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Visceral leishmaniasis (VL) in the Americas is a chronic systemic disease caused by infection with Leishmania infantum parasites. The toxicity of antileishmanial drugs, long treatment course and limited efficacy are significant concerns that hamper adequate treatment against the disease. Studies have shown the promise of an immunotherapeutics approach, combining antileishmanial drugs to reduce the parasitism and vaccine immunogens to activate the host immune system. In the current study, we developed an immunotherapy using a recombinant T cell epitope-based chimeric protein, ChimT, previously shown to be protective against Leishmania infantum, with the adjuvant monophosphoryl lipid A (MPLA) and amphotericin B (AmpB) as the antileishmanial drug. BALB/c mice were infected with L. infantum stationary promastigotes and later they received saline or were treated with AmpB, MPLA, ChimT/Amp, ChimT/MPLA or ChimT/MPLA/AmpB. The combination of ChimT/MPLA/AmpB significantly reduced the parasite load in mouse organs (p < 0.05) and induced a Th1-type immune response, which was characterized by higher ratios of anti-ChimT and anti-parasite IgG2a:IgG1 antibodies, increased IFN-γ mRNA and IFN-γ and IL-12 cytokines and accompanied by lower levels of IL-4 and IL-10 cytokines, when compared to other treatments and controls (all p < 0.05). Organ toxicity was also lower with the ChimT/MPLA/AmpB immunotherapy, suggesting that the inclusion of the vaccine and adjuvant ameliorated the toxicity of AmpB to some degree. In addition, the ChimT vaccine alone stimulated in vitro murine macrophages to significantly kill three different internalized species of Leishmania parasites and to produce Th1-type cytokines into the culture supernatants. To conclude, our data suggest that the combination of ChimT/MPLA/AmpB could be considered for further studies as an immunotherapy for L. infantum infection.

Artificial grammar learning is a popular paradigm to study syntactic ability in nonhuman animals. Subjects are first trained to recognize strings of tokens that are sequenced according to grammatical rules. Next, to test if recognition depends on grammaticality, subjects are presented with grammar-consistent and grammar-violating test strings, which they should discriminate between. However, simpler cues may underlie discrimination if they are available. Here, we review stimulus design in a sample of studies that use particular sounds as tokens, and that claim or suggest their results demonstrate a form of sequence rule learning. To assess the extent of acoustic similarity between training and test strings, we use four simple measures corresponding to cues that are likely salient. All stimulus sets contain biases in similarity measures such that grammatical test stimuli resemble training stimuli acoustically more than do non- grammatical test stimuli. These biases may contribute to response behaviour, reducing the strength of grammatical explanations. We conclude that acoustic confounds are a blind spot in artificial grammar learning studies in nonhuman animals., Netherlands Organization for Scientific Research (NWO) (grant number 024.001.003)

Sleep-related brain activity occurring during non-rapid eye-movement (NREM) sleep is proposed to play a role in processing information acquired during wakefulness. During mammalian NREM sleep, the transfer of information from the hippocampus to the neocortex is thought to be mediated by neocortical slow-waves and their interaction with thalamocortical spindles and hippocampal sharp-wave ripples (SWRs). In birds, brain regions composed of pallial neurons homologous to neocortical (pallial) neurons also generate slow-waves during NREM sleep, but little is known about sleep-related activity in the hippocampus and its possible relationship to activity in other pallial regions. We recorded local field potentials (LFP) and analogue multiunit activity (AMUA) using a 64-channel silicon multi-electrode probe simultaneously inserted into the hippocampus and medial part of the nidopallium (i.e., caudal medial nidopallium; NCM) or separately into the caudolateral nidopallium (NCL) of adult female zebra finches (Taeniopygia guttata) anesthetized with isoflurane, an anesthetic known to induce NREM sleep-like slow-waves. We show that slow-waves in NCM and NCL propagate as waves of neuronal activity. In contrast, the hippocampus does not show slow-waves, nor sharp-wave ripples, but instead displays localized gamma activity. In conclusion, neuronal activity in the avian hippocampus differs from that described in mammals during NREM sleep, suggesting that hippocampal memories are processed differently during sleep in birds and mammals.

Both mammals and birds exhibit two sleep states, slow wave sleep (SWS) and rapid eye movement (REM) sleep. Studying certain aspects of sleep-related electrophysiology in freely behaving animals can present numerous methodological constraints, particularly when even fine body movements interfere with electrophysiological signals. Interestingly, under light general anesthesia, mammals and birds also exhibit slow waves similar to those observed during natural SWS. For these reasons, slow waves occurring under general anesthesia are commonly used in the investigation of sleep-related neurophysiology. However, how spectral properties of slow waves induced by anesthesia correspond to those occurring during natural SWS in birds has yet to be investigated systematically. In this study, we systematically analyzed spectral properties of electroencephalographic (EEG) patterns of pigeons (Columba livia) occurring under two commonly used anesthetics, isoflurane and urethane. These data were compared with EEG patterns during natural sleep. Slow waves occurring during spontaneous SWS, and those induced with isoflurane and urethane all showed greatest absolute power in the slowest frequencies (

Study Objectives: The changes in electroencephalogram (EEG) activity that characterize sleep and its sub-states-slow-wave sleep (SWS) and rapid eye movement (REM) sleep-are similar in mammals and birds. SWS is characterized by EEG slow waves resulting from the synchronous alternation of neuronal membrane potentials between hyperpolarized down-states with neuronal quiescence and depolarized up-states associated with action potentials. By contrast, studies of non-avian reptiles report the presence of high-voltage sharp waves (HShW) during sleep. How HShW relate to EEG phenomena occurring during mammalian and avian sleep is unclear. We investigated the spatiotemporal patterns of electrophysiological phenomena in Nile crocodiles (Crocodylus niloticus) anesthetized with isoflurane to determine whether they share similar spatiotemporal patterns to mammalian and avian slow waves. Methods: Recordings of anesthetized crocodiles were made using 64-channel penetrating arrays with electrodes arranged in an 8 × 8 equally spaced grid. The arrays were placed in the dorsal ventricular ridge (DVR), a region implicated in the genesis of HShW. Various aspects of the spatiotemporal distribution of recorded signals were investigated. Results: Recorded signals revealed the presence of HShW resembling those reported in earlier studies of naturally sleeping reptiles. HShW propagated in complex and variable patterns across the DVR. Conclusions: We demonstrate that HShW within the DVR propagate in complex patterns similar to those observed for avian slow waves recorded from homologous brain regions. Consequently, sleep with HShW may represent an ancestral form of SWS, characterized by up-states occurring less often and for a shorter duration than in mammals and birds.

Sleep in birds is composed of two distinct sub-states, remarkably similar to mammalian slow-wave sleep (SWS) and rapid eye movement (REM) sleep. However, it is unclear whether all aspects of mammalian sleep are present in birds. We examined whether birds suppress REM sleep in response to changes in sleeping conditions that presumably evoke an increase in perceived predation risk, as observed previously in rodents. Although pigeons sometimes sleep on the ground, they prefer to sleep on elevated perches at night, probably to avoid nocturnal mammalian ground predators. Few studies to date have investigated how roosting sites affect sleep architecture. We compared sleep in captive pigeons on days with and without access to high perches. On the first (baseline) day, low and high perches were available; on the second day, the high perches were removed; and on the third (recovery) day, the high perches were returned. The total time spent sleeping did not vary significantly between conditions; however, the time spent in REM sleep declined on the low-perch night and increased above baseline when the pigeons slept on the high perch during the recovery night. Although the amount of SWS did not vary significantly between conditions, SWS intensity was lower on the low-perch night, particularly early in the night. The similarity of these responses between birds and mammals suggests that REM sleep is influenced by at least some ecological factors in a similar manner in both groups of animals.

Auditory systems bias responses to sounds that are unexpected on the basis of recent stimulus history, a phenomenon that has been widely studied using sequences of unmodulated tones (mismatch negativity; stimulus-specific adaptation). Such a paradigm, however, does not directly reflect problems that neural systems normally solve for adaptive behavior. We recorded multiunit responses in the caudomedial auditory forebrain of anesthetized zebra finches (Taeniopygia guttata) at 32 sites simultaneously, to contact calls that recur probabilistically at a rate that is used in communication. Neurons in secondary, but not primary, auditory areas respond preferentially to calls when they are unexpected (deviant) compared with the same calls when they are expected (standard). This response bias is predominantly due to sites more often not responding to standard events than to deviant events. When two call stimuli alternate between standard and deviant roles, most sites exhibit a response bias to deviant events of both stimuli. This suggests that biases are not based on a use-dependent decrease in response strength but involve a more complex mechanism that is sensitive to auditory deviance per se. Furthermore, between many secondary sites, responses are tightly synchronized, a phenomenon that is driven by internal neuronal interactions rather than by the timing of stimulus acoustic features. We hypothesize that this deviance-sensitive, internally synchronized network of neurons is involved in the involuntary capturing of attention by unexpected and behaviorally potentially relevant events in natural auditory scenes.

In this Formal Comment the authors respond to objections to their previous Essay, reiterating that comparative linguistics is not an easy undertaking.

Humans readily distinguish spoken words that closely resemble each other in acoustic structure, irrespective of audible differences between individual voices or sex of the speakers. There is an ongoing debate about whether the ability to form phonetic categories that underlie such distinctions indicates the presence of uniquely evolved, speech-linked perceptual abilities, or is based on more general ones shared with other species. We demonstrate that zebra finches ( Taeniopygia guttata ) can discriminate and categorize monosyllabic words that differ in their vowel and transfer this categorization to the same words spoken by novel speakers independent of the sex of the voices. Our analysis indicates that the birds, like humans, use intrinsic and extrinsic speaker normalization to make the categorization. This finding shows that there is no need to invoke special mechanisms, evolved together with language, to explain this feature of speech perception.

Unlike nonhuman primates, thousands of bird species have articulatory capabilities that equal or surpass those of humans, and they develop their vocalizations through vocal imitation in a way that is very similar to how human infants learn to speak. An understanding of how speech mechanisms have evolved is therefore unlikely to yield key insights into how the human brain is special.

Comparative studies of linguistic faculties in animals pose an evolutionary paradox: language involves certain perceptual and motor abilities, but it is not clear that this serves as more than an input–output channel for the externalization of language proper. Strikingly, the capability for auditory–vocal learning is not shared with our closest relatives, the apes, but is present in such remotely related groups as songbirds and marine mammals. There is increasing evidence for behavioral, neural, and genetic similarities between speech acquisition and birdsong learning. At the same time, researchers have applied formal linguistic analysis to the vocalizations of both primates and songbirds. What have all these studies taught us about the evolution of language? Is the comparative study of an apparently species-specific trait like language feasible? We argue that comparative analysis remains an important method for the evolutionary reconstruction and causal analysis of the mechanisms underlying language. On the one hand, common descent has been important in the evolution of the brain, such that avian and mammalian brains may be largely homologous, particularly in the case of brain regions involved in auditory perception, vocalization, and auditory memory. On the other hand, there has been convergent evolution of the capacity for auditory–vocal learning, and possibly for structuring of external vocalizations, such that apes lack the abilities that are shared between songbirds and humans. However, significant limitations to this comparative analysis remain. While all birdsong may be classified in terms of a particularly simple kind of concatenation system, the regular languages, there is no compelling evidence to date that birdsong matches the characteristic syntactic complexity of human language, arising from the composition of smaller forms like words and phrases into larger ones.

There are remarkable behavioral, neural, and genetic similarities between song learning in songbirds and speech acquisition in human infants. Previously, we have argued that this parallel cannot be extended to the level of sentence syntax. Although birdsong can indeed have a complex structure, it lacks the combinatorial complexity of human language syntax. Recently, this conclusion has been challenged by a report purporting to show that songbirds can learn so-called context-free syntactic rules and then use them to discriminate particular syllable patterns. Here, we demonstrate that the design of this study is inadequate to draw such a conclusion, and offer alternative explanations for the experimental results that do not require the acquisition and use of context-free grammar rules or a grammar of any kind, only the simpler hypothesis of acoustic similarity matching. We conclude that the evolution of vocal learning involves both neural homologies and behavioral convergence, and that human language reflects a unique cognitive capacity.

SUMMARYBirdsong and human speech share many features with respect to vocal learning and development. However, the vocal production mechanisms have long been considered to be distinct. The vocal organ of songbirds is more complex than the human larynx, leading to the hypothesis that vocal variation in birdsong originates mainly at the sound source, while in humans it is primarily due to vocal tract filtering. However, several recent studies have indicated the importance of vocal tract articulators such as the beak and oropharyngeal–esophageal cavity. In contrast to most other bird groups, parrots have a prominent tongue, raising the possibility that tongue movements may also be of significant importance in vocal production in parrots, but evidence is rare and observations often anecdotal. In the current study we used X-ray cinematographic imaging of naturally vocalizing monk parakeets (Myiopsitta monachus) to assess which articulators are possibly involved in vocal tract filtering in this species. We observed prominent tongue height changes, beak opening movements and tracheal length changes, which suggests that all of these components play an important role in modulating vocal tract resonance. Moreover, the observation of tracheal shortening as a vocal articulator in live birds has to our knowledge not been described before. We also found strong positive correlations between beak opening and amplitude as well as changes in tongue height and amplitude in several types of vocalization. Our results suggest considerable differences between parrot and songbird vocal production while at the same time the parrot's vocal articulation might more closely resemble human speech production in the sense that both make extensive use of the tongue as a vocal articulator.

Sustainable agricultural practices are needed to minimize nitrate leaching. The crop-soil simulation model STICS coupled with a geographic information system was used to estimate the amount of NO3 – leaching and to assess the ability of alternative management practices to reduce NO3 – leaching in a nitrate vulnerable zone (NVZ) in La Rioja, Spain. Model performance was examined by comparing the simulations and measurements of irrigated grapevine crops (variety Tempranillo) over various soil types. The measurements were obtained from five pilot plots over a period of three years and included the mineral nitrogen, the water content of the soil profiles and the nitrogen content of the crops. The simulated and measured values were in satisfactory agreement with each other. Then, eight management scenarios were simulated, combining two NO3 – concentrations of irrigation water and four levels of organic manure applications. The simulations identified good agricultural practices (GAP) for mitigating NO3 – pollution. High soil mineral nitrogen (SMN) and water pollution were driven by both the NO3 – concentration of irrigation water and the level of organic manure application. The use of aquifer water for irrigation would lead to diminish aquifer pollution at the expense of maintaining high SMN, non desirable for grape quality production. River water would offer an opportunity for the recovery of soils and the improvement of underground water quality if the application of organic manure was limited according to soil type. Differences in NO3 - leaching of the NVZ soils depended more on their ability to store N than on their annual drainage., Las prácticas agrícolas sostenibles son necesarias para minimizar el lavado de nitratos (NO3 –). Se utilizó el modelo de simulación de cultivos STICS para estimar el lavado de NO3 – y evaluar prácticas de manejo para reducirlo en una zona vulnerable (NZV) en La Rioja, España. Se compararon las simulaciones con las medidas en cultivos de vid en regadío (variedad Tempranillo) en distintos tipos de suelo. Las medidas de nitrógeno mineral, contenido en agua en el suelo y contenido en nitrógeno de los cultivos se obtuvieron en cinco parcelas piloto durante tres años. Las simulaciones mostraron una coincidencia satisfactoria con las medidas. Después, se simularon ocho escenarios de manejo, combinando dos concentraciones de NO3 – en el agua de riego y cuatro niveles de abonado orgánico. Las simulaciones permitieron identificar buenas prácticas agrícolas (GAP) para la mitigación de la contaminación por NO3 –. Tanto la concentración de NO3 – en el agua de riego como el nivel de abonado orgánico determinaron la contaminación del agua y el alto nitrógeno mineral del suelo. Regar con agua del acuífero reduciría la contaminación del acuífero a expensas de mantener alto el nitrógeno mineral del suelo, no deseable para la producción de uva de calidad. Regar con agua del río permitiría recuperar los suelos mejorando la calidad del agua subterránea, si el abonado orgánico se limitase en función del tipo de suelo. Las diferencias en el lavado de NO3 – de los suelos de la NVZ dependieron más de su capacidad para almacenar N que de su drenaje anual.

Research into speech perception by nonhuman animals can be crucially informative in assessing whether specific perceptual phenomena in humans have evolved to decode speech, or reflect more general traits. Birds share with humans not only the capacity to use complex vocalizations for communication but also many characteristics of its underlying developmental and mechanistic processes; thus, birds are a particularly interesting group for comparative study. This review first discusses commonalities between birds and humans in perception of speech sounds. Several psychoacoustic studies have shown striking parallels in seemingly speech-specific perceptual phenomena, such as categorical perception of voice-onset-time variation, categorization of consonants that lack phonetic invariance, and compensation for coarticulation. Such findings are often regarded as evidence for the idea that the objects of human speech perception are auditory or acoustic events rather than articulations. Next, I highlight recent research on the production side of avian communication that has revealed the existence of vocal tract filtering and articulation in bird species-specific vocalization, which has traditionally been considered a hallmark of human speech production. Together, findings in birds show that many of characteristics of human speech perception are not uniquely human but also that a comparative approach to the question of what are the objects of perception--articulatory or auditory events--requires careful consideration of species-specific vocal production mechanisms.

Unlike our primate cousins, many species of bird share with humans a capacity for vocal learning, a crucial factor in speech acquisition. There are striking behavioural, neural and genetic similarities between auditory-vocal learning in birds and human infants. Recently, the linguistic parallels between birdsong and spoken language have begun to be investigated. Although both birdsong and human language are hierarchically organized according to particular syntactic constraints, birdsong structure is best characterized as 'phonological syntax', resembling aspects of human sound structure. Crucially, birdsong lacks semantics and words. Formal language and linguistic analysis remains essential for the proper characterization of birdsong as a model system for human speech and language, and for the study of the brain and cognition evolution.

BACKGROUND: Birdsong and human vocal communication are both complex behaviours which show striking similarities mainly thought to be present in the area of development and learning. Recent studies, however, suggest that there are also parallels in vocal production mechanisms. While it has been long thought that vocal tract filtering, as it occurs in human speech, only plays a minor role in birdsong there is an increasing number of studies indicating the presence of sound filtering mechanisms in bird vocalizations as well. METHODOLOGY/PRINCIPAL FINDINGS: Correlating high-speed X-ray cinematographic imaging of singing zebra finches (Taeniopygia guttata) to song structures we identified beak gape and the expansion of the oropharyngeal-esophageal cavity (OEC) as potential articulators. We subsequently manipulated both structures in an experiment in which we played sound through the vocal tract of dead birds. Comparing acoustic input with acoustic output showed that OEC expansion causes an energy shift towards lower frequencies and an amplitude increase whereas a wide beak gape emphasizes frequencies around 5 kilohertz and above. CONCLUSION: These findings confirm that birds can modulate their song by using vocal tract filtering and demonstrate how OEC and beak gape contribute to this modulation.

The TRiga Accelerator Driven Experiment (TRADE) to be performed in the existing TRIGA reactor of the ENEA Casaccia centre had the objective of coupling a spallation target, a proton accelerator and a sub-critical reactor core. On the way of the transmutation of minor actinides in dedicated facilities, the TRADE experiment represents the intermediate step between the validation of each component of an Accelerator Driven System (ADS), i.e. the spallation target, the high intensity accelerator and the sub-critical core, and the final facility aiming to incinerate radioactive nuclear waste. Measurements have been performed in the TRIGA - RC1 reactor of the ENEA Casaccia centre in the second semester of 2004 to characterize the neutron flux in a critical configuration. Gold and Indium samples were introduced in the core with the help of a "fast rabbit" pneumatic device. Bare samples and samples covered with Cadmium foils were successively irradiated to estimate the thermal and epithermal fluxes. Measurements have been carried out along a traverse of the core. Samples were put inside Aluminium shuttles travelling in stainless steel tubes. Trains composed of five shuttles allowed for simultaneous measurements in five different axial positions. γ spectrometry of the radioactive nuclides has been performed with two HPGe detectors.

The occurrence of coral snake coloration among unrelated venomous and non-venomous New World snake species has often been explained in terms of warning coloration and mimicry. The idea that snake predators would avoid coral snakes in nature seems widely established and is postulated in many discussions on coral snake mimicry. However, the few workers that have tested a potential aposematic function of the conspicuous colour pattern focused exclusively on behaviour of snake predators towards coloured abstract models. Here we report on behaviour of temporarily caged, wild coatis (Nasua narica) when confronted with co-occurring live snakes, among which were two species of venomous coral snakes. Five different types of responses have been observed, ranging from avoidance to predation, yet none of the coatis avoided either of the two coral snake species or other species resembling these. As in earlier studies coatis appeared to avoid coral snake models, our findings show that results from studies with abstract snake models cannot unconditionally serve as evidence for an aposematic function of coral snake coloration.