Search PubMed⌕ Search

Biomedical subjects

N S Cherniack

Publications and source records attributed to N S Cherniack.

At least 55 records · Page 3Linked to original sources
PubMed1993

Phase-dependent dynamic responses of respiratory motor activities following perturbation of the cycle in the cat.

1. Electroneurographical (ENG) activities of a phrenic nerve, a thyroarytenoid (TA) branch of a recurrent laryngeal nerve, and a triangularis sterni (TS) branch of an internal intercostal nerve were recorded in decerebrate, vagotomized and paralysed cats. A superior laryngeal nerve (SLN) was stimulated electrically. Our objective was to evaluate transient changes in motor activity following a brief perturbation of the respiratory cycle by SLN stimulation. 2. Each motor nerve recorded represents a separate phase of the respiratory cycle. We measured the duration of phrenic ENG activity for inspiratory phase duration (TI) and similarly the duration of TA and TS ENG activity for the duration of stages I and II of expiration, respectively. Changes in the duration of the total respiratory cycle (TTOT) were also measured. Therefore, the changes in TTOT were accounted for directly by changes in each phase of the respiratory cycle. 3. Perturbation during the inspiratory phase inhibited phrenic activity either reversibly or irreversibly (premature termination of inspiration) depending on the strength and timing of the stimulus. Reversible inhibition of inspiration was associated with a transient activation (< 100 ms) of the TA nerve followed by a reactivation of the phrenic nerve, but the duration of the subsequent stages I and II of expiration remained the same. Thus, the prolongation of TTOT was completely accounted for by the lengthening of TI. 4. Premature termination of inspiration was followed by either a shortening (the first half of inspiration) or a lengthening (the second half of inspiration) of the duration of stage I expiration and consistently by a shortening of the duration of stage II expiration. The magnitude of these changes in the durations of stages I and II of expiration was phase dependent. Changes in the duration of all three phases of motor activity contributed to the changes in TTOT. 5. Perturbation during stage I expiration prolonged this stage but did not affect the duration of the succeeding stage (stage II expiration). The increase in the duration of stage I expiration appeared constant and not dependent on the time when the perturbation was delivered in stage I expiration. Thus, the change in TTOT was less phase dependent during stage I expiration than during inspiration and stage II expiration and was accounted for by changes in the duration of TA activity alone. 6. Perturbation during stage II expiration inhibited TS activity and evoked TA activity transiently.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1993

Interaction between central pattern generators for breathing and swallowing in the cat.

1. We examined the interaction between central pattern generators for respiration and deglutition in decerebrate, vagotomized, paralysed and ventilated cats (n = 10), by recording activity from the following nerves: hypoglossal, phrenic, thyroarytenoid and triangularis sterni. Fictive breathing was spontaneous with carbon dioxide above the apnoeic threshold (end-tidal PCO2, 32 +/- 4 mmHg) and fictive swallowing was induced by stimulating the internal branch of the left superior laryngeal nerve (SLN) continuously (0.2 ms pulse duration, 10 Hz). 2. In all ten animals, SLN stimulation evoked short bursts of thyroarytenoid and hypoglossal nerve activity indicative of fictive swallowing. In two of ten animals, respiration was inhibited completely during deglutition. In the other eight animals, fictive breathing and swallowing occurred simultaneously. 3. With SLN stimulation below threshold for eliciting swallowing, the respiratory rhythm decreased, the duration of inspiration did not change but the duration of expiration, especially stage II, increased. Integrated nerve activities indicated that the rate of rise and peak of phrenic nerve activity decreased, stage I expiratory activity of the thyroarytenoid and especially that of the hypoglossal nerve increased and stage II expiratory activity of the triangularis sterni nerve was suppressed completely. However, if inspired carbon dioxide was increased, i.e. hypercapnic ventilation, stage II expiratory activity remained partially during continuous SLN stimulation. 4. Fictive-swallowing bursts occurred only at respiratory phase transitions. At the minimal stimulus intensity that evoked repetitive swallowing bursts, the pattern of interaction between breathing and swallowing central pattern generators was consistent for each animal (n = 7) but was different across animals. In four animals, fictive swallows occurred at the phase transition between stage II expiration and inspiration, at the transition between inspiration and stage I expiration in one animal; and in two other animals, at the transition between stage I and II of expiration. 5. The response to SLN stimulation accommodated during the stimulus train. Accommodation was evident in both the interswallow interval (ISI) which lengthened, and the interaction pattern which had fewer swallows per breath as the stimulus period progressed. In contrast to the ISI, characteristics of the fictive swallow did not accommodate. For example, duration of the swallow was constant, distributed over a narrow range throughout the stimulus train. 6. We conclude that the central pattern generators for swallowing and breathing interact. The pattern of interaction supports the three-phase theory of respiratory pattern generation.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1993

Central and spinal effects of sodium cyanide on respiratory activity.

The pharmacological actions of cyanide on respiratory activity have been known for some time and are attributed mainly to effects on peripheral chemoreceptors. In the studies reported here, we have examined the acute central effects of cyanide when applied topically to the ventral surface of the medulla (VMS) and when administered into the spinal intrathecal space at the C5-T3 level on activities of the phrenic nerve, diaphragm, parasternal intercostal, triangularis sterni, and transversus abdominis muscles. Topical application of 10-100 micrograms (10 microliters of 1-10 mg/ml) cyanide to the intermediate area of the VMS decreased respiratory activity by > 50%, and expiratory muscles were more sensitive to inhibition than inspiratory muscles. The onset of depression of phrenic nerve or respiratory muscle activity occurred within 20 s of administration, and the effects reversed after washout. In contrast, intrathecal administration of cyanide in doses of 10-100 micrograms (100 microliters of 0.1-1 mg/ml) increased electrical activity of the respiratory muscles. Diaphragm activity changed from 17 +/- 2 to 42 +/- 8 (SE) units (P < 0.01), parasternal intercostal activity increased from 18 +/- 3 to 46 +/- 9 units (P < 0.01), and expiratory activity of the chest wall and abdominal muscles increased from 9 +/- 2 to 39 +/- 10 units (P < 0.05). Both topical application on the VMS and intrathecal administration of cyanide caused an increase in arterial blood pressure and a slight insignificant acceleration of heart rate. These data suggest that cyanide acting on the VMS causes respiratory depression and enhancement of sympathetic outflow.(ABSTRACT TRUNCATED AT 250 WORDS)

Administration, Topical↗
PubMed1993

Pathways and mechanisms involved in neural control of laryngeal submucosal gland secretion.

The purpose of this study was to define the pathways and mechanisms involved in the neural regulation of laryngeal mucosal gland functions. In anesthetized, paralyzed, and artificially ventilated dogs, the responses of laryngeal submucosal glands to stimulation of laryngeal mechanoreceptors and peripheral chemoreceptors were examined by measuring the number of hillocks and volume of secreted fluid before and after activation of sensory nerve endings. Compared with a control period, the number of hillocks and volume of secreted fluid significantly increased (P < 0.05) with mechanical stimulation of the vocal folds (n = 13) and with chemical activation of peripheral chemoreceptors by systemic administration of sodium cyanide (100 micrograms/kg; n = 11). The reflex responses induced by vocal fold stimulation and activation of peripheral chemoreceptors were slightly decreased by interrupting transmission in the recurrent laryngeal nerves (P > 0.05) and were abolished by subsequent sectioning of superior laryngeal nerves or prior intravenous administration of atropine methylnitrate (P < 0.05). In denervated animals, topical application of nicotine on laryngeal epithelium caused significant activation of submucosal glands (P < 0.05). We conclude that laryngeal secretion can be significantly altered reflexly by stimulation of laryngeal sensory nerve endings and peripheral chemoreceptors, that both superior and recurrent laryngeal nerves convey cholinergic outflow to laryngeal submucosal glands, and that nicotine acting locally activates laryngeal submucosal glands.

Animals↗
PubMed1993

Central effects of somatostatin and atrial natriuretic peptide on tracheal tone.

The effects of somatostatin and atrial natriuretic peptide applied topically to the ventral surface of the medulla (VMS) on tracheal tone and phrenic nerve activity (Phr) were studied in chloralose-anesthetized and paralyzed cats artificially ventilated with 7% CO2 in O2. Surface application of drugs to the chemosensitive areas of the VMS significantly decreased tracheal tension measured by changes in pressure in a balloon placed in a bypassed segment of the trachea (Ptseg). Application of somatostatin (9 cats) caused a mean decrease in Ptseg from 17.3 +/- 1.8 (SE) to 4.3 +/- 1.4 cmH2O (P < 0.01) and a reduction in Phr from 24.9 +/- 3.4 to 10.3 +/- 3.4 units (P < 0.05). Like somatostatin, application of atrial natriuretic peptide to the VMS (5 cats) produced tracheal relaxation (Ptseg decreased from 19.3 +/- 2.6 to 9.9 +/- 1.3 cmH2O, P < 0.01), but in contrast there was an insignificant reduction in Phr (from 18.5 +/- 3.6 to 16.1 +/- 3.8 units, P > 0.05). When parasympathetic activity was abolished by atropine methylnitrate and tracheal tone was restored with 5-hydroxytryptamine, somatostatin and atrial natriuretic peptide applied on the VMS had no effect on tracheal pressure, suggesting that observed changes were not caused by direct action of peptides on tracheal smooth muscle via the bloodstream or by facilitation of inhibitory pathways. Both somatostatin and atrial natriuretic peptide applications were associated with a slight but significant decrease in arterial blood pressure. These data suggest that somatostatin and atrial natriuretic peptide acting on the chemosensitive structure of the VMS may play significant roles in modulating para-sympathetic outflow to airway smooth muscle.

Animals↗
PubMed1992

Salivary secretion elicited by activation of the parabrachial nuclei in the cat.

The lateral pontine tegmentum contains the parabrachial nuclei (NPB) which have been identified as a relay nucleus for cardiovascular, respiratory and gustatory systems, but their role in the regulation of these systems is not well understood. We examined the effects of electrical and chemical stimulation of the NPB on blood pressure, phrenic and hypoglossal nerve activity and salivary secretion. These variables were measured in eight anesthetized (alpha-chloralose/urethane, 30/150 mg/kg, n = 5) or decerebrate (n = 3) cats before, during, and after trains of electrical stimulation (1 ms pulse duration, 10 Hz 5 min train duration, currents as low as 10 microA) delivered unilaterally to NPB. Stimulation of the NPB elicited copious salivary secretion (1100 +/- 270 mg, mean +/- S.D.; P less than 0.001). Secretion was blocked completely by prior administration of atropine. The effects of the stimulus train on the respiratory and cardiovascular systems were variable and inconsistent even though short-latency responses of phrenic and hypoglossal nerve activities to single pulses were consistent. The short-latency response of phrenic nerve activity was biphasic, a decrease followed by an increase in activity; the response of hypoglossal nerve activity was monophasic, a transient increase in activity. Effects of electrical stimulation were replicated by the injection of an excitatory amino acid agonist (kainic acid) into the dorsolateral pons. Injection of 50 nl of 10 mM kainic acid into the NPB evoked salivary secretion, indicating that this response was elicited by stimulation of cell bodies in the region. In addition, chemical excitation increased breathing frequency, peak phrenic nerve activity, and blood pressure.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1992

Cardiorespiratory changes induced by vertebral artery injection of sodium cyanide in cats.

Brain stem hypoxia caused by vertebral artery injection of sodium cyanide (NaCN) (1-20 micrograms) in artificially ventilated cats depressed phrenic and stimulated sympathetic nerve activity with a simultaneous increase in arterial blood pressure. Larger doses of NaCN caused greater effects. Hypercapnia produced by inhalation of 7% CO2 in O2 tended to reduce NaCN-induced responses on phrenic activity but not on blood pressure or sympathetic activity. Infusion into the vertebral artery with hypoxic saline (3% CO2 in N2) altered blood pressure, also affecting phrenic and sympathetic nerves similarly to NaCN administration. However, washout of CO2 by infusion of 100% O2 bubbled saline at high flow rates (3.6 ml/min) depressed phrenic as well as sympathetic activity and blood pressure. Spinal transection at the first cervical level eliminated sympathetic excitatory response to intravertebral cyanide injection. However, a large dose of NaCN (600 micrograms) given intravenously in spinal animal excited sympathetic activity. We conclude that intravertebral injection of NaCN can be used to study the effects of local hypoxia of the brain stem on cardiorespiratory responses and that hypoxia acts at both these sites (brain stem and spinal cord) to stimulate sympathetic excitation.

Animals↗
PubMed1992

Nonvagal modulation of hypoglossal neural activity.

Upper airway dilating muscle activity is characterized by an early-peaking pattern which serves to dilate or stiffen the upper airway at the time when the greatest negative intraluminal pressure is generated by contraction of chest wall muscles. This pattern has been attributed to phasic afferent inputs from pulmonary stretch receptors. The present study examines the hypothesis that nonvagal factors may also influence the discharge pattern and coordination of upper airway and chest wall muscle activities. Therefore, in anesthetized, paralyzed, vagotomized and artificially ventilated cats, we examined the effects of changes in respiratory drive produced by activation of cholinergic and GA-BAergic (gamma-aminobutyric acid) receptors at the ventrolateral aspects of the medulla oblongata on phasic intrabreath discharge patterns of hypoglossal and phrenic nerves. Cholinergic agents (acetylcholine, carbachol, methacholine, physostigmine) applied directly to chemoreceptive areas on the ventral medullary surface increased hypoglossal activity, and in addition converted inspiratory discharge from an augmenting to a decrementing pattern of activity. The reverse effect on the discharge pattern of hypoglossal activity was observed with a decrease in respiratory drive. While the amplitude of the phrenic nerve discharge was also affected by these interventions, the augmenting discharge pattern of phrenic nerve activity did not change. These results suggest that the early peaking pattern of hypoglossal nerve discharge in vagotomized cats also depends on the level of respiratory drive, and is not solely dependent on vagal afferent inputs. In addition, the data suggest that structures near the ventral surface of the medulla are influential in shaping the pattern of hypoglossal nerve activity and maintaining balanced activity of upper airway and chest wall muscles.

Acetylcholine↗
PubMed1991

Alteration of phrenic high frequency oscillation by local cooling of the ventral medullary surface.

Cooling of small sites on the ventral medullary surface of the cat produces a decrease in frequency of phrenic high frequency oscillation (HFO). The effect on HFO frequency of cooling near the hypoglossal rootlets, but not the effect of cooling near the inferior cerebellar artery, can be completely offset by raising arterial pCO2 to restore phrenic activity to its precooling level. Thus, structures near the ventral medullary surface are important for generation or propagation of phrenic HFO. The effect of cooling of the 'intermediate area' cannot be entirely explained as depression of putative central chemoreceptor activity.

Animals↗
PubMed1991

Sensation and control of breathing: a dynamic model.

A dynamic model of the CO2 respiratory control system is proposed, which can provide a qualitative basis for predicting breathing sensations. The discomfort index, which represents breathing sensations, is assumed to be composed of two sources: the arterial CO2 level and the respiratory motor command. The respiratory controller receives inhibitory neuromechanical and excitatory CO2 signals from the plant. The CO2 signal is enhanced by exercise stimuli. This dynamic multiplicative-type controller is used in simulations of key experiments: exercise and CO2 rebreathing with and without resistive loading. The dynamics of the discomfort index, the respiratory motor command, ventilation, and arterial CO2 concentration conform to the experimental data. The perceptual sensitivity to CO2 relative to respiratory effort is significantly correlated with the slope of hypercapnic ventilatory response. This result shows a clear linkage between ventilatory response and breathing sensations. Although it is shown that the automatic controller effectively minimizes the discomfort index for perturbations about an operating point under certain conditions, the discomfort index itself does not seem to be an underlying control principle of the proposed automatic controller model. Rather, breathing sensations may influence ventilatory responses by modifying the output of the automatic controller.

Carbon Dioxide↗
PubMed1991

Role of alpha 2-adrenergic receptors in the carotid body response to isocapnic hypoxia.

Alpha-2 adrenergic receptors have been identified in numerous tissues containing norepinephrine. Carotid bodies are sensory organs that detect changes in partial pressure of arterial oxygen and contain substantial amounts of norepinephrine. However, neither the presence nor the functional significance of alpha 2-receptors in the carotid body is known. The purpose of the present study was two-fold: (1) to determine the alpha 2-receptor density in the carotid body and (2) to assess their participation in the chemoreceptor responses to hypoxia. Experiments were performed on 27 anesthetized, paralyzed and artificially ventilated cats. Alpha-2 receptor binding determined by [125I]p-iodoclonidine averaged 10 +/- 2 fmol/mg of protein (n = 18 carotid bodies). Intracarotid infusion of an alpha 2-agonist (guanabenz; 0.5, 1, and 5 micrograms/min for 5 min) caused a dose-dependent depression of the baseline activity and 5 micrograms/min reduced the magnitude of the chemoreceptor response to isocapnic hypoxia by 61% (n = 8). Systemic administration of an alpha 2-antagonist (SKF-86466, 0.5-2 mg/kg) prevented the effects of guanabenz on the chemoreceptor activity. Furthermore, alpha 2-antagonist (0.5-2 mg/kg, i.v.) alone significantly increased baseline discharge by 68% and potentiated the chemoreceptor response to isocapnic hypoxia by 46% (n = 10). These results demonstrate that (1) alpha 2-adrenergic receptors are present in the carotid body and (2) they exert an inhibitory influence on chemoreceptor response to hypoxia. It is suggested that norepinephrine may tune the carotid body responses to hypoxia in part by its action on alpha 2-receptors.

Action Potentials↗
PubMed1991

Central effects of tachykinin peptide on tracheal secretion.

Tachykinin peptides acting on structures located on the ventral surface of the medulla can increase cholinergic outflow to the tracheal smooth muscles and augment respiratory motor output. In the experiments reported here (performed in anesthetized, paralyzed and artificially ventilated dogs), we examined the effects of tachykinin peptides substance P on secretion from submucosal glands. Changes in secretion were measured in an exposed section of tantalum-coated tracheal epithelium. Substances P was administered intracisternally or applied topically on the intermediate area of the ventral surface of medulla (VMS). Intracisternal infusion and the local medullary administration of tachykinin peptide caused a significant increase in tracheal submucosal gland secretion. Atropine given intravenously prevented the secretory changes induced by central action of tachykinins. In addition, prior application of 2% lidocaine to the medullary surface blocked the responses caused by substance P locally applied on the VMS. These findings suggest that substances P acting centrally can tracheal fluid secretion mainly via cholinergic mechanisms, and that the ventral surface of the medulla is one of the site of these action.

Amino Acid Sequence↗
PubMed1991

Influence of ventrolateral surface of medulla on tracheal gland secretion.

Airway secretion can be modified reflexly as well as locally. Previous studies indicate that neurons in a circumscribed region near the ventral surface of the medulla (VMS) can substantially modify airway tone and reflex responses to vagal inputs. In the present studies we assessed the importance of these neurons on tracheal gland secretion. We examined the changes in the number of hillocks of secretion appearing from submucosal glands in an exposed field of tracheal epithelium (1.2 cm2) coated with tantalum dust before and after interventions on the VMS. Experiments were performed in alpha-chloralose-anesthetized dogs paralyzed and ventilated with 40% O2. Stimulation of nicotinergic receptors by application of a pledget containing nicotine in 11 dogs caused a significant elevation in tracheal gland secretion in the subsequent 60 s, compared with a control period in which buffered saline was applied. Prior application of lidocaine or hexamethonium bromide to the VMS blocked the effect of topically applied nicotine. The central effects of nicotine were diminished by atropine methylnitrate given intravenously. In addition, lidocaine application to the VMS or focal cooling of intermediate areas to between 20 and 15 degrees C significantly decreased secretion rates reflexly produced by capsaicin-induced stimulation of pulmonary C-fiber receptors and by mechanical stimulation of the carina and larynx. These findings suggest that the ventral medulla contains cells near its surface that influence tracheal fluid secretion and modulate reflex responses of airway submucosal glands, probably by altering the level of general excitation within the central respiratory integrating circuits.

Animals↗
PubMed1991

Reflex responses of laryngeal and pharyngeal submucosal glands in dogs.

In dogs tracheal secretion is enhanced reflexly and by locally acting mediators such as substance P (SP). To evaluate the role of these mechanisms on submucosal gland secretion in the larynx (L) and pharynx (Ph), we compared the effects of mechanical stimulation of intrapulmonary irritant receptors and stimulation of pulmonary C-fiber receptors by capsaicin (20 micrograms/kg iv) with the response produced by intravenous SP. In six alpha-chloralose-anesthetized, paralyzed, and artificially ventilated dogs, submucosal gland secretion was monitored by analyzing the areas covered by hillocks of liquid and calculating the volume of secreted liquid (microliter) in the L and Ph. Mechanical stimulation of the carina increased both the number of hillocks and the volume of secreted liquid in the L. Excitation of pulmonary C-fiber receptors also increased the number of hillocks, and total volume of secreted liquid was elevated from 1.9 +/- 0.5 to 8.3 +/- 1.4 microliters (P less than 0.01). These responses were significantly reduced by prior cervical vagotomy and intravenous administration of atropine. Neither stimulation of irritant receptors nor stimulation of pulmonary C-fiber receptors caused discernible effects on Ph submucosal gland secretion. However, intravenous SP increased the number of Ph hillocks and elevated the volume of secreted Ph liquid from 1.0 +/- 0.6 to 10.2 +/- 1 microliters (P less than 0.01); similar responses to intravenous SP were observed in the L. Prior intravenous administration of atropine methylnitrate or bilateral vagotomy did not alter Ph or L secretory responses to intravenous SP.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1990

Occurrence of neutral endopeptidase activity in the cat carotid body and its significance in chemoreception.

The carotid body contains both tachykinins and enkephalins. Neutral endopeptidase (NEP, E.C. 3.4.24.11), has been suggested to involve in the metabolism of these neuropeptides in several organs. In the present study we determined neutral endopeptidase activity of the cat carotid body and assessed its significance in chemoreception. The cytosolic and membrane fractions of the carotid body contained NEP-like activity whereas it occurred only in the membrane fractions of the superior cervical and the nodose ganglia. Phosphoramidon, thiorphan and metal ion chelators inhibited NEP-like activity of all the 3 tissues studied; other protease inhibitors, however, were ineffective. Close carotid body administration of phosphoramidon significantly potentiated the carotid body response to low PO2 but not to hypercapnia. The enhanced response to hypoxia following phosphoramidon was further augmented by naloxone, an enkephalin antagonist. These results demonstrate that the glomus tissue contains detectable amounts of NEP-like activity and its inhibition selectively affects the hypoxic response of the carotid body.

Animals↗
PubMed1990

Effect of adenosine on isolated and superfused cat carotid body activity.

Adenosine is known to increase carotid chemoreceptor discharge in vivo. Since adenosine has powerful vascular effects it is possible that this chemoexcitation is indirectly caused by changes in carotid body blood flow. To evaluate this possibility the effect of adenosine (0.02-2.0 mumol) was assessed on the chemoreceptor activity of the cat carotid bodies in vitro. All three doses of adenosine produced an increase in chemoreceptor discharge which reached its maximum within 10-20 s and subsequently returned to preinjection controls within 1 min. The chemoreceptor excitation caused by adenosine was dose-dependent. These results suggest that adenosine induces chemoexcitation without changes in blood pressure and blood flow.

Action Potentials↗
PubMed1990

Effects of focal cooling in the ventrolateral medulla on chemoresponsiveness in dogs.

Studies in cats and dogs have shown that the ventrolateral region of the medulla participates significantly in the shaping of the respiratory rhythm. The purpose of this study was to examine the effects of unilateral focal cooling (15-20 degrees C) in the ventrolateral medullary region on respiratory responses to hypercapnia and hypoxia in dogs. A cryoprobe was used to cool selected locations in the ventrolateral medulla in 9 anesthetized and vagotomized dogs. Diaphragmatic electromyogram (EMG) was measured with implanted electrodes. The animals were ventilated artificially at a constant rate with 100% O2 and the inspired gas was switched to 7% CO2 in O2 or 10% O2 in N2 to determine the response to hypercapnia or hypoxia. The sites cooled ranged 4.0-8.0 mm rostral to obex, 3.0-5.5 mm lateral to midline, and within 1.5 mm deep from the ventral surface of the medulla. Unilateral focal cooling in this region significantly decreased the responses of both the amplitude and the rate of rise of diaphragmatic EMG to hypercapnia and hypoxia. These results support the hypothesis that neural structures in the ventrolateral medulla are important in the respiratory responses to hypoxia and hypercapnia as well as for the setting of respiratory drive and timing.

Animals↗