text:
Paper submitted for the Special Interest Section on Ultrasonic Vocalizations on February 16, 2010.
Translating mouse vocalizations: prosody and frequency modulation1
Version of Record online: 21 MAY 2010
DOI: 10.1111/j.1601-183X.2010.00603.x
© 2010 The Authors. Genes, Brain and Behavior © 2010 Blackwell Publishing Ltd and International Behavioural and Neural Genetics Society
Genes, Brain and Behavior
Volume 10, Issue 1, pages 4–16, February 2011
Additional Information
How to Cite
Lahvis, G. P., Alleva, E. and Scattoni, M. L. (2011), Translating mouse vocalizations: prosody and frequency modulation. Genes, Brain and Behavior, 10: 4–16. doi: 10.1111/j.1601-183X.2010.00603.x
- †
Publication History
- Issue online: 21 JAN 2011
- Version of Record online: 21 MAY 2010
- Received 16 February 2010 revised 14 April 2010 accepted for publication 05 May 2010
- Abstract
- Article
- References
- Cited By
- Addiction;
- affective disorders;
- animal communication;
- autism;
- bioacoustic communication;
- empathy;
- mood;
- schizophrenia;
- ultrasonic vocalizations
Mental illness can include impaired abilities to express emotions or respond to the emotions of others. Speech provides a mechanism for expressing emotions, by both what words are spoken and by the melody or intonation of speech (prosody). Through the perception of variations in prosody, an individual can detect changes in another's emotional state. Prosodic features of mouse ultrasonic vocalizations (USVs), indicated by changes in frequency and amplitude, also convey information. Dams retrieve pups that emit separation calls, females approach males emitting solicitous calls, and mice can become fearful of a cue associated with the vocalizations of a distressed conspecific. Because acoustic features of mouse USVs respond to drugs and genetic manipulations that influence reward circuits, USV analysis can be employed to examine how genes influence social motivation, affect regulation, and communication. The purpose of this review is to discuss how genetic and developmental factors influence aspects of the mouse vocal repertoire and how mice respond to the vocalizations of their conspecifics. To generate falsifiable hypotheses about the emotional content of particular calls, this review addresses USV analysis within the framework of affective neuroscience (e.g. measures of motivated behavior such as conditioned place preference tests, brain activity and systemic physiology). Suggested future studies include employment of an expanded array of physiological and statistical approaches to identify the salient acoustic features of mouse vocalizations. We are particularly interested in rearing environments that incorporate sufficient spatial and temporal complexity to familiarize developing mice with a broader array of affective states.
Many of life's enriching experiences occur within a social context, with the ability to share enjoyment, to grasp how another individual feels, or to see the world from the perspective of another. In a variety of developmental disabilities and neurological diseases, these capacities are diminished or lost (Abdi & Sharma 2004, Freeman et al. 2009, Hagerman et al. 1986, Lord et al. 2000, O'Keeffe et al. 2007, Orbelo et al. 2005, Ruby & Decety 2004, Testa et al. 2001). To elucidate how these deficits emerge in development, adulthood or aging, emotional experiences cannot be directly measured. Rather, changes in behaviors are used as a metric to infer changes in emotion. For example, the level of fear experienced by a person cannot be measured but the quiver of a fearful voice can be detected. Scientists can also measure behavioral indications of empathy; how one individual responds to the emotional expressions of another individual. For instance, in response to hearing the quiver in a voice, an individual might feel compassion, fearful, vindicated, frustrated or angry. In turn, these emotions can engender various behavioral responses, such as expression of consoling words, flight from the scene, pursed lips, or a confrontation with the dangerous stimulus. The purpose of this review is to explore how mouse vocalizations convey emotional information, how mice respond to these vocalizations, and how we can infer affective states from changes in the mouse vocal repertoire.
Vocalizations provide highly salient cues that can indicate changes in emotion. Emotional information can be provided by what words are spoken (from the meanings of the words themselves). 'That feels great!''You're making me frustrated'. 'That dude gives me the creeps'. We know the meaning of these statements and we often express an emotional response to them. But emotions are also expressed by how words are spoken. This quality of speech, which is called 'prosody' and is often referred to as intonation, is central to communication. Prosody is measured by variations in the pitch (notes), energy (loudness), and the duration of spoken words and the timing between them.
Prosody can be classified into three functional groups. Grammatical prosody entails changes in emphasis on particular syllables, or changes in pitch, within a sentence that can help us distinguish meanings. For example, applications of stress on one of two syllables in a word can help us to differentiate a dry, treeless ecosystem (des'-ert) from the pleasant plate of food at the end of a meal (des-sert'). An upward trend in pitch through a sentence enables us to discern a question from an assertion. The soup bowl is in the kitchen. The soup bowl is in the kitchen? Pragmatic prosody is used to help us put emphasis on an important idea, sentence or word in a sentence, and thus shows our focus and intentions. For example, enhanced duration and loudness of a particular word can indicate its importance within a sentence. 'I don't want the brown coat, I want the DOWN coat'. A different function of pragmatic prosody is that it signals the context, roles, and relationships between speakers. People speak to infants in 'motherese' or 'infant-directed talk' in a voice that they do not use with older children (Fernald 1989). People speak with one tone to their employer and a different tone with their intimate partner. Affective prosody is responsive to internal emotional experiences, such as sadness, anger, fatigue, annoyance, concern, contentment, relief, excitement or victory. For instance, high levels of frequency modulation, in which there are broad variations in the level of pitch, from syllable to syllable, can be associated with excitement. A minimal level of frequency modulation, or monotone, can indicate boredom or fatigue. Prosody is also responsive to social context. Adults talk with old friends and new acquaintances with different vocal patterns. The variability of speech within the context of changes in emotion or social context is referred to as emotional or affective prosody.
Affective prosody and mental disorders
Autistic individuals have been shown to assign stress to the wrong syllables of a word (Baltaxe 1984, Baltaxe & Guthrie 1987, Shriberg et al. 2001) and children with high-functioning autism can have difficulties modulating the pitch and controlling the volume of their speech (Shriberg et al. 2001). Some aspects of autistic speech incorporate all three features of the diagnosis: repetitive behaviors, deficits in the ability to express emotions and deficits in communication. For instance, some children with autism repeat the use of certain sounds, syllables or words, more than typically developing children (Shriberg et al. 2001), or fail to use appropriate patterns of intonation to communicate. Prosody can be monotonic, minimally pitched or energy modulated, or it can be amplified in pitch range or even singsong-like (Amorosa 1992), masking dynamics in emotional status. Autistic language may not be appropriately attuned to context, or 'machine-like' (Fine et al. 1991). Overall, speech impairments in autism are generally more prevalent in the expression of pragmatic or affective prosody than grammatical prosody (Shriberg et al. 2001). Individuals with an autism diagnosis can have impairments in the ability to understand the spoken language and gestures of others, even as infants (Golan et al. 2007, Kuhl et al. 2005, Rutherford et al. 2002). The ability to respond to the emotional expressions of others can be deficient among people with autism (Silani et al. 2008, Smith 2009). In this regard, autistic children also show deficits in their abilities to use prosodic cues to disambiguate sentence meaning (Diehl et al. 2008). These deficits can extend to inabilities to interpret intonations that denote liking or disliking (McCann et al. 2007).
Schizophrenia, mania and depression can also be significantly impaired in the ability to comprehend affective prosody, while schizophrenia can additionally feature substantial deficits in the ability to express affective prosody (Murphy & Cutting 1990). Schizophrenia can feature deficits in expressive and receptive prosody (Bach et al. 2009, Leentjens et al. 1998, Leitman et al. 2006), especially in paranoid (Bach et al. 2009) and flat affect (Alpert et al. 2000) schizophrenia. For instance, individuals with flat affect schizophrenia employ normal word patterns to express emotions (frequency of words describing pleasurable or distressful experiences), but they communicate these experiences with less prosodic inflection (Alpert et al. 2000). Like autism disorders, there are many forms of schizophrenia which do not all show deficits in the expressive prosody (Cohen & Docherty 2005, Wan et al. 2008). Receptivity to emotional expressions of others can also be impaired in schizophrenia (Derntl et al. 2009, Fujiwara et al. 2008, Haker & Rossler 2009), particularly in males who are responding to emotions that have negative valence (Bozikas et al. 2006, Pijnenborg et al. 2007, Scholten et al. 2008). In addition to schizophrenia, responsiveness to affective prosody can be impaired in depression (Emerson et al. 1999, Uekermann et al. 2008), multiple sclerosis (Beatty et al. 2003), Parkinson's disease (Benke et al. 1998), alcoholism (Monnot et al. 2001, 2002, Uekermann & Daum 2008, Uekermann et al. 2005) and following exposure to MDMA or 'ecstasy' (Yip & Lee 2006).
Mouse vocalizations
To model deficits in expressive and receptive prosody, there must be evidence that features of mouse vocalizations typically associate with specific affective states, such as distress, or reward, or anticipation of reward and whether these features can be sensitive to differences in specific alleles or genetic background. Mice have the capability to vocalize across a broad range of frequencies that extend from as low as the human-audible range (when we hear squeaks) to well into the ultrasound range, above the limit of human hearing [20 000 cycles per second (hertz) = 20 kHz]. Audible squeaks are produced by laboratory mice in stressful and painful situations (Whitney & Nyby 1983), such as during handling and restraint (Whitney 1969), grid-shock test (Winter 1965) or during aggressive encounters (Gourbal et al. 2004, Houseknecht 1968). In reproductive contexts, human-audible squeaks are produced by females when a sexually motivated male is interacting with a non-receptive female (Sales 1972).
Over the last 40 years, mouse vocalizations have been studied mostly within two social contexts: adult male vocalizations emitted in response to the cues of estrous females and vocalizations emitted when pups are separated from the nest. Initial analyses of these calls were limited to rate of call production within a particular bandwidth of frequencies. More recently, analyses include classification of discrete vocalizations based upon their differences in frequency (pitch) and energy (loudness).
Adult male–female interactions
Adult vocalizations have been primarily studied within reproductive contexts. Adult males emit ultrasonic vocalizations (USVs) when exposed to a female partner or to her odor cues (Maggio et al. 1983, Nyby 2001, Whitney & Nyby 1979). During courtship, the male vocalizes and approaches the female (Maggio et al. 1983, Whitney & Nyby 1979). The relationship between male USV production and female behavior suggests that USVs predict mating opportunities rather than aggression. Female responses to these USVs include attenuated aggressive behaviors, approach and more frequent expression of mating postures (Sewell 1969). In a two-compartment choice test, females approach tethered males that are vocalizing vs. males that are surgically rendered incapable of vocalizing. If both males are rendered unable to vocalize, but one of the males is placed next to an ultrasound generator producing 70 kHZ ultrasounds, females prefer to reside in the compartment containing the male with the sound generator (Pomerantz et al. 1983).
During these male–female social interactions, males emit two types of vocalizations (Maggio et al. 1983, Nyby & Whitney 1978, Wang et al. 2008, Whitney & Nyby 1979). One is a highly modulated pure tone with a mean frequency of around 70 kHz and an intensity of 40 dB (Sales & Pye 1974). The second is a pure tone around 40 kHz with an intensity of more than 100 dB. The male emits the 70 kHz calls almost continuously prior to the first mount, while he actively sniffs and investigates the female. After the first mount, the number of calls declines, although during the latter stages of copulatory behavior, the male intermixes both 70 and 40 kHz calls immediately before and between mounting bouts (Nyby 1983, Sales & Pye 1974, White et al. 1998).
The development of digital sound spectrographic analysis of USVs has allowed researchers to expand analyses of mouse vocalizations to include dynamic changes in mean frequency (pitch). Individual calls can then be classified according to changes in dominant frequency, as a function of time. This analytical approach was used to assess USVs of adult males that were elicited by odors of receptive adult females (Holy & Guo 2005). Recordings were collected and calls were classified. For each call, pitch changes were compared for sudden punctuations in pitch that occur within a syllable. This analysis showed distinct clusters of pitch changes, such as upward and downward jumps between comparatively low frequencies (35–50 kHz) and similar transitions from between 70–90 kHz and 55–70 kHz (see Fig. 2e,f). There were also calls that lacked any abrupt changes in pitch. Often these calls were emitted in repeated temporal sequences (Holy & Guo 2005) and changed reproducibly with the succession of approach, mounting, and release behaviors that constitute the mating encounter. Before mounting, males usually called in either flat or continuous frequency-modulated USVs. However, when males mounted females, males shifted to frequency-modulated broken 'step-like' USVs with 70-kHz and 40/80 harmonic frequencies until the end of mounting (Wang et al. 2008).
Could these male vocalizations be relevant to affective prosody? Inferences that males derive a feeling of reward from social interactions with estrous females are warranted, based upon studies that use conditioned place preference (CPP) testing (and see below for discussion of CPP). Specifically, male mice prefer environments that have been associated with either mating (Kudwa et al. 2005, Popik et al. 2003) or even access to (Pankevich et al. 2006, Pierman et al. 2006) estrous female mice. The changing repertoire of male USVs through the mating process could reflect a shift from anticipation to consumption of sexual reward. In other words, these studies suggest that male mice may emit different peak frequencies that correlate with incentive salience vs. hedonic pleasure (see Berridge 2007).
Adult same-sex interactions
During male–male agonistic encounters, two types of calls can be expressed. V-shaped USVs encompass a broad frequency range and are emitted during naso-nasal contacts and body sniffing behaviors that precede fighting. A second set of calls consists of harmonics, or overtones, which are recorded during fighting behavior (Gourbal et al. 2004). These results are in agreement with the previous studies (Scott 1966) including observations that the number of audible vocalizations emitted during a male–male encounter correlates with time fighting and number of attacks (Brain et al. 1980, Morgret & Dengerink 1972). These audible vocalizations emitted during solicitation and mating, as there were similar to the ultrasonic vocalizations, are differences in USV structure that are associated with the anticipation vs. participation in social interactions. Taken together, the differences in frequency modulation associated with male anticipation and consumption of sexual vs. aggressive encounter indicate that these call patterns respond to differences in the affective nature of the social encounter.
There are fewer studies of vocalizations among female mice. Females express audible low-frequency vocalizations during copulation, particularly when they are not in estrous, consistent with the observation that they tend to avoid sexual encounters with males unless they are reproductively receptive. By contrast, female–female encounters engender USV production, especially when they were engaged in olfactory investigation of the other (Sewell 1970). Female mice emit a large number of USVs, typically emitted at 70 kHz, during the first minutes of social interaction (Gourbal et al. 2004, Moles et al. 2007). The rate of USV production among females appears to be responsive to levels of social motivation, experimentally imposed by food deprivation, estrous cycle, pregnancy and aging, such that USV rates positively correlate with levels of social investigation (Moles et al. 2007). Interestingly, just as estrous females produce fewer calls during same-sex interactions, brief exposure to a male also inhibits USV production (Maggio & Whitney 1985).
USVs of infant mice and the maternal response
Infant mice of postnatal day (PD) 1–4 are born blind and deaf (Fox 1965), have limited motor abilities, lack fur and subcutaneous fat and cool rapidly when displaced from the nest. When removed from the nest, they emit separation calls that elicit female retrieval. Under conditions of stress, cold and hunger, pups emit calls that engender maternal nest building, licking behaviors, and crouching behaviors that allow for access to teats (Noirot 1969). Rates of separation calls change with age and peak between PD 3 and PD 7, depending upon background strain (Branchi et al. 1998, Elwood & Keeling 1982, Hennessy et al. 1980, Scattoni et al. 2008b). By two weeks of age, juvenile mice no longer emit separation calls (Elwood & Keeling 1982, Noirot 1966).
As early as 1970, Sewell showed that lactating female wood mice (Apodemus sylvaticus) more frequently enter a compartment containing a loudspeaker emitting the recorded USVs vs. background noise or an artificial stimulus (Sewell 1970). Early studies also showed that lactating Mus musculus females preferentially approach ultrasounds within the frequency range of the natural calls (ranging from 40 to 80 kHz) vs. other ultrasonic sounds (Ehret & Haack 1981). More recently, a strain-dependent difference in the dynamic relationship between maternal responsiveness and pup calling rate was found. Using a three-compartment test structure, in which the mother could only reach her pups by crossing the central chamber which contained olfactory cues from a potentially infanticidal male, C57BL/6 mothers expressed higher maternal responsiveness vs. BALB/c females, and their pups emitted fewer calls than BALB/c pups (D'Amato et al. 2005). These data suggest that maternal responsiveness to pup calls, in turn, affects the rate that pups emit USVs.
Several studies show that USV production in response to separation and isolation can be a physiological response to a thermal challenge, e.g. a reflexive abdominal compression reaction in response to the cold that helps return venous blood to the heart (Blumberg & Alberts 1990, Blumberg & Sokoloff 2001). Additionally, pups vocalize when placed in a warm location (Branchi et al. 2004, Shair et al. 2003, Wöhr et al. 2008). In these cases, studies that employ knockout mice with alleles relevant to social bonding and separation distress suggest that separation calls are generated in response to affective changes. For instance, mice that lack a functional oxytocin allele, a molecule that plays a role in social bonding (Ferguson et al. 2000, 2002, Kavaliers et al. 2004, Mahler & Berridge 2009, Petrovic et al. 2008), emit fewer separation calls than their wild-type littermates (Winslow et al. 2000). Rates of separation calls are also influenced by the presence or absence of a mu-opioid receptor (Moles et al. 2004), which is a well-defined mediator of reward physiology and separation distress in rodents (Mahler & Berridge 2009, Panksepp et al. 1980).
Spectrographic analysis has showed that neonatal laboratory mice emit different kinds of USVs in response to varied environmental conditions to which the pup is exposed, such as odor from the nest, social isolation, low temperature, tactile stimulation or odor of an unfamiliar conspecific adult male (Branchi et al. 1998). For instance, isolated pups emit a high percentage of calls that contain abrupt breaks in frequency steps but an equal number of gradually modulated frequency calls relative to pups exposed to the odor of an unfamiliar, potentially infanticidal, adult male (Branchi et al. 1998). Variations in USV frequency modulation are also sensitive to the age of the pups, such that calls become stereotyped with maturation but remain acoustically distinct from adult USV patterns (Liu et al. 2003). See Fig. 1 for comparison of mouse vocalizations across development.
Mouse pups also produce 'wriggling' calls when struggling in the nest, mainly when pushing for the teats during suckling by the mother (Ehret 1975). These calls release at least three types of maternal behavior that include licking of pups, changes of suckling position, and nest building (Ehret & Bernecker 1986). Wriggling calls usually consist of a fundamental frequency near 4 kHz and overtones that can extend to a maximum frequency of about 20 kHz (Ehret & Bernecker 1986, Geissler & Ehret 2002, 2004). These calls elicit a specific neural activity. There is a differential response of C-fos immunoreactivity within the auditory cortex to synthetic wriggling calls played back to nursing females vs. similar synthetic wriggling USVs with a temporally displaced harmonic (Geissler & Ehret 2004). Interestingly, differences in immunoreactivity were more pronounced in the left hemisphere (Geissler & Ehret 2004), consistent with electrophysiological studies (Stiebler et al. 1997) and sharing similarities with human auditory processing (Ehret & Riecke 2002).
Call rates and frequency modulation of male USVs are responsive to genetic manipulations of endogenous reward pathways. For instance, targeted deletion of muscarinic receptor M5 results in an 80% decrease of male USVs production during sexual interaction. Male exposure to amphetamine, a direct ligand of D2 receptors, results in increased USV production, also responsive to targeted disruption of muscarinic receptors (Wang et al. 2008). Amphetamine can modulate dopamine release in the nucleus accumbens (Yeomans et al. 2000, 2001), thereby exerting a substantial role in appetitive behaviors (Berridge 2007, Ikemoto & Panksepp 1999, Kelley & Berridge 2002). Similarly, infant mice lacking functional mu-opioid receptors express reduced numbers of USVs during maternal separation (Moles et al. 2004), consistent with the evidence that enkephalin interactions with mu-opioid receptor can mediate social reward and isolation distress (Bertrand et al. 1997, Kalin et al. 1988, Panksepp et al. 1980, 1994, Vanderschuren et al. 1995).
Genetic manipulations that do not directly interact with incentive salience or hedonia can also modify call rate and call repertoire. For example, mice lacking functional components of circuits involved in social recognition, which include oxytocin and vasopressin receptor 1b, emit fewer calls (Scattoni et al. 2008a, Winslow & Insel 2002). Mice that lack functional alleles of genes associated with autistic-like social deficits, such as MECP2 and neuroligin 4 knockout mice, can express a diminished call rate (Jamain et al. 2008, Picker et al. 2006). Classification analysis has also showed that a spontaneous mutation (BTBR) and a targeted allelic replacement (foxp2 R552H) can influence the pup repertoire (Fujita et al. 2008, Scattoni et al. 2008b, Shu et al. 2005). This recent demonstration that ultrasound vocalizations can be modified by manipulations of foxp2, a gene involved in a familial form of speech impairment in humans and required for normal vocal development in songbirds, suggests that rodents may also be an important model for studying the neural and genetic basis of speech learning.
Mouse USVs and affect: theoretical considerations
Changes in subjective affective experience can be a proximal cause for changes in behavior. At a basic level, fear can induce freezing behavior, discomfort can induce withdrawal behavior, and anticipation of reward can induce an approach behavior. To the extent that specific aspects of the USV repertoire are associated with particular affective states, USV studies can provide insight to the genetic mechanisms that underlie affect regulation in normal psychological development and in mental illness.
Among humans, individuals make inferences to the quality of another person's experience based upon verbal communication or based upon observations of an individual's behavior. One means of communicating personal experience is through use of words. Self-report is an essential analytical tool for human research and clinical diagnosis, the basis of tremendous advances in psychological and psychiatric medicine. However, self-report has scientific and philosophical limitations. Bertrand Russell (1912) identified the problem: while words or labels are used to describe universal subjective states, it remains unknown whether these labels identify identical internal experiences. For example, two individuals can use the label 'red' to describe an apple but the red apple may appear as red-orange to one person and red-purple to another. It is inherently unknown whether both individuals have identical internal experiences, because of perceptual variations, such as differences in vision or brain function. Differences in sex, age, cultural context and mental illness further contribute to variation in how words label subjective experiences. For instance, post-traumatic stress disorder might impose an atypical sensitivity to stimuli that may be inadequately described with the existing verbal repertoire. Discrete labels used to communicate individual perceptions, no matter how basic or complicated, have limitations in terms of describing internal experience.
To gain greater insight to the affective experience of individuals, new assessment tools have been developed that do not depend upon verbal report. Directionality of eye-gaze (Klin et al. 2002, Wolf et al. 2008) and patterns of brain activity provide insights to how autistic individuals attend to social interactions (Baron-Cohen et al. 1999). Assessments of heart rate variability provide insight to how children experience distress of others (Eisenberg et al. 1988). To gain insight to the etiology of developmental disorders, there are ongoing efforts to identify the psychological experiences that are experiences during mental illness. For example, there is ongoing controversy about whether some forms of autism can result from social anxiety (Kuusikko et al. 2008), deficits in temporal cognition (Boucher et al. 2007) or central coherence (Briskman et al. 2001, Happe et al. 2001). Just as efforts to infer psychological experience can be useful for elucidating mechanisms underlying mental illness, the same issues are important for mouse research. If a knockout mouse expresses low levels of social approach, how do we know whether the targeted allele models autism, vs. depression, vs. shyness?
Such questions are rather new to behavioral neuroscience. Considerable precedence in the history of philosophy, extending back to the work of Rene Descartes (1637), maintains that animals are 'automata' or machine-like, in contrast to 'sentient' or soulful humans. With the advent of cognitive and affective neuroscience, these Cartesian distinctions are no longer relevant, but it is important to be careful about inferences to rodent affective experience (Panksepp 1998, Schneirla 1959). Such inferences require consideration of the theoretical framework of affective neuroscience, including operant, fear and preference conditioning paradigms, as well as studies of functional anatomy and brain/systems physiology (Panksepp 1998). Such an approach provides working hypotheses for comparative studies of affective prosody that can be subject to falsification.
Operant and place preference conditioning are used to establish anticipation and consumption of rewards (Bardo & Bevins 2000, Kelley et al. 2002, Moles et al. 2004), and aversive conditions can be identified by conditioned fear learning paradigms (Falls et al. 1997, LeDoux et al. 1988, Paylor et al. 1994). Emotional responses to stimuli can also be inferred from studies of functional anatomy, particularly the highly conserved limbic structures in the brain (MacLean 1990) and the underlying physiological systems that mediate affect regulation, including dopamine, serotonin, corticosterone and endogenous opiates (Panksepp 1998). Drugs of abuse, such as opiates and amphetamine, are also useful, because they can influence natural reward systems (Abarca et al. 2002, Bardo & Bevins 2000, Kelley & Berridge 2002, Reith & Selmeci 1992, van Ree et al. 1999). Pharmaceuticals that modulate affective states in humans can be employed that influence analogous behaviors in mouse models (Holsboer 2001, Hunsberger et al. 2007, Kato 2006, Pezet & Malcangio 2004, Rupniak et al. 2001).
In summary, assessments of prosody in mice along with measures of neural and behavioral correlates of motivated behavior, allow for greater reliability in inferences to subjective experience. Such assessments are important for translational applications of mouse research to mental illness. While human vocal patterns are vastly more complex with regard to frequency modulation, timing, overtones, and range of sound produced, and may provide more nuanced emotional content, and studies of human language can assess the temporal associations between prosodic content and referential aspects of language, basic elements of prosodic content in mice are measurable and have been associated with rewarding and aversive conditions. Thus, studies of prosody in humans and mice may yield apples-to-apples comparisons useful for subjective inference.
Mouse USVs and affect: practical considerations
Behavioral associations with USV production
Juvenile mice emit USVs when meeting a littermate after 24 h of social isolation. When USVs of juvenile C57BL/6J (B6) and BALBc/J (BALB) mice are compared during reunion, there are clear differences in the kinds of calls that they emit. For example, there are more downward-modulated calls among reunited B6 juveniles and more upward-modulated and chevron calls among BALB juveniles (see Fig. 2 for example spectrograms).
Differences in USV production between genetic variants of mice could be explained as products of discrete genetic contributions to the anatomies of the larynx or labia. Alternatively, there may be particular calls that are associated with state, such as the motivation to solicit social interaction. To assess the likelihood of these opposing hypotheses, we compared the usage of various call types by different individuals within a strain that express differences in social approach behaviors. Individual B6 and BALB juveniles express more USVs when they are engaged in more vigorous social approach behaviors (Panksepp et al. 2007). During more vigorous social approach behaviors, B6 mice disproportionally express calls that are of a higher pitch and more downward-modulated. These differences in B6 vocal repertoire suggest the possibility that different call types correspond to different degrees of social motivation or function to engender discrete levels of motivation in others. Also important, selective B6 use of particular call types during higher levels of social interaction was specific to B6 mice. BALB mice that engaged in vigorous social interactions expressed all call types at a higher rate. This finding indicates that the degree of repertoire flexibility within a strain is also dependent upon genetic background.
Rodents prefer social encounter to isolation, consistent with classical theories of motivated behavior (Bardo & Bevins 2000, Berridge & Robinson 2003, Tzschentke 1998) that underscore a role for reward and punishment in behavioral approach and withdrawal (Glickman & Schiff 1967, Schneirla 1959, Young 1959). Mouse responses to conditioned place preference (CPP) tests show that they can find social encounters rewarding and social isolation aversive. In a standard CPP experiment, mice are placed in one of two distinct environments. For instance, the beddings of the two environments may be different (one is aspen and the other is paper), the walls may have horizontal vs. vertical lines, or the floors may have circular holes in them or exist as a parallel grid. During the conditioning phases of these experiments, the test mouse is housed successively in either one or the other environment. The mouse has access to a putative reward (e.g. peanut butter, chocolate, cocaine, morphine) in one environment but not in the alternate environment. If the mouse finds the test stimulus rewarding, then through repeated association, the environment associated with access to the reward eventually elicits approach behavior. At the end of the conditioning phase, the mouse is placed in a testing structure that does not contain the reward but contains both of the 'conditioned' environments. The dependent measure of CPP is the difference in the amount of time spent in the environment associated with the presence of the putative reward vs. the environment paired with its absence. If the stimulus is rewarding, the mouse should spend more time in the environment that is paired with it. Among rodents, CPP tests have showed reward processes during different kinds of social encounters, including play (Calcagnetti & Schechter 1992, Douglas et al. 2004), sexual interactions (Camacho et al. 2004, Jenkins & Becker 2003), mother–infant bonding (Mattson et al. 2001, Weller et al. 2001) and even aggression (Martinez et al. 1995).
CPP experiments can also be helpful in determining whether mouse strains that express different USV repertoires experience different levels of social motivation. For instance, B6 mice show a strong place preference for bedding environments that have been associated with access to age-matched juveniles vs. beddings that are paired with social isolation (Panksepp & Lahvis 2007). These results suggest the possibility that the vocal repertoire of B6 mice may be associated with the degree of social reward; that there is a relationship between call pattern and various levels of social motivation. We can also gain insight to useful approaches for studying mouse USVs from insightful experiments of rat vocalizations. Rats emit high-frequency (50 kHz) calls when they anticipate a future reward or when they are experiencing the reward, and emit lower frequency (22 kHz) USVs under aversive conditions (Burgdorf et al. 2008, Knutson et al. 1999, Schwarting et al. 2007, Wöhr & Schwarting 2009). In addition to conditioning experiments, studies with operant tasks indicate that rats will self-administer playbacks of recordings of 50 kHZ 'trill' calls but not 22 kHz calls (Burgdorf et al. 2008). Similar kinds of evaluations might be useful for interpreting particular mouse USVs. It would be very useful to know whether specific call types associate with different CPP responses to specific drugs. Measures of mouse USVs have the potential to provide insight to real-time fluctuations in affect that we cannot obtain through CPP or fear conditioning testing. Experiments that take advantage of the differences in social motivation that accompany variations in the duration of social isolation or the time of day when social encounter occurs (Panksepp et al. 2008) could be used to discern how call patterns and responses to them change with social conditions.
Behavioral and systemic responses to vocalizations
Studies of animal acoustic communication utilize behavioral responses to recordings or 'playbacks' of particular kinds of animal calls to ascertain the meaning of various signals. In a series of experiments, we found that mice can become differentially fearful of a tone that is temporally associated with the vocalizations of a distressed mouse. This experiment involves a cue-conditioned fear learning paradigm (LeDoux et al. 1988). A standard fear conditioning procedure entails presenting an animal with a neutral stimulus, such as a tone (conditioned stimulus, CS), forward paired with an aversive stimulus, such as an electrical shock (unconditioned stimulus, UCS). Upon repeated administration of the paired CS–UCS, mice acquire a robust freezing behavior in response to presentation of the CS only. When B6 and BALB mice observe others undergo this fear conditioning procedure, only B6 mice are capable of learning from the distress of others. B6 also acquire the freezing response to the tone if they simply hear playbacks of the vocalizations of other distressed mice (played back 2-second recordings of the vocalizations of shocked mice) in association with the 30-second tone. Control experiments indicate that the BALB mice can hear the calls, because they respond to an environmental cue (the tone) of an overlapping frequency and energy level (approximately 85 dB). Importantly, heart rate deceleration is expressed in B6 when demonstrator mice undergo fear conditioning, but BALB mice do not show heart rate deceleration. Depression of heart rate can be associated with the empathic response to other's distress and this occurs among children more capable of detecting distress among others (Eisenberg et al. 2006). These studies indicate that fear can be perceived through vocal signals under the appropriate genetic background.
Brain activity in response to vocalizations
To assess the salience of different call types, researchers are beginning to examine freezing behavior, heart rate changes or other behavioral end-points in response to playbacks of auditory cues, such as distress vocalizations (Jeon et al. 2010, Knapska et al. 2006, Liu et al. 2006, Sadananda et al. 2008). For instance, variations in USV frequency modulation are sensitive to the age of the pups, with distinct influences on activity of the auditory cortex (Liu et al. 2003). In fact, experience appears to change neural processing of auditory stimuli; maternal auditory processing of pup USVs is distinct from that of naïve females (Liu et al. 2006, Liu & Schreiner 2007).
Future directions
USV analysis
There are a variety of approaches to vocal analysis, ranging from classification schemes to principal components analysis to hidden-Markov approaches. Various software programs have been developed to serve different purposes and each has its own benefits. Software programs can be employed to directly measure USV s, such as Avisoft SASLabPro (Panksepp et al. 2007, Scattoni et al. 2008a, Wöhr et al. 2008), others are designed to dissociate calls based upon highly nuanced spectral features, such as continuity and entropy (Tchernichovski et al. 2001), or identify calls in low signal-to-noise conditions (Mellinger 2009). Investigators also utilize custom MATLAB software programs to analyze frequency modulation in mouse USVs (Liu et al. 2006). Pattern recognition in USV sequence is also aided by the application of advanced statistical approaches (Ren et al. 2009). In this regard, future studies will likely utilize a variety of software approaches to obtain different kinds of information from calls, ranging from simple classifications of relatively similar call types (Panksepp et al. 2007) to discriminations of quite subtle changes in frequency modulation within a single call type. Differences in affective content might then be discerned by use of playbacks in combination with measures of brain activity, heart rate changes, behavioral responses, or other parameters that allow us to make inferences to affective response.
Complex environments
The rodent has long been considered a mammalian substitute for the human patient (Beach 1950), but when concentrating on a single, increasingly exploited, lab species, it has to be taken in mind that such a species is the 'ultimate' product (a variable genetic pool fragmented in a variety of local populations) of species-specific phylogenetic and epigenetic processes. Laboratory mice have been strongly selected for husbandry in a small cage, where there are limited opportunities for temporal and geographic variations in food access, social encounter, temperature, diurnal rhythms, encounter with predators, or exploration. By contrast, feral mice, by definition strictly associated to human settlements (Bronson 1979), show highly variable adaptive strategies that are not promoted by standard cage housing. It is likely that laboratory mice are thus not raised to vocalize in response to the more complex variety of social-emotional states that accompany a multifaceted environment. In this regard, wild-derived mice provide potentially useful opportunities to dissociate USV patterns (Musolfa et al. 2010, Kalcounis-Rueppell et al. 2010). A truly, genuine, comparative approach should be based on a much broader range of mouse experiences and representative species: the search for functional and/or structural invariance could be a fruitful 'zoo-semeiotic' perspective (sensu Sebeok) (Sebeok 1972). Studies with feral mice, as well as other feral rodent species, might provide a high degree of fidelity between call type and affective state.
More substantive vocal behaviors might also be showed by naturalistic or semi-naturalistic settings (Alleva et al. 1994, Portfors 2007). While mouse vocalizations are sensitive to targeted loss-of-function mutations in genes involved in affect regulation, language development, social bonding and autism, we may gain greater insight to how these mutations modify call patterns by expanding mouse developmental histories. Genetic variants can be raised in complex settings to expand their emotional experience and vocal repertoire. Rearing could entail geographic and temporal variation, such as random temporal access to food rewards, such as peanut butter, and stressors, such as cat urine, within a complex three-dimensional environment. Mice raised in environments that incorporate rewarding and aversive stimuli, in the context of more complex social paradigms, might provide a useful model for delineating relationships between affective states and specific call types.
- •Assessments of frequency modulation provide researchers with an ability to study USVs in a fashion that has translational relevance to prosodic deficits in mental illness. Measures of mouse USVs have the potential to provide a window to real-time changes in affective state that are not accessible by CPP or fear conditioning tests.
- •Inferences about call meaning should be considered within the context of ethologically relevant behaviors or traditional assessments of reward and aversion (e.g. conditioning experiments) and through the use of heart rate monitoring, assessments of brain activity, and other physiological indicators of affective response.
- •Mutant genetic models that are relevant to reward neurobiology, language, autism and social bonding can express abnormal USV patterns. Use of multiple statistical approaches for bioacoustic analysis, including clustering and hidden-Markov approaches, can help elucidate associations between USV structure and function.
- •By using wild-derived mice as rearing mice in more complex environments, researchers might also gain greater insight to the discrete functions, individual calls or call sequences within the diversity of the vocal repertoire.
- 2002) Cocaine sensitization and reward are under the influence of circadian genes and rhythm. Proc Natl Acad Sci U S A 99, 9026–9030. , & (
- 2004) Social cognition and its neural correlates in schizophrenia and autism. CNS Spectrums 9, 335–343. & (
- 1994) Behavioural Brain Research in Naturalistic and Semi-naturalistic Settings. Kluwer Academic Publishers, Dordrecht, The Netherlands. , , , & (
- 2000) Prosody and lexical accuracy in flat affect schizophrenia. Psychiatry Res 97, 107–118. , , & (
- 1992) Disorders of vocal signaling in children. In Papousek, U., Jurgens, U. & Papousek, M. (eds), Nonverbal Vocal Communication: Comparative and Developmental Approaches. Cambridge University Press, Cambridge, UK, pp. 192–204. (
- 2009) The influence of emotion clarity on emotional prosody identification in paranoid schizophrenia. Psychol Med 39, 927–938. , , & (
- 1984) Use of contrastive stress in normal, aphasic, and autistic children. J Speech Hear Res 27, 97–105. (
- 1987) The use of primary sentence stress by normal, aphasic, and autistic children. J Autism Dev Disord 17, 255–271. & (
- 2000) Conditioned place preference: what does it add to our preclinical understanding of drug reward? Psychopharmacology 153, 31–43. & (
- 1999) Social intelligence in the normal and autistic brain: an fMRI study. Eur J Neurosci 11, 1891–1898. , , , , , & (
- 1950) The snark was a boojum. Am Psychol 5, 115–124. (
- 2003) Comprehension of affective prosody in multiple sclerosis. Mult Scler 9, 148–153. , , & (
- 1998) A study of emotional processing in Parkinson's disease. Brain Cogn 38, 36–52. , & (
- 2007) The debate over dopamine's role in reward: the case for incentive salience. Psychopharmacology 191, 391–431. (
- 2003) Parsing reward. Trends Neurosci 26, 507–513. & (
- 1997) Social interaction increases the extracellular levels of [Met]enkephalin in the nucleus accumbens of control but not of chronic mild stressed rats. Neuroscience 80, 17–20. , , , & (
- 1990) Ultrasonic vocalizations by rat pups in the cold: an acoustic by-product of laryngeal braking? Behav Neurosci 104, 808–817. & (
- 2001) Do infant rats cry? Psychol Rev 108, 83–95. & (
- 2007) Temporal cognition in children with autistic spectrum disorders: tests of diachronic thinking. J Autism Dev Disord 37, 1413–1429. , , & (
- 2006) Impaired perception of affective prosody in schizophrenia. J Neuropsychiatry Clin Neurosci 18, 81–85. , , , , & (
- 1980) A device for recording submissive vocalizations of laboratory mice. Physiol Behav 24, 1003–1006. , , & (
- 1998) Ultrasonic vocalizations by infant laboratory mice: a preliminary spectrographic characterization under different conditions. Dev Psychobiol 33, 249–256. , , & (
- 2004) Neonatal behaviors associated with ultrasonic vocalizations in mice (Mus musculus): a slow-motion analysis. Dev Psychobiol 44, 37–44. , , & (
- 2001) Exploring the cognitive phenotype of autism: Weak "central coherence" in parents and siblings of children in autism: II. Real-life skills and preferences. J Child Psychol Psychiatry 42, 309–316. , & (
- 1979) The reproductive ecology of the house mouse. Q Rev Biol 54, 265–299. (
- 2008) Ultrasonic vocalizations of rats (Rattus norvegicus) during mating, play, and aggression: behavioral concomitants, relationship to reward, and self-administration of playback. J Comp Psychol 122, 357–367. , , , , & (
- 1992) Place conditioning reveals the rewarding aspect of social interaction in juvenile rats. Physiol Behav 51, 667–672. & (
- 2004) Sexual experience and conditioned place preference in male rats. Pharmacol Biochem Behav 78, 419–425. , & (
- 2005) Effects of positive affect on speech disorder in schizophrenia. J Nerv Ment Dis 193, 839–842. & (
- 2010) Observational fear learning involves affective pain system and Cav1.2 Ca2+ channels in ACC. Nat Neurosci 13, 482–488. , , , , , , , & (
- 2005) Pups call, mothers rush: does maternal responsiveness affect the amount of ultrasonic vocalizations in mouse pups? Behav Genet 35, 103–112. , , & (
- 2009) Discourse on the Method. Oxford University Press, Oxford. (
- 2009) Generalized deficit in all core components of empathy in schizophrenia. Schizophr Res 108, 197–206. , , , , , & (
- 2008) Resolving ambiguity: a psycholinguistic approach to understanding prosody processing in high-functioning autism. Brain Lang 106, 144–152. , , , & (
- 2004) Rewarding properties of social interactions in adolescent and adult male and female rats: impact of social versus isolate housing of subjects and partners. Dev Psychobiol 45, 153–162. , & (
- 1975) Schallsignale der Hausmaus (Mus Musculus). Behaviour 52, 38–56. (
- 1986) Low-frequency sound communication by mouse pups (Mus musculus): wriggling calls release maternal behaviour. Anim Behav 34, 821–830. & (
- 1981) Categorical perception of mouse pup ultrasound by lactating females. Naturwissenschaften 68, 208–209. & (
- 2002) Mice and humans perceive multiharmonic communication sounds in the same way. Proc Natl Acad Sci U S A 99, 479–482. & (
- 1988) Differentiation of vicariously induced emotional reactions in children. Dev Psychol 24, 237–246. , , , , & (
- 2006) Prosocial development. In Eisenberg, N. (ed), Handbook of Child Psychology. John Wiley & Sons Inc., New Jersey, pp. 646–718. , & (
- 1982) Temporal organization of ultrasonic vocalizations in infant mice. Dev Psychobiol 15, 221–227. & (
- 1999) Investigation of receptive affective prosodic ability in school-aged boys with and without depression. Neuropsychiatry Neuropsychol Behav Neurol 12, 102–109. , & (
- 1997) Fear-potentiated startle in two strains of inbred mice. Behav Neurosci 111, 855–861. , , & (
- 2000) Social amnesia in mice lacking the oxytocin gene. Nat Genet 25, 284–288. , , , , & (
- 2002) The neuroendocrine basis of social recognition. Front Neuroendocrinol 23, 200–224. , & (
- 1989) Intonation and communicative intent in mothers' speech to infants: is the melody the message? Child Dev 60, 1497–1510. (
- 1991) The use of intonation to communicate in pervasive developmental disorders. J Child Psychol Psychiatry 32, 771–782. , , & (
- 1965) Reflex-ontogeny and behavioural development of the mouse. Anim Behav 13, 234–241. (
- 2009) Comprehension of affective prosody in veterans with chronic posttraumatic stress disorder. The Journal of Neuropsychiatry and Clinical Neurosciences 21, 52–58. , , & (
- 2008) Ultrasonic vocalization impairment of Foxp2 (R552H) knockin mice related to speech-language disorder and abnormality of Purkinje cells. Proceedings of the National Academy of Sciences of the United States of America 105, 3117–3122. , , , , , & (
- 2008) Female specific anterior cingulate abnormality and its association with empathic disability in schizophrenia. Prog Neuropsychopharmacol Biol Psychiatry 32, 1728–1734. , , , , , , , & (
- 2002) Time-critical integration of formants for perception of communication calls in mice. Proc Natl Acad Sci U S A 99, 9021–9025. & (
- 2004) Auditory perception vs. recognition: representation of complex communication sounds in the mouse auditory cortical fields. Eur J Neurosci 19, 1027–1040. & (
- 1967) A biological theory of reinforcement. Psychol Rev 74, 81–109. & (
- 2007) The 'Reading the Mind in the Voice' test-revised: a study of complex emotion recognition in adults with and without autism spectrum conditions. J Autism Dev Disord 37, 1096–1106. , , & (
- 2004) Spectrographic analysis of the ultrasonic vocalisations of adult male and female BALB/c mice. Naturwissenschaften 91, 381–385. , , & (
- 1986) An analysis of autism in fifty males with the fragile X syndrome. American Journal of Medical Genetics 23, 359–374. , , , & (
- 2009) Empathy in schizophrenia: impaired resonance. Eur Arch Psychiatry Clin Neurosci 259, 352–361. & (
- 2001) Exploring the cognitive phenotype of autism: weak "central coherence" in parents and siblings of children with autism: I. Experimental tests. J Child Psychol Psychiatry 42, 299–307. , & (
- 1980) Maternal behavior, pup vocalizations, and pup temperature changes following handling in mice of 2 inbred strains. Dev Psychobiol 13, 573–584. , , & (
- 2001) Prospects for antidepressant drug discovery. Biol Psychol 57, 47–65. (
- 2005) Ultrasonic songs of male mice. PLoS Biol 3, e386. & (
- 1968) Sonographic analysis of vocalizations of three species of mice. J Mammal 49, 555–560. (
- 2007) Antidepressant actions of the exercise-regulated gene VGF. Nat Med 13, 1476–1482. , , , , , & (
- 1999) The role of nucleus accumbens dopamine in motivated behavior: a unifying interpretation with special reference to reward-seeking. Brain Res Rev 31, 6–41. & (
- 2008) Reduced social interaction and ultrasonic communication in a mouse model of monogenic heritable autism. Proc Natl Acad Sci U S A 105, 1710–1715. , , , , , , , , , , , , , & (
- 2003) Female rats develop conditioned place preferences for sex at their preferred interval. Horm Behav 43, 503–507. & (
- 2010) Differences in ultrasonic vocalizations between wild and laboratory California mice (Peromyscus californicus). PLoS One 5: DOI: 10.1371/journal.pone.0009705, 1–10. , , , , , , , & (
- 1988) Opiate modulation of separation-induced distress in non-human primates. Brain Res 440, 285–292. , & (
- 2006) The role of mitochondrial dysfunction in bipolar disorder. Drug News Perspect 19, 597–602. (
- 2004) Olfactory-mediated parasite recognition and avoidance: linking genes to behavior. Horm Behav 46, 272–283. , , & (
- 2002) The neuroscience of natural rewards: relevance to addictive drugs. J Neurosci 22, 3306–3311. & (
- 2002) Opioid modulation of taste hedonics within the ventral striatum. Physiol Behav 76, 365–377. , , , , & (
- 2002) Visual fixation patterns during viewing of naturalistic social situations as predictors of social competence in individuals with autism. Arch Gen Psychiatry 59, 809–816. , , , & (
- 2006) Between-subject transfer of emotional information evokes specific pattern of amygdala activation. Proc Natl Acad Sci U S A 103, 3858–3862. , , , , , & (
- 1999) High-frequency ultrasonic vocalizations index conditioned pharmacological reward in rats. Physiol Behav 66, 639–643. , & (
- 2005) Dopamine D5 receptor modulates male and female sexual behavior in mice. Psychopharmacology 180, 206–214. , , , & (
- 2005) Links between social and linguistic processing of speech in preschool children with autism: behavioral and electrophysiological measures. Dev Sci 8, F1–F12. , , & (
- 2008) Social anxiety in high-functioning children and adolescents with autism and Asperger syndrome. J Autism Dev Disord 38, 1697–1709. , , , , , , & (
- 1988) Different projections of the central amygdaloid nucleus mediate autonomic and behavioral correlates of conditioned fear. J Neurosci 8, 2517–2529. , , & (
- 1998) Disturbances of affective prosody in patients with schizophrenia; a cross sectional study. J Neurol Neurosurg Psychiatry 64, 375–378. , , & (
- 2006) Theory of Mind (ToM) and counterfactuality deficits in schizophrenia: misperception or misinterpretation? Psychol Med 36, 1075–1083. , , & (
- 2007) Auditory cortical detection and discrimination correlates with communicative significance. PLoS Biol 5, e173. & (
- 2003) Acoustic variability and distinguishability among mouse ultrasound vocalizations. J Acoust Soc Am 114, 3412–3422. , , & (
- 2006) Improved cortical entrainment to infant communication calls in mothers compared with virgin mice. Eur J Neurosci 23, 3087–3097. , & (
- The autism diagnostic observation schedule-generic: a standard measure of social and communication deficits associated with the spectrum of autism. Journal of Autism & Developmental Disorders 30, 205–223. , , , , , , & (2000)
- 1990) The Triune Brain in Evolution: Role in Paleocerebral Functions. Plenum Press, New York. (
- 1985) Ultrasonic vocalizing by adult female mice (Mus musculus). J Comp Psychol 99, 420–436. & (
- 1983) Experience-based vocalization of male mice to female chemosignals. Physiol Behav 31, 269–272. , & (
- 2009) Which cue to "want?" Central amygdala opioid activation enhances and focuses incentive salience on a prepotent reward cue. J Neurosci 29, 6500–6513. & (
- 1995) Successful intermale aggression and conditioned place preference in mice. Physiol Behav 58, 323–328. , , & (
- 2001) Comparison of two positive reinforcing stimuli: pups and cocaine throughout the postpartum period. Behav Neurosci 115, 683–694. , , & (
- 2007) Prosody and its relationship to language in school-aged children with high-functioning autism. Int J Lang Commun Disord 42, 682–702. , , , & (
- 2009) Noise-resistant acoustic measurements implemented in user-friendly software. J Acoust Soc Am 125, 2737. (
- 2004) Deficit in attachment behavior in mice lacking the mu-opioid receptor gene. Science 304, 1983–1986. , & (
- 2007) Ultrasonic vocalizations emitted during dyadic interactions in female mice: a possible index of sociability? Behav Brain Res 182, 223–230. , , , & (
- 2001) Altered emotional perception in alcoholics: deficits in affective prosody comprehension. Alcohol Clin Exp Res 25, 362–369. , , & (
- 2002) Neurological basis of deficits in affective prosody comprehension among alcoholics and fetal alcohol-exposed adults. J Neuropsychiatry Clin Neurosci 14, 321–328. , , & (
- 1972) The squeal as an indicator of aggression in mice. Behav Res Methods Instrumentation 4, 138–140. & (
- 1990) Prosodic comprehension and expression in schizophrenia. J Neurol Neurosurg Psychiatry 53, 727–730. & (
- 2010) Ultrasonic courtship vocalizations in wild house mice, Mus musculus musculus. Anim Behav 79, 757–764. , & (
- 1966) Ultra-sounds in young rodents. I. Changes with age in albino mice. Anim Behav 14, 459–462. (
- 1969) Changes in responsiveness to young in the adult mouse. V. Priming. Anim Behav 17, 542–546. (
- 1983) Ultrasonic vocalizations during sex behavior of male house mice (Mus musculus): a description. Behav Neural Biol 39, 128–134. (
- 2001) Auditory communication among adults. In Willott, J.F. (ed), Handbook of Mouse Auditory Research: From Behavior to Molecular Biology. CRC Press, Boca Raton, Florida, pp. 3–18. (
- 1978) Ultrasonic communication of adult myomorph rodents. Neurosci Biobehav Rev 2, 1–14. & (
- 2007) Loss of insight in frontotemporal dementia, corticobasal degeneration and progressive supranuclear palsy. Brain: A Journal of Neurology 130, 753–764. , , , , , , & (
- 2005) Impaired comprehension of affective prosody in elderly subjects is not predicted by age-related hearing loss or age-related cognitive decline. Journal of Geriatric Psychiatry & Neurology 18, 25–32. , , & (
- 2006) Accessory olfactory neural Fos responses to a conditioned environment are blocked in male mice by vomeronasal organ removal. Physiol Behav 87, 781–788. , & (
- 1998) Affective Neuroscience: The Foundations of Human and Animal Emotions. Oxford University Press, Oxford. (
- 2007) Social reward among juvenile mice. Genes Brain Behav 6, 661–671. & (
- 1980) Endogenous opioids and social behavior. Neurosci Biobehav Rev 4, 473–487. , , , & (
- 1994) Brain opioids and mother-infant social motivation. Acta Paediatr Suppl 397, 40–46. , & (
- 2007) Affiliative behavior, ultrasonic communication and social reward are influenced by genetic variation in adolescent mice. PLoS ONE 2: DOI: 10.1371/journal.pone.0000351, 1–13. , , , , , , & (
- 2008) Differential entrainment of a social rhythm in adolescent mice. Behav Brain Res 195, 239–245. , , & (
- 1994) DBA/2 and C57BL/6 mice differ in contextual fear but not auditory fear conditioning. Behav Neurosci 108, 810–817. , , & (
- 2008) Oxytocin attenuates affective evaluations of conditioned faces and amygdala activity. J Neurosci 28, 6607–6615. , , & (
- 2004) Brain-derived neurotrophic factor as a drug target for CNS disorders. Expert Opin Ther Targets 8, 391–399. & (
- 2006) An altered neonatal behavioral phenotype in Mecp2 mutant mice. Neuroreport 17, 541–544. , , & (
- 2006) Male aromatase knockout mice acquire a conditioned place preference for cocaine but not for contact with an estrous female. Behav Brain Res 174, 64–69. , , , & (
- 2007) Impaired perception of negative emotional prosody in schizophrenia. Clin Neuropsychol 21, 762–775. , , & (
- 1983) Female behavior is affected by male ultrasonic vocalizations in house mice. Physiol Behav 31, 91–96. , & (
- 2003) Effects of memantine, an NMDA receptor antagonist, on place preference conditioned with drug and nondrug reinforces in mice. Behav Pharmacol 14, 237–244. , , , & (
- 2007) Types and functions of ultrasonic volcaizations in laboratory rats and mice. J Am Assoc Lab Anim Sci 46, 28–34. (
- 1992) Cocaine binding sites in mouse striatum, dopamine autoreceptors, and cocaine-induced locomotion. Pharmacol Biochem Behav 41, 227–230. & (
- 2009) A Framework for Bioacoustic Vocalization Analysis Using Hidden Markov Models Algorithms 2, 1410–1428. , , , , , & (
- 2004) How would you feel versus how do you think she would feel? A neuroimaging study of perspective-taking with social emotions. Journal of Cognitive Neuroscience 16, 988–999. & (
- 2001) Comparison of the phenotype of NK1R-/- mice with pharmacological blockade of the substance P (NK1) receptor in assays for antidepressant and anxiolytic drugs. Behav Pharmacol 12, 497–508. , , , , , , , , & (
- 2002) Reading the mind in the voice: a study with normal adults and adults with Asperger syndrome and high functioning autism. J Autism Dev Disord 32, 189–194. , & (
- 2008) Playback of 22-kHz and 50-kHz ultrasonic vocalizations induces differential c-fos expression in rat brain. Neurosci Lett 435, 17–23. , & (
- 1972) Ultrasound and mating behaviour in rodents with some observations on other behavioural situations. J Zool 168, 149–164. (
- 1974) Ultrasonic Communication by Animals. Chapman & Hall Ltd., London. & (
- 2008a) Reduced ultrasonic vocalizations in vasopressin 1b knockout mice. Behav Brain Res 187, 371–378. , , , , , & (
- 2008b) Unusual repertoire of vocalizations in the BTBR T+tf/J mouse model of autism. PLoS ONE 3, e3067. , , & (
- 1959) An evolutionary and developmental theory of biphasic processes underlying approach and withdrawal. In Jones, M.R. (ed), Nebraska Symposium on Motivation. University of Nebraska Press, Lincoln, pp. 1–42. (
- 2008) The processing of emotional prosody and semantics in schizophrenia: relationship to gender and IQ. Psychol Med 38, 887–898. , & (
- 2007) Situational factors, conditions and individual variables which can determine ultrasonic vocalizations in male adult Wistar rats. Behav Brain Res 182, 208–222. , & (
- 1966) Agonistic behavior of mice and rats: a review. Am Zool 6, 683–701. (
- 1972) Perspectives in Zoosemiotics. Mouton, The Hague. (
- 1969) Ultrasound in Small Mammals. Unpublished doctoral thesis, University of London, London. (
- 1970) Ultrasonic communication in rodents. Nature 227, 410. (
- 2003) Social, thermal, and temporal influences on isolation-induced and maternally potentiated ultrasonic vocalizations of rat pups. Dev Psychobiol 42, 206–222. , , , & (
- 2001) Speech and prosody characteristics of adolescents and adults with high-functioning autism and Asperger syndrome. J Speech Lang Hear Res 44, 1097–1115. , , , , & (
- 2005) Altered ultrasonic vocalization in mice with a disruption in the Foxp2 gene. Proceedings of the National Academy of Sciences of the United States of America 102, 9643–9648. , , , , , , , , , , , , & (
- 2008) Levels of emotional awareness and autism: an fMRI study. Soc Neurosci 3, 97–112. , , , , & (
- 2009) The empathy imbalance hypothesis of autism: a theoretical approach to cognitive and emotional empathy in autistic development. Psychol Record 59, 273–294. (
- 1997) The auditory cortex of the house mouse: left-right differences, tonotopic organization and quantitative analysis of frequency representation. J Comp Physiol A Sens Neural Behav Physiol 181, 559–571. , , & (
- 2001) Dynamics of the Vocal Imitation Process: How a Zebra Finch Learns Its Song. Science 291, 2564–2569. , , & (
- 2001) Impaired affective prosody in AD: Relationship to aphasic deficits and emotional behaviors. Neurology 57, 1474–1481. , , , & (
- 1998) Measuring reward with the conditioned place preference paradigm: a comprehensive review of drug effects, recent progress and new issues. Prog Neurobiol 56, 613–672. (
- 2008) Social cognition in alcoholism: a link to prefrontal cortex dysfunction? Addiction 103, 726–735. & (
- 2005) Processing of affective stimuli in alcoholism. Cortex 41, 189–194. , , & (
- 2008) Perception of affective prosody in major depression: a link to executive functions? J Int Neuropsychol Soc 14, 552–561. , , , & (
- 1999) Opioids, reward and addiction: an encounter of biology, psychology, and medicine. Pharmacol Rev 51, 341–396. , & (
- 1995) Social play alters regional brain opioid receptor binding in juvenile rats. Brain Res 680, 148–156. , , & (
- 2008) Content and style of speech from mothers with schizophrenia towards their infants. Psychiatry Res 159, 109–114. , , & (
- 2008) Ultrasonic vocalizations induced by sex and amphetamine in M2, M4, M5 muscarinic and D2 dopamine receptor knockout mice. PLoS ONE [Electronic Resource] 3, e1893. , , , & (
- 2001) Examining the role of cholecystokinin in appetitive learning in the infant rat. Peptides 22, 1317–1323. , , & (
- 1998) 40- and 70-kHz vocalizations of mice (Mus musculus) during copulation. Physiol Behav 63, 467–473. , , & (
- 1969) Vocalization of mice: a single genetic unit effect. J Hered 60, 337–340. (
- 1979) Cues that elicit ultrasounds from adult male mice. Am Zool 19, 457–463. & (
- 1983) Sound communication among adults. In Willot, J.F. (ed),The Auditory Psychology of the Mouse. Charles C Thomas, Springfield, pp. 98–129. & (
- 2002) The social deficits of the oxytocin knockout mouse. Neuropeptides 36, 221–229. & (
- 2000) Infant vocalization, adult aggression, and fear behavior of an oxytocin null mutant mouse. Horm Behav 37, 145–155. , , , , & (
- 1965) The physiology and pharmacology of pain. In de Stevens, G. (ed), Analgesics. Academic, New York. (
- 2009) Ultrasonic communication in rats: effects of morphine and naloxone on vocal and behavioral responses to playback of 50-khz vocalizations. Pharmacol Biochem Behav 94, 285–295. & (
- 2008) Effects of genetic background, gender, and early environmental factors on isolation-induced ultrasonic calling in mouse pups: an embryo-transfer study. Behav Genet 38, 579–595. , , , , & (
- 2008) Specific impairment of face-processing abilities in children with autism spectrum disorder using the Let's Face It! skills battery. Autism Res 1, 329–340. , , , , , , , , , & (
- 2000) Brain-stimulation reward thresholds raised by an antisense oligonucleotide for the M5 muscarinic receptor infused near dopamine cells. J Neurosci 20, 8861–8867. , , , , , , & (
- 2001) M5 muscarinic receptors are needed for slow activation of dopamine neurons and for rewarding brain stimulation. Life Sci 68, 2449–2456. , & (
- 2006) Selective impairment of sadness and disgust recognition in abstinent ecstasy users. Neuropsychologia 44, 959–965. & (
- 1959) The role of affective processes in learning and motivation. Psychol Rev 66, 104–125. (
The authors wish to thank Jan Van Santen and Lois Black of Oregon Health and Science University for communicating their insights to prosodic communication. We also want to acknowledge the Waisman Center at the University of Wisconsin for support (P30 HD03352). This work was supported by R01 funding from the National Institute of Drug Abuse (DA022543).


text:
'California Gurls,' 'OMG' vie for song of summer
Published: Thursday, July 8, 2010 at 5:31 p.m.
Last Modified: Thursday, July 8, 2010 at 5:31 p.m.
NEW YORK — The summer's barely begun, but Katy Perry's "California Gurls" is already on pace to be the song of the season.
Billboard is tracking the summer's most popular songs and, so far, Perry's No. 1 hit is leading, followed by another ubiquitous song: Usher's "OMG" featuring will.i.am.
In third place is B.o.B's "Airplanes" featuring Hayley Williams. Travie McCoy's "Billionaire" with Bruno Mars is fourth, and Ke$ha's "Your Love Is My Drug" rounds out the top five.
Billboard started collecting data for the "summer" season Memorial Day Week and will crown the winner after Sept. 9.
Reader comments posted to this article may be published in our print edition. All rights reserved. This copyrighted material may not be re-published without permission. Links are encouraged.

subset: commoncrawl


text:
OLYMPIA, Wash. (AP) — U.S. Sen. Maria Cantwell is well ahead of her Republican challenger in fundraising for her re-election bid.
According to filings this week with the Federal Election Commission, Cantwell raised $1.2 million during the first three months of the year compared to the $188,000 raised by state Sen. Michael Baumgartner of Spokane. Overall, Cantwell's fundraising report shows she has $4.6 million in cash to spend, compared to Baumgartner's $144,600.
Cantwell is seeking her third term in office.
Information from: The Seattle Times, http://www.seattletimes.com


text:
5 years in jail for heroin chargesFebruary 09,2013Five-year jail term
for heroin charges
BRATTLEBORO A Connecticut man has been sentenced to serve five years in prison for selling heroin.
Jesse Anderson of Hartford pleaded guilty last month to three counts of heroin sale or delivery. He was arrested in September following an investigation that resulted in police-controlled purchases of 91 bags of heroin.
Court documents say police used a cooperating individual to set up an initial drug buy in August from a man they knew only as Country, according to the Brattleboro Reformer. He was later identified as the 33-year-old Anderson.
Police said the deal was supposed to be for 20 bags of heroin, but the informant returned with only 18. The informant then returned to Country, who apologized by offering three more bags.MORE IN This Just InMONTPELIER The races might be new, but many of the faces are the same. Full StoryRutland Town officials want to ensure businesses pay the towns 1 percent sales tax. Full StoryThe sole Democrat in the Rutland City delegation is not seeking another term.
Rep. Full Story
- Most Popular
- Most Emailed


text:
Trelstar LA
FDA Expands Zytiga for Late-Stage Prostate Cancer »
"The U.S. Food and Drug Administration today expanded the approved use of Zytiga (abiraterone acetate) to treat men with late-stage (metastatic) castration-resistant prostate cancer prior to receiving chemotherapy.
The FDA initially appr"...
Read the FDA Expands Zytiga for Late-Stage Prostate Cancer article »
Trelstar LA
Trelstar LA Consumer (continued)
OVERDOSE: If overdose is suspected, contact a poison control center or emergency room immediately. US residents can call their local poison control center at 1-800-222-1222. Canada residents can call a provincial poison control center.
NOTES: Laboratory and/or medical tests (such as testosterone blood levels, PSA blood tests, blood glucose) should be performed periodically to monitor your progress or check for side effects. Consult your doctor for more details.
MISSED DOSE: It is very important that you do not miss any doses. However, if you do miss a dose, contact your doctor promptly to establish a new schedule.
STORAGE: Not applicable. This medication is given in a doctor's office and will not be stored at home.
Information last revised July 2014. Copyright(c) 2014 First Databank, Inc.
Trelstar LA Consumer Overview
Additional Trelstar LA Information
Trelstar LA - User Reviews
Trelstar LA User Reviews
Now you can gain knowledge and insight about a drug treatment with Patient Discussions.
Here is a collection of user reviews for the medication Trelstar LA sorted by most helpful.
Report Problems to the Food and Drug Administration
You are encouraged to report negative side effects of prescription drugs to the FDA. Visit the FDA MedWatch website or call 1-800-FDA-1088.
Get the latest treatment options.


text:
Just as important as being able to take the CPA Exam is whether you can pass the CPA Exam. The CPA Exam is very rigorous and the pass rate is significantly lower than the pass rates for college accounting courses. Students having successfully completed 30 college credits in accounting courses (plus 120 credits of other college courses) find it very difficult to pass the CPA exam. As a result, students with accounting degrees often enroll in CPA review courses to increase their likelihood of passing the CPA Exam. In other words, you need to ask yourself whether you have the accounting knowledge to pass the CPA exam in addition to meeting your state or jurisdiction's educational requirements.
Our Accounting Career Center provides links to each state's board of accountancy and more details on the difficulty of passing the CPA Exam.
Studies show that exam questions are a great way to learn and retain important information. Gain access to our 1,700 accounting exam questions (and answers) when you upgrade to PRO.


text:
Editors' Note: The "Download Now" link directs you to the iTunes App Store, where you must continue the download process. You must have iTunes installed in order to open the link, and you must have an active iTunes account to download the application. This download may not be available in some countries.
Publisher's Description+
What's new in this version:
+ New option to hide expired deal+ Expired deal will show in italic font and gray color+ Reply function implemented


text:
Readout of the Secretary-General's phone call with H.E. Mr. Victor Yanukovich, President of Ukraine
Paris, France, 8 December 2013
The Secretary-General spoke today with the President of Ukraine, H.E. Mr. Victor Yanukovich.
He congratulated the President on Ukraine's successful Organization for Security and Co-operation in Europe (OSCE) Chairmanship and Ministerial Meeting.
The Secretary-General expressed his grave concern about the situation in Ukraine, emphasised that there must be no resort to violence, and appealed for peaceful dialogue amongst all parties concerned.
The Secretary-General welcomed the President's assurances that consultations would be initiated to defuse the situation.


text:
Coming up in the next {{countdown}} {{countdownlbl}}
Coming up next:
Skip to this video now
Now Playing:
Play Video
This transcript has been automatically generated and may not be 100% accurate.
Missing Malaysia Plane: Radar Hits of 'Significant Size' in Indian Ocean
The U.S. Navy's P-8 Poseidon is searching an area where two objects have been spotted by satellites.
Objects in Indian Ocean Are 'Best Lead' in Plane Search
The search for a missing Malaysia Airlines plane picks up off Australia's coast.
EXCLUSIVE: David Wright Aboard Navy Search and Rescue Plane
ABC News' David Wright gets an exclusive look at U.S. Navy's search for missing plane off coast of Australia.
Now Playing: Missing Malaysia Plane: Radar Hits of 'Significant Size' in Indian Ocean
Now Playing: EXCLUSIVE: David Wright Aboard Navy Search and Rescue Plane
Now Playing: EXCLUSIVE: David Wright Reports as US Navy Plane Zeroes in on Debris
Now Playing: Khloe Kardashian Files to Divorce Lamar Odom--Again
Now Playing: World Record Set For Speed-Hugging
Now Playing: Friday Rewind: Baltimore Officer Acquitted in Freddie Gray Case
Now Playing: Michelle Obama Hosts Turnaround Talent Show at the White House
Now Playing: Amber Heard and Johnny Depp are Calling it Quits
Now Playing: Weekend Work Emails Are Illegal In France
Now Playing: Obama Attends G-7 Summit
Now Playing: Kid Gets Same Haircut as His Neighbor
Now Playing: 'Dancing With the Stars' Finale Recap
Now Playing: Obama Wraps Up Vietnam Trip
Now Playing: Skinny 'The Bachelorette' 2016 Premiere
Now Playing: President Obama Sends Human Rights Message to Vietnam Government
Now Playing: The History of Comedy
Now Playing: Tap Dancers' Tribute to Prince
Now Playing: Nile Crocodiles Found In Florida
Now Playing: Obama Visits Vietnam, Seeks to Strengthen Ties
Now Playing: Kesha Will Perform at the Billboard Music Awards
Now Playing: High School Valedictorian Banned at Graduation
Now Playing: The Pentagon Will Refund Tax Dollars Used for 'Paid Patriotism'
Now Playing: {{itm.title}}


text:
Juan Luis Cifuentes Lemus: Doctor Taxista: The enthusiasm and passion of Dr. Juan Luis Cifuentes Lemus, Honoris Causa of CUCosta (Centro Universitario de la Costa, UDG), precedes this marine biologist, public activist and environmentalist. With a motto of "living beings well-evaluated" (los seres vivos bien evaluados), he focuses his prime attention on the youth, believing that "what we know, we take care of."
Surfing: For decades, surfers of all ages have flocked to Mexico's Pacific coast, long considered to be a destination of choice, in order to benefit from its more than 1,500 miles of beaches, points and reefs. Well-known surf spots from the northern state of Sinaloa all the way down to Oaxaca are awaiting exploration year round.
Cravings: A Guayaba: A very rich source of vitamin C (even more so than the ubiquitous orange), guayabas – or guavas, as they are known up north – are as delicious fresh as they are in their many widely available incarnations, from thick rolls of rich guava paste to bottled guava nectar. And since they can be harvested year round in warm climates, they are considered one of Mexico's leading fruits.
Living with Art in a Tropical Environment: The paper and chemicals traditionally used in photography make it especially vulnerable to damage, such as fading, mold and disintegration, from exposure to light, humidity and heat. Special technology and products used in the printing and mounting processes, along with temperature and humidity-control of the space where photos are displayed or stored, can circumvent the risk.
Molcajete Salsa at Your Table: A basalt (volcanic rock) tool traditionally used at home to grind spices and prepare salsas and guacamole, the molcajete, or Mexican mortar, is the big enchilada of kitchen gadgets here, dating back to Aztec times.
Earnestly Speaking Nonsense — In a Spanish-Speaking Land: I doubt any gringos actually set out to make fools of themselves here. But speaking "Spanglish" certainly adds to the likelihood that it's going to happen now and then. Personally, since my livelihood depends on a facility with language, I'm mortified to have an IQ that apparently slumps 80% when speaking what I've come to call "Silly Spanish."
La Piñata: The soul of a Mexican fiesta: Piñatas are the very soul of children's parties in Mexico. All youngsters wait impatiently for the moment they can try to break this model made of papier mache and metal, its belly a clay pot filled with candies. This folk art attracts not only children; adults also fall under its colorful spell when it appears at a Christmas party or even sometimes at a wedding.
The Huichol Universe: They are easily spotted at diverse shops around town, painstakingly attaching multi-colored beads to hand-carved wooden sculptures depicting a variety of fantastic creatures.
Empowering the Youth of Banderas Bay: Today's young people hold the future in their hands. Yet the Mexican educational system alone is not equipped to prepare them to benefit from and cope with the challenges and opportunities inherent in Vallarta, which is experiencing largescale tourism and development.
Marvel of the Sea: Long before jet skis and condominiums, ATVs and fishing trawlers, female sea turtles crawled from the oceans and bays along the Pacific coast of Mexico to fling away loose sand and construct their nests.
Hot Spot: Olas Altas: Bustling with activity day and night, this trendy street has such a formidable draw that many tourists and locals actually refer to the entire neighborhood by its name. Saunter about and you will be delighted to have scheduled ample time to discover its many restaurants, gift stores, outdoor cafés and nightclubs.
Mexlend Closes the First Refinancing Loan in Puerto Vallarta: Puerto Vallarta-based MEXLend, an industry-leading mortgage broker in Mexico, has closed the first GE refinancing loan in Vallarta, and only the second in Mexico, marking the beginning of many more financing options to fund in Banderas Bay.
Title Insurance: A home is usually your largest single investment. When you purchase a home, you purchase homeowner's or hazard insurance to protect against loss from fire, theft or wind damage.
Music: Movers and Shakers: We cannot possibly do justice to the increasingly vast and varied live music scene here in Puerto Vallarta in a single installment. Like waves on the sand, every year seems to come and go, leaving behind new and memorable performances by talented performers from all walks of life.
Birds of the Bay: These are two of more than 300 different bird species that can be spotted locally. Search for them on your own or visit www.birdinginmexico.com to learn about professionally guided birding tours.
Head to the Mountains!: If you are new to Banderas Bay or on a short-term vacation, chances are that you will want to remain fairly close to your home or hotel, as you will find a plethora of things to do in Puerto Vallarta and its immediate surroundings.
Mazunte, Empowering Villagers to Protect the Planet: What happens when you provide turtle hunters with an alternative, more profitable modus vivendi, one they can pursue without further endangering a protected species?
Elote: Widely cultivated around the world today, this grain was originally domesticated more than 9,000 years ago in central Mexico and to this day remains one of our country's staple foods.
Tortilla Soup: The tortilla has been a staple in Mexico since, as the Mayan legend goes, a peasant made the first dozen as a gift for his king in 10,000 BC. And ever since the 16th century, when the Spaniards came to Mexico bringing their tradition of soup, as well as the chickens and spices from which to make flavorful stock, there has been wonderful tortilla soup.
Hot Spot: Pulpito Street: A funky South Side block with a bohemian feel stretching, for the sake of argument, from landmark beachfront restaurants El Dorado and La Palapa to happening Olas Altas and Picnic restaurant 21 stairs above it, this place to see and be seen offers a bit of everything.


text:
Renowned Riverfront lodge on the world famous Kenai River. World class guided fishing and adventures. Anchorage pickup. 2-7 day, all inclusive experiences. All Meals, nightly Happy hour and all guides/equipment included.
Deluxe Fly-in Alaska fishing lodge offering inclusive trip package and fully outfitted, guided fishing for all Alaska salmon and rainbow trout on our scenic clear-water river. We offer the best value and customer service all-inclusive from Anchorage
Minimum 5 heads (partridge, grouse, pheasant) with breakfast included: 170 € (all day long). € 20, 00 is available the use of the dog Discover our 3 days package " Living Tuscany" Hunting, cooking class, truffle hunting & guided tour of Arez
FBA sets itself apart from our competitors with a superior customer service. Since we are a highly experienced Golden Dorado fishing tour operator, we can handle all of your fishing needs. That is why we look into every detail of your fishing trip.
Be the first to discover the new lodge for Bass fishing El Palmar Lake Huites! If you enjoy lake fishing and a quiet relaxing landscape you'll love this place. Make advance reservations with an atractive discount. now open
Dana Wharf has the BEST Sportfishing and Whale Watching tours in Orange County, Ca. We have been serving the public since 1971 from Dana Point. We also have FREE PARKING for our customers!
Fishing trips in S. Miguel - Azores, spend some hours relaxing and fishing. Inshore techniques: trolling, jigging and bottom fishing and Big Game for tuna and Marlin. Also available Shore Excursions tours around the island on a Jeep tours or van.
Fishing charters from Port Credit Mississauga aboard a 40 foot Luhrs SF loaded with the lattest electronics and fishing gear. We start May 15 till Oct 15..Besides group specials we also have individual fishing packages for those which cant get groups
Charter the " HOT ROD " at 34 foot Express Sport Fishermen with tower for 3 to view from above. This is a luxury charter for the same price the others charge. When the water gets ruff the others follow HOT ROD back to the harbor.
One Day Get-a-way Guide Catch Hybrids Stripers Crappies Catfish Deep Sea Catch Groupers Amberjacks, Snappers, Georgia Coast, Panama City Beach Florida Transportation Depart From All Georgia Metro Areas To Fishing Vessel All Inclusive Fishing Packages
Browse thousands of listings for Sports.


text:
If you look around you
- Artists Under Break, Bartdon
- Release Date 2013-07-08
- Labels Spektra Recordings
- Catalog SPK099
- Description Bartdon's new track "If you look around you", is a incredible track with a fantastic melodies & sounds. Includes a great remix created by Under Break. This reference is supported from the best Radio Stations & DJs dedicated to Breaks sound.


text:
Your comment is being published.
Thanks for keeping alive the conversation.
5 search results for "Bellevue, NE"
Hip Hop Indie Rap RnB
Bellevue, WA, United States
Bellevue, NE, United States
Christian Gospel Religious
Bellevue, NE, United States
Entertainment Gaming News
Bellevue, WA, United States
Christian Education Religious
Bellevue, NE, United States
Bellevue, NE - Radio Stations on your iPhone, iPad, iPod Touch, Android, Blackberry, and other app-enabled mobile phones. Get in touch via the Contact Us below if you're interested in these apps. Also, find the top new songs, playlists, and music on our website!


text:
Welcome to the Wisconsin Airsoft Association Forums.
This is a temporary forum for questions, suggestions or comments with the new forums. Be aware that if you are having problems registering you may still post in this forum. *ANY UNRELATED POSTS WILL BE CLEANSED*
15 Nov 12, 02:33 PM
Area for public discussion. Please remember to keep it airsoft based. This is not a general topic forum. Thanks.
Postings for the teams that play here and information about them.
20 Jun 15, 06:51 AM
This forum is for the new players to post a few words about themselves and let the rest of the players here know who they are.
25 Feb 16, 04:03 PM
This section is for new players to be able to ask questions. Please post an introduction in the above section before you start posting in this section.
Commando Paintball - Average...
15 Apr 15, 05:33 PM
Skirmishes that are available to the players on this forum are posted here. Regional and National events are welcome as well as Local events.
WAA BASE RULESET - Operation Vigilance 11 - Apoc...
22 May 16, 03:41 PM
Discussion limited to the guns (AEGs) and gear we use in airsoft.
07 Apr 15, 07:56 PM
Just like the Title says: Buy and Sell. This is a marketplace for airsoft items. The main section here is for private sales, a special section for retailers is inside and will detail their sales and such.
WTS/WTT G96, Masada, And more!
23 May 16, 05:17 PM
Welcome to our newest member, brownaustin17


text:
Efficacy Expert Witnesses testifying in Litigation
Efficacy experts in this Bio Engineering directory provide witness opinion and testimony in all types of Efficacy accidents including Automotive Dealer Management, Operation of Automotive Dealer Dealerships, Termination or Non renewal Manufacturer Automotive Dealer Relations, Automotive Dealer Valuation and assigned dealer area.
Efficacy accidents can occur in OpenTV, Liberate, JavaTV, O S and Workers Compensation.
In litigation, Efficacy accident experts attempt to use forensic engineering and biomechanical engineering research to reconstruct the accident and damages or event by providing witness testimony, computer animation or graphical exhibits to opine in a compelling way to the jury
Bio Engineering Expert Witnesses for Attorneys
Search results for: Efficacy
1 result found.
Provides Opinion & Testimony In:
Medical Device R D Engineer, Biomedical Engineering, Cardiovascular Signal Analysis, Medical Device Design, Communications Strategies, Interventional Cardiovascular Devices, Cardiac Pacing Devices, Cardiac Stents, Artificial Organ Design, Neuro Modulation, Biocompatibility Studies, FDA Device Controls Regulatory Issues, CNS Epilepsy, Neurology, Cardiology, Cardiac Echo Imaging, Nuclear Medicine, MRI, CT Imaging, Cardiac Echocardiography, Patent Prosecution, Echocardiography, patent testimony experience federal court, bio synch case vs nautilus Medical device heart rate monitor, design el...
Click to view this experts CV Click to request this expert
Click to view this experts CV Click to request this expert


text:
Lot 163 View Catalog
Two Hardwood Plank Carved Food Molds. A cookie mold in a heart design with serrated edge and a decorated pot with saucer filled with flowers. Stamped "O.R.W." on each end. 10 5/8"h. X 6 1/2"w. X 7/8" thick. Along with a maple sugar candy mold. One side with 25 star shaped molds on one side and 20 decorated rectangular molds on the other side. 23 3/4"l. X 3 1/2"w. X v7/8" thick.
Sign In to see what this sold for
|$100 – $200
|Starting Bid
Your maximum bid will remain private.


text:
What's New
We are making monthly updates to improve performance and fix issues we identify from your feedback.
App Description
The BBC News app has been completely redesigned with a fresh look and feel to bring you more content, greater personalisation and better overall performance. So you can get more of the news that matters to you.
Key features:
Top Stories brings you the latest, breaking news from our trusted global network of journalists.
My News is a newly introduced feature allowing you to personalise your news offering.
Add topics wherever you see the + icon for a quick route straight to the stories you care about.
Stories from topics you add will appear in your personalised My News feed. Here you can order stories by topic or time, and use the Manage Topics screen to stay on top of the news of the day.
You can also switch the My News by Topic screen to a more traditional carousel view from the Settings screen.
Find content fast
The app can suggest topics based on stories you've recently viewed and what's in the news now.
And you can quickly find topics that interest you using the Topic Search. Choose from the familiar list of BBC News indexes such as Business, Technology, Entertainment and Health, or search for your own subjects of interest, whether they're people, places, things or themes.
More of the news you care about
We now offer a fuller selection of stories from the newsroom, including:
- Most Read and Most Watched. Keep on top of popular stories across the BBC, regularly updated throughout the day
- Live BBC World Service Radio. Expertise and insight from our journalists available 24-hours a day and ready to listen to in the background as you browse
- Read the best journalism from BBC correspondents
- Access to our full range of video and audio
- Photo galleries and full-screen images
Apple Watch Support:
Story summaries, glanceable headlines and breaking news alerts delivered straight to your wrist
Other features:
- Push notifications for breaking news stories
- Video over cellular and wifi
- Improved offline experience. Download the latest stories when you have a signal and then read them at your leisure
- Viewing modes. On iPhone see more stories per index by choosing 'Compact layout' from the Settings screen
- Settings to allow the app to automatically update in the background
- Share stories to your social networks, or email/SMS to a friend
- Support for dynamic text. The app will respect the font size in your device settings so you can increase or decrease the story text size as suits your preference
- Related stories, related topics. Go deeper into a storyline with suggestions for further reading and viewing and a range of topics related to each story
If you choose to receive push notifications, a unique identifier relating to your device will be stored by ProcessOne on behalf of the BBC to provide you with the service.
No other personal data relating to you (such as a username or email address) is processed. The BBC will keep your information secure and not share it with anyone else in accordance with the BBC's Privacy and Cookies Policy, available at http://www.bbc.com/privacy/
You can choose to unsubscribe from push notifications from your device's Notifications screen: locate the BBC News app icon from the list and turn off 'Allow Notifications'.
App Changes
- May 26, 2011 New version 1.7.1
- February 01, 2011 New version 1.6.9
- December 02, 2010 New version 1.6.6
- November 18, 2010 New version 1.6.5
- October 05, 2010 New version 1.6.4
- September 27, 2010 New version 1.6.3
- August 03, 2010 New version 1.4.1
- July 30, 2010 New version 1.4
- June 28, 2010 New version 1.3
- June 06, 2010 New version 1.2.1
- May 21, 2010 New version 1.2
- June 24, 2011 New version 1.7.1
- October 12, 2011 New version 1.7.2
- December 20, 2011 New version 1.7.3
- September 21, 2012 New version 1.7.5
- October 25, 2012 New version 1.7.6
- December 11, 2012 New version 1.7.7
- March 28, 2013 New version 2.0
- May 10, 2013 New version 2.0.1
- May 23, 2013 New version 2.0.2
- June 13, 2013 New version 2.0.3
- July 05, 2013 New version 2.1
- August 16, 2013 New version 2.1.1
- September 18, 2013 New version 2.1.2
- October 08, 2013 New version 2.1.3
- November 12, 2013 New version 2.1.4
- July 11, 2015 New version 3.1.0
- July 12, 2015 New version 3.1.1
- August 06, 2015 New version 3.2.0
- September 02, 2015 New version 3.3.0
- September 26, 2015 New version 3.4.0
- October 17, 2015 New version 3.4.1
- December 15, 2015 New version 3.6.0
- January 27, 2016 New version 3.6.2
- March 05, 2016 New version 3.7.0
- March 12, 2016 New version 3.7.1


text:
Funding via VisionFund Cambodia
Kum P. has asked for a loan of US$1000 to meet some of her families needs and improve her standard of living. She would like to purchase a water pump for her home, gas for the family's motorbike, and if possible, livestock and/or fertilizer. In the future, she would like to open up a grocery stand in the market. She believes this loan will increase her daily income and help expand her business in the future.
Success!! The loan was 100% repaid


text:
Select the red location marker for directions to this station.
251 Hungerford Drive
Rockville, MD 20850
Daily: $5.10 all day
22 short-term metered spots.
Reserved: Reserved rate: $65
Availability: Yes
See full parking details for this station.
|Bikes: There are 69 bike racks and 40 lockers at this station. Bikesharing is available near or at this station. See map
|Carsharing: Available at this station. Read more.
|Bus Maps: Bus service from this station
Show full list of elevators & escalators at this station.
Show list of Bikeshare locations at this station.
Station Opens at 4:53 AM (M-F)
|First Trains
Trains leave exactly 2 hours later on Saturdays and Sundays.
|Last Trains
Trains leave exactly 3 hours later on Friday and Saturday nights.
Glenmont - 5:03 AM
Shady Grove - 5:41 AM
Glenmont - 11:34 PM
Shady Grove - 12:36 AM


text:
see any connect between terrorism and any religion. The party has earlier also made it clear that terror has no religion or colour. Congress never uses word like saffron terror or Hindu terror," party general secretary Janardhan Dwivedi told reporters here.
Dwivedi's comments came amid a raging political controversy over Shinde's remarks at the AICC meeting at Jaipur on Sunday accusing BJP and RSS of conducting terror training camps and promoting "Hindu terrorism", setting off an angry reaction from the saffron parties which demanded an apology from Congress President Sonia Gandhi and Prime Minister Manmohan Singh.
Asked why the Home Minister used the word "Hindu terrorism", Dwivedi said "that must not have been the intention of the Home Minister. No Congress leader can say such thing with intention. Sometimes it happens that some words come out of the mouth of an individual unintentionally."
Dwivedi also recalled that saffron is also one of the colours in the national flag as well as in the party flag of Congress.
"How can we have an objection to the saffron colour. Kesaria has been part of old tradition of a large part of the country.
"Saffron is also one of the colours of our national flag as well as the flag of the Congress party...this dispute should be brought to an end now. It is not appropriate to blame Congress for this," he said.
Addressing the AICC meeting at Jaipur on Sunday, Shinde had said, "...we have got an investigation report that be it RSS or BJP, their training camps are promoting Hindu terrorism. We are keeping a strict vigil on all this".


text:
About This FirmThe firm of Crary Buchanan originated when former State Senator Evans Crary, Sr. began practicing law in Martin County, Florida, in 1927. The firm's solid reputation for service and excellence continues in the community today. Presently, the firm's client base includes the largest and oldest bank in the community, Seacoast National Bank, as well as the community's only hospital, Martin Memorial Medical Center and the largest local newspaper, The Stuart News.
Areas of Law
- General Civil Trial
- Appellate Practice
- Trial Practice
- Federal Practice
- Corporate Law
- Real Estate
- Probate
- Estate Planning
- Health Care
- Hospital Law
- Personal Injury
- Wrongful Death
- Family Law
- Criminal Law
- Employment Law
- Construction Law
Overall Client Ratingin Trusts and Estates5.0 out of 5.0Communication Ability5.0Responsiveness5.0Quality of Service5.0Value for Money5.0
Posted by a Corporation on 02/21/105.0 out of 5.0This the most well respected firm in the Stuart area. They have earned that reputation with an incredible group of attorneys. The firms work is second to none. They serve this community on a number of different levels, from business to philanthropy. I would recommend this firm to anyone in need of legal advice.
- Data based on 1 reviews
- Recommended by 1 Client(s)
- Last reviewed on 02/21/10
Average Peer RatingBased on this firm's rated lawyers4.6/5.0Peer Rating
- Lawyers rated at this firm meet very high criteria of general ethical standards
People at This Firm
Legal Community ActivityShow:
- All activity
- Google+
- RSS Feed
Firm DetailsYear Established1927Firm Size12LanguagesEnglish.ClientsSeacoast National Bank; Martin Memorial Medical Center; Scripps-Howard Publishing Co.; Martin County Industrial Development Authority; Chicago Title Insurance Co.; Westinghouse Electric Corp.; The Town of Ocean Breeze Park; The Stuart News; Seacoast Banking Corp.ClientsAPPROVED ATTORNEYS FOR: Chicago Title Insurance Co.; Attorney's Title Insurance Fund.
Community ContributionsArticles Blogs Q&A
Published on {{entry.postDate}} by {{defaultAuthor}}
Published on {{entry.postDate}} by {{author.name}} {{author.name}} and ,
... Read more
Legal Topics: {{legalTopic}}
Contact InformationFirm AddressCrary Buchanan
759 SW Federal Highway, Suite 106,
Stuart, FL 34994
U.S.A.Phone772-287-2600Fax772-223-4370Firm Website