This statement provides practical guidelines and suggestions for methacholine and exercise challenging testing. Specifically, it reviews indications for these challenges, details factors that influence the results, presents brief step-by-step protocols, outlines safety measures, describes proper patient preparation and procedures, provides an algorithm for calculating results, and offers guidelines for clinical interpretation of results. The details are important because methacholine and exercise challenge tests are, in effect, dose-response tests and delivery of the dose and measurement of the response must be accurate if a valid test is to be obtained. These guidelines are geared to patients who can perform good-quality spirometry tests; they are not appropriate for infants or preschool children. They are not intended to limit the use of alternative protocols or procedures that have been established as acceptable methods. We do not discuss the general topic of bronchial hyperresponsiveness (BHR). 100 mg 100 mg 6.25 ml A: 16 mg/ml 3 ml of dilution A 9 ml B: 4 mg/ml 3 ml of dilution B 9 ml C: 1 mg/ml 3 ml of dilution C 9 ml D: 0.25 mg/ml 3 ml of dilution D 9 ml E: 0.0625 mg/ml * Schedule obtained from Methapharm (Brantford, ON, Canada). * This list is provided to simplify access to some sources of equipment. It is not a complete list of all possible sources of acceptable equipment.

The efficacy of adenovirus-mediated gene therapy is attenuated by the host immune responses to both vector and transgene products. Even for helper-dependent adenoviral (HD-Ad) vectors, which have all viral-coding sequences deleted, the viral capsid proteins still cause immune reactions. In order to improve the efficiency in transgene expression during HD-Ad vector readministration, we administered cyclophosphamide to transiently modulate the mouse immune system. We delivered a high dose (5Â10 10 vector particles (vp) per mouse) of empty HD-Ad to the mouse airway to induce an initial immune response. After 4 weeks, the mice were readministered with an HD-Ad vector containing either the reporter gene, LacZ, or the gene for the human cystic fibrosis transmembrane conductance regulator (CFTR) (1.5Â10 10 vp per mouse). We found that the expression of both transgenes was greatly improved by the administration of cyclophosphamide when compared with the expression in mice without the immunosuppressing drug. We also found that the high dose of the empty HD-Ad vector administered intranasally does not induce an acute systemic immune response, but it does elicit an acute local response of proinflammatory cytokine production. Antibodies against Ad vector, including the neutralizing antibodies, were greatly reduced by the presence of cyclophosphamide in vector readministratiton. Moreover, cyclophosphamide reduced the infiltration of inflammatory cells, including total leukocytes, lymphocytes, CD4+ and CD8+T cells. These results indicate that transient administration of immunosuppressive agent can be used to extend transgene expression as well as attenuating immunogenicity to HD-Ad vectors in airway readministration.

In all rapidly changing systems, an appropriate response time of all sensing devices is essential for the accurate conversion of a physiological parameters to a proportional electrical signal. The frequency response of equipment is the ability to accurately reflect both the magnitude and temporal relationship of dynamic events over a defined frequency range. The phase lag expresses the temporal delay between the physical event being measured and the output signal. The attenuation expresses diminution of the amplitude ratio of the output signal in relation to the input signal over a range of frequencies. We have developed a method that specifically addresses the measurement of frequency response and attenuation of pneumotachographs and low pressure transducers. The system consists of a 81 liter rectangular box separated in the middle by a 30.5 cm acoustic loudspeaker, the cone sealed by latex, and driven by a signal generator coupled to a low frequency amplifier. This system can produce an undistorted sinusoidal signal between 0 and 20 Hz, and the peak flow through the pneumotachograph is only minimally affected by changes in frequency. Rapid analysis is possible using an oscilloscope to produce Lissajou loops. There is no measurable attenuation between the electrical signal and the pressure generated over frequencies from 1 to 20 Hz. The system is accurate at low frequencies and can generate appropriate signals over the frequency range of interest for respiratory applications, and it can be inexpensively constructed.