Localized delivery of therapeutics impact laryngeal mechanics, local inflammatory response, and respiratory microbiome following upper airway intubation injury in swine.

BACKGROUND: Laryngeal injury associated with traumatic or prolonged intubation may lead to voice, swallow, and airway complications. The interplay between inflammation and microbial population shifts induced by intubation may relate to clinical outcomes. The objective of this study was to investigate laryngeal mechanics, tissue inflammatory response, and local microbiome changes with laryngotracheal injury and localized delivery of therapeutics via drug-eluting endotracheal tube. METHODS: A simulated traumatic intubation injury was created in Yorkshire crossbreed swine under direct laryngoscopy. Endotracheal tubes electrospun with roxadustat or valacyclovir- loaded polycaprolactone (PCL) fibers were placed in the injured airway for 3, 7, or 14 days (n = 3 per group/time and ETT type). Vocal fold stiffness was then evaluated with normal indentation and laryngeal tissue sections were histologically examined. Immunohistochemistry and inflammatory marker profiling were conducted to evaluate the inflammatory response associated with injury and ETT placement. Additionally, ETT biofilm formation was visualized using scanning electron microscopy and micro-computed tomography, while changes in the airway microbiome were profiled through 16S rRNA sequencing. RESULTS: Laryngeal tissue with roxadustat ETT placement had increasing localized stiffness outcomes over time and histological assessment indicated minimal epithelial ulceration and fibrosis, while inflammation remained severe across all timepoints. In contrast, vocal fold tissue with valacyclovir ETT placement showed no significant changes in stiffness over time; histological analysis presented a reduction in epithelial ulceration and inflammation scores along with increased fibrosis observed at 14 days. Immunohistochemistry revealed a decline in M1 and M2 macrophage markers over time for both ETT types. Among the cytokines, IL-8 levels differed significantly between the roxadustat and valacyclovir ETT groups, while no other cytokines showed statistically significant differences. Additionally, increased biofilm formation was observed in the coated ETTs with notable alterations in microbiota distinctive to each ETT type and across time. CONCLUSION: The injured and intubated airway resulted in increased laryngeal stiffness. Local inflammation and the type of therapeutic administered impacted the bacterial composition within the upper respiratory microbiome, which in turn mediated local tissue healing and recovery.

Routine restaging after primary non-surgical treatment of laryngeal squamous cell carcinoma-a review.

PURPOSE: Treatment of patients with laryngeal squamous cell carcinoma with radiotherapy or chemoradiation is an established alternative to laryngeal surgery in many cases, but particularly for advanced tumors without cartilage invasion. Imaging modalities face the challenge of distinguishing between posttherapeutic changes and residual disease in the complex anatomic subsite of the larynx. Guidelines concerning restaging of head and neck squamous cell carcinomas (HNSCC) are presented by the National Comprehensive Cancer Network (NCCN) and other national guidelines, but clearly defined recommendations for routine restaging particularly for laryngeal cancer are lacking. METHODS: A systematic search was carried out in PubMed to identify studies evaluating routine restaging methods after primary non-surgical treatment of laryngeal squamous cell carcinoma from 2009 to 2020. RESULTS: Only three studies were deemed eligible, as they included at least ≥50% patients with laryngeal squamous cell carcinoma and evaluated imaging modalities to detect residual cancer. The small number of studies in our review suggest restaging with fluoro-deoxy-glucose positron-emission tomography/computed tomography (FDG PET/CT) 3 months after initial treatment, followed by direct laryngoscopy with biopsy of the lesions identified by FDG PET/CT. CONCLUSION: Studies evaluating restaging methods after organ-preserving non-surgical treatment of laryngeal carcinoma are limited. As radiotherapy (RT), chemoradiotherapy (CRT), systemic therapy followed by RT and radioimmunotherapy are established alternatives to surgical treatment, particularly in advanced laryngeal cancers, further studies are needed to assess and compare different imaging modalities (e.g. PET/CT, MRI, CT, ultrasound) and clinical diagnostic tools (e.g., video laryngoscopy, direct laryngoscopy) to offer patients safe and efficient restaging strategies. PET or PET/CT 3 months after initial treatment followed by direct laryngoscopy with biopsy of the identified lesions has the potential to reduce the number of unnecessary laryngoscopies.

Vocal fold injection material does not preclude interpretation of laryngeal electromyography.

INTRODUCTION/AIMS: Temporary vocal fold injection (VFI) is a common treatment for acute and subacute vocal fold paralysis (VFP). Laryngeal electromyography (LEMG) is useful for diagnosing neurogenic causes of VFP. This study evaluated whether the presence of VFI material prevents interpretation of LEMG in patients with acute and subacute VFP. METHODS: Patients with acute and subacute unilateral VFP (onset ≤6 mo) who underwent temporary VFI within 3 mo preceding LEMG were evaluated. A matched control group that did not undergo VFI was also studied. The LEMG team (laryngologist and electromyographer) performed and interpreted LEMG using a pre-specified protocol, including qualitative and quantitative motor unit analysis. RESULTS: Eighteen patients with VFI underwent LEMG successfully with interpretation of spontaneous activity and motor unit recruitment. Fourteen patients were seen in follow-up to determine accuracy of established LEMG prognosis. Seven of seven subjects with poor LEMG prognosis did not recover vocal fold motion. Five of seven subjects with fair LEMG prognosis recovered vocal fold motion. Findings were similar for the control group. DISCUSSION: VFI augmentation material did not prevent interpretation of meaningful LEMG data in patients with acute and subacute VFP, and accurate prognoses of vocal fold motion recovery were established.

Liquiritin apioside attenuates laryngeal chemoreflex but not mechanoreflex in rat pups.

Liquiritin apioside (LA), a main flavonoid component of licorice, reportedly suppresses cough responses to inhalation of aerosolized capsaicin [CAP; a stimulant to transient receptor potential vanilloid 1 (TRPV1)] in conscious guinea pigs via acting on peripheral nerves. However, the evidence of LA having a direct effect on airway sensory fibers is lacking. Considering the important role laryngeal chemoreceptors and mechanoreceptors play in triggering apnea and cough, we studied whether LA suppressed the apneic responses to stimulation of these receptors via directly acting on the superior laryngeal nerve (SLN). Intralaryngeal delivery of chemical [CAP, HCl, and distilled water (DW)] and mechanical [an air-pulse (AP)] stimulations was applied in anesthetized rat pups to evoke the apnea. These stimuli were repeated after intralaryngeal LA treatment or peri-SLN LA treatment to determine the direct effect of LA on the SLN. Our results showed that all stimuli triggered an immediate apnea. Intralaryngeal LA treatment significantly attenuated the apneic response to chemical but not mechanical stimulations. The same attenuation was observed after peri-SLN LA treatment. Owing that TRPV1 receptors of laryngeal C fibers are responsible for the CAP-triggered apneas, the LA impact on the activity of laryngeal C neurons retrogradely traced by DiI was subsequently studied using a patch-clamp approach. LA pretreatment significantly altered the electrophysiological kinetics of CAP-induced currents in laryngeal C neurons by reducing their amplitudes, increasing the rise times, and prolonging the decay times. In conclusion, our results, for the first time, reveal that LA suppresses the laryngeal chemoreceptor-mediated apnea by directly acting on the SLN (TRPV1 receptors of laryngeal C fibers).

Laryngeal reflex responses in pediatric anesthesia.

Laryngeal and respiratory reflexes are vitally important defense mechanisms against foreign body aspiration, safeguarding airway patency, and ventilation. These highly preserved automatisms easily overrule external influences like willpower or (anesthetic) medication. Prevention and anticipation are, therefore, the essential strategies to avoid adverse events and damage, and treatment is most effective in the early stage of the reflex response. The physiology and pathophysiology of the various defensive reflexes as well as a comprehensive anesthetic approach to prevention and treatment are outlined in this review.

Laryngeal inflammatory response to smoke and vape in a murine model.

PURPOSE: To build a murine model for tobacco smoke and electronic cigarette vapor exposure to characterize the inflammatory and immune responses in the larynx. MATERIALS AND METHODS: In this pilot study, twenty-four wild-type C57BL/6 mice were divided into four groups: smoke, vapor with nicotine, vapor without nicotine, and air only. Following daily exposure for 4 months, larynges were dissected and processed with cytokine detection arrays. Each laryngeal cytokine level between the four different groups was analyzed statistically by using statistical analysis software (SAS) to calculate the analysis of variance (ANOVA). RESULTS: IL-4 was the only cytokine found to achieve statistically significant different levels in this study, with elevated levels of IL-4 in the tobacco smoke and vapor with nicotine groups compared to the levels found in the vapor without nicotine and air only groups (p = 0.0418). While statistically non-significant, prominent findings revealed up-regulation of TGF-ß2 and TGF-ß3 in the smoke group, but near-normal levels of TGF-ß2 and TGF-ß3 and suppression of IL-10 in the vapor groups (p > 0.05). CONCLUSION: The potential utility of the murine model is established for studying the inflammatory and immune effects of tobacco smoke and vapor on the mammalian larynx. IL-4 levels in mice larynges were significantly elevated in the tobacco smoke and vapor with nicotine groups.

Comparison of methohexital and propofol as induction agents for evaluation of laryngeal function in healthy dogs.

OBJECTIVE: To determine the influence of propofol or methohexital, with and without doxapram, on the examination of laryngeal function in dogs. STUDY DESIGN: Experimental study. ANIMALS: Forty healthy dogs randomly assigned to 4 groups: propofol with saline (n = 10), propofol with doxapram (n = 10), methohexital with saline (n = 10), or methohexital with doxapram (n = 10). METHODS: Propofol and methohexital were administered to effect. Investigators examined laryngeal function (initial) simultaneously with video laryngoscopy. Doxapram or saline was administered, and laryngeal function was reevaluated (second). Laryngeal motion, quality of laryngeal exposure, and the degree of swallowing, laryngospasm, and jaw tone were scored at each evaluation. Adverse events were recorded. Initial and second videos were evaluated by a masked observer, and still images obtained from both evaluations were evaluated for change in rima glottidis size by 2 masked observers. RESULTS: Administration of doxapram and saline was delayed with propofol (P = .001). Laryngeal function did not differ between dogs receiving propofol or methohexital, irrespective of doxapram administration. Doxapram improved breathing scores in both groups (P < .001). Jaw tone increased with propofol during the second evaluation (P = .049). Swallowing was more prevalent at initial examination (P = .020). Methohexital resulted in an increased heart rate (P < .001) compared with propofol. Twenty-five percent of dogs receiving methohexital developed seizure-like activity (n = 5/20). CONCLUSION: Evaluation of laryngeal function did not differ between healthy dogs anesthetized with propofol or methohexital. Methohexital provided shorter examination times with less jaw tone but was associated with adverse events. CLINICAL SIGNIFICANCE: This study provides evidence to recommend propofol over methohexital as an induction agent for laryngeal function examination.

Effects of chemical and mechanical stimulation on laryngeal motion during alfaxalone, thiopentone or propofol anaesthesia in healthy dogs.

OBJECTIVE: To compare the effect of chemical and mechanical stimulation on arytenoid cartilage motion during anaesthetic induction with alfaxalone, thiopentone or propofol. STUDY DESIGN: Masked, randomized, crossover study. ANIMALS: A group of eight adult Beagle dogs. METHODS: Anaesthesia was induced with thiopentone (7.5 mg kg-1), propofol (3 mg kg-1) or alfaxalone (1.5 mg kg-1) intravenously (IV), which were concurrently paired with either chemical (doxapram at 2.5 mg kg-1 IV) or mechanical (gentle pressure to the corniculate process of the right arytenoid cartilage using a cotton bud) stimulation for enhanced assessment of laryngeal motion, in random order, with a 1 week wash-out period between treatments. If deemed inadequately anaesthetized, supplemental boli of thiopentone (1.8 mg kg-1), propofol (0.75 mg kg-1) or alfaxalone (0.4 mg kg-1) were administered. Assessment of number of arytenoid motions and vital breaths, among others, was initiated immediately after induction. Chemical (doxapram) and mechanical stimulation were begun 2 minutes after anaesthetic induction. Data were collected at 2, 3 and 5 minutes after anaesthetic induction and the Friedman rank-sum or repeated-measures analysis of variance tests were used when applicable for statistical analysis. RESULTS: The duration of examination time was shorter among treatments combined with chemical stimulation (p=0.001). Examination time during induction was longer for alfaxalone-chemical (8.9 minutes) and -mechanical (10.9 minutes) compared to both induction with thiopentone-chemical (3.8 minutes) and propofol-chemical (4.0 minutes). The median number of arytenoid motions for both thiopentone (67) and propofol (59) induction combined with chemical stimulation was significantly higher in comparison to that of alfaxalone (1), thiopentone (2) and propofol (2), when combined with mechanical stimulation at 3 minutes after induction. CONCLUSION AND CLINICAL RELEVANCE: Among the regimens for assessing laryngeal motion assessed in the present study, combinations of thiopentone or propofol with doxapram are the most effective means of stimulating arytenoid motion and could improve the accuracy of diagnosis of laryngeal paralysis in dogs.

Comparison of morphological changes and tactile sensitivity of the pharynx and larynx between four standing sedative and analgesic protocols in eight adult healthy horses.

OBJECTIVE: To compare the topographic modifications and tactile sensitivity of the pharynx and larynx after administration of four sedative and analgesic protocols in standing horses. STUDY DESIGN: Experimental, observer-blinded, crossover study. ANIMALS: Eight healthy mares. METHODS: Five protocols were evaluated: 1) xylazine and butorphanol administered intravenously (IV); 2) detomidine and butorphanol administered IV; 3) xylazine administered IV and lidocaine topically; 4) detomidine administered IV and lidocaine topically and 5) no analgesia or sedation (control). Quality of sedation, head height and sudden head movements were recorded. The degree of arytenoid cartilage displacement, the degree of pharyngeal collapse and the occurrence of soft palate displacement were scored using standardized scales. Tactile sensitivity was tested on 10 different pharyngeal and laryngeal regions using an atraumatic transendoscopic probe. Statistical analysis was performed using linear or generalized mixed-effects models. RESULTS: Head height was significantly decreased in protocols with xylazine (p = 0.002). Head movements were significantly increased in protocols with butorphanol (p = 0.0001). No changes in abduction grade or degree of soft palate displacement were observed between all sedative protocols and the control group. Pharyngeal collapse was significantly more frequent in protocols with lidocaine (p < 0.001) or xylazine (p = 0.017). For the pharyngeal regions, no tactile sensitivity difference was observed between the control and treatment protocols. All treatment protocols led to greater desensitization of all the laryngeal regions compared with the control protocol. CONCLUSION AND CLINICAL RELEVANCE: All the protocols provided adequate sedation and analgesia for the manipulation of the larynx and pharynx but significant differences were noted. Xylazine produces a more profound sedation compared with detomidine, but can induce dorsal pharyngeal collapse. Lidocaine caused pharyngeal collapse and its use should be limited to the target area. Butorphanol can be added to improve analgesia in the other regions but frequent head jerking can be expected.

A comparison of the effect of propofol and alfaxalone on laryngeal motion in nonbrachycephalic and brachycephalic dogs.

OBJECTIVE: To compare the effect of propofol and alfaxalone on laryngeal motion under a light plane of anaesthesia in nonbrachycephalic and brachycephalic dogs anaesthetized for nonemergency procedures. STUDY DESIGN: Prospective, randomized clinical trial. ANIMALS: A total of 48 client-owned dogs (24 nonbrachycephalic and 24 brachycephalic). METHODS: A standardized premedication of methadone (0.2 mg kg-1) and acepromazine (0.01 mg kg-1) was administered intramuscularly. Dogs were randomly assigned to be induced with increments of propofol (1-4 mg kg-1) or alfaxalone (0.5-2 mg kg-1). Laryngeal assessment was performed under a light plane of anaesthesia by a surgeon (GTH) who was unaware of the induction protocol. Laryngeal movement was assessed as either being present when abduction of the laryngeal cartilages upon inspiration was identified, or absent when abduction was not recognized. Simultaneously, a 60-second video was recorded. The same surgeon (GTH) and an additional surgeon (NK) re-evaluated the videos 1 month later. Categorical comparisons were studied using Chi square and Fisher's exact test where appropriate. Pairwise evaluation of agreement between scorers was undertaken with the kappa statistic (κ). RESULTS: There were no significant differences (p > 0.05) identified between the presence or absence of laryngeal motion between dogs administered propofol or alfaxalone, as well as when analysing nonbrachycephalic and brachycephalic dogs separately. The majority of dogs (>75%) maintained some degree of laryngeal motion with both protocols. Agreement between assessors was excellent (κ = 0.822). CONCLUSIONS: Alfaxalone maintained laryngeal motion similarly to propofol in nonbrachycephalic and brachycephalic dogs. CLINICAL RELEVANCE: Both agents would appear appropriate for allowing assessment of laryngeal motion in nonbrachycephalic and brachycephalic dogs. The assessment technique of subjective evaluation of laryngeal motion via peroral laryngoscopy under a light plane of anaesthesia produced consistent results amongst assessors, regardless of the induction agent used.

Comparison of the effects of alfaxalone and propofol with acepromazine, butorphanol and/or doxapram on laryngeal motion and quality of examination in dogs.

OBJECTIVE: To compare the effects of alfaxalone and propofol, with and without acepromazine and butorphanol followed by doxapram, on laryngeal motion and quality of laryngeal examination in dogs. STUDY DESIGN: Randomized, crossover, blinded study. ANIMALS: Ten female Beagle dogs, aged 11-13 months and weighing 7.2-8.6 kg. METHODS: The dogs were administered four intravenous (IV) treatments: alfaxalone (ALF), alfaxalone+acepromazine and butorphanol (ALF-AB), propofol (PRO) and propofol+AB (PRO-AB). AB doses were standardized. Dogs were anesthetized 5 minutes later by administration of alfaxalone or propofol IV to effect. Arytenoid motion during maximal inspiration and expiration was captured on video before and after IV doxapram (0.25 mg kg-1). The change in rima glottidis surface area (RGSA) was calculated to measure arytenoid motion. An investigator blinded to the treatment scored laryngeal examination quality. RESULTS: A 20% increase in RGSA was the minimal arytenoid motion that was detectable. RGSA was significantly less in ALF before doxapram compared with all other treatments. A <20% increase in RGSA was measured in eight of 10 dogs in PRO and in all dogs in ALF before doxapram. After doxapram, RGSA was significantly increased for PRO and ALF; however, 20% of dogs in PRO and 50% of dogs in ALF still had <20% increase in RGSA. A <20% increase in RGSA was measured in five of 10 dogs in PRO-AB and ALF-AB before doxapram. All dogs in PRO-AB and ALF-AB with <20% increase in RGSA before doxapram had ≥20% increase in RGSA after doxapram. Examination quality was significantly better in PRO-AB and ALF-AB. CONCLUSIONS AND CLINICAL RELEVANCE: The use of acepromazine and butorphanol improved the quality of laryngeal examination. Any negative impact on arytenoid motion caused by these premedications was overcome with doxapram. Using either propofol or alfaxalone alone is not recommended for the evaluation of arytenoid motion.

A Comparison of Rodent and Nonrodent Laryngeal and Tracheal Bifurcation Sensitivities in Inhalation Toxicity Studies and Their Relevance for Human Exposure.

In inhalation toxicity studies, drug-induced lesions are frequently reported in the larynx and sometimes at the tracheal bifurcation (carina) in the rat, but less so in the dog or monkey, bringing into question the relevance of these rodent findings for humans. The rat larynx is widely considered to be more sensitive than that of the dog and monkey in its response to inhaled xenobiotics, although we could find no published data to support this. In this review, data from 52 inhalation studies involving rodent and nonrodent species were collated and reviewed. These data showed that the rodent larynx, and to a lesser extent the carina, was far more commonly affected by treatment than those of the nonrodent. This review indicates the greater susceptibility of the rodent larynx and carina and emphasizes their lack of relevance for man. Observations and data suggest that the human larynx is much closer to the beagle dog and cynomolgus monkey in its response to inhaled xenobiotics and that greater clinical relevance should be placed on any specific findings in these animal models.

Impact of high concentrations of sevoflurane on laryngeal reflex responses.

BACKGROUND: Exaggerated defensive upper airway reflexes, particularly laryngospasm, may cause hypoxemic damage, especially in children. General clinical experience suggests that laryngeal reflex responses are more common under light levels of anesthesia, and previous clinical studies have shown an inverse correlation between laryngeal responsiveness and depth of hypnosis. However, this seems to be less obvious in children anesthetized with sevoflurane. The aim of this study was to assess the impact of high concentrations of sevoflurane on laryngeal and respiratory reflex responses in spontaneously breathing children. Accordingly, we tested the hypothesis that laryngeal and respiratory reflex responses were completely suppressed in spontaneously breathing children when anesthetized with sevoflurane 4.7% (=MACED95Intubation ) as compared with sevoflurane 2.5% (=1 MAC). METHODS: In this prospective observational study, we tested the hypothesis that the incidence of laryngospasm evoked by laryngeal stimulation is diminished under high concentrations of sevoflurane. Following Ethics approval, trial registration, and informed consent, 40 children (3-7 years) scheduled for elective surgery participated in the trial. All children received sevoflurane 2.5% (1 MAC) and 4.7% (ED95Intubation ) in random order with 5-min equilibration between the states. Under both conditions, distilled water was sprayed under bronchoscopic view onto the larynx. Potential laryngeal and respiratory reflex responses were assessed offline by a blinded reviewer. RESULTS: Laryngospasm (episodes lasting >10 s) occurred in 12/38 (32%) of the patients anesthetized with sevoflurane 2.5%, vs 7/38 (18%) in those anesthetized with sevoflurane 4.7% (difference: OR 3.5; 95% CI [0.72-16.84], P = 0.18). All other reflex responses (coughing, expiration reflexes, and spasmodic panting) were infrequent and were similar among the examined concentrations. CONCLUSION: Against our hypothesis, laryngospasm could still be observed in 18% of children under the higher concentration of sevoflurane (4.7%, ED95Intubation ).

Comparative pulmonary toxicity of inhaled metalworking fluids in rats and mice.

Metalworking fluids (MWFs) are complex formulations designed for effective lubricating, cooling, and cleaning tools and parts during machining operations. Adverse health effects such as respiratory symptoms, dermatitis, and cancer have been reported in workers exposed to MWFs. Several constituents of MWFs have been implicated in toxicity and have been removed from the formulations over the years. However, animal studies with newer MWFs demonstrate that they continue to pose a health risk. This investigation examines the hypothesis that unrecognized health hazards exist in currently marketed MWF formulations that are presumed to be safe based on hazard assessments of individual ingredients. In vivo 13-week inhalation studies were designed to characterize and compare the potential toxicity of four MWFs: Trim VX, Cimstar 3800, Trim SC210, and Syntilo 1023. Male and female Wistar Han rats or Fischer 344N/Tac rats and B6C3F1/N mice were exposed to MWFs via whole-body inhalation at concentrations of 0, 25, 50, 100, 200, or 400 mg/m3 for 13 weeks, after which, survival, body and organ weights, hematology and clinical chemistry, histopathology, and genotoxicity were assessed following exposure. Although high concentrations were used, survival was not affected and toxicity was primarily within the respiratory tract of male and female rats and mice. Minor variances in toxicity were attributed to differences among species as well as in the chemical components of each MWF. Pulmonary fibrosis was present only in rats and mice exposed to Trim VX. These data confirm that newer MWFs have the potential to cause respiratory toxicity in workers who are repeatedly exposed via inhalation.

Recovery from rocuronium-induced neuromuscular block was longer in the larynx than in the pelvic limb of anesthetized dogs.

OBJECTIVE: To determine if neuromuscular monitoring at the pelvic limb accurately reflects neuromuscular function in the larynx after administration of rocuronium in anesthetized dogs. STUDY DESIGN: Prospective experimental study. ANIMALS: Six healthy Beagle dogs. METHODS: Anesthesia was maintained in dogs with isoflurane and a continuous infusion of dexmedetomidine. Rocuronium (0.6 mg kg-1) was administered intravenously to induce neuromuscular block. Train-of-four (TOF) impulses were applied to the left recurrent laryngeal nerve (RLn) and the peroneal nerve (Pn). The evoked TOF ratio (TOFR; T4:T1) was measured with electromyography (EMG) simultaneously at the larynx and at the pelvic limb. Spontaneous recoveries of T1 to 25% (T125%) and 75% (T175%) of twitch height, and to TOFR of 0.70 and 0.90 (TOFR0.90) at each EMG site were compared. RESULTS: Data from five dogs were analyzed. Times to T125% were similar at the pelvic limb and larynx when measured by EMG; time to T175% was slower at the larynx by 6±4 minutes (p=0.012). The larynx had a slower recovery to TOFR0.70 (41±13 minutes) and TOFR0.90 (45±13 minutes) than did the pelvic limb [29±8 minutes (p=0.011) and 33±9 minutes (p=0.003), respectively]. When the pelvic limb EMG returned to TOFR0.70 and TOFR0.90, the larynx EMG TOFR0.70 and TOFR0.90 values were 0.32±0.12 (p=0.001) and 0.38±0.13 (p=0.001), respectively. CONCLUSIONS AND CLINICAL RELEVANCE: After administration of rocuronium, neuromuscular function assessed by EMG recovered approximately 36% slower at the larynx than at the pelvic limb. The results in these dogs suggest that quantitative neuromuscular monitoring instrumented at a pelvic limb may be unable to exclude residual block at the larynx in anesthetized dogs.

Effects of thiopentone, propofol and alfaxalone on laryngeal motion during oral laryngoscopy in healthy dogs.

OBJECTIVE: To compare the effects of thiopentone, propofol and alfaxalone on arytenoid cartilage motion and establish the dose rates to achieve a consistent oral laryngoscopy examination. STUDY DESIGN: Randomised crossover study. ANIMALS: Six healthy adult Beagle dogs. METHODS: Each dog was randomly administered three induction agents with a 1-week washout period between treatments. Thiopentone (7.5 mg kg-1), propofol (3 mg kg-1) or alfaxalone (1.5 mg kg-1) was administered over 1 minute for induction of anaesthesia. If the dog was deemed inadequately anaesthetised, then supplemental boluses of 1.8, 0.75 and 0.4 mg kg-1 were administered, respectively. Continual examination of the larynx, using a laryngoscope, commenced once an adequate anaesthetic depth was reached until examination end point. The number of arytenoid motions and vital breaths were counted during three time periods and compared over time and among treatments. Data were analysed using Friedman and Mann-Whitney U tests, Spearman rho and a linear mixed model with post hoc pairwise comparison with Tukey correction. RESULTS: The median (range) induction and examination times were 2.8 (2.0-3.0), 2.7 (2.0-3.3) and 2.5 (1.7-3.3) minutes (p = 0.727); and 14.1 (8.0-41.8), 5.4 (3.3-14.8) and 8.5 (3.8-31.6) minutes (p = 0.016) for thiopentone, propofol and alfaxalone, respectively. The median dose rates required to achieve an adequate anaesthetic depth were 6.3 (6.0-6.6), 2.4 (2.4-2.4) and 1.2 (1.2-1.2) mg kg-1 minute-1, respectively. There was no significant difference for the total number of arytenoid motions (p = 0.662) or vital breaths (p = 0.789) among induction agents. CONCLUSION AND CLINICAL RELEVANCE: The number of arytenoid motions were similar among the induction agents. However, at the dose rates used in this study, propofol provided adequate conditions for evaluation of the larynx with a shorter examination time which may be advantageous during laryngoscopy in dogs.

Stabilization of G-quadruplex DNA and inhibition of Bcl-2 expression by a pyridostatin analog.

The G-quadruplexes located in the P1 promoter of B-cell lymphoma-2 (Bcl-2) gene are implicated to regulate Bcl-2 expression. Here, we designed a new pyridostatin analog named PDF, which exhibited high specificity and stabilizing effect toward G-quadruplexes. The luciferase assay demonstrated that PDF could significantly suppress Bcl-2 transcriptional activation in human laryngeal squamous carcinoma cells (Hep-2) cells. Besides, PDF also induced cell apoptosis in vitro assays. These results provide an excellent G-quadruplex specific ligand as an efficient Bcl-2 inhibitor. These results also implicate that PDF may be a potential anticancer drug to head neck cancer.

Effects of intravenous and topical laryngeal lidocaine on heart rate, mean arterial pressure and cough response to endotracheal intubation in dogs.

OBJECTIVE: To compare the effects of intravenous (IV) and topical laryngeal lidocaine on heart rate (HR), mean arterial pressure (MAP) and cough response to endotracheal intubation (ETI) in dogs. STUDY DESIGN: Prospective, randomized, blinded clinical study. ANIMALS: Forty-two client-owned dogs (American Society of Anesthesiologists class I and II status) undergoing elective orthopaedic surgery. METHODS: Dogs were randomized to three groups. Dogs in group SALIV received 0.1 mL kg(-1) IV saline. Dogs in group LIDIV received 2 mg kg(-1) IV 2% lidocaine. Dogs in group LIDTA received 0.4 mg kg(-1) topically sprayed laryngeal 2% lidocaine. All dogs were premedicated with methadone (0.2 mg kg(-1) IV). After 30 minutes, IV propofol was administered to abolish the lateral palpebral reflex and produce jaw relaxation. The allocated treatment was then administered and, after 30 seconds, further propofol was administered to abolish the medial palpebral reflex and facilitate ETI. HR and MAP were measured at four time-points using cardiac auscultation and automated oscillometry, respectively. The cough response at ETI was recorded. One-way anova and post hoc Tukey adjustment were used to analyse parametric data. The Kruskal-Wallis test was used to analyse non-parametric data. Odds ratios were calculated for the cough response. A p-value of ≤0.05 was considered to indicate statistical significance. RESULTS: In response to ETI, changes in MAP differed significantly between groups. In SALIV, MAP increased (4 ± 6 mmHg), whereas it decreased in LIDIV (6 ± 13 mmHg) (p = 0.013) and LIDTA (7 ± 11 mmHg) (p = 0.003). Dogs in SALIV were almost 10 times more likely to cough than dogs in LIDIV (odds ratio 9.75, 95% confidence interval 0.98-96.60; p = 0.05). CONCLUSIONS AND CLINICAL RELEVANCE: In propofol-anaesthetized dogs, IV and topical laryngeal lidocaine attenuated the pressor response to ETI, whereas IV lidocaine reduced the cough response.

Retinoic acid-inducible gene-I-like receptor (RLR)-mediated antiviral innate immune responses in the lower respiratory tract: Roles of TRAF3 and TRAF5.

Upon viral infection, the cytoplasmic viral sensor retinoic acid-inducible gene-I (RIG-I) recognizes viral RNA to activate antiviral signaling to induce type I interferon (IFN). RIG-I-like receptors (RLRs) activate antiviral signaling in a tissue-specific manner. The molecular mechanism underlying antiviral signaling in the respiratory system remains unclear. We studied antiviral signaling in the lower respiratory tract (LRT), which is the site of many harmful viral infections. Epithelial cells of the LRT can be roughly divided into two groups: bronchial epithelial cells (BECs) and pulmonary alveolar epithelial cells (AECs). These two cell types exhibit different phenotypes; therefore, we hypothesized that these cells may play different roles in antiviral innate immunity. We found that BECs exhibited higher antiviral activity than AECs. TNF receptor-associated factor 3 (TRAF3) has been shown to be a crucial molecule in RLR signaling. The expression levels of TRAF3 and TRAF5, which have conserved domains that are nearly identical, in the LRT were examined. We found that the bronchus exhibited the highest expression levels of TRAF3 and TRAF5 in the LRT. These findings suggest the importance of the bronchus in antiviral innate immunity in the LRT and indicate that TRAF3 and TRAF5 may contribute to RLR signaling.