Case-case analyses of cryptosporidiosis and giardiasis using routine national surveillance data in the United States - 2005-2015.

Understanding endemic infectious disease risk factors through traditional epidemiological tools is challenging. Population-based case-control studies are costly and time-consuming. A case-case analyses using surveillance data addresses these limitations by using resources more efficiently. We conducted a case-case analyses using routine surveillance data reported by 16 U.S. states (2005-2015), wherein reported cases of salmonellosis were used as a comparison group to identify exposure associations with reported cases of cryptosporidiosis and giardiasis. Odds ratios adjusted for age and reporting state (aOR) and 95% confidence intervals (95% CI) were calculated. A total of 10 704 cryptosporidiosis cases, 17 544 giardiasis cases, and 106 351 salmonellosis cases were included in this analyses. When compared with cases of salmonellosis, exposure to treated recreational water (aOR 4.7, 95% CI 4.3-5.0) and livestock (aOR: 3.2; 95% CI: 2.9-3.5) were significantly associated with cryptosporidiosis and exposure to untreated drinking (aOR 4.1, 95% CI 3.6-4.7) and recreational water (aOR 4.1, 95% CI 3.7-4.5) were associated with giardiasis. Our analyses shows that routine surveillance data with standardised exposure information can be used to identify associations of interest for cryptosporidiosis and giardiasis.

Central pattern generators and the control of rhythmic movements.

Central pattern generators are neuronal circuits that when activated can produce rhythmic motor patterns such as walking, breathing, flying, and swimming in the absence of sensory or descending inputs that carry specific timing information. General principles of the organization of these circuits and their control by higher brain centers have come from the study of smaller circuits found in invertebrates. Recent work on vertebrates highlights the importance of neuro-modulatory control pathways in enabling spinal cord and brain stem circuits to generate meaningful motor patterns. Because rhythmic motor patterns are easily quantified and studied, central pattern generators will provide important testing grounds for understanding the effects of numerous genetic mutations on behavior. Moreover, further understanding of the modulation of spinal cord circuitry used in rhythmic behaviors should facilitate the development of new treatments to enhance recovery after spinal cord damage.

GABA enhances transmission at an excitatory glutamatergic synapse.

GABA mediates both presynaptic and postsynaptic inhibition at many synapses. In contrast, we show that GABA enhances transmission at excitatory synapses between the lateral gastric and medial gastric motor neurons and the gastric mill 6a and 9 (gm6a, gm9) muscles and between the lateral pyloric motor neuron and pyloric 1 (p1) muscles in the stomach of the lobster Homarus americanus. Two-electrode current-clamp or voltage-clamp techniques were used to record from muscle fibers. The innervating nerves were stimulated to evoke excitatory junctional potentials (EJPs) or excitatory junctional currents. Bath application of GABA first decreased the amplitude of evoked EJPs in gm6a and gm9 muscles, but not the p1 muscle, by activating a postjunctional conductance increase that was blocked by picrotoxin. After longer GABA applications (5-15 min), the amplitudes of evoked EJPs increased in all three muscles. This increase persisted in the presence of picrotoxin. beta-(Aminomethyl)-4-chlorobenzenepropanoic acid (baclofen) was an effective agonist for the GABA-evoked enhancement but did not increase the postjunctional conductance. Muscimol activated a rapid postsynaptic conductance but did not enhance the amplitude of the nerve-evoked EJPs. GABA had no effect on iontophoretic responses to glutamate and decreased the coefficient of variation of nerve-evoked EJPs. In the presence or absence of tetrodotoxin, GABA increased the frequency but not the amplitude of miniature endplate potentials. These data suggest that GABA acts presynaptically via a GABA(B)-like receptor to increase the release of neurotransmitter.

Global structure, robustness, and modulation of neuronal models.

The electrical characteristics of many neurons are remarkably robust in the face of changing internal and external conditions. At the same time, neurons can be highly sensitive to neuromodulators. We find correlates of this dual robustness and sensitivity in a global analysis of the structure of a conductance-based model neuron. We vary the maximal conductance parameters of the model neuron and, for each set of parameters tested, characterize the activity pattern generated by the cell as silent, tonically firing, or bursting. Within the parameter space of the five maximal conductances of the model, we find directions, representing concerted changes in multiple conductances, along which the basic pattern of neural activity does not change. In other directions, relatively small concurrent changes in a few conductances can induce transitions between these activity patterns. The global structure of the conductance-space maps implies that neuromodulators that alter a sensitive set of conductances will have powerful, and possibly state-dependent, effects. Other modulators that may have no direct impact on the activity of the neuron may nevertheless change the effects of such direct modulators via this state dependence. Some of the results and predictions arising from the model studies are replicated and verified in recordings of stomatogastric ganglion neurons using the dynamic clamp.

Modulators with convergent cellular actions elicit distinct circuit outputs.

Six neuromodulators [proctolin, Cancer borealis tachykinin-related peptide Ia, crustacean cardioactive peptide (CCAP), red pigment-concentrating hormone, TNRNFLRFamide, and pilocarpine] converge onto the same voltage-dependent inward current in stomatogastric ganglion (STG) neurons of the crab C. borealis. We show here that each of these modulators acts on a distinct subset of pyloric network neurons in the STG. To ask whether the differences in cell targets could account for their differential effects on the pyloric rhythm, we systematically compared the motor patterns produced by proctolin and CCAP. The motor patterns produced in proctolin and CCAP differed quantitatively in a number of ways. Proctolin and CCAP both act on the lateral pyloric neuron and the inferior cardiac neuron. Proctolin additionally acts on the pyloric dilator (PD) neurons, the pyloric (PY) neurons, and the ventricular dilator neuron. Using the dynamic clamp, we introduced an artificial peptide-elicited current into the PD and PY neurons, in the presence of CCAP, and converted the CCAP rhythm into a rhythm that was statistically similar to that seen in proctolin. This suggests that the differences in the network effects of these two modulators can primarily be attributed to the known differential distributions of their receptors onto distinct subsets of neurons, despite the fact that they activate the same current.

The roles of co-transmission in neural network modulation.

Neuromodulation provides considerable flexibility to the output of neural networks. In spite of the extensive literature documenting the presence of modulatory peptide co-transmitters in many neurons, considerably less is known about the specific roles of co-transmission in circuit function. This review describes some of the potential consequences of peptide co-transmission in functional circuits, using specific examples from recent work on the actions of identified peptidergic projection neurons acting on the multifunctional neural network within the crustacean stomatogastric ganglion. This system reveals that co-transmission provides projection neurons with a rich assortment of strategies for eliciting multiple outputs from a multifunctional network.

Activity-dependent modification of inhibitory synapses in models of rhythmic neural networks.

The faithful production of rhythms by many neural circuits depends critically on the strengths of inhibitory synaptic connections. We propose a model in which the strengths of inhibitory synapses in a central pattern-generating circuit are subject to activity-dependent plasticity. The strength of each synapse is modified as a function of the global activity of the postsynaptic neuron and by correlated activity of the pre- and postsynaptic neurons. This allows the self-assembly, from random initial synaptic strengths, of two cells into reciprocal oscillation and three cells into a rhythmic triphasic motor pattern. This self-assembly illustrates that complex oscillatory circuits that depend on multiple inhibitory synaptic connections can be tuned via simple activity-dependent rules.

Multiple peptides converge to activate the same voltage-dependent current in a central pattern-generating circuit.

The stomatogastric ganglion of the crab, Cancer borealis, is modulated by >20 different substances, including numerous neuropeptides. One of these peptides, proctolin, activates an inward current that shows strong outward rectification (Golowasch and Marder, 1992). Decreasing the extracellular Ca(2+) concentration linearizes the current-voltage curve of the proctolin-induced current. We used voltage clamp to study the currents evoked by proctolin and five additional modulators [C. borealis tachykinin-related peptide Ia (CabTRP Ia), crustacean cardioactive peptide, red pigment-concentrating hormone, TNRNFLRFamide, and the muscarinic agonist pilocarpine] in stomatogastric ganglion neurons, both in the intact ganglion and in dissociated cell culture. Subtraction currents yielded proctolin-like current-voltage relationships for all six substances, and the current-voltage curves of all six substances showed linearization in low external Ca(2+). The lateral pyloric neuron responded to all six modulators, but the ventricular dilator neuron only responded to a subset of them. Bath application of saturating concentrations of proctolin occluded the response to CabTRP and vice versa. N-(6-Aminohexyl)-5-chloro-1-napthalensulfonamide, a calmodulin inhibitor, increased the amplitude and altered the voltage dependence of the responses elicited by CabTRP and proctolin. Together, these data indicate that all six substances converge onto the same voltage-dependent current, although they activate different receptors. Therefore, differential network responses evoked by these substances may primarily depend on the receptor distribution on network neurons.

The actions of crustacean cardioactive peptide on adult and developing stomatogastric ganglion motor patterns.

The motor patterns produced by the stomatogastric ganglion (STG) are strongly influenced by descending modulatory inputs from anterior ganglia. With these inputs intact, in control saline, the motor patterns produced by the stomatogastric nervous system of embryonic and larval lobsters are slower and less regular than those of adult lobsters. We studied the effects of the hormonal modulator, crustacean cardioactive peptide (CCAP) on the discharge patterns of STG motor patterns in embryos, larvae, and adult Maine lobsters, Homarus americanus, with the anterior inputs present and absent. In adults, CCAP initiated robust pyloric rhythms from STGs isolated from their descending control and modulatory inputs. Likewise, CCAP initiated robust activity in isolated embryonic and larval STGs. Nonetheless, quantitative analyses revealed that the frequency and regularity of the STG motor neuron discharge seen in the presence of CCAP in isolated STGs from embryos were significantly lower than those seen late in larval life and in adults under the same conditions. In contrast, when the descending control and modulatory pathways to the STG were left intact, the embryonic and larval burst frequency seen in the presence of CCAP was increased by CCAP, whereas the burst frequency in adults was decreased by CCAP, so that in CCAP the frequencies at all stages were statistically indistinguishable. These data argue that immature embryonic motor patterns seen in the absence of CCAP are a function of immaturity in both the STG and in the descending and modulatory pathways.

GABA and responses to GABA in the stomatogastric ganglion of the crab Cancer borealis.

The multifunctional neural circuits in the crustacean stomatogastric ganglion (STG) are influenced by many small-molecule transmitters and neuropeptides that are co-localized in identified projection neurons to the STG. We describe the pattern of gamma-aminobutyric acid (GABA) immunoreactivity in the stomatogastric nervous system of the crab Cancer borealis and demonstrate biochemically the presence of authentic GABA in C. borealis. No STG somata show GABA immunoreactivity but, within the stomatogastric nervous system, GABA immunoreactivity co-localizes with several neuropeptides in two identified projection neurons, the modulatory proctolin neuron (MPN) and modulatory commissural neuron 1 (MCN1). To determine which actions of these neurons are evoked by GABA, it is necessary to determine the physiological actions of GABA on STG neurons. We therefore characterized the response of each type of STG neuron to focally applied GABA. All STG neurons responded to GABA. In some neurons, GABA evoked a picrotoxin-sensitive depolarizing, excitatory response with a reversal potential of approximately -40 mV. This response was also activated by muscimol. In many STG neurons, GABA evoked inhibitory responses with both K(+)- and Cl(-)-dependent components. Muscimol and beta-guanidinopropionic acid weakly activated the inhibitory responses, but many other drugs, including bicuculline and phaclofen, that act on vertebrate GABA receptors were not effective. In summary, GABA is found in projection neurons to the crab STG and can evoke both excitatory and inhibitory actions on STG neurons.

Motor pattern generation.

Recent work on the circuits that generate rhythmic movements illustrates the role of cotransmitter complement in motor pattern selection and demonstrates that many principles first established in invertebrates also hold in vertebrates. Major new areas of investigation include the development of central pattern generating networks, and the use of mouse mutants.

Development of the peptidergic modulation of a rhythmic pattern generating network.

The stomatogastric ganglion (STG) of adult lobsters and crabs receives dense aminergic and peptidergic projections. The neuropeptides are found in sensory neurons and in descending interneurons that modulate the output of the rhythmic central pattern generating networks in the STG. We describe the presence of these peptidergic projections in the adult Homarus americanus, and the effects of some of these neuropeptides on the motor patterns of the adult STG. We describe the developmental acquisition of these neuropeptides during embryonic and larval times and demonstrate that the immature STG networks are already sensitive to a variety of neuromodulators.

Neural signalling: Does colocalization imply cotransmission?

Recent results suggest that neurons that contain multiple neurotransmitters may make synaptic connections with different target neurons that are mediated by only a subset of their transmitter complement.

Encoding of muscle movement on two time scales by a sensory neuron that switches between spiking and bursting modes.

The gastropyloric receptor (GPR) neurons of the stomatogastric nervous system of the crab Cancer borealis are muscle stretch receptors that can fire in either a spiking or a bursting mode of operation. Our goal is to understand what features of muscle stretch are encoded by these two modes of activity. To this end, we characterized the responses of the GPR neurons in both states to sustained and rapidly varying imposed stretches. The firing rates of spiking GPR neurons in response to rapidly varying stretches were directly related to stretch amplitude. For persistent stretches, spiking-mode firing rates showed marked adaptation indicating a more complex relationship. Interspike intervals of action potentials fired by GPR neurons in the spiking mode were used to construct an accurate estimate of the time-dependent amplitude of stretches in the frequency range of the gastric mill rhythm (0.05-0.2 Hz). Spike trains arising from faster stretches (similar to those of the pyloric rhythm) were decoded using a linear filter to construct an estimate of stretch amplitude. GPR neurons firing in the bursting mode were relatively unaffected by rapidly varying stretches. However, the burst rate was related to the amplitude of long, sustained stretches, and very slowly varying stretches could be reconstructed from burst intervals. In conclusion, the existence of spiking and bursting modes allows a single neuron to encode both rapidly and slowly varying stimuli and thus to report cycle-by-cycle muscle movements as well as average levels of muscle tension.

Maturation of lobster stomatogastric ganglion rhythmic activity.

The stomatogastric ganglion of the adult lobster, Homarus americanus generates extremely regular pyloric rhythms with a characteristic period of 0.5-1.5 Hz. To study the changes in the pyloric rhythm during embryonic and larval development, we recorded excitatory junctional potentials evoked by lateral pyloric (LP) neuron activity. Early in development the motor discharge of the LP neuron was often irregular, preventing use of conventional analysis methods that rely on extracting burst times to calculate cycle frequency and its variability. Instead, cycle frequency was determined for the LP neuron from the peak of the power spectrum obtained from the occurrence times of excitatory junctional potentials in the p1 muscle. The ratio of the power in the peak to the power from 0 to 3 Hz was used as a relative measure of the regularity of the rhythm. Throughout embryonic and the first larval stage, LP neuron activity is slow, irregular, and only weakly periodic. The regularity of the rhythm increased during midlarval stages, and both the frequency and regularity increased considerably by the postlarval stage LIV.

Activity-dependent regulation of potassium currents in an identified neuron of the stomatogastric ganglion of the crab Cancer borealis.

Identified neurons of the stomatogastric ganglion of the crab Cancer borealis were voltage-clamped, and the current densities of three K+ currents were measured. The current densities of each of the three K+ currents varied twofold to fivefold in inferior cardiac (IC) neurons from different animals. Conventionally, this degree of variability has been attributed to experimental artifacts. Instead, we suggest that it reflects a natural variability that may be related to an underlying process of plasticity. First, we found that there is no fixed ratio among the three K+ currents. Second, we found that several hours of stimulation with depolarizing current pulses (0.5 sec duration at 1 Hz) altered the current density of the Ca2+-dependent outward current, IK(Ca), and the transient outward current, IA. This stimulation paradigm mimics the normal pattern of activity for these neurons. The effect of stimulation on the IA current density was eliminated when Ca2+ influx was blocked by extracellular Cd2+. In contrast, the K+ current densities of the lateral pyloric (LP) neuron were unaffected by the same pattern of stimulation, and the currents expressed by both the IC and the LP neurons were insensitive to hyperpolarizing pulses at the same frequency. We conclude that the conductance densities expressed by neurons may vary continually depending on the recent history of electrical activity in the preparation, and that intracellular Ca2+ may play a role in the processes by which activity influences the regulation of current densities in neurons.