, 2005). Supraspinal centers are also the target for diverse information channels from the spinal cord, reporting on action programs to the brain. Aspects featured

here will include a handful of specific examples for which defined subcircuits are implicated in certain behavioral aspects and/or molecular entry points have been elucidated. Cross-regulatory transcription factor networks are involved click here in developmental specification of cortical pyramidal neurons. They instruct the establishment of subcortical projections to pons, tectum, and spinal cord and distinguish this cortical population from callosal projection neurons with trajectories to contralateral cortical territory. In this transcriptional network, Fezf2 acts through Ctip2 to program corticospinal axonal trajectories (Arlotta et al., 2005, Chen et al., 2008 and Molyneaux et al., 2005), whereas SatB2 represses Ctip2 and promotes callosal projections (Alcamo et al., 2008 and Britanova et al., 2008) (Figure 7A). Subcortical projection neurons establish synaptic connections with many different postsynaptic targets. Direct connections between cortical neurons and motor neurons are subject to evolutionary adaptation, and their existence and weight selleckchem correlate with the degree of skilled motor performance involving distal forelimb muscles used during object manipulation tasks (Lemon, 2008). Cortical

neurons also exhibit pronounced indirect influence on motor neurons through connections to brainstem centers and spinal interneurons (Lemon, 2008 and Orlovsky et al., 1999), but it is difficult to assess the relative contributions of these diverse connections to motor behavior. Recent work has put forward the the provocative idea that descending cortical

control of motor behavior may not be restricted to motor cortex but, at least in the whisker system, is in part mediated by somatosensory cortical territory (Matyas et al., 2010). In this system, pyramidal neurons in motor cortex M1 connect to the reticular formation in the brainstem, which in turn controls the activity of facial motor neurons regulating whisker protraction (Figure 7A). The antagonistic movement of whisker retraction is initiated by descending input from somatosensory cortex S1 connecting to motor neurons via the spinal trigeminal nucleus (SPV), without M1 involvement in this pathway (Figure 7A). These findings suggest that fundamentally different descending cortical pathways influence specific motor behaviors. Given that both motor- and somatosensory cortex project to spinal levels, these observations raise the possibility that spinal motor circuits may also be differentially regulated by similar mechanisms. Whether distinct molecular programs of the kind observed for different cortical projection neurons (Arlotta et al.

Single-unit activity in the caudate nucleus, a primary input station of the basal ganglia, can encode a number of decision-related signals in monkeys performing the visual motion saccade task (Ding and Gold, 2010). fMRI studies revealed striatal activation in human subjects performing visual motion discrimination tasks

(Forstmann et al., 2008; van Veen et al., 2008). In contrast, the frequency of clinically observed perceptual impairments is much lower than that of motor deficits for diseases associated with basal ganglia dysfunction (e.g., Parkinson’s disease). This observation seems to argue against a major role of the basal ganglia in perceptual decision-making, selleck chemicals llc although non-motor symptoms are often under-reported or unrecognized

by clinicians (Chaudhuri et al., 2006). In this study, we used electrical microstimulation in the caudate nucleus in monkeys performing a visual motion discrimination task (Figure 1) to address three questions: (1) is there a causal link between caudate activity and perceptual decision behavior? find more (2) What are the specific decision-related computations that are influenced by caudate activity? (3) How do the basal ganglia’s roles in perceptual decisions relate to their roles in movement control? The results indicate that the basal ganglia can bias perceptual decisions toward particular alternatives. These effects are distinct from their role in movement execution. Thus, the basal ganglia STK38 appear to make multiple causal contributions to simple decisions that link sensory input to motor output. As described in previous reports (Ding and Gold, 2010, 2012), the performance of the two monkeys on the RT dots task depended critically on the strength (coherence) of the motion stimulus. Both monkeys achieved near-perfect accuracy and had the shortest RTs for coherences >20%, with steadily

decreasing accuracy and increasing RT at lower coherences (Figure 2). We fit choice and RT performance simultaneously with a drift-diffusion model (DDM; see Experimental Procedures and curves in Figure 2), and we fit choice data alone using logistic functions (see Figure S2 available online). We quantified performance using two measures estimated from the fits: choice bias, corresponding to the horizontal position of the psychometric curve (Figure 2, top panels), and discrimination threshold, corresponding to the steepness of the psychometric curve. We examined the effects of electrical microstimulation on performance in 43 sessions (n = 29 and 14, for monkey C and F, respectively). The microstimulation sites were within the general regions sampled in our previous recording study (Figure S1; Ding and Gold, 2010). The motion directions used were similar to our previous recoding studies for the caudate nucleus and FEF (Table S1; Ding and Gold, 2010, 2012).

The synaptogenic activity of LRRTM4, but not of LRRTM2, requires HS. Knockdown

of LRRTM4 in vivo decreases the strength of glutamatergic synaptic transmission and the density of dendritic spines, indicating that LRRTM4 controls synapse development in vivo. These results identify glypican as a receptor for LRRTM4 and highlight the diversity in ligand-receptor interactions that regulate excitatory synapse development. Glypican binding to LRRTM4 requires HS, and HS is required for LRRTM4 function. Binding of GAGs to LRR proteins is not unprecedented: a recent study identified chondroitin sulfate (CS) proteoglycans as ligands for the Nogo receptor family members NgR1 and NgR3 (Dickendesher et al., 2012). Interestingly, NgR1 and NgR3 showed strong selectivity toward specific CS GAG types, suggesting that differences in GAG sulfation

patterns may regulate compound screening assay NgR binding. Synaptic transmission at the Drosophila neuromuscular junction is differentially affected by knockdown of two different enzymes that regulate HSPG sulfation ( Dani et al., 2012), suggesting that HS modifications are also important for synapse development. Whether LRRTM4 displays any selectivity with regard to modifications of HS chains is unknown. Glypicans are widely expressed throughout the body and bind many secreted and surface-bound selleck compound proteins (Bernfield et al., 1999 and Van Vactor et al., 2006). Based on mRNA and protein expression patterns, it appears likely that LRRTM4 is not the only endogenous binding partner of GPC4, as LRRTM4 expression is much more restricted than that of GPC4. The full complement of synaptic GPC4 interactors is not yet known. In addition to LRRTM4, our GPC4-Fc pulldown experiment also identified LRRTM3, a largely uncharacterized LRRTM family member. LRRTM3 and LRRTM4 are more closely related to each other than to LRRTM1 and LRRTM2 (Laurén et al., 2003), and this evolutionary

relationship appears to be reflected in LRRTM-receptor interactions. Our experiments suggest that GPC4 needs to aggregate on the cell surface before it can induce LRRTM4 clustering and postsynaptic differentiation. Although GPC4 released from the cell surface was able to bind LRRTM4 in solution, bath-applied soluble GPC4 did not affect LRRTM4 clustering or postsynaptic differentiation. heptaminol In RGCs, soluble GPC4 induces clustering of the glutamate receptor subunit GluR1 and promotes excitatory synapse formation (Allen et al., 2012). Cultured RGCs are more reluctant to form synapses than hippocampal neurons, and soluble GPC4 may have more pronounced effects on RGC synaptogenesis. Alternatively, soluble GPC4 levels in hippocampal cultures may already be saturating or secreted GPC4 may induce GluR1 clustering through an LRRTM4-independent mechanism. It will be of interest to determine whether GPC4 exerts these effects through LRRTM4 in RGCs.

5% (148/155) of animals were females and 71.6% (111/155)

were over one year old. Serum samples were stored at −20 °C until being tested for the presence of antibodies against T. gondii and N. caninum. All procedures were performed according to the Ethical Principles in Animal Research adopted by the Brazilian College of Animal Experimentation and this study received approval from the Ethical Committee of the Institution. T. gondii tachyzoites (RH strain) were maintained by intraperitoneal serial passages in outbred Swiss mice at regular 48 h intervals ( Mineo et al., 1980). Mouse peritoneal exudates were harvested when the majority of tachyzoites were extracellular, and then washed twice (720 × g, 10 min, 4 °C) in phosphate-buffered saline (PBS, pH 7.2). The resulting pellet was resuspended GSK126 in PBS for antigen preparation. N. caninum tachyzoites (Nc-1 strain) were maintained in Vero cells cultured in RPMI 1640 medium supplemented with 2% heat-inactivated calf fetal Talazoparib datasheet serum in a 5% CO2 atmosphere at 37 °C and harvested by scraping off the cell monolayer after 2–3 days of infection ( Silva et al., 2007). Tachyzoites were purified

by forcible extrusion through a 26-gauge needle to lyse any remaining intact host cells, which were also removed by centrifugation at low speed (45 × g) for 1 min at 4 °C. The supernatant containing parasite suspension was collected and then washed twice (720 × g, 10 min, 4 °C) in PBS and the resulting pellet was resuspended in PBS for antigen preparation. N. caninum and T. gondii whole antigens were prepared according to Camargo (1964). Parasite suspensions were treated with 1% formaldehyde for 30 min at room temperature. After washing in PBS, parasites were dry-fixed in microscopic slides and stored at −20 °C until being used in IFAT. N. caninum and T. gondii soluble antigens were prepared as described elsewhere ( Silva et al., 2007). Parasite suspensions

were treated with protease inhibitors (aprotinin, 10 μg/mL; leupeptin, 50 μg/mL; phenyl-methylsulfonyl fluoride [PMSF], 1.6 mM) and then lysed by five freeze–thaw (liquid nitrogen Adenosine and water bath at 37 °C) cycles and further by ultrasound (six 60 Hz cycles for 1 min each) on ice. After centrifugation (10,000 × g, 30 min, 4 °C), supernatants were collected and the protein concentration was determined ( Lowry et al., 1951). Different batches were done for antigen preparation and pooled together to obtain the required protein concentration. Soluble antigen aliquots were stored at −20 °C until being used in ELISA. IFATs were carried out to detect IgG antibodies to T. gondii (IFAT-Tg) and N. caninum (IFAT-Nc) as described elsewhere ( Figliuolo et al., 2004). Slides containing formolyzed tachyzoites were incubated with sheep sera at serial twofold dilutions starting from 1:64 to 1:8192 (for IFAT-Tg) or 1:50 to 1:3200 (for IFAT-Nc) in PBS.

Interestingly, PKA inhibitors reduce efficiency of mitochondrial ATP production in starved cells by blocking autophagy-induced mitochondrial elongation (Gomes et al., 2011), opening the possibility that the BAD complex is also a hub where morphological and metabolic cues meet. We are just beginning to unravel the complex loop between mitochondrial metabolism and seizures. For example, although Giménez-Cassina et al. (2012)

convincingly show that the activity of KATP channels is enhanced in Bad−/− and BadS155A mice, the molecular link between BAD and opening time of KATP channels is still obscure. In cardiac cells, the intracellular pool of KATP channels is mobile and can relocate to the sarcolemma after ischemia, increasing their surface density ( Bao et al., 2011). Similarly, it is conceivable that in Bad−/− and BadS155A neurons, the density of STI571 mw KATP channels on the plasma membrane might be enhanced, following a yet unknown mechanism of BAD-dependent regulation of endocytic recycling. Alternatively, the KATP channels might be activated by a signal emanating from mitochondria only when they preferentially use fatty acids: a similar

“second messenger” has been identified in glutamate, released from beta-cells mitochondria upon glucose stimulation to promote insulin secretion ( Maechler and Wollheim, 1999). In conclusion, the work of Giménez-Cassina et al. (2012) paves the way toward the understanding of the molecular mechanisms see more of mitochondrial and metabolic control of seizures. “
“Visualizations are vital tools for neuroscientists of every discipline, affording the ability to reveal relationships in large data sets and communicate information to a broad audience. But with the great power of graphs, one might say, comes great responsibility. Graphs can be fundamentally misleading about underlying

data, and design choices can skew viewers’ perceptions, leading them toward incorrect conclusions out (Jones, 2006). For example, recent studies suggest that results rendered on aesthetically pleasing brain images are perceived as more persuasive and credible than identical information presented in other formats (Keehner et al., 2011 and McCabe and Castel, 2008). Beyond the attractiveness of displays, readers may also be misled by the frequent errors that plague scientific figures (Cleveland, 1984) or a lack of sufficient information. In the words of statistician and graphic design expert Howard Wainer, effective data visualization must “remind us that the data being displayed do contain some uncertainty” and “characterize the size of that uncertainty as it pertains to the inferences we have in mind” (Wainer, 1996). It is our impression that such descriptions (along with more basic elements) are often lacking from published figures. In this NeuroView, we perform a survey of figures from leading neuroscience journals with an eye toward clarity and the portrayal of uncertainty.

The SC and the dLGN are the dominant targets of retinal projections in mammals. Despite its relatively small size in rodents, RGC projections to the dLGN are segregated with respect to eye of origin and display sharp retinotopic organization (Lund et al., 1974, Godement et al., 1984 and Pfeiffenberger et al., 2006). We examined retinotopy and eye segregation in the dLGN of β2(TG) mice and observed conditions analogous to that in the SC. In particular, we found that the retinotopy of projections to the dLGN from the dorsal monocular zone of the retina are normal (Figures 4A and 4B; 12% ±

14%, mean ± SD for WT; 29% ± 11%, mean ± SD for β2(KO); 17% ± 9%, mean ± SD for β2(TG); p < 0.001 for comparison between β2(KO) and both WT and β2(TG)), but RGC projections from the ventral-temporal binocular zone of the retina remain unrefined Metabolism inhibitor (Figures 4C and 4D; 18% ± 5%, mean ± SD for WT; 40% ± 10%, mean ± SD for β2(KO); 41% ± 9%, mean ± SD for β2(TG); p < 0.001 for comparison between WT and both β2(KO) LY294002 and β2(TG)),

unless binocular competition is removed through monocular enucleation (Figures 4E, 4F, and S4; 22% ± 5%, mean ± SD for WT; 42% ± 8%, mean ± SD for β2(KO); 25% ± 8%, mean ± SD for β2(TG); p < 0.001 for comparison between β2(KO) and WT; p = 0.005 between β2(KO) and β2(TG); p = 0.52 for comparison between β2(TG) and WT). Eye-specific segregation is also completely disrupted in the dLGN of β2(TG) mice, like in β2(KO) mice (Figures 4G–4K; Rossi et al., 2001, Muir-Robinson et al., 2002, Grubb et al., 2003, Pfeiffenberger et al., 2005 and Pfeiffenberger et al., 2006). These

data demonstrate that normal levels of spontaneous neuronal activity and “small” retinal waves are not sufficient to mediate the segregation of retinal afferents with respect to eye of origin in the dLGN and SC but are sufficient to mediate normal retinotopy (in the absence of binocular competition) throughout the dLGN and SC. We tested whether the abnormal spatiotemporal properties of waves in the β2(TG) mice are responsible for their visual map defects by manipulating β2(TG) retinal waves pharmacologically in vivo. Spontaneous retinal activity, retinal wave dynamics, and size are modulated by cAMP levels (Stellwagen and Shatz, 2002, Stellwagen et al., 1999 and Zheng Dichloromethane dehalogenase et al., 2006). Acute application of CPT-cAMP and other cAMP signaling agonists increases retinal wave size and frequency (Stellwagen and Shatz, 2002 and Stellwagen et al., 1999). Daily binocular intravitreal injection of CPT-cAMP, a nonhydrolyzable membrane-permeable analog of cAMP, beginning at P2 in β2(TG) mice significantly improves eye-specific segregation in both the dLGN and SC in comparison to saline (control) injections (Figure 5). This strengthens the assertion that the altered spatiotemporal properties of retinal waves in β2(TG) mice are responsible for their visual map defects, and demonstrates that expression of β2-nAChRs in the dLGN and SC is not necessary for eye-specific RGC axon segregation.

It is clear that neuroscientists must recognize the importance, both symbolic and real, of “replacement, reduction, and refinement” whenever animals are used. However, they may be most persuaded of this through realizing that rational implementation of the 3Rs will improve their science and help enable them to strive for “relevance, robustness, and reliability” in their investigations. The IOM Forum was a useful step in the honest and nuanced dialog that must continue as scientists, lawmakers, regulators, welfare organizations, and the public

define the path forward for realizing the huge potential of neuroscience while supporting the proliferation Selleckchem MG132 of sensible, ethical, and balanced legal and regulatory systems. “
“Activity-dependent plasticity of neurotransmission is central

to memory Selleckchem Sunitinib encoding and also plays a key role in the development of the nervous system. Persistent changes in communication among neurons also probably represent both adaptive and maladaptive responses to many forms of injury to the CNS. Plasticity in all its forms is thus inextricably intertwined with almost all aspects of brain function. Until recently, most efforts to understand the cellular and molecular mechanisms of plasticity of neurotransmission in the CNS were overwhelmingly directed at long-term potentiation (LTP) of excitatory synapses on pyramidal neurons and, to a much lesser extent, long-term depression (LTD) in pyramidal neurons and at parallel fiber synapses on cerebellar Purkinje cells. Plasticity of inhibition has received less attention. Although progress in one or the other aspect of this topic has recently been reviewed (Castillo et al., 2011; Kullmann and Lamsa, 2011; Luscher et al., 2011), this article has a broader scope: to consider the diversity of inhibitory plasticity in the context

of circuit development and function. The most obvious impediment to understanding inhibitory plasticity is the diversity of interneurons, loosely defined as locally projecting cells that release Carnitine palmitoyltransferase II GABA from their terminals. Even classifying interneurons as exclusively inhibitory is problematic, because GABA can depolarize targets early in development (Ben-Ari et al., 2007), and axo-axonic synapses may even retain this ability into adulthood (Szabadics et al., 2006). Although a definitive taxonomy of interneurons is still some way off, recent advances in identifying the time and birthplace of GABAergic neurons in the ganglionic eminences, and the transcription factors that are active early on, are helping to classify them (Ascoli et al., 2008). It remains to be determined to what extent they exist as discrete nonoverlapping types, as opposed to unique outcomes of combinatorial transcription factor expression and stochastic interactions as they migrate through the cortical mantle.

, 1957). Although there is some experimental

evidence for AP initiation in the node (Clark et al., 2005 and Colbert and Johnston, 1996), more recent electrophysiological investigations showed that the AIS initiates all APs, and the first node faithfully follows spike frequencies with a ∼100 μs delay (Foust et al., 2010, Khaliq and Raman, 2006, Doxorubicin mouse Palmer et al., 2010 and Palmer and Stuart, 2006). It remains an open question whether the first node can influence the generation of APs. In mechanosensory leech axons, a direct role of branchpoints to neural computation has been demonstrated by a facilitation of transmitter release after the AP fails at a Depsipeptide molecular weight branchpoint and subsequently propagates in both reverse and forward directions (Baccus, 1998 and Debanne et al., 2011). Also, in mammalian axons there is evidence that the strategic placing of Na+ channels at axonal branchpoints exerts computational roles by counteracting impedance mismatches when APs invade

daughter collaterals at axonal bifurcations and thereby increase the safety factor for high-frequency spike propagation toward the presynaptic terminals (Goldstein and Rall, 1974, Khaliq and Raman, 2006, Manor et al., 1991 and Monsivais et al., 2005). To address whether the first node in mammalian axons plays a role in the input-output function, electrophysiological recordings of rat neocortical L5 neurons were made in combination with targeted inactivation of visually identified nodes. The results show that the over first node facilitates the initiation

of APs in the AIS selectively during high-frequency bursts (≥100 Hz). Inactivation of nodal Na+ channels demonstrated that the first node generates a TTX-sensitive persistent Na+ current, which lowers the axosomatic AP voltage threshold and amplifies the afterdepolarization (ADP). These results unveil a role for the first node of Ranvier in the temporal encoding of synaptic inputs into high-frequency APs in axons. Branchpoints in neocortical axons contain ultrastructural markers of nodes (Khattab, 1968 and Sloper and Powell, 1979). Furthermore, previous work showed that the first branchpoint in rodent L5 axons is physiologically characterized by AP-mediated Na+ influx and acts as an acceleration point in the saltatory propagation of APs (Fleidervish et al., 2010 and Palmer and Stuart, 2006). In the present study, it is therefore assumed that the first node is localized at the first branchpoint of the primary axon. To obtain detailed insight into the location and geometrical properties of the branchpoint, whole-cell patch-clamp recordings were made from large L5 pyramidal neurons in parasagittal slices in combination with two-photon laser scanning microscopy (n = 13) or post hoc biocytin staining (n = 9).

However, exposure to this bitter tastant had no impact on the fluorescence (Figure 8I). We did not detect signals when we expressed OBP49a-t-YFP(2) with YFP(1):GR64f (Figure 8J). The combination click here of YFP(1):GR64a with SNMP1-YFP(2) also did not produce fluorescence (Figure 8K). These findings support the conclusion that OBP49a either interacts with or is adjacent to GR64a. Many bitter-tasting chemicals are toxic (Glendinning, 2007). Therefore, the ability of animals to suppress their attraction to sugars and

other nutritious foods that are laced with bitter tastants is critical for survival. Consequently, this avoidance behavior is conserved throughout the animal kingdom. Nevertheless, the molecules and molecular mechanisms through which positive feeding behavior is inhibited by deterrent compounds are poorly unexplored in most animals, such as the fruit fly. Potentially, there are multiple neural mechanisms that could explain how aversive tastants suppress the otherwise stimulatory effects of sweeteners and other attractive compounds. Animals ranging from flies to humans have separate taste receptor cells devoted to

Vorinostat sensing bitter and sweet tastants, and the suppression of sweet by bitter compounds could take place through integration of separate inputs in the brain. There could also be lateral interactions in the periphery between separate sweet- and bitter-responsive afferent receptor cells. Alternatively, a bitter compound might directly suppress sweet-activated taste receptor cells. In this study, we

unexpectedly identified a mechanism through which an array of bitter compounds inhibited the stimulatory effects of sucrose in flies. This work emerged from a functional analysis of OBPs in Drosophila taste, and was motivated by the finding that multiple Obp genes were highly enriched in taste sensilla ( McKenna et al., 1994, Pikielny et al., 1994, Ozaki et al., 1995, Galindo and Smith, 2001, Shanbhag et al., 2001, Koganezawa and Shimada, 2002, Sánchez-Gracia et al., 2009 and Yasukawa et al., 2010), but their roles in the gustatory system were largely unexplored. We found that OBP49a was required for avoiding bitter-tasting compounds in a standard two-way choice assay consisting of 1 mM sucrose alone versus 5 mM sucrose plus bitter tastants. Because Cytidine deaminase wild-type flies find bitter tastants aversive, they prefer the lower concentration of sucrose, when the higher concentration of sucrose is laced with tastants such as berberine, quinine, or denatonium. However, the Obp49a mutant animals were impaired in this avoidance behavior. The phenotype was similar to that resulting from elimination of GRs, such as GR33a and GR66a, which are broadly expressed in avoidance GRNs, and are necessary in these GRNs for induction of bitter-induced action potentials ( Moon et al., 2006, Moon et al., 2009 and Lee et al., 2009).

Other studies have also argued for a multi-component model of the TPB in the exercise

domain [26] and [27]. An extended model that incorporates insights from interviews, as well as sociodemographic characteristics, may provide a clearer picture of parents’ immunisation intentions. Indeed, the views of interviewees incorporated as items within the belief composites proved to be informative in this context: scores differed markedly between parents with maximum intentions and those who had intention scores below the possible maximum. Despite the controversy surrounding MMR, there was no significant difference between parents’ intentions to take their child for MMR CX-5461 mouse compared with dTaP/IPV. This may be explained partly by the fact that both are normally given at the same appointment and so parents’ beliefs and intentions are www.selleckchem.com/products/Perifosine.html likely to be similar. This may also reflect the possibility that there are now fewer concerns about MMR. Research published since this study has shown that there has been an increase in the proportion of mothers saying that MMR is ‘completely safe’ or ‘posing just a slight risk’ [28]. Whilst mean intention scores were generally high (1.96 for MMR and 2.30 for dTaP/IPV), only 44.2% of parents had maximum intentions to immunise their child with MMR and only

52.8% of parents had maximum intentions to immunise their child with dTaP/IPV (52.8%). Whilst direct comparisons are not possible, these figures are less than the 2006–2007 NHS reported uptake rates for MMR (73%) and isothipendyl dTaP/IPV (79%) [29]. It may be that some parents with less than maximum intentions will actually go on to have their child immunised e.g. following advice from a trusted healthcare professional. Nonetheless, potential barriers to parental uptake of both vaccinations need to be addressed in future interventions. The

finding that parental attitude was the best predictor of intention for both vaccinations is consistent with other TPB-based studies. For example, Paulussen et al. [13] and Prislin et al. [14] have demonstrated the role of parental attitude in immunisation status. In the present research, examination of the beliefs underpinning parents’ attitudes about MMR and dTaP/IPV (behavioural beliefs) revealed that parents with maximum immunisation intentions had more positive beliefs that this would prevent their child from getting the associated diseases and that this would help to eradicate them from the country. This supports research in America, where belief in the protective value of immunisation was found to Modulators contribute to positive attitudes among parents considering primary vaccinations [14].