños en las diversas dimensiones aportan evidencia de la validez de la escala.

The Matson Evaluation of Social Skills with Youngsters assesses specific social skills involved in adaptive behaviors, as well as non-adaptive behaviors. The objective of this study is to test the psychometric properties of the scale from teacher report, particularly the factor structure and the internal consistency. Participants were 26 teachers who tested 311 children of 7 to 13 years old (X = 9.16, SD = 1.48) of both sexes (52.7% males). Teachers participated voluntarily and completed the scale regarding to 60-80% of children whose parents send the signed informed consent. We conducted exploratory studies and then we studied the factor structure, we obtained three dimensions: Aggressiveness/Antisocial Behavior, Appropriate Social Skills and Overconfidence/Jealousy/Pride. The total scale and each of the dimensions showed adequate internal consistency indices. Furthermore, differences between men and women in the various dimensions provide evidence for the validity of the scale.