Influence of preload and afterload on genioglossus muscle length in awake goats

The genioglossus is an upper airway dilator muscle, the length of which is directly related to patency in the oropharyngeal region. We hypothesized that genioglossal length (Lgg) is dynamically influenced by the afterload exerted by negative upper airway pressure during inspiration and by the intrinsic length-tension characteristics of the muscle (preload). Seven awake goats were chronically instrumented with electrodes for EMGgg and sonomicrometry for Lgg. We examined the Lgg-EMGgg relationship during hypercapnia and inspiratory resistive loading (18 cm H2O/L/s). The goats breathed through the upper airway (TC) or airflow was diverted through a tracheostomy (TO). We found that: (1) passive inspiratory lengthening was observed with negative upper airway pressure (UAP) but not when UAP = 0 (TO breathing), (2) Lgg shortening for a given EMGgg was significantly decreased with negative inspiratory UAP, and (3) phasic Lgg shortening per unit EMGgg was greatest when Lgg was near optimal length (Lo). We conclude that genioglossal length is substantially influenced by afterload exerted by negative UAP and that genioglossal active shortening may be limited if the muscle operates at a length significantly greater or less than the optimal length.     

Airway occlusion pressures in awake and anesthetized goats

The ability of intramuscular injections of gonadal steroids to exert a positive feedback action on LH secretion was investigated in the ovariectomized hen. Plasma LH was measured by radioimmunoassay. Single injections of progesterone (dose range: 0.05-10 mg/kg) or oestradiol benzoate (dose range: 0.01-1 mg/kg) did not result in an increase in plasma LH concentration. After priming with 0.1 mg oestradiol benzoate/kg on alternate days for 7 days and with 0.5 mg progesterone/kg on days 5, 6 and 7, a single injection of progesterone on day 8 (dose range: 0.1-2 mg/kg) caused the plasma LH concentration to start increasing after 15 to 30 min. Peak LH concentration was reached around 1.5-2 h after injection. The magnitude of LH response to progesterone was dose related. In contrast, a single injection of oestradiol benzoate (dose range: 0.01-1 mg/kg) failed to stimulate LH release in the oestrogen-progesterone primed ovariectomized (O-P-OVX) hen. A single injection of testosterone (dose range: 0.1-2.0 mg/kg) failed to stimulate LH release in ten out of 12 O-P-OVX hens. A small increase in LH secretion was observed in the two remaining birds. When oestrogen or progesterone was omitted from the priming schedule, a LH positive feedback response to a single injection of progesterone was not observed. Increasing or decreasing the mount of oestrogen or progesterone in the priming schedule modified the LH response to a single injection of progesterone on the day following the last priming injection. This suggested that a critical oestrogen to progesterone ratio was required to prime the LH positive feedback mechanism. It is suggested that, in the hen, the release of LH is facilitated by the positive feedback effect of a combination of oestrogen and progesterone in a two-phase process. The first is the priming phase, which depends on the presence in the blood of oestrogen and progesterone; the second is the ind .uctive phase, which depends only on an incremental change in plasma progesterone concentration. Oestrogen is not involved in the induceive phase.     

Free radical induced respiratory muscle dysfunction

1,3-Butadiene (BD) is carcinogenic and mutagenic in B6C3F1 mice. We determined the lacI mutant frequency and mutational spectrum in spleen following inhalation exposure to BD at levels that are known to induce tumors. B6C3F1 lacI transgenic mice were exposed to air or to 62.5, 625, or 1250 ppm BD for 4 weeks (6 h/day, 5 days/week) and euthanized 14 days after the last exposure. BD increased the lacI mutant frequency in spleen at all levels of BD examined. In BD-exposed mice, an increased frequency of G:C-->A:T transitions occurred at non-5'-CpG-3' sites. Exposure to BD in B6C3F1 lacI transgenic mice also increased the frequency of base substitution mutations that occurred at A:T base pairs when compared to air controls. The increased frequency of specific mutations at G:C base pairs in spleen was not observed in our previous studies in bone marrow and indicates tissue-specific differences in the BD-induced mutational spectrum. These data demonstrate that in vivo transgenic mouse mutagenicity assays can identify tissue-specific mutagenicity and mutational spectrum responses of genotoxic carcinogens at exposure levels that are known to induce tumors.     

Motor-unit recruitment in self-reinnervated muscle

1. Recruitment order of motor units in self-reinnervated medial gastrocnemius (MG) muscles was studied in decerebrate cats 16 mo after surgical reunion of the cut MG nerve. Pairs of MG motor units were isolated by dual microelectrode penetration of ventral roots to measure their recruitment sequence during cutaneous reflexes in relation to their physiological properties. 2. Physiological properties of reconstituted motor units appeared normal, as expected. Also normal were the relationships among these properties: twitch and tetanic tension tended to increase with axonal conduction velocity and decrease with twitch contraction time. A small fraction of motor units (10/116) in reinnervated muscles produced either no measurable tension or unusually large amounts of tension compared with controls. This was the only distinct feature of the sample of reconstituted units. 3. In muscles reinnervated after nerve section, stretch was notably ineffective in eliciting reflex contraction of MG muscles or their constituent motor units (only 5/116 units). Incomplete recovery from nerve section was probably the cause of this impairment, because stretch reflexes were readily evoked in adjacent untreated muscles and in one reinnervated MG muscle that was studied 16 mo after nerve crush. In contrast with the ineffectiveness of muscle stretch, sural nerve stimulation succeeded in recruiting 49/116 units, a proportion fairly typical of normal MG muscles. 4. The contractions of the first unit recruited in cutaneous reflexes tended to be slower and less forceful than those of the other unit in a pair. By these measures, recruitment obeyed the size principle. This recruitment order with respect to unit contractile properties was not significantly different (P > 0.05) between untreated and reinnervated muscles but was significantly (P < 0.005) different from random order in both groups. The same recruitment pattern was observed for pairs of motor units sampled from the muscle reinnervated after nerve crush, whether units were recruited by muscle stretch or sural nerve stimulation. 5. The usual tendency for motor units with slower conduction velocity (CV) to be recruited in sural nerve reflexes before those with faster CV was not strong in reinnervated muscles. After nerve section the proportion of units exhibiting the usual recruitment pattern was not significantly different (P > 0.05) from a random pattern for CV. 6. The central finding is that the normal recruitment patterns recover from nerve injury in a muscle that is reinnervated by its original nerve. By contrast, stretch reflexes do not recover well from nerve section, and this deficiency may contribute to motor disability.     

Enhancement of pressor response to intravenous phenylephrine following oral clonidine medication in awake and anaesthetized patients

Clonidine, an alpha 2-adrenergic agonist, augments the pressor response to intravenous ephedrine. If this effect is partly due to clonidine-induced potentiation of alpha 1-adrenoceptor-mediated vasoconstriction, it is also assumed that clonidine would enhance the pressor effect of phenylephrine as an alpha 1-adrenergic agonist. The authors studied haemodynamic responses to intravenous phenylephrine in 80 patients who received either preanaesthetic medication with clonidine approximately 5 micrograms.kg-1 po (clonidine group, n = 40), or no medication (control group, n = 40). Each group was further divided into either awake subjects (n = 20) or subjects anaesthetized with enflurane and nitrous oxide in oxygen (n = 20). Haemodynamic measurements were made at one-minute intervals for ten minutes after phenylephrine 2 micrograms.kg-1 iv was injected as a bolus. The magnitudes of maximal mean blood pressure increases in the clonidine group (26 +/- 7% (mean +/- SD) for awake and 32 +/- 15% for anaesthetized subjects) were greater (P < 0.05) than in the control group (13 +/- 7% for awake and 18 +/- 7% for anaesthetized subjects). However, there was no difference in the pressor effect of phenylephrine between awake and anaesthetized patients in both groups. Oral clonidine preanaesthetic medication, 5 micrograms.kg-1, augments the pressor responses to phenylephrine 2 micrograms.kg-1 iv in awake and anaesthetized patients. These results suggest that the enhancement of the pressor responses to phenylephrine following oral clonidine may be due to clonidine-induced potentiation of alpha 1-adrenoceptor-mediated vasoconstriction. This implies that restoration of blood pressure can be achieved effectively by phenylephrine in hypotensive patients with clonidine premedication.     

Assessment of the respiratory compliance in awake subjects using pressure support

Pressure support (PS), a new mode of ventilatory assistance, is known to induce respiratory muscle relaxation. It was used to obtain reliable measurements of the compliance of the respiratory system (Crs) in awake subjects. PS was applied, through a mouthpiece, at four successive levels (0, 0.75, 1 and 1.25 kPa) to 30 healthy subjects. At the highest PS level, the subject's relaxation was obtained as assessed by a decrease in the occlusion pressure from 0.10 +/- 0.06 to 0.05 +/- 0.04 kPa, whereas the minute ventilation increased (from 7.5 +/- 1.5 to 13.8 +/- 3.3 l.min-1), and the end-tidal carbon dioxide tension (PCO2) decreased (from 5.0 +/- 0.4 to 3.2 +/- 0.5 kPa) below its apnoea threshold. In three subjects, respiratory muscle relaxation was confirmed by a fall in diaphragmatic electromyographic activity. Crs was calculated as the ratio of the tidal volume to the corresponding end-inspiratory airway pressure (i.e. PS level) since, at end inspiration, a zero-flow period was obtained. Crs was highly correlated (r = 0.77) to the height (Ht) of the subjects: Crs (l.kPa-1) = 3.56 x Ht (m) -4.86 (+/- 0.23), allowing normal values to be determined. In order to evaluate the applicability of the method to patients, Crs was measured in four patients with scoliosis, and was found to range from 45-82% of the predicted values. It is suggested that this simple method of Crs determination may be used to characterize various chest wall or pulmonary diseases.     

Effect on breathing of surface ventrolateral medullary cooling in awake, anesthetized and asleep goats

In adult and neonatal goats, we chronically implanted thermodes on the ventrolateral (VLM) medullary surface to create reversible neuronal dysfunction and thereby gain insight into the role of superficial VLM neurons in control of breathing in anesthetized, awake and asleep states. Consistent with data of others, cooling caudal area M and rostral area S caused sustained apnea under anesthesia. However, in the awake and NREM sleep states, cooling at this site caused only a modest reduction in breathing, indicating that neurons at this site are not critical for respiratory rhythm in these states. Moreover, data in the awake state over multiple conditions suggest neurons at this site do not integrate all intracranial and carotid chemoreception. The data suggest though that neurons at this site have a facilitatory-like effect on breathing both unrelated and related to intracranial chemoreception. We believe that this facilitation serves a function similar to the facilitation provided by the carotid chemoreceptors and by sources associated with wakefulness. Accordingly, elimination/attenuation of any one of these three influences (caudal M rostral S VLM, wakefulness, carotid chemoreception) results in a slight decrease in breathing, removal of two of the three results in a greater decrease in breathing, and removal of all three results in sustained apnea.     

Adrenergic reactivity after inhibition of nitric oxide synthesis in the cerebral circulation of awake goats

The interaction between nitric oxide (NO) and adrenergic reactivity in the cerebral circulation was studied using in vivo and in vitro preparations. Blood flow to one brain hemisphere (cerebral blood flow) was electromagnetically measured in conscious goats, and the effects of norepinephrine, tyramine and cervical sympathetic nerve stimulation were recorded before (control) and after inhibition of NO formation with Nw-nitro-l-arginine methyl ester (l-NAME). The responses to norepinephrine, tyramine and electrical field stimulation were also recorded in segments, 4 mm in length, from the goat's middle cerebral artery under control conditions and after l-NAME. In vivo, l-NAME (10 goats, 47 mg kg-1 administered i.v.) reduced resting cerebral blood flow by 37+/-2%, increased mean systemic arterial pressure by 24+/-3%, reduced heart rate by 35+/-2%, and decreased cerebrovascular conductance by 52+/-2% (all P<0.01). Norepinephrine (0.3-9 microgram), tyramine (50-500 microgram), and supramaximal electrical sympathetic cervical nerve stimulation (1. 5-6 Hz) decreased cerebrovascular conductance, and these decreases were significantly higher after l-NAME than under control conditions, remaining higher for about 48 h after this treatment. Norepinephrine (10-8-10-3 M), tyramine (10-6-10-3 M) and electrical field stimulation (1.5-6 Hz) contracted isolated cerebral arteries, and the maximal contraction, but not the sensitivity, was significantly higher in the arteries treated than in non-treated with l-NAME (10-4 M). Therefore, the reactivity of cerebral vasculature to exogenous and endogenous norepinephrine may be increased after inhibition of NO synthesis. This increase might be related, at least in part, to changes at postjunctional level in the adrenergic innervation of the vessel wall, and it might contribute to the observed decreases in resting cerebral blood flow after inhibition of NO synthesis.     

Sternocleidomastoid muscle inhibition induced by trigeminal stimulation

Among numerous reports of anatomical and functional coupling between the trigeminal and cervical systems is the demonstration that the sternocleidomastoid (SCM) muscles may become activated along with the masseter muscles during forceful abrupt biting maneuvers. Whether the co-activated SCM is also inhibited by stimuli that produce masseter inhibition is not known. This study evaluated the SCM for the presence of inhibition during mechanically-elicited (chin or forehead tap) and electrically-elicited (anterior maxillary gingiva stimulation) inhibition of the masseter muscle in ten healthy men. Surface EMG data were recorded bilaterally from the masseter and SCM muscles. The data for each muscle were converted to ratios of the pre-stimulus maximum voluntary contraction activity for each subject and averaged across subjects. Means of these percentages were determined at several defined pre- and post-stimulus intervals. The results indicate that masseter inhibition was clearly elicited by the electrical and both forms of mechanical stimulation. SCM co-inhibition could be evoked by electrical and chin tap stimulation but not by forehead tap. The responses to these stimuli varied among subjects, from trial to trial, and within subjects depending on the experimental condition. The fact that it was possible for this co-inhibition to be evoked is presented as further indication of the functional coupling of the trigeminal and cervical systems.     

The optimal test dose of epinephrine for epidural injection with lidocaine solution in awake patients premedicated with oral clonidine

We attempted to determine the optimal test dose of epinephrine for use with epidural anesthesia in awake patients premedicated with clonidine. Eighty-eight adult patients were randomized into two groups [oral premedication with clonidine 5 microg/kg (CLON) or no premedication (CONT)]. Before induction of general anesthesia, heart rate (HR) and blood pressure (BP) were measured for 3 min after the i.v. injection of 3 mL of 1.5% lidocaine containing epinephrine (0, 1.25, 2.5, 5, 7.5, or 15 microg) in a randomized, double-blind manner. We calculated 95% confidence intervals for the peak HR and BP increases induced by each dose of epinephrine. At 7.5 microg, epinephrine induced a significantly greater increase in HR and BP in CLON than in CONT. The 95% confidence interval for the HR change induced by 7.5 microg of epinephrine in CLON was nearly the same as the accepted standard dose of epinephrine (15 microg) in CONT. We conclude that premedication with clonidine enhances HR and BP responses to the i.v. administration of epinephrine-containing epidural test solutions. Consequently, 7.5 microg of epinephrine may be sufficient to enable detection of accidental injection into a blood vessel in awake patients premedicated with clonidine 5 microg/kg. Implications: Clonidine, a commonly used preanesthetic medication, alters patients' cardiovascular responses to drugs such as epinephrine. Our randomized, double-blind study suggests that, in awake patients receiving oral clonidine premedication, 7.5 microg of epinephrine (half the usual dose) is adequate as an indicator of accidental injection into the epidural vessels during epidural anesthesia.     

Quantitative research for improving respiratory muscle contraction by breathing exercise

Mycobacterium gordonae is an atypical mycobacterium of very low pathogenic potential. It is widely distributed in soil and water and often detected on the mucous membranes of healthy persons. In recent years, there have been numerous reports of infections by M. gordonae in immunocompromised patients. In contrast, only four cases of skin infections by M. gordonae in immunocompetent patients have been published. We report on another patient without evidence of immunodeficiency who developed an atypical mycobacteriosis after a thorn injury during gardening. M. gordonae was isolated by tissue culture. The skin lesion cleared completely after treatment with doxycycline for three months.     

Changes in stiffness induced by hindlimb suspension in rat soleus muscle

Soleus muscle atrophy was induced by hind-limb suspension of rats for 3 weeks with the intention of inducing a relative increase in the percentage of fast-twitch fibres and assessing modifications in muscle stiffness. A method of dual controlled releases was used to obtain tension/extension curves and force/velocity relationships characterizing the mechanical behaviour of the soleus. Fibre typing was achieved by myofibrillar adenosine 5'-triphosphatase staining. Results showed that hindlimb suspension decreased the percentage of slow-twitch fibres (-31%) to the profit of fast-twitch fibres (+370%) and intermediate fibres (+255%). This led to an increase in maximal shortening velocity. Tension/extension curves indicated a decrease in soleus stiffness after 3 weeks of unloading. Changes in elastic properties are interpreted in terms of modifications occurring in the active part and the passive part of the so-called series elastic component. These changes also suggest that the parameters derived from a twitch are inappropriate to account for modifications in speed-related properties of muscle.     

Differential cerebrovascular responsiveness in spontaneously hypertensive rats following antihypertensive treatment with clonidine and verapamil

Numerous studies have been reported examining the effects of antihypertensive treatment on peripheral vascular responsiveness in spontaneously hypertensive rats (SHR). This study was conducted to determine the effects of chronic treatment with 2 antihypertensive agents on cerebrovascular responsiveness in male SHR and Wistar-Kyoto (WKY) rats. SHR and WKY (3-4 weeks old) received either placebo, clonidine (CLON, 10 mg pellet) or verapamil (VER, 5 mg pellet). Vascular reactivity studies on the basilar artery, using standard smooth muscle bath techniques, were conducted following 6 weeks of treatment. Both CLON and VER significantly attenuated the rise in blood pressure in SHR. Basilar artery responsiveness to KCl, serotonin (5-HT), and calcium were significantly increased whereas responses to acetylcholine (ACH), isoproterenol (ISO) and sodium nitroprusside (SNP) were significantly reduced in SHR compared to WKY. CLON had no effect on basilar artery responsiveness to either the contractile or relaxation agents in SHR. However, although responses to KCl, 5-HT and calcium were not affected by VER in SHR, VER significantly increased the responses to ACH, ISO and SNP. Neither CLON nor VER treatment affected basilar artery responsiveness to any of the agents in WKY. These data demonstrate that, even though CLON and VER have similar antihypertensive effects, differential effects of the 2 agents on cerebrovascular responsiveness in the SHR are apparent. This would suggest that the vascular effects of VER and CLON are dependent upon the mechanism of action of the agents and not simply due to prevention of the elevation in blood pressure.     

Cellular mediation of respiratory distress syndrome induced by fragment D

The serum concentration of fibrinogen degradation product D (Fg D) is elevated after injury and sepsis. Purified human Fg D infused into awake rabbits causes progressive thrombocytopenia, complement depletion, hypoxia, vascular permeability to albumin and neutrophil congestion. In previous work experimentally induced thrombocytopenia protected lungs of rabbits against the effects of Fg D. The present study was designed to determine the role of the neutrophil in the development of Fg D-induced respiratory distress by rendering rabbits neutropenic with antiserum. Neutrophil depletion offered some, but not total, protection of the lungs against the toxic effects of Fg D infusion. Only one neutropenic rabbit became hypoxic. Vascular leak to albumin and water was diminished. Platelet and white blood cell counts decreased. However, complement activity was unaffected. The results suggest that neutrophils, like platelets, also contribute to the onset of respiratory failure in this model.     

A novel phasic contraction induced by dithiothreitol in frog skeletal muscle

1. Dithiothreitol (DTT), at 50-100 mM, induced a phasic reversible contraction of frog skeletal muscle. 2. Exposure of single fibers to nifedipine (20 microM), an L-type Ca2+ antagonist, blocked the twitch and tetanus tensions but never affected the DTT-induced contraction. 3. DTT also produced a phasic contraction in fibers where voltage sensors were inactivated in the presence of high K+ concentration (190 mM). 4. A fiber was mechanically skinned after observation of DTT-induced contraction. The skinned fiber contracted in response to a DTT concentration similar to that required to produce contraction in intact fibers before skinning. 5. In skinned fibers, DTT, at 100 or 200 mM, inhibited the accumulation of Ca2+ by SR, but not Ca2+ ATPase activity. 6. These results suggest that a high concentration of DTT triggers Ca2+ efflux from the SR through action on the Ca2+ release channel and/or closely associated proteins, such as triadin and FK-506 binding protein.     

Skeletal muscle fiber degeneration in mdx mice induced by electrical stimulation

We present an in vitro model in which mouse skeletal muscle fibers undergo degeneration by increasing the current strength of tetanic stimulation. To understand the mechanisms of muscle fiber necrosis in Duchenne muscular dystrophy patients, the process of fiber degeneration was compared between mdx and control mice. The process consisted of four steps, beginning with muscle fiber contraction and extending to onset of myofibril disruption. The four processes were not observed in fibers in Krebs-HEPES (-Ca2+) buffer, nor in the presence of L-type Ca2+ channel blockers. These results suggest that this degenerative phenomenon is regulated by intracellular Ca2+, which moved into fibers mainly through voltage-dependent L-type Ca2+ channels. With the exception of myofibril disruption, mdx mice also exhibited the three other steps, but at a significantly lower current strength than in the fibers in the control mice. We postulate that excess Ca2+ flux occurs in fibers, mainly through abnormal L-type Ca2+ channels, and that the excessively accumulated calcium results in premature degeneration of the fibers by tetanic contraction. This study would provide a clue to investigate and prevent the degeneration processes in Duchenne muscular dystrophy.     

Differential withdrawal of retinal axons induced by a secreted factor

To understand the development of the topographic map in the chick retinotectal projection, we studied the long-term interactions between retinal axons and tectal cell processes using a novel coculture system, the ryomen chamber. Both nasal and temporal retinal axons initially grew equally well on a substrate consisting of posterior tectal cell processes; however, subsequently most temporal axons withdrew from this surface, whereas most nasal axons did not. Experiments using conditioned media indicate that posterior tectal cells induced withdrawal of the temporal axons by secreting a soluble factor. This withdrawal seems to be distinct from the immediate repulsive effect of ephrin-A2 (ELF-1) and ephrin-A5 (RAGS) seen in the stripe assay because (1) the withdrawal-inducing factor was diffusible, whereas ephrin-A2 and -A5 are membrane-bound, and (2) the withdrawal-inducing factor appeared later in development than ephrin-A2 and -A5. Furthermore, sensitivity to the withdrawal-inducing factor decreased continuously from the temporal to nasal retina. These results suggest that target cell-induced axonal withdrawal may be involved during a late stage of the development of the retinotectal map.     

Differential recruitment of leukocyte populations and alteration of airway hyperreactivity by C-C family chemokines in allergic airway inflammation

Allergic airway inflammation is characterized by peribronchial leukocyte accumulation within the airway. Subsequent tissue damage leading to airway hyperreactivity is a result of activation of multiple leukocyte populations. Using an established model of allergic airway inflammation induced by intratracheal challenge with parasite (Schistosoma mansoni) egg Ag in presensitized mice, we have examined differential leukocyte recruitment. These studies have identified key chemokines involved in the accumulation of specific subsets of cells and the induction of airway hyperreactivity. In this study we have examined three C-C family chemokines, MCP-1, MIP-1alpha, and RANTES, which promote mononuclear cell- and eosinophil-specific recruitment to the airway. The in vivo neutralization of either MIP-1alpha or RANTES, but not MCP-1, significantly reduced the intensity of the eosinophil recruitment to the lung and airway during the allergic airway response by >50 and >60%, respectively. In contrast, neutralization of MCP-1 significantly reduced total leukocyte migration (>50% reduction), whereas neutralization of RANTES and MIP-1alpha had no significant affect on the overall leukocyte migration. Further examination of the effect of MCP-1 depletion indicated that both CD4+ and CD8+ lymphocyte subsets were decreased. Depletion of MCP-1 significantly reduced the airway hyperreactivity to near control levels, whereas depletion of MIP-1alpha or RANTES did not affect the intensity of airway hyperreactivity. These data indicate that multiple C-C chemokines are involved in the recruitment of particular leukocyte populations and that neutralization of MCP-1, but not RANTES or MIP-1alpha, significantly reduced airway hyperreactivity.