RESUMO
AIMS: This study investigated the efficacy of hydrogen peroxide vapour (HPV) at inactivating hazard group 3 bacteria that have been presented dried from their growth medium to present a realistic challenge. METHODS AND RESULTS: Hydrogen peroxide vapour technology (Bioquell) was used to decontaminate a class III microbiological safety cabinet containing biological indicators (BIs) made by drying standard working suspensions of the following agents: Bacillus anthracis (Ames) spores, Brucella abortus (strain S99), Burkholderia pseudomallei (NCTC 12939), Escherichia coli O157 ST11 (NCTC 12079), Mycobacterium tuberculosis (strain H37Rv) and Yersinia pestis (strain CO92) on stainless steel coupons. Extended cycles were used to expose the agents for 90 min. The HPV cycle completely inactivated B. anthracis spores, B. abortus, B. pseudomallei, E. coli O157 and Y. pestis when BIs were processed using quantitative and qualitative methods. Whilst M. tuberculosis was not completely inactivated, it was reduced by 4 log10 from a starting concentration of 106 colony-forming units. CONCLUSIONS: This study demonstrates that HPV is able to inactivate a range of HG3 agents at high concentrations with associated organic matter, but M. tuberculosis showed increased resistance to the process. SIGNIFICANCE AND IMPACT OF THE STUDY: This publication demonstrates that HPV can inactivate HG3 agents that have an organic load associated with them. It also shows that M. tuberculosis has higher resistance to HPV than other agents. This shows that an appropriate BI to represent the agent of interest should be chosen to demonstrate a decontamination is successful.
Assuntos
Contenção de Riscos Biológicos/métodos , Descontaminação/métodos , Desinfetantes/farmacologia , Gases/farmacologia , Peróxido de Hidrogênio/farmacologia , Bactérias/efeitos dos fármacos , Contenção de Riscos Biológicos/instrumentação , Farmacorresistência Bacteriana , Biomarcadores Ambientais/efeitos dos fármacos , Aço InoxidávelRESUMO
New vaccines against biodefense-related and emerging pathogens are being prepared for licensure using the US Federal Drug Administration's "Animal Rule." This allows licensure of drugs and vaccines using protection data generated in animal models. A new acellular plague vaccine composed of two separate recombinant proteins (rF1 and rV) has been developed and assessed for immunogenicity in humans. Using serum obtained from human volunteers immunised with various doses of this vaccine and from immunised cynomolgus macaques, we assessed the pharmacokinetic properties of human and cynomolgus macaque IgG in BALB/c and the NIH Swiss derived Hsd:NIHS mice, respectively. Using human and cynomolgus macaque serum with known ELISA antibody titres against both vaccine components, we have shown that passive immunisation of human and nonhuman primate serum provides a reproducible delay in median time to death in mice exposed to a lethal aerosol of plague. In addition, we have shown that Hsd:NIHS mice are a better model for humoral passive transfer studies than BALB/c mice.
Assuntos
Soros Imunes/imunologia , Imunização Passiva , Macaca fascicularis/imunologia , Peste/imunologia , Peste/prevenção & controle , Especificidade da Espécie , Yersinia pestis/imunologia , Animais , Anticorpos Antibacterianos/administração & dosagem , Anticorpos Antibacterianos/imunologia , Modelos Animais de Doenças , Feminino , Humanos , Soros Imunes/administração & dosagem , Camundongos , Peste/mortalidade , Vacina contra a Peste/administração & dosagem , Vacina contra a Peste/imunologia , Virulência , Yersinia pestis/patogenicidadeRESUMO
Ovine airway epithelial explants, cultured at an air-liquid interface, were used to determine whether endothelin (ET-1) acts via ET(A)- or ET(B)-receptors to increase ciliary beat frequency (CBF). Further, the role of prostanoids and nitric oxide (NO) downstream of receptor activation was explored. CBF was measured using an image analysis system with a sampling rate of 224 frames s(-1). At 37 °C, baseline CBF was 14.90 ± 3.49 Hz (n = 116 cells). ET-1 dose-dependently stimulated CBF (EC(50) = 60.98 ± 27.99 nM). The stimulatory effect of ET-1 (1 µM) could be mimicked by the ET(A)-receptor agonist sarafotoxin S6b (100 nM) but not the ET(B)-receptor agonist sarafotoxin S6c (100 nM) and was completely abolished by the ET(A)-receptor antagonist BQ-123 (1 µM). Thus the ciliostimulatory effect of ET-1 appears to be via its interaction with ET(A) receptors. Application of a combination of the cyclo-oxygenase (COX) inhibitors indomethacin (30 µM) and ibuprofen (10 µM) did not alter baseline CBF but prevented ciliostimulation by ET-1. Incubation of the explants with the nitric oxide synthase (NOS) inhibitor L-NMMA (100 µM) caused a significant decrease in CBF relative to baseline (p < 0.01) which developed over 20 min and remained stable thereafter. Subsequent perfusion of the explant with ET-1 was unable to increase CBF above the new baseline. These results suggest a role for both prostanoids and nitric oxide in ET-1-stimulated CBF. This study has shown, for the first time, that ET-1 stimulates CBF via its interaction with ET(A)-receptors. Inhibition of both COX and NOS inhibited the effect of ET-1 suggesting a role for both these enzymes in the ET-1 response downstream from the ET(A)-receptor.
Assuntos
Cílios/metabolismo , Endotelina-1/farmacologia , Receptor de Endotelina A/metabolismo , Animais , Relação Dose-Resposta a Droga , Antagonistas do Receptor de Endotelina A , Endotelina-1/administração & dosagem , Óxido Nítrico/metabolismo , Óxido Nítrico Sintase/metabolismo , Prostaglandina-Endoperóxido Sintases/metabolismo , Prostaglandinas/metabolismo , Receptor de Endotelina A/agonistas , Mucosa Respiratória/metabolismo , OvinosRESUMO
The minimum inhibitory concentration (MIC) of a homologous series of alcohol ethoxylates with the same head group size (E6) but differing in the number of carbon atoms in their 'tail group' from 10 to 16 was determined for Staphylococcus aureus NCTC 4163 and Escherichia coli NCTC 8196 using a turbidimetric assay. All the surfactants tested demonstrated bacteriostatic activity against both organisms. A tetrazolium assay showed that C14E6 and C16E6 had little effect on the membrane-bound dehydrogenase enzyme activity of E. coli NCTC 8196 compared with C10E6 and C12E6. C10E6 caused leakage both of K(+) and nucleotides in a concentration-dependent manner above its MIC of 0.2 mM. C12E6 caused some leakage at concentrations below its MIC (0.12 mM).
Assuntos
Álcoois/farmacologia , Antibacterianos/farmacologia , Membrana Celular/efeitos dos fármacos , Escherichia coli/efeitos dos fármacos , Tensoativos/farmacologia , Permeabilidade da Membrana Celular/efeitos dos fármacos , Testes de Sensibilidade Microbiana , Staphylococcus aureus/efeitos dos fármacosRESUMO
Peritonitis is a frequent complication of continuous ambulatory peritoneal dialysis (CAPD), with patients suffering recurrent attacks. The microorganisms most frequently implicated in the infection are the skin microflora, in particular, the coagulase-negative staphylococci such as Staphylococcus epidermidis. These microorganisms gain access to the peritoneal cavity via the in-dwelling silicone rubber catheter in the abdominal wall and often persist as biofilms on the surface of the catheter. The surface characteristics of S. epidermidis were monitored during growth in a CAPD in-vitro model together with their ability to adhere to silicone rubber substrata. Fresh dialysis fluid exerted an injurious effect on the cells leading to a decrease in cell numbers but during the simulated dialysis period the cells adapted to the applied stresses. Over a 96-h period in the model both a clinical isolate and a skin isolate of S. epidermidis adopted a more hydrophobic phenotype. The data presented here show that the bacteria grown in this in-vivo reflective CAPD model continually adapt to their environment and become more tolerant to the stresses imposed. The adapted cells were seen to colonise silicone rubber substrata.
Assuntos
Biofilmes/crescimento & desenvolvimento , Diálise Peritoneal Ambulatorial Contínua/efeitos adversos , Staphylococcus epidermidis , Aderência Bacteriana , Cateteres de Demora/microbiologia , Contagem de Colônia Microbiana , Soluções para Diálise , Humanos , Interações Hidrofóbicas e Hidrofílicas , Técnicas In Vitro , Modelos Biológicos , Diálise Peritoneal Ambulatorial Contínua/instrumentação , Peritonite/etiologia , Peritonite/microbiologia , Elastômeros de Silicone , Pele/microbiologia , Staphylococcus epidermidis/isolamento & purificaçãoRESUMO
Polyoxyethylene alkyl ether surfactants have been shown to have excellent penetration enhancing abilities although they are associated with a high level of local toxicity. We have compared the toxicity of a range of polyoxyethylene alkyl ethers (Brij 96, Brij 76, Brij 56, 10 lauryl ether and 9 lauryl ether) to an anionic surfactant (sodium dodecyl sulphate (SDS)), an ampholytic surfactant (lysophosphatidylcholine) and a cationic surfactant (tetradecyltrimethylammonium bromide (TTAB)), in the presence and absence of egg phosphatidylcholine. The toxicity of the surfactants or phospholipid/surfactant mixtures was assessed by measuring haemolytic activity. The test samples were incubated with a suspension of red blood cells for 30 min and Drabkin's reagent was used to indicate the amount of haemoglobin released. All of the polyoxyethylene alkyl ethers, SDS, TTAB and lysophosphatidylcholine exhibited haemolytic activity at concentrations between 0.10 and 0.25 mM. The addition of egg phosphatidylcholine reduced the toxicity for all of the surfactants, with the toxicity of Brij 96 being mitigated to a greater extent than the toxicity of the other polyoxyethylene surfactants examined. The rate of haemolysis induced by Brij 96 or 10 lauryl ether was also reduced by increasing concentrations of phosphatidylcholine. As the phosphatidylcholine content of a mixed surfactant system comprising egg phosphatidylcholine: Brij 96 was replaced by lysophosphatidylcholine and fatty acid, the haemolytic action of the mixture increased markedly. The results from this study show that the toxicity of surfactants to erythrocytes can be mitigated by the addition of egg phosphatidylcholine. Synthetic surfactants combined with phosphatidylcholine may generate drug delivery systems worthy of more extensive investigation.
Assuntos
Eritrócitos/efeitos dos fármacos , Hemólise/efeitos dos fármacos , Fosfatidilcolinas/toxicidade , Óleos de Plantas , Polietilenoglicóis/toxicidade , Tensoativos/toxicidade , Sistemas de Liberação de Medicamentos , Ovos , Eritrócitos/fisiologia , Feminino , Hemólise/fisiologia , Humanos , Dodecilsulfato de Sódio/toxicidade , Estatísticas não ParamétricasRESUMO
The small volume of airway lining fluid renders it susceptible to alteration by the deposition of inhaled formulations. The increasing popularity of the pulmonary route for drug delivery has led to an increasing number of pharmaceutical excipients being incorporated into inhaled dosage forms. The effects of drug delivery vehicle on airway epithelial cells can be studied with the aid of cell culture models of the respiratory epithelium. The effects of pH, osmolarity, and lactose on epithelial cell layers were studied using 16HBE14o- cells. Mannitol flux was used to assess epithelial permeability, enzymatic conversion of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) was used as a measure of epithelial cell metabolism, and release of the cytosolic enzyme lactate dehydrogenase was used as a measure of cell integrity. The effect of buffer composition on epithelial cell mucus secretion was studied using HT29-clone H cells, with mucus secretion measured using an enzyme-linked lectin assay. The permeability of 16HBE14o- cell layers was increased by apical fluid of pH 5, 6, and 9 as well as osmolarities of 96 and 545 mOsm. MTT conversion was reduced by apical fluid of pH 5 and 6 and osmolarity of 96 mOsm. Lactate dehydrogenase release was only increased by apical fluid of pH 9. No effect of lactose solution (100 mM) on the epithelial cells was observed. Mucus secretion by HT29-clone H cells was lowest in Dulbecco's modified Eagle medium (2.92 +/- 0.23 ng per cell layer) and was increased in phosphate-buffered saline with magnesium and calcium (4.28 +/- 0.38 ng per cell layer) and phosphate-buffered saline without magnesium and calcium (6.56 +/- 0.72 ng per cell layer). These results suggest that the physicochemical properties of inhaled formulations should be carefully controlled. The effect of buffer composition on mucus secretion suggests that even the application of "physiological" solutions may affect the epithelium. These cell models represent an opportunity to investigate the interaction of drug delivery vehicles with the epithelium.
Assuntos
Células Epiteliais/efeitos dos fármacos , Excipientes/farmacologia , Análise de Variância , Brônquios/citologia , Células Cultivadas , Colo/citologia , Células Epiteliais/metabolismo , Humanos , Concentração de Íons de Hidrogênio , L-Lactato Desidrogenase/metabolismo , Manitol/metabolismo , Muco/metabolismo , Concentração Osmolar , PermeabilidadeRESUMO
The formation of oil-in-water (o/w) microemulsions stabilized by the nonionic surfactants, polyoxyethylene-10-dodecyl ether, polyoxyethylene-10-oleyl ether, N,N-dimethyldodecylamine-N-oxide and N,N-dimethyloleylamine-N-oxide and containing a variety of pharmaceutically acceptable oils, namely ethyl butyrate, ethyl caprylate, ethyl oleate and the triglycerides, soybean oil, Miglyol 812 and tributyrin, has been examined at 298 K. The effect on microemulsion formation of replacing water with phosphate buffered saline (PBS) and complete PBS has been established. In addition, the effect of changing temperature (from 298 to 310 K) on the phase behaviour of microemulsions formulated using PBS as continuous phase has been determined. Although some small differences in phase behaviour were noted when altering the continuous phase, the greatest difference in phase behaviour was observed when changing the experimental temperature, particularly for microemulsions stabilized by polyoxyethylene-10-oleyl ether. Regardless of the temperature and aqueous phase used, however the larger molecular volume oils (soybean oil, Miglyol 812 and ethyl oleate) were solubilized to a lower extent than the smaller molecular volume oils (namely, ethyl butyrate and ethyl caprylate). The only exception to this rule was when polyoxyethylene-10-oleyl ether was used as surfactant, particularly at 298 K, where it was the larger molecular volume oils that were solubilized to the greatest extent. Cloud point/phase inversion temperature experiments suggested that the higher molecular volume oils were incorporated into the microemulsions prepared using the polyoxyethylene-based surfactants in a different way than the smaller molecular volume oils and suggest that the smaller molecular volume oils are acting in much the same way as a cosurfactant in that they interchelate with their hydrophilic group interspersed in the surfactant head group region. As N,N-dimethyldodecylamine-N-oxide does not exhibit a cloud point it was not possible to determine the mode of oil incorporation in microemulsions prepared with this surfactant.
Assuntos
Emulsões/química , Óleos/química , Aminas , Fenômenos Químicos , Físico-Química , Cromatografia em Camada Fina , Eletrólitos , Ésteres , Polietilenoglicóis , Tensoativos , Temperatura , ÁguaRESUMO
PURPOSE: To determine how the structures of peptides influence their alveolar permeability. METHODS: The studies were performed using 14 synthetic 'model' peptides, labelled with a novel, non-intrusive amino acid fluorophore, and their transport studied using rat alveolar cell monolayers cultured on permeable supports. RESULTS: The passage of the peptides across the epithelial cell monolayers is shown to be primarily paracellular, with an inverse dependence on molecular size, and an enhanced flux observed for cationic peptides. The apparent permeability coefficients (Papp) for the peptides (together with those for other organic solutes, taken from the literature) are shown to be well-modelled assuming two populations of 'pores' in the monolayers, modelled as cylindrical channels of radii 15 A and 22 nm. The former pores are shown to be numerically equitable with the monolayer tri-junctional complexes, and the latter are taken as monolayer defects. CONCLUSIONS: The various monolayer Papp values correlate well with the results from in vivo transport experiments, and the conclusion is drawn that the pulmonary delivery of peptide drugs is perfectly exploitable.
Assuntos
Peptídeos/farmacocinética , Alvéolos Pulmonares/metabolismo , Animais , Transporte Biológico , Células Cultivadas , Masculino , Permeabilidade , Alvéolos Pulmonares/citologia , Ratos , Ratos Sprague-Dawley , Relação Estrutura-AtividadeRESUMO
The apparent permeability coefficient (P(app)) of two fluorescently tagged model hydrophilic peptides, acXASNH(2) and acXAS(GAS)(7)NH(2), and (14)C-mannitol across monolayers of cultured rat alveolar epithelial cells of varying transepithelial electrical resistance (TER) has been examined. In line with their design features, the peptides were not degraded under the conditions of the test. Furthermore, no concentration dependence of transport of the tripeptide acXASNH(2) was observed over the concentration range studied, nor was any directional transport seen for either of the model peptides, indicating that under the conditions of the test they were not substrates for any transporters or efflux pumps. From the hydrophilic nature of the peptides (as assessed by their log P), and their inverse dependence of transport with molecular weight and TER, it was assumed that the peptides were transported across the cell monolayer passively via the paracellular route. The observed P(app) for the transport of (14)C-mannitol and the peptides across rat alveolar epithelial cell monolayers were found to be inversely (though not linearly) related to the measured TER and could be well-modeled assuming the presence of two populations of "pores" in the cell monolayer, namely, cylindrical pores of diameter 1.5 nm and large pores of diameter 20 nm. The relative populations of the two types of pores varied with the TER of the monolayer, with the number of large pores decreasing with an increase in TER (and the number of small pores taken as fixed). These results suggest that if the cell monolayer is well characterized with respect to the passage of a range of probe molecules across monolayers of varying electrical resistance, it should be possible to predict the P(app) of any hydrophilic peptide or drug crossing the membrane by the paracellular route at any desired TER using a monolayer of any electrical resistance, above a minimum value.
Assuntos
Peptídeos/farmacocinética , Alvéolos Pulmonares/metabolismo , Alvéolos Pulmonares/fisiologia , Alanina/análogos & derivados , Animais , Transporte Biológico , Radioisótopos de Carbono , Permeabilidade da Membrana Celular , Células Cultivadas , Cumarínicos , Impedância Elétrica , Células Epiteliais/metabolismo , Células Epiteliais/fisiologia , Corantes Fluorescentes , Masculino , Manitol/farmacocinética , Ratos , Ratos Sprague-DawleyRESUMO
Dilute 3-component nonionic oil-in-water microemulsions formulated with either a polyoxyethylene surfactant (C18:1E10 or C12E10) or the alkylamine-N-oxide surfactant, DDAO (C12AO), and containing either a triglyceride or an ethyl ester oil have been examined using dynamic and static light-scattering techniques. Analysis of the results showed distinct differences in the tested oil's mode of incorporation into the microemulsion droplets, with both the molecular volume of the oil and the hydrophobic chain length of the surfactant being important. For example, microemulsions formulated by C18:1E10 and containing one of the larger molecular volume oils (that is, either a triglyceride, Miglyol 812, or soybean oil) or the ethyl ester of fatty acid oil, ethyl oleate, exhibited first a decrease and then an increase in hydrodynamic size and surfactant aggregation number, suggesting that the asymmetric C18:1E10 micelles became spherical upon the addition of a small amount of oil and grew thereafter because of further oil being incorporated into the core of the spherical microemulsion droplet. A similar conclusion of sphericity could not be drawn for microemulsions stabilized by C18:1E10 and containing one of the oils smaller in molecular volume (namely tributyrin, ethyl butyrate, or ethyl caprylate) where neither the aggregation number nor the hydrodynamic radius changed much upon the addition of oil. This result suggested that these oils were preferentially located in the interfacial surfactant monolayer, behaving in much the same way as a cosurfactant. A different trend of results, however, was seen for microemulsions prepared using C12E10 and C12AO, most likely because these surfactants produced approximately spherical micelles. In this case, the microemulsions containing the oils larger in molecular volume tended to exhibit an increase in surfactant aggregation number and hydrodynamic size, suggesting the growth of spherical micelles, while the smaller oils (in particular ethyl butyrate) caused a significant decrease in surfactant aggregation number incompatible with their being incorporated into the centre of the droplet, suggesting that the oils were being located in the interfacial surfactant monolayer. These results suggest that the various oils are incorporated into the microemulsions in very different ways.
Assuntos
Óleos/química , Tensoativos/química , Água , Dimetilaminas/química , Emulsões , Ésteres/química , Luz , Polietilenoglicóis/química , Espalhamento de Radiação , Triglicerídeos/químicaRESUMO
Airway ciliary activity is influenced by [Ca2+]i, but this mechanism is not fully understood. To investigate this relationship, ciliary activity and [Ca2+]i were measured simultaneously from airway epithelial ciliated cells. Ciliary beat frequency was determined, for each beat cycle, with phase-contrast optics and high-speed video imaging (at 240 images s-1) and correlated with [Ca2+]i determined, at the ciliary base, by fast imaging (30 images s-1) of fura-2 fluorescence. As a mechanically induced intercellular Ca2+ wave propagated through adjacent cells, [Ca2+]i was elevated from a baseline concentration of 45 to 100 nM, to a peak level of up to 650 nM. When the Ca2+ wave reached the ciliary base, the beat frequency rapidly increased, within a few beat cycles, from a basal rate of 6.4 to 11.6 Hz at 20-23 degrees C, and from 17.2 to 26.7 Hz at 37 degrees C. Changes in [Ca2+]i, above 350 nM, had no effect on the maximum beat frequency. We suggest that airway ciliary beat frequency is 1) controlled by a low range of [Ca2+]i acting directly at an axonemal site at the ciliary base and 2) that a maximum frequency is induced by a change in [Ca2+]i of approximately 250-300 nM.
Assuntos
Cálcio/farmacologia , Cílios/efeitos dos fármacos , Traqueia/efeitos dos fármacos , Animais , Cálcio/metabolismo , Células Cultivadas , Cílios/metabolismo , Fluorescência , Fura-2 , Microscopia de Contraste de Fase/instrumentação , Microscopia de Vídeo/instrumentação , Periodicidade , Coelhos , Estresse Mecânico , Traqueia/metabolismoRESUMO
The synthesis of racemic and l-(6,7-dimethoxy-4-coumaryl)alanine (Dmca) is described and some spectral and physicochemical properties are reported. The utility of Dmca as a highly sensitive and specific label for the quantitative detection of synthetic peptides in HPLC and in minimal essential media (MEM) is also described.
Assuntos
Alanina/análogos & derivados , Cumarínicos/química , Cumarínicos/síntese química , Corantes Fluorescentes/síntese química , Alanina/síntese química , Alanina/química , Corantes Fluorescentes/química , Isomerismo , Modelos Químicos , Espectrometria de Fluorescência , Espectrofotometria UltravioletaRESUMO
Cefuroxime axetil, the orally active prodrug of cefuroxime is marketed as a 1:1 mixture of two diastereomers designated as R (1'R, 6R, 7R) and S (1'S, 6R, 7R). Prodrug hydrolysis is thought to occur during intestinal absorption, however little is known concerning the relative availability of cefuroxime from each isomeric form. The Caco-2 cell monolayer model was used to examine the possible stereoselectivity of absorption by measuring the accumulation and epithelial transport rate in the apical to basolateral direction of cefuroxime and cefuroxime axetil following application of the mixture (1.0 mM) or individual diastereomers (0.5 mM0 of cefuroxime axetil. Cefuroxime appearance in the basolateral chamber was in the order: mixture > R > S following application of the prodrug. The accumulation of unchanged cefuroxime axetil was S > R irrespective of the form applied, i.e. individual diastereomer or the mixture. Such stereoselective differences in both absorption and/or hydrolysis may contribute to the observed oral bioavailability (30-50%) of cefuroxime in vivo.
Assuntos
Cefuroxima/análogos & derivados , Cefalosporinas/farmacocinética , Pró-Fármacos/farmacocinética , Adsorção , Transporte Biológico , Células CACO-2 , Cefuroxima/química , Cefuroxima/farmacocinética , Cefalosporinas/química , Humanos , Hidrólise , Pró-Fármacos/química , EstereoisomerismoRESUMO
Beat frequency and the duration of the constituent recovery, effective, and rest phases of the beat cycle of respiratory tract cilia were measured photoelectronically before and after manipulation with ionomycin or isoproterenol. Both ionomycin, acting by increasing intracellular Ca2+, and isoproterenol, acting by elevating intracellular adenosine 3',5'-cyclic monophosphate (cAMP), increased beat frequency by reducing the duration of the three phases of the ciliary beat cycle in a similar manner. The addition of increasing concentrations of ATP to ciliated cells permeabilized by exposure to saponin caused a pattern of phase reduction indistinguishable from that observed in whole cells. The beat frequency of permeabilized cells was slower than that of whole cells and insensitive to changes in Ca2+ and cAMP. Ca2+ and cAMP may regulate ciliary beat frequency by acting at a common site within intact cells, possibly regulating the rate at which the axoneme can use ATP or the availability of ATP to the axoneme.