Quantifying the Effects of Two Local Anesthetics on the Crayfish Stretch Receptor Organ: An Integrated Neurophysiology Lab.

The crayfish stretch receptor organ (SRO) preparation represents a robust experimental model for undergraduate laboratory experiences. For example, this preparation may be included as part of a course-based undergraduate research experience (CURE), where students work independently to plan and carry out their own experiments. In the current paper, we provide an example of how local anesthetics may be used to manipulate the SRO preparation and to perform quantitative analyses of SRO action potential firing rates. Local anesthetics provide interesting tools for manipulating physiological responses within the nervous system. A variety of inexpensive anesthetics are available for student use and each of these is expected to inhibit neurophysiological responses. While specific anesthetics exhibit subtle differences in chemical organization, they are generally understood to block voltage gated sodium channels. In the current study, we investigated the effects of two local anesthetics, MS-222 and procaine, on the action potential firing rate from the crayfish SRO. Using quantitative analyses of SRO action potential generation, we determined that each anesthetic has unique inhibitory effects on action potential firing rate that may be explained by their neuropharmacological properties. This manipulation may thus be utilized as an interesting experimental tool in undergraduate teaching laboratories. Local anesthetics applied to crayfish SRO preparations can thus be used to deepen student understanding of local anesthetics, exercise quantitative analyses, and provide experimental tools for independent experimental design.

Inhaled furosemide for relief of air hunger versus sense of breathing effort: a randomized controlled trial.

BACKGROUND: Inhaled furosemide offers a potentially novel treatment for dyspnoea, which may reflect modulation of pulmonary stretch receptor feedback to the brain. Specificity of relief is unclear because different neural pathways may account for different components of clinical dyspnoea. Our objective was to evaluate if inhaled furosemide relieves the air hunger component (uncomfortable urge to breathe) but not the sense of breathing work/effort of dyspnoea. METHODS: A randomised, double blind, placebo-controlled crossover trial in 16 healthy volunteers studied in a university research laboratory. Each participant received 3 mist inhalations (either 40 mg furosemide or 4 ml saline) separated by 30-60 min on 2 test days. Each participant was randomised to mist order 'furosemide-saline-furosemide' (n- = 8) or 'saline-furosemide-saline' (n = 8) on both days. One day involved hypercapnic air hunger tests (mean ± SD PCO2 = 50 ± 3.7 mmHg; constrained ventilation = 9 ± 1.5 L/min), the other involved work/effort tests with targeted ventilation (17 ± 3.1 L/min) and external resistive load (20cmH2O/L/s). Primary outcome was ratings of air hunger or work/effort every 15 s on a visual analogue scale. During saline inhalations, 1.5 mg furosemide was infused intravenously to match the expected systemic absorption from the lungs when furosemide is inhaled. Corresponding infusions of saline during furosemide inhalations maintained procedural blinding. Average visual analogue scale ratings (%full scale) during the last minute of air hunger or work/effort stimuli were analysed using Linear Mixed Methods. RESULTS: Data from all 16 participants were analysed. Inhaled furosemide relative to inhaled saline significantly improved visual analogues scale ratings of air hunger (Least Squares Mean ± SE - 9.7 ± 2%; p = 0.0015) but not work/effort (+ 1.6 ± 2%; p = 0.903). There were no significant adverse events. CONCLUSIONS: Inhaled furosemide was effective at relieving laboratory induced air hunger but not work/effort in healthy adults; this is consistent with the notion that modulation of pulmonary stretch receptor feedback by inhaled furosemide leads to dyspnoea relief that is specific to air hunger, the most unpleasant quality of dyspnoea. FUNDING: Oxford Brookes University Central Research Fund. TRIAL REGISTRATION: ClinicalTrials.gov Identifier: NCT02881866 . Retrospectively registered on 29th August 2018.

Effect of interstimulus interval on cortical proprioceptive responses to passive finger movements.

Shortening of the interstimulus interval (ISI) generally leads to attenuation of cortical sensory responses. For proprioception, however, this ISI effect is still poorly known. Our aim was to characterize the ISI dependence of movement-evoked proprioceptive cortical responses and to find the optimum ISI for proprioceptive stimulation. We measured, from 15 healthy adults, magnetoencephalographic responses to passive flexion and extension movements of the right index finger. The movements were generated by a movement actuator at fixed ISIs of 0.5, 1, 2, 4, 8, and 16 s, in separate blocks. The responses peaked at ~ 70 ms (extension) and ~ 90 ms (flexion) in the contralateral primary somatosensory cortex. The strength of the cortical source increased with the ISI, plateauing at the 8-s ISI. Modeling the ISI dependence with an exponential saturation function revealed response lifetimes of 1.3 s (extension) and 2.2 s (flexion), implying that the maximum signal-to-noise ratio (SNR) in a given measurement time is achieved with ISIs of 1.7 s and 2.8 s respectively. We conclude that ISIs of 1.5-3 s should be used to maximize SNR in recordings of proprioceptive cortical responses to passive finger movements. Our findings can benefit the assessment of proprioceptive afference in both clinical and research settings.

Vagal nerve endings in visceral pleura and triangular ligaments of the rat lung.

The inner thoracic cavity is lined by the parietal pleura, and the lung lobes are covered by the visceral pleura. The parietal and visceral plurae form the pleural cavity that has negative pressure within to enable normal respiration. The lung tissues are bilaterally innervated by vagal and spinal nerves, including sensory and motor components. This complicated innervation pattern has made it difficult to discern the vagal vs. spinal processes in the pulmonary visceral pleura. With and without vagotomy, we identified vagal nerve fibres and endings distributed extensively in the visceral pleura ('P'-type nerve endings) and triangular ligaments ('L'-type nerve endings) by injecting wheat germ agglutinin-horseradish peroxidase as a tracer into the nucleus of solitary tract or nodose ganglion of male Sprague-Dawley rats. We found the hilar and non-hilar vagal pulmonary pleural innervation pathways. In the hilar pathway, vagal sub-branches enter the hilum and follow the pleural sheet to give off the terminal arborizations. In the non-hilar pathway, vagal sub-branches run caudally along the oesophagus and either directly enter the ventral-middle-mediastinal left lobe or follow the triangular ligaments to enter the left and inferior lobe. Both vagi innervate: (i) the superior, middle and accessory lobes on the ventral surfaces that face the heart; (ii) the dorsal-rostral superior lobe; (iii) the dorsal-caudal left lobe; and (iv) the left triangular ligament. Innervated only by the left vagus is: (i) the ventral-rostral and dorsal-rostral left lobe via the hilar pathway; (ii) the ventral-middle-mediastinal left lobe and the dorsal accessory lobe that face the left lobe via the non-hilar pathway; and (iii) the ventral-rostral inferior lobe that faces the heart. Innervated only by the right vagus, via the non-hilar pathway, is: (i) the inferior (ventral and dorsal) and left (ventral only) lobe in the area near the triangular ligament; (ii) the dorsal-middle-mediastinal left lobe; and (iii) the right triangular ligament. Other regions innervated with unknown vagal pathways include: (i) the middle lobe that faces the superior and inferior lobe; (ii) the rostral-mediastinal inferior lobe that faces the middle lobe; and (iii) the ventral accessory lobe that faces the diaphragm. Our study demonstrated that most areas that face the dorsal thoracic cavity have no vagal innervation, whereas the interlobar and heart-facing areas are bilaterally or unilaterally innervated with a left-rostral vs. right-caudal lateralized innervation pattern. This innervation pattern may account for the fact that the respiratory regulation in rats has a lateralized right-side dominant pattern.

Regulation of heart rate in vertebrates during hypoxia: A comparative overview.

Acute exposure to low oxygen (hypoxia) places conflicting demands on the heart. Whilst an increase in heart rate (tachycardia) may compensate systemic oxygen delivery as arterial oxygenation falls, the heart itself is an energetically expensive organ that may benefit from slowing (bradycardia) to reduce work when oxygen is limited. Both strategies are apparent in vertebrates, with tetrapods (mammals, birds, reptiles, and amphibians) classically exhibiting hypoxic tachycardia and fishes displaying characteristic hypoxic bradycardia. With a richer understanding of the ontogeny and evolution of the responses, however, we see similarities in the underlying mechanisms between vertebrate groups. For example, in adult mammals, primary bradycardia results from the hypoxic stimulation of carotid body chemoreceptors that are overwhelmed by mechano-sensory feedback from the lung associated with hyperpnoea. Fish-like bradycardia prevails in the mammalian foetus (which, at this stage, is incapable of pulmonary ventilation), and in fish and foetus alike, the bradycardia ensues despite an elevation of circulating catecholamines. In both cases, the reduced heart rate may primarily serve to protect the heart. Thus, the comparative perspective offers fundamental insight into how and why different vertebrates regulate heart rate in different ways during periods of hypoxia.

Interaction between the pulmonary stretch receptor and pontine control of expiratory duration.

Medial parabrachial nucleus (mPBN) neuronal activity plays a key role in controlling expiratory (E)-duration (TE). Pulmonary stretch receptor (PSR) activity during the E-phase prolongs TE. The aims of this study were to characterize the interaction between the PSR and mPBN control of TE and underlying mechanisms. Decerebrated mechanically ventilated dogs were studied. The mPBN subregion was activated by electrical stimulation via bipolar microelectrode. PSR afferents were activated by low-level currents applied to the transected central vagus nerve. Both stimulus-frequency patterns during the E-phase were synchronized to the phrenic neurogram; TE was measured. A functional mathematical model for the control of TE and extracellular recordings from neurons in the preBötzinger/Bötzinger complex (preBC/BC) were used to understand mechanisms. Findings show that the mPBN gain-modulates, via attenuation, the PSR-mediated reflex. The model suggested functional sites for attenuation and neuronal data suggested correlates. The PSR- and PB-inputs appear to interact on E-decrementing neurons, which synaptically inhibit pre-I neurons, delaying the onset of the next I-phase.

[Different skin-stretching devices for the treatment of skin and soft tissue defects].

OBJECTIVE: To compare clinical effects of tension-relief system(TRS) and self-made skin-stretching devices in treating skin and soft tissue defects caused by trauma. METHODS: Totally 41 patients with skin and soft tissue defects caused by trauma treated by skin-stretching devices were retrospectively analyzed from June 2015 to October 2016, including 28 males and 13 females, aged from 22 to 64 years old with an average of(43.2±10.4) years old. The patients were divided into two groups according to therapeutic methods, 18 patients were treated by TRS, including 11 males and 7 females, aged from 22 to 61 years old with an average of (41.7±9.5) years old; 5 patients injured on the upper limb, 11 patients on lower limb and 2 patients on the back; the area of defect ranged from 42 to 160 cm² with an average of(78.6±17.4) cm². Twenty-three patients were treated by self-made skin stretching device, included 17 males and 6 females, aged from 25 to 64 years old with an average of (44.4±12.7) years old; 6 patients injured on the upper limb, 13 patients on lower limb and 4 patients on the back; the area of defect ranged from 54 to 175 cm² with an average of(75.2±14.3) cm². Primary closure at stage I, blood loss, operation time, healing time of wound, complications between two groups were compared, VAS score was used to evaluate pain relief, vancouver scar scale(VSS) was used to assess recover of postoperative scars. RESULTS: Forty-one patients were followed up ranged from 3 to 12 months with an average of(16.2±3.7) months. There were no significant difference between two groups in blood loss and operation time(P>0.05). In TRS group, 16 patients' injury were closured directly, wound healing time of 16 patients were over 3 about weeks, VAS score was 3.9±1.1, VSS score was 3.5±1.2, and 1 patient occurred complication; In self-made skin stretch group, 12 patients' injury were closured directly, wound healing time of 9 patients were over 3 about weeks, VAS score was 4.8±1.4, VSS score was 5.3±1.6, and 9 patients occurred complication; there were no statistical differences between two groups in these items. CONCLUSIONS: Compared with self-made skin stretch, TRS has advantages of good effect of wound, minimal trauma, short healing time, less pain, good outlook, and less complication.

The course of lung inflation alters the central pattern of tracheobronchial cough in cat-The evidence for volume feedback during cough.

The effect of volume-related feedback and output airflow resistance on the cough motor pattern was studied in 17 pentobarbital anesthetized spontaneously-breathing cats. Lung inflation during tracheobronchial cough was ventilator controlled and triggered by the diaphragm electromyographic (EMG) signal. Altered lung inflations during cough resulted in modified cough motor drive and temporal features of coughing. When tidal volume was delivered (via the ventilator) there was a significant increase in the inspiratory and expiratory cough drive (esophageal pressures and EMG amplitudes), inspiratory phase duration (CTI), total cough cycle duration, and the duration of all cough related EMGs (Tactive). When the cough volume was delivered (via the ventilator) during the first half of inspiratory period (at CTI/2-early over inflation), there was a significant reduction in the inspiratory and expiratory EMG amplitude, peak inspiratory esophageal pressure, CTI, and the overlap between inspiratory and expiratory EMG activity. Additionally, there was significant increase in the interval between the maximum inspiratory and expiratory EMG activity and the active portion of the expiratory phase (CTE1). Control inflations coughs and control coughs with additional expiratory resistance had increased maximum expiratory esophageal pressure and prolonged CTE1, the duration of cough abdominal activity, and Tactive. There was no significant difference in control coughing and/or control coughing when sham ventilation was employed. In conclusion, modified lung inflations during coughing and/or additional expiratory airflow resistance altered the spatio-temporal features of cough motor pattern via the volume related feedback mechanism similar to that in breathing.

Sugar concentration and timing of feeding affect feeding characteristics and survival of a parasitic wasp.

The availability of food sources is important for parasitoid survival, especially for those that inhabit ecosystems where nectar and honeydew are spatially or temporally scarce. Therefore, the value of even a single meal can be crucial for survival. Psyttalia lounsburyi is a parasitoid, and biological control agent, of the olive fruit fly, Bactrocera oleae. In order to improve our understanding of the basic nutritional ecology of P. lounsburyi and its role in survival we evaluated the effect of a single sucrose meal on the longevity of female and male wasps. We measured the duration of feeding, volume ingested, sucrose consumption, energy content, and longevity of wasps provided with different concentrations of sucrose (0.5, 1, and 2M) at different times after emergence (0, 1, 2 or 3 days after emergence). Our results showed that longevity was significantly influenced by sucrose concentration and timing of feeding. For females, feeding on sucrose increased the likelihood of survival to varying degrees, ranging from 32.3% to 95.4%, compared to water-only controls. The longest duration of feeding was observed for the highest sucrose concentrations and oldest wasps. The amount of sugar ingested and energy uptake increased, up to a point, as sugar concentration increased. Our results suggest that P. lounsburyi derived greatest benefit from the intermediate concentration (1M) of sucrose provided 2 or 3 days after emergence. Our study emphasizes the importance of finding balance between increasing longevity and limiting the duration of feeding, and concomitant uptake of nutrients, that is fundamental for survival of the wasp in nature.

Effects of prolonged lung inflation or deflation on pulmonary stretch receptor discharge in the alligator (Alligator mississippiensis).

The American alligator (Alligator mississippiensis) is a semi-aquatic diving reptile that has a periodic breathing pattern. Previous work identified pulmonary stretch receptors, that are rapidly and slowly adapting, as well as intrapulmonary chemoreceptors (IPC), sensitive to CO2, that modulate breathing patterns in alligators. The purpose of the present study was to quantify the effects of prolonged lung inflation and deflation (simulated dives) on pulmonary stretch receptors (PSR) and/or IPC discharge characteristics. The effects of airway pressure (0-20 cm H2O), hypercapnia (7% CO2), and hypoxia (5% O2) on dynamic and static responses of PSR were studied in juvenile alligators (mean mass=246 g) at 24°C. Alligators were initially anesthetized with isoflurane, cranially pithed, tracheotomized and artificially ventilated. Vagal afferent tonic and phasic activity was recorded with platinum hook electrodes. Receptor activity was a mixture of slowly adapting PSR (SAR) and rapidly adapting PSR (RAR) with varying thresholds and degrees of adaptation, without CO2 sensitivity. Receptor activity before, during and after 1 min periods of lung inflation and deflation was quantified to examine the effect of simulated breath-hold dives. Some PSR showed a change in dynamic response, exhibiting inhibition for several breaths after prolonged lung inflation. Following 1 min deflation, RAR, but not SAR, exhibited a significant potentiation of burst frequency relative to control. For SAR, the post-inflation receptor inhibition was blocked by CO2 and hypoxia; for RAR, the post-inflation inhibition was potentiated by CO2 and blocked by hypoxia. These results suggest that changes in PSR firing following prolonged inflation and deflation may promote post-dive ventilation in alligators. We hypothesize that PSR in alligators may be involved in recovery of breathing patterns and lung volume during pre- and post-diving behavior and apneic periods in diving reptiles.

Vagal afferent control of abdominal expiratory activity in response to hypoxia and hypercapnia in rats.

In the present study, we tested the hypothesis that vagal afferent information modulates the pattern of expiratory response to hypercapnia and hypoxia. Simultaneous recordings of airflow, diaphragmatic (DIA) and oblique abdominal muscle (ABD) activities were performed in anesthetized (urethane, 1.2g/kg), tracheostomized, spontaneously breathing male Wistar rats (290-320g, n=12). The animals were exposed to hypercapnia (7 and 10% CO2 for 5min) and hypoxia (7% O2 for 1min) before and after bilateral vagotomy. We verified that the percentage increase in DIA burst amplitude elicited by hypercapnia and hypoxia episodes was similar between intact and vagotomized rats (P>0.05). In contrast, hypercapnia and hypoxia promoted a marked increase in ABD activity in vagotomized, but not in intact rats (P<0.01). These amplified expiratory motor changes after vagotomy were associated with enhanced expiratory airflow (P<0.01) and augmented tidal volume responses (P<0.01). Our data indicates that, in anesthetized conditions, the removal of peripheral afferent inputs facilitates the processing of active expiration in response to hypercapnia and hypoxia in rats.

RR interval-respiratory signal waveform modeling in human slow paced and spontaneous breathing.

Our aim was to model the dependence of respiratory sinus arrhythmia (RSA) on the respiratory waveform and to elucidate underlying mechanisms of cardiorespiratory coupling. In 30 subjects, RR interval and respiratory signal were recorded during spontaneous and paced (0.1Hz/0.15Hz) breathing and their relationship was modeled by a first order linear differential equation. This model has two parameters: a0 (related to the instantaneous degree of abdominal expansion) and a1 (referring to the speed of abdominal expansion). Assuming that a0 represents slowly adapting pulmonary stretch receptors (SARs) and a1 SARs in coordination with other stretch receptors and central integrative coupling; then pulmonary stretch receptors relaying the instantaneous lung volume are the major factor determining cardiovagal output during inspiration. The model's results depended on breathing frequency with the least error occurring during slow paced breathing. The role of vagal afferent neurons in cardiorespiratory coupling may relate to neurocardiovascular diseases in which weakened coupling among venous return, arterial pressure, heart rate and respiration produces cardiovagal instability.