The sample is a representation of the NP microbiome, which contai

The sample is a representation of the NP microbiome, which contains numerous bacterial species [67] and may include close relatives of pneumococci such as

S. pseudopneumoniae, Streptococcus mitis and other streptococcal species that also inhabit this niche [68]. The ideal method for non-culture identification in NP swabs should unequivocally detect the pneumococcus with high sensitivity and specificity; it should also be rapid, easy to perform, inexpensive, and deployable on a large scale. In the last decade, several non-culture methods aiming to detect pneumococci in biological samples have been developed including PCR-based strategies targeting specific DNA markers such as rpoA [69], sodA [70], tuf [71], recA [72], selleck piaA [73], Spn9802 [74], ply [75], a 181-bp pneumococcal-specific fragment [76], 16S-rDNA [77], LY2109761 ic50 psaA [78], and lytA [79], [80] and [81]. For many of these methods specificity problems have been detected [64], [65], [82] and [83]. For others, there has been insufficient validation against diverse collections of close relatives of pneumococci. In addition, there is an increasing body of more sophisticated

methods that, although promising, may not be easily applied in routine analysis of NP samples [84], [85], [86] and [87]. While there is currently no gold standard method for non-culture identification of pneumococci from NP swabs [63], [88] and [89], the lytA real-time PCR assay described by Carvalho et al. [81] is widely used and appears to be species-specific. However, given the capacity of pneumococci to exchange genes with other oral streptococci [88] and [90] a multilocus approach such as used in multilocus sequence typing (MLST), microarray or whole genome-sequencing may prove valuable [64], [91] and [92].

Culture should remain the gold standard for detection of pneumococci in NP swab samples. Investigators may wish to complement culture detection with a non-culture technique; the method we currently recommend is lytA real-time PCR [81]. A systematic laboratory validation of non-culture methods against large collections of nasopharyngeal and non-classical isolates is needed to guide future recommendations. Studies that are designed to determine the clinical over relevance of pneumococcal culture-negative but DNA-positive samples are needed. The current standard method for serotyping of pneumococcal isolates is the capsular reaction/swelling test (Quellung reaction or Neufeld test) [1]. The traditional method described by Lund [93], Austrian [94] and the Statens Serum Institut [95] using ×100 magnification with oil immersion, is still widely used in Europe and North America. In Australia and Papua New Guinea, the ‘dry’ method using ×40 magnification without oil [96] has been in use since at least the 1970s (M. Gratten, personal communication).

However, this does not appear to provide a solid explanation for

However, this does not appear to provide a solid explanation for the lack of physiotherapy-led presentations

at national conferences identified in recent years. It find more also fails to explain the imbalance between representation of physiotherapists and other health professionals in this arena. Physiotherapy organisations, academic institutions, and therapists could develop strategies to increase the engagement of physiotherapists in cardiology research. Some simple strategies could include the implementation of a mentoring system designed to link physiotherapists with established research backgrounds and clinicians working in the management or prevention of cardiac disease. Greater mentorship of postgraduate physiotherapy research on cardiac topics is also needed in physiotherapy schools. The establishment of more frequent communication between clinical and research physiotherapists, via bodies such as Cardiorespiratory Physiotherapy Australia, CSANZ, and ACRA may also inspire clinicians to consider research in this area. Funding and academic opportunities in the area of cardiovascular disease management are JNK inhibitor extensive. Exploration of these opportunities by physiotherapists would be fruitful for individual physiotherapists, the profession and, ultimately and most importantly, for patients. Research opportunities are widely available and physiotherapists

are ideally positioned to take a leadership role in the future evolution of cardiac management. In summary,

cardiac disease is a leading international health problem. Despite physiotherapists being ideally trained with relevant clinical experience there appears to be a general lack of engagement with cardiology research. The problem manifests across a range of domains including professional membership, active participation in national conferences, and publication of research in the area of cardiovascular disease. The expertise and capacity of physiotherapists coupled with extensive career opportunities in the area of cardiology research presents a range of opportunities for physiotherapists to explore. “
“Mechanical ventilation temporarily replaces or supports spontaneous breathing in critically ill patients in intensive care units. Weaning is the withdrawal of mechanical ventilation Astemizole to re-establish spontaneous breathing. Patients are considered to have successfully weaned from ventilatory support when they can breathe on their own for at least 48 hours (Sprague and Hopkins, 2003). Weaning typically comprises 40–50% of the total duration of mechanical ventilation, with almost 70% of patients in intensive care weaning without difficulty on the first attempt (Boles et al 2007). Other patients have a more difficult or prolonged period of weaning, which is associated with a poorer prognosis (Vallverdu et al 1998, Esteban et al 1999).

Few analytical methods have been reported for the verification of

Few analytical methods have been reported for the verification of steroidal hormone drugs, especially for those with similar selleck screening library chemical properties. In this paper, our aim was to develop a set of simple High-performance liquid chromatographic (HPLC) with evaporative light scattering detection12, 13, 14 and 15 (ELSD) and with dual

ESI ionization mass spectrometry (LCMS) methods are presented to distinguish and qualitatively analyze used to identify of Dexamethasone, Testosterone and Estrone (E1) in the combination form. Pure standards of Dexamethasone, Testosterone and Estrone (E1) were obtained from the Sigma–Aldrich, India. Organic solvents for chromatography were purchased in LCMS grade, ACS grade Acetonitrile was purchased from Honeywell-Burdick & Jackson (USA), water was obtained from ultra-purified from Elix Advantage 5 system equipped with Milli-Q Biocel (Millipore), all the chemicals used were of analytical reagent grade, and the solvents were of ACS. The purity of each reference standard was determined by HPLC PDA, ELSD detectors and dual ESI (LCMS). All solvents and samples were filtered through MILLEX FG (Millipore),

13 mm, 0.2 μM, fluoropore, non-sterile membrane sample filter paper before injecting into system. The analyses were performed using an Agilent 1200 Series HPLC system, equipped with a binary pump, an auto-sampler, a column oven, PDA detector and a mass hunter software version B.02.01 (B2116.20) over (Agilent Technologies, USA). Agilent 1260 Infinity Evaporative Light Scattering Detector (ELSD) instrument, operated by the Agilent 35900E multichannel interface which converts analog signal to digital (A/D) (Agilent Technologies, USA), was connected to the liquid chromatography for detection of steroids. The separation was carried out on a reverse phase Shodex C18, 3 μm, 4.6 × 100 mm at ambient temperature. The isocratic elution mode with a mobile phases

Acetonitrile and 0.1% formic acid in water and eluted by the following program at the flow 1 mL/min, runtime 6 min. The drift tube temperature for ELSD was set at 50 °C and the nitrogen flow rate was 53 psi. Agilent 6520 Quadrupole time-of-flight (Q-TOF) mass spectrometer. Coupled to an Agilent 1200 series HPLC system (Agilent Technologies, USA) is equipped with binary pump, auto sampler, thermostatted column compartment, variable wavelength detector, auto sampler thermostatted (G 1330B). The Agilent Q-TOF (6520) mass spectrometer is equipped with dual electrospray ionization (ESI) ion source, and the HPLC conditions were identical to those used for HPLC–ELSD analyses mentioned above. Mass spectra were acquired in positive mode with scan range from m/z 100 to 500 Da. The conditions of dual ESI source were as followed: drying gas (N2) flow rate, 30.

Notably, a Beijing-based JE-MB vaccine is not available for inter

Notably, a Beijing-based JE-MB vaccine is not available for international travelers and was thus not included in the present study. The study population consisted of JE vaccinees whose early immune responses were reported in the two former studies. In this follow-up we included subjects who had received (1) a JE-VC primary

series (group VC), (2) a JE-MB primary series followed by a single booster dose of JE-VC (group MB-VC), and (3) a JE-MB primary learn more series followed by a single booster dose of JE-MB (group MB-MB). In the booster groups, the median intervals between primary and booster vaccinations were 5.2 (range 1.1–20.5) years (group MB-VC) and 3.7 (range 1.0–12.2) years (group MB-MB). Eligibility criteria for the participants have been described previously [5] and [16]. Briefly, the subjects were adult volunteers who received JE primary or booster vaccination as part of their pre-travel consultation at two travel clinics in Finland and Sweden. The following exclusion criteria JQ1 were used: age <18 years, acute disease at the time of enrollment, pregnancy or lactation, clinically significant immunosuppression, known history of JE, alcohol or drug abuse, or suspected hypersensitivity to any

of the vaccine components. The initial study comprised 31 volunteers in group VC, 42 in MB-VC and 32 in MB-MB [5]. For this research project, we collected follow-up serum samples from all volunteers available around two years after their last vaccine dose: 15/31 participants (48%) in group VC, 19/42 (45%) in group MB-VC, and 14/32 (44%) in group MB-MB. The samples were evaluated for persistence and cross-reactivity of the JEV neutralizing antibodies. Of the subjects in the JE-VC primary vaccination group (group VC), only those were included in the analyses who showed no antibodies against the JEV strains prior to administering the vaccine series. The these study (EudraCT: 2010-023300-27) was approved by the appropriate ethics

committees and registered in the databases required. All volunteers provided informed consent. Titers of neutralizing antibodies were determined by the plaque-reduction neutralization test (PRNT), which is currently regarded the method of choice for assessment of seroprotection elicited by JE vaccines [17]. The neutralization tests were performed as described previously [5] and [18]. All serum samples were tested against seven different JEV strains representing genotypes I–IV: SM-1 (GI; isolated in Thailand 2002), 1991 (GI; Korea 1991), B 1034/8 (GII; Thailand 1983), Nakayama (GIII; Japan 1935, strain in JE-MB), SA14-14-2 (attenuated GIII strain, strain in JE-VC; parental strain China 1954), Beijing-3 (GIII, China 1949), and 9092 (GIV; Indonesia 1981). The analyses were performed in a blinded manner.

Cold-chain storage cost per dose was estimated using the 2012 WHO

Cold-chain storage cost per dose was estimated using the 2012 WHO vaccine volume calculator [18]. This estimates that the cold chain costs for a 10-dose vial

is $0.03 per dose and 5-dose vials costs $0.05 per dose. The model specified in Eqs. (4) and (5) was used to depict two policy options: (1) offering IPV in 10-dose vials and (2) offering IPV in 5-dose vials. For each country and each policy option the model ran 1000 replications drawing independently from the statistical GSK1349572 clinical trial distributions of session size for all of the various types of clinics in the country as specified in Eqs. (4) and (5). The baseline cost per dose of the vaccine was assumed to be $2.48 per dose in 10-dose vials, using the mean of the price range released by UNICEF [19], and $2.98 per dose in 5-dose vials, which is a procurement price gap of $0.50. As no price information is available for IPV 5-dose vials, we carried out a univariate sensitivity

analysis to vary the price gap from zero to a $1.00 per dose between 10- and 5-dose vials. Our study found that session size varied significantly within and across all four countries included in the analysis. Table 3 lists SB203580 datasheet the median session size and 25th to 75th percentile for different types of healthcare centers in Bangladesh, India (Uttar Pradesh), Mozambique, and Uganda. Depending on whether the clinic setting was urban, rural, outreach or fixed, the median session size varied between 3 and 15 children. To predict session size in different clinical settings, session size field data were used for statistical distribution fitting. Fig. 1 shows the Akaike Information Criteria (AICs) score associated with the best fitting parameters medroxyprogesterone within each statistical distribution family—the lower the AIC, the better the fit. The negative binomial family offered the greatest number of best-fit results compared to the other three families, though as seen in Fig. 1, the AIC score of the second best-fit did not

differ greatly from the best-fit in some cases. The best-fit distributions were parameterized for each clinic type in each country and applied in the calculation of vaccine wastage. Wastage in both 10-dose vials and 5-dose vials presentations was calculated, indicating a lower wastage rate for using 5-dose vials. Table 4 shows that by switching from 10-dose vials to 5-dose vials, the wastage rate was reduced in all four countries. While using 5-dose vials produced a lower wastage rate, it also triggered an increase in the per-dose fully loaded cost, which included the procurement costs, cold-chain costs, and cost of open vial wastage. Fig. 2 shows the distributions of the present values of fully loaded per dose costs in a 10-year analytical horizon for IPV with a procurement price of $2.48 per dose in 10-dose vials and a price gap of $0.50 per dose in 5-dose vials in Bangladesh, India (Uttar Pradesh), Mozambique, and Uganda.

11 in Kinnell (2014)

11 in Kinnell (2014) Selleck PCI-32765 was incorrect. They suggested that it should be equation(12) b1(QR30EI)c1=b1(Ve30EIPe−1)c1b1QREI30c1=b1VeEI30Pe−1c1where

b1 and c1 are the empirical coefficients, QR is the runoff ratio, E is the storm kinetic energy, I30 is the maximum 30-minute intensity, Ve is the runoff amount, and Pe is the rainfall amount. While their Eq. (12) was mathematically correct, Eq. 11 in Kinnell (2014) was presented in the context of modelling soil loss in terms of runoff and sediment concentration with the expression for sediment concentration enclosed in square brackets. Consequently, Eq. 11 in Kinnell (2014) should have been written as equation(13) b1(QR30EI)c1=Ve[b1Vec1–1(30EIPe−1)c1].b1QREI30c1=Veb1Vec1–1EI30Pe−1c1. The term Vec1–1Vec1–1 was inadvertently omitted from Eq. 11 in Kinnell (2014). Eq. (13) is a mathematically correct rearrangement of Eq. (12). Eq. (13) indicates that sediment concentration varies nonlinearly with both the runoff amount and the product of the kinetic energy per unit quantity of rain (E Pe− 1) and I30. The relevance of the discussion about the effect of runoff on sediment concentration that followed Eq. 11 in Kinnell (2014) is more obvious from Eq. (13) than Eq. (12). However, the discussion in Kinnell (2014) about Ae Pe (EI30)− 1 increasing with Ve to a

power of 1.48 on 22 m long plots at Sparacia followed the observation in Bagarello et al. (2011) that nonlinear relationships between sediment concentration and the product of the kinetic energy per unit quantity of rain and BLU9931 mw I30 did not second definitely exist in experimental data obtained from runoff and soil loss plots at Masse and Sparacia when both runoff and the product of the kinetic energy per unit quantity of rain and I30 were used as independent variables in the prediction of sediment concentration. Although not stated explicitly, the discussion in Kinnell (2014) about Ae Pe (EI30)− 1 increasing with Ve to a power of 1.48 on 22 m long plots at Sparacia focussed on equation(14) b1(QR30EI)c1=Ve[b1Vec2(30EIPe−1)]b1QREI30c1=Veb1Vec2EI30Pe−1where c2 = 0.48

on 22 m long plots at Sparacia, being an alternative to Eq. (13). Given that c2 was greater than c1 − 1 at Sparacia, the conclusion by Kinnell (2014) that runoff had a significant effect on sediment concentration at Sparacia followed more from Eq. (14) than Eq. (13). “
“The authors regret that there were errors in the units for total carbon and total nitrogen in Fig. 5. The corrected version of the figure is shown below. The authors would like to apologise for any inconvenience caused. Figure options Download full-size image Download as PowerPoint slide Fig. 5. Concentrations of carbon, nitrogen, and phosphorus in the organic horizon and the upper mineral soil (0–20 cm) along the Haast dune sequence, New Zealand. Values are the mean ± standard error of three replicate plots located along the dune crest at each site.

Pentylenetetrazol (50 mg/mL) was administered at a dose rate of 4

Pentylenetetrazol (50 mg/mL) was administered at a dose rate of 40 mg/kg/h by intravenous (IV) infusion at a rate of 10 mL/h in twelve (12) monkeys, until convulsion onset and the infusion was stopped immediately. Diazepam (Sandoz, Boucherville,

QC, Canada; Everolimus research buy 1.0 mg/kg) was administered IV at seizure onset. A caffeine challenge was conducted using a prospective, randomized, controlled, crossover study to illustrate applications of qEEG in drug development. Caffeine (10 mg/kg, IM) was administered approximately 10 min prior to lights off to ten (10) animals (i.e. at 18:00). Sterile saline USP was administered as a control. A wash-out of at least 3 days was allowed between each treatment. Pentylenetetrazol (50 mg/mL) was administered at a dose rate of 100 mg/kg/h by IV infusion to five (5) dogs, until the start of convulsions and was then stopped immediately. Diazepam (1 mg/kg,

IV) was administered immediately following seizure onset. Both EEG and EMG were recorded following saline IV infusion in rats (n = 49) Autophagy Compound Library price or IV PTZ infusion (cross over design with n = 8). Three (3) rats were used to illustrate qEEG effects of diazepam (3 mg/kg, IM) and amphetamine (3.75 mg/kg, PO). Twenty-four (24) rats received a PTZ (8 mg/mL) IV infusion at a dose rate of 288 mg/kg/h. Diazepam (2–6 mg/kg) was administered by intra-peritoneal (IP) injection following tonic convulsion onset. – Sixteen (16) Sprague–Dawley rats were used to illustrate the effect of yohimbine (18 mg/kg, SC, 8 min prior to PTZ infusion) as a positive control (n = 8) in the PTZ seizure threshold model, with an equal number of animals (n = 8) receiving either yohimbine or saline (control). The EEG and EMG (when applicable) traces were analyzed using manual and

automated detection (NeuroScore, Data Science International, St.-Paul, MN, USA). Spectral analysis was performed on 60-s epochs to quantify the absolute and relative amplitude of EEG frequency bands (delta [0.5–4 Hz], theta [4–8 Hz], alpha [8–12 Hz], sigma [12–16 Hz], beta [16–24 Hz] and gamma [24–50 Hz]) and individual frequencies [0.5–127 Hz]. For each parameter, a one-way analysis-of-variance (ANOVA) was conducted and the residuals were saved. Gaussian distribution was evaluated using the Astemizole Shapiro–Wilk test on residuals. Whenever the Shapiro–Wilk test was found to be significant (p ≤ 0.001) then the data were transformed and re-submitted to the analysis. The Levene test was used to examine the homogeneity of group variances. For the ANOVA model, if the overall group differences were shown significant (F-Test), then pair-wise comparisons were conducted using Tukey’s test. Data are presented as mean (SD). The EEG and EMG (when applicable) electrodes were well tolerated in all animals. Premonitory seizure signs observed during the pre-ictal period with associated average PTZ doses are listed in Table 1. Emesis and decreased activity were the most common clinical signs followed by hypersalivation and ataxia.

The control saponin R, was as expected the most hemolytic (HD50 =

The control saponin R, was as expected the most hemolytic (HD50 = 35 μg/ml). Furthermore, the safety analysis detected neither lethality nor local pain or swelling ( Table 1) for any of the C. alba vaccines. AZD6738 datasheet Only loss of hair at the local of injection was detected in the 5 mice treated

with the QS21 containing saponin R. The increase in hemolytic activities of C. alba saponins was not correlated to the increase in the size of the C-28 attached carbohydrate chain. In contrast, the CA3 and CA3X saponins that both have three sugar units in that chain strongly differed in their hemolytic capabilities. Saponin CA3X which has a xylose terminal unit induced strong hemolysis while saponin CA3 that shows an apiose unit instead was much less hemolytic. In correlation with our findings, the QS21 adjuvant is composed of two isomers that include either apiose (QS21-Api) or xylose (QS21-Xyl) as the terminal sugar residue within the linear find more tetrasaccharide segment, in a ratio of 65:35, respectively [34]. The saponin QS21-Xyl was marginally more toxic than QS21-Api or the QS21 mixture. Overall mice weight loss was greatest in the SQS21-Xyl groups and although one mouse of both groups died over the course of immunizations, the mice

in the QS21-Xyl group showed the worst clinical status. On the other hand, the QS21-Xyl treated mice induced a higher IgM and IgG response [34]. In our investigation we demonstrated that the adjuvant potential

of C. alba saponins Idoxuridine is correlated to the increase of their C-28 attached sugar chain. We also demonstrated that the addition of an extra apiose unit in CA4 saponin is determinant of its enhanced adjuvant potential. Both the CA3 and CA3X saponins have three sugar chains and three exposed hydroxyl groups on the terminal sugar unit, therefore sharing the same HLB. However the spatial configuration and exposition of the HO groups on the apiose terminal sugar unit is optimized when compared to the configuration of the same groups in xylose. This would explain also the reason for the increased adjuvant potential of CA4 which has an additional apiose unit. The CA4 saponin of C. alba in formulation with FML induced a higher response after challenge, significant increases in IgG and IgG2a anti-FML antibodies which were absent in the CA3-saponin. These results confirm the relevance of the addition of a fourth unit of apiose 1 → 3 linked to the rhamnose residue of the C-28 attached sugar chain in the induction of the anti-FML humoral response. As expected for a positive adjuvant control, the global humoral response induced by the saponin QS21 containing saponin R vaccine was the highest. The intensity of the humoral response generated by saponins has been shown to be related to the presence of carbohydrate moieties attached to the triterpene nucleus [14], [17] and [25] and this response increases in direct proportion to their length [22].

Several trials indicate that reducing immobilisation time alone a

Several trials indicate that reducing immobilisation time alone after an upper limb fracture without therapy intervention could be beneficial (Davis and Buchanan 1987, Dias et al 1987, McAuliffe

et al 1987). A theme that emerged from the review was that the trials that reported contrary findings or lack of effect included more severe fractures that had been surgically managed (Agorastides et al 2007, Krischak et al 2009). In these trials the group that SNS-032 nmr received more exercise (ie, supervised exercise in addition to home exercise program or earlier commencement of exercise) had poorer observed outcomes than the group that received less exercise (ie, home exercise program alone or delayed exercise). These results lead to the speculation that the amount of inflammation and tissue damage from the severity of the fracture and surgery might mean that a period of relative rest or controlled movement NVP-AUY922 manufacturer may be an important part of recovery during rehabilitation. However, further research that controls for co-interventions and closely monitors the amount of exercise completed would be needed to confirm this. Another theme that emerged was that exercise may be more likely to lead to reduction in impairment,

particularly range of movement, than improvements in activity limitations. A number of trials reported short-term improvements in range of movement in the group receiving more exercise (Lefevre-Colau et al 2007, Wakefield and McQueen, 2000, Watt et al 2000), but there were few examples whatever where the improvements carried over into an improved ability to complete daily activities. Given the principle of specificity of training, it is perhaps not surprising that exercises for upper limb fracture rehabilitation that focus on repeated movements or repeated contractions

might lead, when effective, to increased range of movement and increased strength. A couple of trials attempted to address this possible limitation by implementing ‘activity-focused’ exercises, but the content of the interventions were not well described and the investigators did not detect any beneficial effect (Christensen et al 2001, Maciel et al 2005). The findings of this review are similar to two previously published systematic reviews that concluded there was insufficient evidence to determine which rehabilitation interventions may be useful for the management of distal radial fractures (Handoll et al 2006) and proximal humeral fractures (Handoll et al 2003). The current systematic review adds to the literature by focusing on exercise and including recently published studies (Agorastides et al 2007, Hodgson et al 2007, Kay et al 2008, Krischak et al 2009). A strength of this systematic review was its comprehensive search strategy which included eight electronic databases, citation tracking, and manual reference list checks with no included trials identified outside the database searches.

, 2012) Like other CPPW communities, the SNHD used a portion of

, 2012). Like other CPPW communities, the SNHD used a portion of their grant funds to support PA. The SNHD’s strategies to

increase PA included buy Staurosporine the promotion and improvement of local trails. We have previously reported on the characteristics and effect of its media campaign promoting trail use, where we observed a 52% increase in mean users per hour over six months (Clark et al., in press). This portion of the project involves the same trails but a longer time period and also includes an alteration to the trail environment. A recent review of trails and PA completed by Starnes et al. (2011) reports that trail use has been both positively and negatively associated with age, racial and ethnic minority status, and gender. The reviewers

also reported mixed results from studies investigating access to trails and levels of PA, and called for further Smad inhibitor research to investigate the relationship between trails and PA. Price et al. (2013) recently studied correlates of trail use in Michigan and reported higher levels of use among males, those with higher levels of education, and White race/ethnicity. Most previously published studies of trail usage are cross-sectional and rely on self-reported behaviors (Starnes et al., 2011). Few studies have reported on objective measures of trail use or changes in trail usage over time. Evenson et al. (2005) analyzed PA among those living near a new trail, before and after construction, but their study showed no significant increase in PA. Another study of the promotion of a newly constructed trail in Australia over used data from telephone surveys and objective counts to assess PA changes among people living nearby (Merom et al., 2003). The authors reported both an increase in cycling traffic and an increase in PA among one subgroup (Merom et al., 2003). Fitzhugh et al.

(2010) reported a positive effect on PA in adults when trail access was improved, but they did not report on the effect of signage. Price et al. (2012) studied seasonal variations in trail use among older adults, but they did not assess the effect of changing the trail environment. Although the presence of trail signage is noted in trail environment assessment tools (Troped et al., 2006), to our knowledge there are no published articles on the effect of trail signage on trail usage. Accordingly, the purpose of our study was to assess the longer term effects of the marketing campaign and to compare usage on trails which were altered by adding way-finding and incremental distance signage to usage on control trails which were not altered, using longitudinal data obtained from objective measures of trail use. We employed a quasi-experimental design with a comparison group to assess the effect of signage additions on trail use in Southern Nevada.