Beat Frequency Research Articles

Characterizing PFAS hazards and risks: a human population-based in vitro cardiotoxicity assessment strategy

Per- and poly-fluoroalkyl substances (PFAS) are emerging contaminants of concern because of their wide use, persistence, and potential to be hazardous to both humans and the environment. Several PFAS have been designated as substances of concern; however, most PFAS in commerce lack toxicology and exposure data to evaluate their potential hazards and risks. Cardiotoxicity has been identified as a likely human health concern, and cell-based assays are the most sensible approach for screening and prioritization of PFAS. Human-induced pluripotent stem cell (iPSC)-derived cardiomyocytes are a widely used method to test for cardiotoxicity, and recent studies showed that many PFAS affect these cells. Because iPSC-derived cardiomyocytes are available from different donors, they also can be used to quantify human variability in responses to PFAS. The primary objective of this study was to characterize potential human cardiotoxic hazard, risk, and inter-individual variability in responses to PFAS. A total of 56 PFAS from different subclasses were tested in concentration-response using human iPSC-derived cardiomyocytes from 16 donors without known heart disease. Kinetic calcium flux and high-content imaging were used to evaluate biologically-relevant phenotypes such as beat frequency, repolarization, and cytotoxicity. Of the tested PFAS, 46 showed concentration-response effects in at least one phenotype and donor; however, a wide range of sensitivities were observed across donors. Inter-individual variability in the effects could be quantified for 19 PFAS, and risk characterization could be performed for 20 PFAS based on available exposure information. For most tested PFAS, toxicodynamic variability was within a factor of 10 and the margins of exposure were above 100. This study identified PFAS that may pose cardiotoxicity risk and have high inter-individual variability. It also demonstrated the feasibility of using a population-based human in vitro method to quantify population variability and identify cardiotoxicity risks of emerging contaminants.

The lysogenic filamentous Pseudomonas bacteriophage phage Pf slows mucociliary transport

Abstract Pseudomonas aeruginosa is a major pulmonary pathogen causing chronic pulmonary infections in people with cystic fibrosis (CF). The P. aeruginosa filamentous and lysogenic bacteriophage, Pf phage, is abundant in the airways of many people with CF and has been associated with poor outcomes in a cross-sectional cohort study. Previous studies have identified roles for Pf phage in biofilm formation, specifically forming higher-order birefringent, liquid crystals when in contact with other biopolymers in biofilms. Liquid crystalline biofilms are more adherent and viscous than those without liquid crystals. A key feature of biofilms is to enhance bacterial adherence and resist physical clearance. The effect of Pf phage on mucociliary transport is unknown. We found that primary CF and non-CF nasal epithelial cells cultured at air–liquid interface treated with Pf phage exhibit liquid crystalline structures in the overlying mucus. On these cell cultures, Pf phage entangles cilia but does not affect ciliary beat frequency. In both these in vitro cell cultures and in an ex vivo porcine trachea model, introduction of Pf phage decreases mucociliary transport velocity. Pf phage also blocks the rescue of mucociliary transport by CF transmembrane conductance regulator modulators in CF cultures. Thus, Pf phage may contribute to the pathogenesis of P. aeruginosa-associated CF lung disease via induction of liquid crystalline characteristics to airway secretions, leading to impaired mucociliary transport. Targeting Pf phage may be useful in treatment CF as well as other settings of chronic P. aeruginosa infections.

P01-68 Quantification of ciliary beating frequency in differentiated bronchial epithelial cells

P01-68 Quantification of ciliary beating frequency in differentiated bronchial epithelial cells

Stanovenie mukociliárneho klírensu u pacientov so symptómami laryngofaryngeálneho refl uxu – preliminárne výsledky

Summary: Introduction: Mucociliary clearance represents an important protective mechanism of the respiratory tract. Results of multiple studies indicate that gastroesophageal reflux can participate in worsening mucociliary clearance leading to laryngopharyngeal reflux symptoms including recurrent upper respiratory tract infections. Thereby, we decided to provide ciliary beat frequency of the upper respiratory tract epithelium in patients with laryngopharyngeal reflux. Materials and methods: We enrolled 23 patients with laryngopharyngeal reflux symptoms. Control group consisted of 9 healthy volunteers. Patients were enrolled upon a positive questionnaire reflux symptom index (RSI >13 points), with no proton pump inhibitor treatment. We obtained a nasopharyngeal swab and evaluated ciliary beat frequency of the respiratory epithelium using highspeed videomicroscopy and special sofware Ciliary Analysis. We compared the values of ciliary beat frequency with values from healthy volunteers. Results: We provided median, minimal and maximum values of ciliary beat frequency. Median ciliary beat frequency in healthy volunteers was significantly higher than in patients with laryngopharyngeal reflux symptomatology (10.86 ± 1.67 vs. 6.6 ± 2.2; P <0.001). Maximum ciliary beat frequency was also significantly higher in healthy volunteers than in patients with LPR (17.9 ± 0.95 vs. 15.3 ± 2.39; P <0.001). Minimal value had no significant difference. Correlation analysis showed an inverse correlation between median ciliary beat frequency of the respiratory epithelium and LPR symptoms (RSI value) (P <0.04). Conclusion: Patients with laryngopharyngeal reflux symptoms had lower ciliary beat frequency compared to healthy volunteers. The degree of frequency defect correlates with the degree of their laryngopharyngeal symptoms. More studies are needed to confirm direct evidence of gastroesophageal reflux etiology in mucociliary clearance worsening. Key words: mucociliary clearance – laryngopharyngeal reflux – cilia

MACSFeD-a database of mosquito acoustic communication and swarming features.

Acoustic communication plays an important role during the courtship of many mosquito species. Male mosquitoes show strong attraction to female wing beat frequencies, mediated via spectral matching between female wing beat frequency and male ear mechanical tuning frequency. Such acoustic communication typically occurs within swarms, male-dominated aggregations with species-specific properties. Despite hundreds of relevant publications being available, the lack of a central platform hosting all associated data hinders research efforts and limits cross-species comparisons. Here, we introduce MACSFeD (Mosquito Acoustic Communication and Swarming Features Database), an interactive platform for the exploration of our comprehensive database containing 251 unique reports focusing on different aspects of mosquito acoustic communication, including hearing function, wing beat frequency and phonotaxis, as well as male swarming parameters. MACSFeD serves as an easily accessible, efficient, and robust data visualization tool for mosquito acoustic communication research. We envision that further in-depth studies could arise following the use of this new platform. Database URL: https://minmatt.shinyapps.io/MACSFeD/.

The molecular mechanism under the behavioral and physiological changes of large yellow croaker (Larimichthys crocea) in response to hypoxia

The large yellow croaker (Larimichthys crocea) displays pronounced behavioral changes in response to low oxygen environments, including restlessness, constant floating, and increases in respiratory rate and tail beat frequency. However, the underlying molecular mechanisms governing these behavioral and physiological alterations remain unclear. To address this knowledge gap, the present study employed transcriptome analysis to investigate the molecular responses in the large yellow croaker muscle tissue after the fish were exposed to hypoxic treatment. A total of 4553, 1797, 3222, 3148, and 4815 differentially expressed genes (DEGs) were identified at 1 h, 6 h, 9 h, 24 h, and 72 h after hypoxia treatment, respectively. Subsequent analysis utilizing Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichments unveiled a notable up-regulation of genes related to muscular movement-related pathways, specifically the calcium pathway. This up-regulation coincided with an observed increase in the intracellular Ca2+ concentration and Ca2+ transport enzyme activity, suggesting that hypoxia-induced muscle contraction and relaxation occurred by elevating the intracellular Ca2+ concentration within the muscle tissue. Key DEGs associated with the glycolysis and the TCA cycle pathways were also found to be significantly increased in expression, indicating an activation of energy generation pathways in the muscle of hypoxic large yellow croakers. Conversely, genes related to protein synthesis and cell growth were significantly down-regulated, suggesting an inhibition of energy-consuming processes. Additionally, hypoxia was observed to impact various physiological processes such as oxygen transport, vascular control, and apoptosis. These processes may have been mediated by the up-regulation of HIF-1α and its target genes. Consequently, these results offer novel insights into comprehending the molecular basis of the changes in behavior and physiology of large yellow croaker in response to the stress induced by low oxygen levels.

Ciliary Motility Decreased by a CO2/HCO3--Free Solution in Ciliated Human Nasal Epithelial Cells Having a pH Elevated by Carbonic Anhydrase IV.

An application of CO2/HCO3--free solution (Zero-CO2) did not increase intracellular pH (pHi) in ciliated human nasal epithelial cells (c-hNECs), leading to no increase in frequency (CBF) or amplitude (CBA) of the ciliary beating. This study demonstrated that the pHi of c-hNECs expressing carbonic anhydrase IV (CAIV) is high (7.64), while the pHi of ciliated human bronchial epithelial cells (c-hBECs) expressing no CAIV is low (7.10). An extremely high pHi of c-hNECs caused pHi, CBF and CBA to decrease upon Zero-CO2 application, while a low pHi of c-hBECs caused them to increase. An extremely high pHi was generated by a high rate of HCO3- influx via interactions between CAIV and Na+/HCO3- cotransport (NBC) in c-hNECs. An NBC inhibitor (S0859) decreased pHi, CBF and CBA and increased CBF and CBA in c-hNECs upon Zero-CO2 application. In conclusion, the interactions of CAIV and NBC maximize HCO3- influx to increase pHi in c-hNECs. This novel mechanism causes pHi to decrease, leading to no increase in CBF and CBA in c-hNECs upon Zero-CO2 application, and appears to play a crucial role in maintaining pHi, CBF and CBA in c-hNECs periodically exposed to air (0.04% CO2) with respiration.

Donepezil Nanoemulsion Induces a Torpor-like State with Reduced Toxicity in Nonhibernating Xenopus laevis Tadpoles.

Achieving a reversible decrease of metabolism and other physiological processes in the whole organism, as occurs in animals that experience torpor or hibernation, could contribute to increased survival after serious injury. Using a Bayesian network tool with transcriptomic data and chemical structure similarity assessments, we predicted that the Alzheimer's disease drug donepezil (DNP) could be a promising candidate for a small molecule drug that might induce a torpor-like state. This was confirmed in a screening study with Xenopus laevis tadpoles, a nonhibernator whole animal model. To improve the therapeutic performance of the drug and minimize its toxicity, we encapsulated DNP in a nanoemulsion formulated with low-toxicity materials. This formulation is composed of emulsified droplets <200 nm in diameter that contain 1.250 mM DNP, representing ≥95% encapsulation efficiency. The DNP nanoemulsion induced comparable torpor-like effects to those produced by the free drug in tadpoles, as indicated by reduced swimming motion, cardiac beating frequency, and oxygen consumption, but with an improved biodistribution. Use of the nanoemulsion resulted in a more controlled increase of DNP concentration in the whole organism compared to free DNP, and to a higher concentration in the brain, which reduced DNP toxicity and enabled induction of a longer torpor-like state that was fully reversible. These studies also demonstrate the potential use of Xenopus tadpoles as a high-throughput in vivo screen to assess the efficacy, biodistribution, and toxicity of drug-loaded nanocarriers.

Evaluation of Open-Source Ciliary Analysis Software in Primary Ciliary Dyskinesia: A Comparative Assessment.

Primary Ciliary Dyskinesia (PCD) is a rare genetic disorder characterized by alterations in motile cilia function. The diagnosis of PCD is challenging due to the lack of standardized methods in clinical practice. High-speed video microscopy analysis (HSVA) directly evaluates ciliary beat frequency (CBF) in PCD. Recently, open-source ciliary analysis software applications have shown promise in measuring CBF accurately. However, there is limited knowledge about the performance of different software applications, creating a gap in understanding their comparative effectiveness in measuring CBF in PCD. We compared two open-source software applications, CiliarMove (v219) and Cilialyzer (v1.2.1-b3098cb), against the manual count method. We used high-speed videos of nasal ciliary brush samples from PCD RSPH4A-positive (PCD (RSPH4A)) patients and healthy controls. All three methods showed lower median CBF values for patients with PCD (RSPH4A) than in healthy controls. CiliarMove and Cilialyzer identified lower CBF in patients with PCD (RSPH4A), similarly to the manual count. Cilialyzer, CiliarMove, and manual count methods demonstrated statistical significance (p-value < 0.0001) in the difference of median CBF values between patients with PCD (RSPH4A) and healthy controls. Correlation coefficients between the manual count values against both software methods demonstrated positive linear relationships. These findings support the utility of open-source software-based analysis tools. Further studies are needed to validate these findings with other genetic variants and identify the optimal software for accurate CBF measurement in patients with PCD.

Magnetic field enhanced terahertz generation from shape-dependent metallic nanoparticles

Abstract This communication deals with the analytical study of terahertz (THz) generation via frequency-difference mechanism using two circularly symmetric Gaussian laser beams with slightly different frequencies ω 1 and ω 2 and wave vectors k → 1 and k → 2 simultaneously propagating through a mixture of spatially corrugated noble-metal nanoparticles. The mixture, consisting of spherical nanoparticles (SNPs) and cylindrical nanoparticles (CNPs), is placed in a host medium under the influence of an externally applied static magnetic field. The two co-propagating laser beams impart a nonlinear ponderomotive force on the electrons of the NPs, causing them to experience nonlinear oscillatory velocity. Furthermore, the consequent nonlinear current density excites THz radiation at the beat frequency ω ( = ω 1 − ω 2 ) . Magnetic fields influence the surface plasmon resonance condition associated with electrons of the nanoparticles due to enhancement in ponderomotive nonlinearities, thereby causing an increment in the amplitude of the generated THz field. It is observed that the generated THz radiation has a strong dependence on the shape and size of the NPs in addition to the magnetic field strength. CNPSs provide greater THz amplitude than SNPs due to additional resonance modes, and combining both kinds of nanostructures further enhances the amplitude. THz radiation plays an important role in biomedical and pharmaceutical fields, communications, security and THz spectroscopy.

Characterization of the ciliary beating efficiency in primary diffuse chronic rhinosinusitis.

Ciliary dysfunction may result in chronic airway inflammation and infection causing injury and structural changes to the airway epithelium, leading to a variety of diseases, like bronchiectasis and primary diffuse chronic rhinosinusitis (CRS). Currently, ciliary beating analysis has mainly been studied through the measure of the ciliary beating frequency (CBF) by high-speed digital video microscopy (HSDV). However, a normal CBF has been described in different forms of primary and acquired ciliary dyskinesia.

Analysis of beat frequencies of transverse mode degeneracy in diode-pumped Nd:YVO4 laser

Analysis of beat frequencies of transverse mode degeneracy in diode-pumped Nd:YVO4 laser

Obesity impairs ciliary function and mucociliary clearance in the murine airway epithelium.

Obesity is a risk factor for increased morbidity and mortality in viral respiratory infection. Mucociliary clearance (MCC) in the airway is the primary host defense against viral infections. However, the impact of obesity on MCC is unclear, prompting this study. Using murine tracheal tissue culture and in vitro influenza A virus (IAV) infection models, we analyzed cilia-driven flow and ciliary beat frequency (CBF) in the airway epithelium to evaluate MCC. Short-term IAV infection increased cilia-driven flow and CBF in control mice, but not in high-fat diet-induced obese mice. Basal cilia-driven flow and CBF were also lower in obese mice than in control mice. Mechanistically, the increase of extracellular adenosine triphosphate (ATP) release during IAV infection, which was observed in the control mice, was abolished in the obese mice; however, the addition of ATP increased cilia-driven flow and CBF both in control and obese mice to a similar extent. In addition, RNA sequencing and reverse transcription-polymerase chain reaction revealed the downregulation of several cilia-related genes, including Dnah1, Dnal1, Armc4, and Ttc12 (the dynein-related genes); Ulk4 (the polychaete differentiation gene); Cep164 (the ciliogenesis and intraflagellar transport gene); Rsph4a, Cfap206, and Ppil6 (the radial spoke structure and assembly gene); and Drc3(the nexin-dynein regulatory complex genes) in obese murine tracheal tissues compared with their control levels. In conclusion, our studies demonstrate that obesity attenuates MCC under basal conditions and during IAV infection by downregulating the expression of cilia-related genes and suppressing the release of extracellular ATP, thereby increasing the susceptibility and severity of IAV infection.NEW & NOTEWORTHY Our study shows that obesity impairs airway mucociliary clearance (MCC), an essential physical innate defense mechanism for viral infection. Mechanically, this is likely due to the obesity-induced downregulation of cilia-related genes and attenuation of extracellular ATP release. This study provides novel insights into the mechanisms driving the higher susceptibility and severity of viral respiratory infections in individuals with obesity.

Low-loss compact chalcogenide microresonators for efficient stimulated Brillouin lasers.

Chalcogenide glasses (ChGs) possess a high elasto-optic coefficient, making them ideal for applications in microwave photonics and narrow-linewidth lasers based on stimulated Brillouin scattering (SBS). However, current As2S3-based integrated devices suffer from poor stability and low laser-induced damage threshold, and planar ChG devices feature limited quality factors. In this Letter, we propose and demonstrate a high-quality integrated GeSbS ChG Brillouin photonic device. By introducing Euler bending structures, we suppress high-order optical modes and reduce propagation losses in a finger-shaped GeSbS microresonator, resulting in a compact footprint of 3.8 mm2 and a high intrinsic quality factor of 5.19 × 106. The combination of GeSbS material's high Brillouin gain and the resonator's high-quality factor enables the generation of stimulated Brillouin lasers with a low threshold of 0.96 mW and a fundamental linewidth of 58 Hz. Moreover, cascaded stimulated Brillouin lasers can be realized up to the seventh order, yielding microwave beat frequencies up to 40 GHz.

Body length determines flow refuging for rainbow trout (Oncorhynchus mykiss) behind wing dams.

Complex hydrodynamics abound in natural streams, yet the selective pressures these impose upon different size classes of fish are not well understood. Attached vortices are produced by relatively large objects that block freestream flow, which fish routinely utilize for flow refuging. To test how flow refuging and the potential harvesting of energy (as seen in Kármán gaiting) vary across size classes in rainbow trout (Oncorhynchus mykiss; fingerling, 8 cm; parr, 14 cm; adult, 22 cm; n=4 per size class), we used a water flume (4100 l; freestream flow at 65 cm s-1) and created vortices using 45deg wing dams of varying size (small, 15 cm; medium, 31 cm; large, 48 cm). We monitored microhabitat selection and swimming kinematics of individual trout and measured the flow field in the wake of wing dams using time-resolved particle image velocimetry (PIV). Trout of each size class preferentially swam in vortices rather than the freestream, but the capacity to flow refuge varied according to the ratio of vortex width to fish length (WV:LF). Consistent refuging behavior was exhibited when WV:LF≥1.5. All size classes exhibited increased wavelength and Strouhal number and decreased tailbeat frequency within vortices compared with freestream, suggesting that swimming in vortices requires less power output. In 17% of the trials, fish preferentially swam in a manner that suggests energy harvesting from the shear layer. Our results can inform efforts toward riparian restoration and fishway design to improve salmonid conservation.

Deformation theory and nonlinear dynamic behavior of PVC gel actuators

Polyvinyl chloride (PVC) gel generates complex nonlinear vibration behavior under an alternating current voltage excitation, which has potential application as a dynamic electromechanical actuator. However, there are few studies on the deformation theory of PVC gel actuators, especially the dynamic nonlinear response theory. In this paper, a complex dynamic model is established according to the electrodeformation mechanism of PVC film, and the nonlinear dynamic behavior of the actuator is numerically studied by a differential equation. The effects of applied voltage amplitude, voltage frequency, dibutyl adipate content, mechanical tension, and bias voltage on the dynamic properties of PVC film were analyzed under the condition of equal biaxial tension. The variation of amplitude and the generation and disappearance of the beat frequency during vibration are analyzed by using time-domain characteristics. The degree of PVC actuator nonlinearity as well as vibration stability and periodicity is also reflected based on the phase path and Poincare map. Finally, the law of influence of external condition parameters on the dynamic response of the PVC actuator is obtained.

The Increase in the Frequency and Amplitude of the Beating of Isolated Mouse Tracheal Cilia Reactivated by ATP and cAMP with Elevation in pH.

Single cilia, 100 nm in diameter and 10 µm in length, were isolated from mouse tracheae with Triton X-100 (0.02%) treatment, and the effects of pH on ciliary beating were examined by measuring the ciliary beat frequency (CBF) and the ciliary bend distance (CBD-an index of amplitude) using a high-speed video microscope (250 fps). ATP (2.5 mM) plus 8Br-cAMP (10 µM) reactivated the CBF and CBD in the isolated cilia, similar to the cilia of in vivo tracheae. In the reactivated isolated cilia, an elevation in pH from 7.0 to 8.0 increased the CBF from 3 to 15 Hz and the CBD from 0.6 to 1.5 µm. The pH elevation also increased the velocity of the effective stroke; however, it did not increase the recovery stroke, and, moreover, it decreased the intervals between beats. This indicates that H+ (pHi) directly acts on the axonemal machinery to regulate CBF and CBD. In isolated cilia priorly treated with 1 µM PKI-amide (a PKA inhibitor), 8Br-cAMP did not increase the CBF or CBD in the ATP-stimulated isolated cilia. pH modulates the PKA signal, which enhances the axonemal beating generated by the ATP-activated inner and outer dyneins.

Investigations into the role of platelet-activating factor in the peri-conception period of the mare.

In many mammals, the lipid platelet-activating factor (PAF) has important functions in female reproduction and fertility. This study shows that PAF is present in the reproductive tissues of mares and is involved in processes related to ovulation and early pregnancy. Platelet-activating factor (PAF) has been implicated in a number of reproductive processes ranging from ovulation to embryo motility but has not been widely explored in the mare. To identify the presence and examine the role of PAF in the equine periconception processes, targeted mass spectrometry coupled with chromatographic separation was performed on equine follicular fluid (FF), and PAF was quantitatively detected. Subsequently, untargeted high-resolution mass spectrometry-based lipidomic analysis was carried out to quantify PAF in different-sized pre-ovulatory follicles, whereby different molecular species of PAF, PAF (14:0) and PAF (16:1), were both seen to be increasing with follicle diameter. These findings suggest that PAF within FF is increasing as preovulatory follicles approach ovulation. Additionally, immunofluorescence staining identified the PAF receptor in the luminal pericellular, apical, and basal aspect of equine oviductal epithelial cells. Lastly, an equine oviductal epithelial organoid model was generated and showed that the addition of PAF significantly increased the ciliary beat frequency (CBF) (Hz), an action consistent with a role for PAF in embryo migration. It is proposed that the local action of PAF on the ciliated cells of the oviduct propels both the oocyte and the conceptus towards the uterus. In the mare, it appears that PAF is a contributor during the periconception period, potentially being a mediator in the mechanisms of ovulation and in the dialogue of very early pregnancy.

Beat frequency induced transitions in synchronization dynamics

In neurosciences, the brain processes information via the firing patterns of connected neurons operating across a spectrum of frequencies. To better understand the effects of these frequencies in the neuron dynamics, we have simulated a neuronal network of Izhikevich neurons to examine the interaction between frequency allocation and intermittent phase synchronization dynamics. As the synchronized population of neurons passes through a bifurcation, an additional frequency mode emerges, enabling a match in the mean frequency while retaining distinct most probable frequencies among neurons. Subsequently, the network intermittently transits between two patterns, one partially synchronized and the other unsynchronized. Through our analysis, we demonstrate that the frequency changes on the network lead to characteristic transition times between synchronization states. Moreover, these transitions adhere to beat frequency statistics when the neurons’ frequencies differ by multiples of a frequency gap. Finally, our results can improve the performance in predicting transitions on problems where the beat frequency strongly influences the dynamics.

Promoting successful healing of artificial trachea by intratracheal administration of basic fibroblast growth factor†.

This study evaluated the effect of intratracheal administration of basic fibroblast growth factor (bFGF) on tracheal healing following implantation of a novel layered polyglycolic acid (PGA) material to replace a critical-size defect in rat trachea. A critical-size defect in the rat cervical trachea was covered with PGA. Distilled water (DW) or 3.125, 6.25, 12.5 or 25 µg bFGF was administered into the trachea for 2 weeks (n = 6 for each of 5 groups). Regenerated areas of cilia, ciliary beat frequency and ciliary transport function (CTF) in the centre of the PGA were measured. To examine potential side effects of intratracheal administration of bFGF, the right lower lobe was pathologically evaluated. All rats survived during the study period. Histological examination showed ciliated epithelization on the PGA material after 2 weeks. Bronchoscopy revealed stenosis due to granulation following administration of high concentrations of bFGF (12.5 and 25 µg). Compared with the DW group, groups administered 3.125, 6.25, 12.5 and 25 µg bFGF had significantly larger areas of regenerated cilia (15.2%, 27.0%, 41.3%, 33.1% and 31.0%, respectively; P = 0.00143), improved ciliary beat frequency (7.10, 8.18, 10.10, 9.50 and 9.50 Hz, respectively), and improved CTS (6.40, 9.54, 16.89, 16.41 and 14.29 µm/sec, respectively). Pathological examination of the right lower lobe revealed pulmonary fibrosis and hyperplasia with high concentrations of bFGF (12.5 and 25 µg). Intratracheal administration of bFGF effectively promoted tracheal regeneration at an optimal dose of 6.25 µg following implantation of an artificial trachea.