Conidia from all colonies, which were incubated for 10 days, were photographed under a phase-contrast microscope (400×) at the same light exposure. To compare the levels of light-penetrating activity of conidia, which was observed under a phase-contrast microscope, a densitometric analysis was

used to generate a relative densitometric value (RDV) of conidia. In each observation of the photographed conidia, the densitometric values at four spots of background (DV0) and four spots of conidia (DV1), randomly taken, were converted to RDV as follows: RDV = DV1/DV0. The highest RDV was arbitrarily given to 1.00 to compare it with the other RDVs. All conidial suspensions (c. 5 × 106 conidia mL−1) were transferred to fresh Eppendorf tubes (500 μL per Selleckchem Alectinib tube) and held in a water bath at 45 °C for 30, 60, 90 and 120 min. For each strain treatment (non-paired and paired), controls (non-exposed Selleckchem MI-503 conidial suspensions) were kept at room temperature (c. 25 °C). A 10-μL sample

was taken from each tube and dropped on ¼SDAY medium for a germination test prior to and after the exposures. After incubation of all plates at 20 °C for 24 h, percent germination was determined by randomly counting the number of germinated and ungerminated conidia among 100 counts microscopically (400×). A conidium was considered germinated if a germ tube was longer than the length of a conidium (Avery et al., 2004). In addition, the length of hyphae possibly including germ tubes was measured (10 hyphae per plate) after 24 h incubation. Each treatment was replicated three times (three tubes per treatment) and the entire test was repeated twice using different cultures. The virulence of conidia from the isolated colonies against WFT

larvae was investigated using a leaf dipping method in laboratory conditions (Butt & Goettel, 2000). Conidia from the non-paired ERL1578 and ERL1576 colonies Tyrosine-protein kinase BLK served as positive controls. Conidial suspensions were adjusted to 1 × 106 conidia mL−1 using 0.08% siloxane solution as a wetting agent. A siloxane solution (0.08%) served as a negative control. WFT were continuously reared on green beans, Phaseolus vulgaris L. at 25 ± 1 °C and a 16:8 (L/D) photoperiod with 40–50% relative humidity in wooden chambers (45 × 30 × 30 cm) in an insectary at the Entomology Research Laboratory, University of Vermont. Fresh green bean leaves were aseptically cut into 35 mm diam. circles using a cork borer sterilized with 70% ethanol. Three leaf discs were dipped for 10 s in a conidial suspension (15 mL) in a 35-mm Petri dish and dried at room temperature (c. 25 °C) for 20 min. All discs were placed on moistened filter papers (50 μL sterile distilled water per 35 mm diameter paper) in the lids of 35-mm Petri dishes (one disc/lid). Using an aspirator, 15 thrips 2 days old were placed on each leaf disc in the lid of a Petri dish.

Conidia from all colonies, which were incubated for 10 days, were photographed under a phase-contrast microscope (400×) at the same light exposure. To compare the levels of light-penetrating activity of conidia, which was observed under a phase-contrast microscope, a densitometric analysis was

used to generate a relative densitometric value (RDV) of conidia. In each observation of the photographed conidia, the densitometric values at four spots of background (DV0) and four spots of conidia (DV1), randomly taken, were converted to RDV as follows: RDV = DV1/DV0. The highest RDV was arbitrarily given to 1.00 to compare it with the other RDVs. All conidial suspensions (c. 5 × 106 conidia mL−1) were transferred to fresh Eppendorf tubes (500 μL per click here tube) and held in a water bath at 45 °C for 30, 60, 90 and 120 min. For each strain treatment (non-paired and paired), controls (non-exposed this website conidial suspensions) were kept at room temperature (c. 25 °C). A 10-μL sample

was taken from each tube and dropped on ¼SDAY medium for a germination test prior to and after the exposures. After incubation of all plates at 20 °C for 24 h, percent germination was determined by randomly counting the number of germinated and ungerminated conidia among 100 counts microscopically (400×). A conidium was considered germinated if a germ tube was longer than the length of a conidium (Avery et al., 2004). In addition, the length of hyphae possibly including germ tubes was measured (10 hyphae per plate) after 24 h incubation. Each treatment was replicated three times (three tubes per treatment) and the entire test was repeated twice using different cultures. The virulence of conidia from the isolated colonies against WFT

larvae was investigated using a leaf dipping method in laboratory conditions (Butt & Goettel, 2000). Conidia from the non-paired ERL1578 and ERL1576 colonies Interleukin-3 receptor served as positive controls. Conidial suspensions were adjusted to 1 × 106 conidia mL−1 using 0.08% siloxane solution as a wetting agent. A siloxane solution (0.08%) served as a negative control. WFT were continuously reared on green beans, Phaseolus vulgaris L. at 25 ± 1 °C and a 16:8 (L/D) photoperiod with 40–50% relative humidity in wooden chambers (45 × 30 × 30 cm) in an insectary at the Entomology Research Laboratory, University of Vermont. Fresh green bean leaves were aseptically cut into 35 mm diam. circles using a cork borer sterilized with 70% ethanol. Three leaf discs were dipped for 10 s in a conidial suspension (15 mL) in a 35-mm Petri dish and dried at room temperature (c. 25 °C) for 20 min. All discs were placed on moistened filter papers (50 μL sterile distilled water per 35 mm diameter paper) in the lids of 35-mm Petri dishes (one disc/lid). Using an aspirator, 15 thrips 2 days old were placed on each leaf disc in the lid of a Petri dish.

OMV components synergistically modulate the host immune response. The single most abundant immune stimulating component in OMVs is LPS. Munford et al. (1982) showed that purified LPS from bacteria and vesicular LPS have the highest biological activity, whereas bacteria-associated LPS is less active. In addition to LPS, OMVs contain immune-stimulating PAMPs such as outer membrane porins, flagellins and peptidoglycans (Renelli et al., 2004; Bauman & Kuehn, 2006). These immune activating ligands in Gram-negative pathogens interact with host cells and promote proinflammatory activities (Tufano et al., 1994; Galdiero et al., click here 1999; Ellis & Kuehn, 2010; Kulp

& Kuehn, 2010). However, whether the innate immune response induced by OMVs from different bacterial species stimulates the clearance of bacteria or enhances pathogen virulence remains to be determined. Klebsiella

pneumoniae OMVs did not induce direct cytotoxicity in HEp-2 or U937 cells, but induced a proinflammatory response in vitro. Neutropenic mice were inoculated intratracheally with 20 μg of K. pneumoniae OMVs to determine whether K. pneumoniae OMVs induced lung pathology in vivo. Immunocompromised mice were used, because K. pneumoniae usually infects critically ill or immunocompromised patients. As a control, 1 × 107 CFU of K. pneumoniae ATCC 13883 were inoculated. The control mice treated with PBS showed normal lung histology (Fig. 4a), whereas live bacteria induced pathological changes in lung tissues, including congestion, oedema, collapse of alveoli Ivacaftor in vitro and a mild lymphocytic infiltration (Fig. 4b). Klebsiella pneumoniae OMVs induced more severe pathological changes as compared with live bacterial

infection (Fig. 4c). These results suggest that K. pneumoniae OMVs can induce lung pathology in vivo. The present study demonstrated that K. pneumoniae OMVs induce the innate immune response in vitro and induce lung pathology in vivo. Klebsiella pneumoniae OMVs induced neither cytotoxicity in both HEp-2 and U937 cells in vitro nor cell death in lung tissues in vivo. Instead, K. pneumoniae until OMVs induced expression of proinflammatory cytokine genes. Proinflammatory cytokines, IL-1β and IL-8, function as a mediator of local inflammation and recruit neutrophils and monocytes to sites of infection. Inflammatory cell infiltration was not prominent in mice treated with K. pneumoniae OMVs, because neutropenic mice were used. However, pathological changes of lung tissues were seen following intratracheal inoculation of K. pneumoniae OMVs. These results suggest that K. pneumoniae OMVs induce a strong innate immune response. In conclusion, we have shown that K. pneumoniae OMVs serve as a strong immune modulator to induce an inflammatory response, but do not serve as a transport system for toxic elements to host cells. Our results extend the role of OMVs in the pathogenesis of K.

OMV components synergistically modulate the host immune response. The single most abundant immune stimulating component in OMVs is LPS. Munford et al. (1982) showed that purified LPS from bacteria and vesicular LPS have the highest biological activity, whereas bacteria-associated LPS is less active. In addition to LPS, OMVs contain immune-stimulating PAMPs such as outer membrane porins, flagellins and peptidoglycans (Renelli et al., 2004; Bauman & Kuehn, 2006). These immune activating ligands in Gram-negative pathogens interact with host cells and promote proinflammatory activities (Tufano et al., 1994; Galdiero et al., BVD-523 1999; Ellis & Kuehn, 2010; Kulp

& Kuehn, 2010). However, whether the innate immune response induced by OMVs from different bacterial species stimulates the clearance of bacteria or enhances pathogen virulence remains to be determined. Klebsiella

pneumoniae OMVs did not induce direct cytotoxicity in HEp-2 or U937 cells, but induced a proinflammatory response in vitro. Neutropenic mice were inoculated intratracheally with 20 μg of K. pneumoniae OMVs to determine whether K. pneumoniae OMVs induced lung pathology in vivo. Immunocompromised mice were used, because K. pneumoniae usually infects critically ill or immunocompromised patients. As a control, 1 × 107 CFU of K. pneumoniae ATCC 13883 were inoculated. The control mice treated with PBS showed normal lung histology (Fig. 4a), whereas live bacteria induced pathological changes in lung tissues, including congestion, oedema, collapse of alveoli buy Sorafenib and a mild lymphocytic infiltration (Fig. 4b). Klebsiella pneumoniae OMVs induced more severe pathological changes as compared with live bacterial

infection (Fig. 4c). These results suggest that K. pneumoniae OMVs can induce lung pathology in vivo. The present study demonstrated that K. pneumoniae OMVs induce the innate immune response in vitro and induce lung pathology in vivo. Klebsiella pneumoniae OMVs induced neither cytotoxicity in both HEp-2 and U937 cells in vitro nor cell death in lung tissues in vivo. Instead, K. pneumoniae Lonafarnib price OMVs induced expression of proinflammatory cytokine genes. Proinflammatory cytokines, IL-1β and IL-8, function as a mediator of local inflammation and recruit neutrophils and monocytes to sites of infection. Inflammatory cell infiltration was not prominent in mice treated with K. pneumoniae OMVs, because neutropenic mice were used. However, pathological changes of lung tissues were seen following intratracheal inoculation of K. pneumoniae OMVs. These results suggest that K. pneumoniae OMVs induce a strong innate immune response. In conclusion, we have shown that K. pneumoniae OMVs serve as a strong immune modulator to induce an inflammatory response, but do not serve as a transport system for toxic elements to host cells. Our results extend the role of OMVs in the pathogenesis of K.

, 2009), and N. devanaterra was cultured in acidic (pH 4.5) freshwater medium as described by Lehtovirta-Morley et al. (2011). The media for AOA contained ammonium chloride at concentrations of 1 mM for N. maritimus and 0.5 mM for N. devanaterra. Media were inoculated with 1% or 10% (v/v) of exponential-phase cultures of AOB or AOA, respectively. Bacterial cultures were sampled (1 mL) at intervals of 8 h for 5 days, and archaeal cultures were sampled daily for 10 days. Photoinhibition was investigated in controlled temperature chambers maintained at 26 °C and illuminated by compact fluorescent lights (55 W) and clear strip lights (30 W) (International Lamps Ltd, Hertford, UK) emitting

light with a wavelength spectrum of 400–680 nm with a maximum selleck chemicals intensity at approximately 580 nm. Ammonia-oxidizing activity of the different cultures was measured under continuous illumination at an intensity of either 15, 60 or 500 μE m−2 s−1 and with diurnal cycles of 8-h light (15 or 60 μE m−2 s−1) and 16-h dark conditions. Control cultures were incubated in the dark in the same incubator. Triplicate cultures were grown for all light treatments and controls. Light intensities were selected

BMN-673 to reflect conditions prevailing in riparian zones of rivers and lakes, with highest light intensity (500 μE m−2 s−1) simulating naturally occurring conditions during a clear summer day in open areas and the lower intensities (60 and 15 μE m−2 s−1) simulating conditions in shaded areas. Ammonia-oxidizing activity was determined by measuring Megestrol Acetate increases in nitrite () concentration over time for each particular culture and light exposure treatment. Specific growth rate was estimated by linear regression during the linear phase of semi-logarithmic plots of nitrite concentration vs. time, as in previous studies (Powell & Prosser, 1992; Könneke et al., 2005; Lehtovirta-Morley et al., 2011). Estimated specific growth rates in control and illuminated cultures were compared using the Student’s t-test (two-sample

assuming unequal variances). All AOA and AOB strains grew exponentially during incubation in the dark. Initial increases in nitrite concentration were sometimes non-exponential, because of carryover of nitrite with inocula, but subsequent increases in nitrite concentration were exponential. Typical nitrite production kinetics are exemplified in Fig. 1 for cultures of N. multiformis and N. devanaterra under continuous light at 60 μE m−2 s−1 and dark controls. Nitrite production kinetics were analysed prior to limitation by reduction in pH (all strains except N. devanaterra) or high nitrite concentration (N. devanaterra). Continuous illumination at 60 μE m−2 s−1 reduced the specific growth rate of N. multiformis from 1.05 (±0.07) day−1 to 0.62 (±0.01) day−1 and completely inhibited that of N. devanaterra. Effects of illumination and associated statistical analysis are summarized in Fig. 2 and Table 1, respectively. AOA were more sensitive to illumination than AOB.

Viewed under a scanning electron microscope, the infiltrant material appeared to cover the adjacent apparently sound enamel more thickly and evenly compared with the MIH lesion surface, and although some surface porosities were still evident, these were less frequent and narrower than those on non-infiltrated MIH lesions (Fig. 2). These initial results demonstrate that caries infiltrant materials are capable of penetrating developmentally hypomineralised MAPK inhibitor enamel; however, this occurs in an inconsistent manner and is not as extensive as reported in carious lesions[7]. Based

on MIH characterisation studies, the pattern of infiltration is not explained easily by mineral content or porosity variation, indicating different lesion characteristic/s determine penetrability; with protein content a probable candidate. The failure of NaOCl pre-treatment to produce consistent or significantly improved results means consideration Tigecycline clinical trial must be given to other enamel properties but could also reflect that only the surface proteins are removed,

that this is not the most efficacious agent for the particular proteins present or, be a result of cross-linking by formaldehyde during sterilisation inhibiting protein removal. The recommended etch time is based on that required to penetrate the relatively hypermineralised surface layer of carious lesions: in MIH, this surface layer may have different properties, and the standard etching may be insufficient to allow full access to the lesion. The clinical history these of the teeth used in this study is unknown but use of remineralising agents, common in MIH management, and time in the oral environment

may influence surface layer properties or enamel penetrability. The inherent variability of MIH lesions may also be a confounding factor in achieving significant differences, particularly in terms of microhardness and given the small sample size. Similarly, given reports of higher protein content in brown lesions[13], different colour grouping of the lesions may yield different results; however, there were insufficient brown lesions for statistical analysis in this study. The surface changes observed under SEM confirm that microporosities in defective enamel can be occluded, although perhaps only partially. The sealing of surface defects and inter-rod diffusion pathways could reduce the susceptibility of the enamel to caries. This improved enamel seal may also reduce irritation to the pulp which may in turn decrease pulpal inflammation and sensitivity to evaporative, thermal, and osmotic stimuli common in MIH.

07 to 1.40). After the AED training, 70 officers absolved a resuscitation drill with all 4 AEDs (in total 280 drills). The mean time period between switching on the device and shocking was 75.8 seconds

(SD: ±21.8 seconds). The mean time from switch on until start of ECG analysis ranged from 51.1 seconds (HeartSave AED-M) to 63.8 seconds (AED Plus) (Figure 2). According to the questionnaire, the officers were pleased with the user-friendliness of the AEDs; it was easier to open the cover of HeartStart FR2+ and Defi FRED easy than of the other two; furthermore, the officers had no problems switching on the AEDs (mean from 1.07 to 1.62), recognizing Crizotinib molecular weight the shock button (mean from 1.07 to 1.39), and pressing the shock button (mean from 1.11 to 1.24). The comprehensibility of the AEDs Selleckchem Sirolimus was also favorably evaluated; the seafarers

had no problems understanding the voice prompts acoustically (mean from 1.14 to 1.50), the meaning of the German voice prompts (mean from 1.43 to 1.87), or the screen messages (mean from 1.44 to 1.87). The seafarers found the electrodes easy to unwrap (mean from 1.33 to 2.00). The electrodes’ illustrations of AED Plus were unclear and caused problems to find the correct anatomical positioning (mean 3.6). Furthermore, some officers had problems connecting the electrodes with the HeartSave AED-M (mean 2.9). In the free text in the questionnaire, the seafarers stated the strengths and weaknesses of the different AEDs. The major aspects of criticism given by at least 10% of the officers are summarized in Table 1. While 25 seafarers appreciated the pictogram instructions

of AED Plus, 19 regarded them as confusing. Concerning the one-piece electrode of AED Plus, 23 seafarers noted having problems finding the correct anatomical position on the basis of the AED’s figure drawing (mean 2.06). Compared with two-piece electrodes, 40 seafarers (57.1%) preferred the one-piece one for cardiopulmonary resuscitation because the feedback on the depth and frequency of thorax compressions was regarded as helpful. Germany is the first flag state that legally requires merchant seagoing ships to carry an AED. Thus, it is of interest to the community of scientists and health care providers in maritime medicine to get information from the German experience. BCKDHB Our results demonstrate that 81.7% of the nautical officers delivered an effective defibrillation shock without training in the handling of AEDs. After resuscitation training, all ship officers shocked effectively and none of the participants touched the manikin during shocking. Our results in nautical officers are comparable with other study populations. In a recent study of 236 laypersons, 85.6% were able to deliver a shock by a mean time to shock of 77.5 seconds. After minimal training, 92.8% were able to deliver a shock. The time to shock decreased to 55.0 seconds after training.

07 to 1.40). After the AED training, 70 officers absolved a resuscitation drill with all 4 AEDs (in total 280 drills). The mean time period between switching on the device and shocking was 75.8 seconds

(SD: ±21.8 seconds). The mean time from switch on until start of ECG analysis ranged from 51.1 seconds (HeartSave AED-M) to 63.8 seconds (AED Plus) (Figure 2). According to the questionnaire, the officers were pleased with the user-friendliness of the AEDs; it was easier to open the cover of HeartStart FR2+ and Defi FRED easy than of the other two; furthermore, the officers had no problems switching on the AEDs (mean from 1.07 to 1.62), recognizing Trichostatin A supplier the shock button (mean from 1.07 to 1.39), and pressing the shock button (mean from 1.11 to 1.24). The comprehensibility of the AEDs LBH589 concentration was also favorably evaluated; the seafarers

had no problems understanding the voice prompts acoustically (mean from 1.14 to 1.50), the meaning of the German voice prompts (mean from 1.43 to 1.87), or the screen messages (mean from 1.44 to 1.87). The seafarers found the electrodes easy to unwrap (mean from 1.33 to 2.00). The electrodes’ illustrations of AED Plus were unclear and caused problems to find the correct anatomical positioning (mean 3.6). Furthermore, some officers had problems connecting the electrodes with the HeartSave AED-M (mean 2.9). In the free text in the questionnaire, the seafarers stated the strengths and weaknesses of the different AEDs. The major aspects of criticism given by at least 10% of the officers are summarized in Table 1. While 25 seafarers appreciated the pictogram instructions

of AED Plus, 19 regarded them as confusing. Concerning the one-piece electrode of AED Plus, 23 seafarers noted having problems finding the correct anatomical position on the basis of the AED’s figure drawing (mean 2.06). Compared with two-piece electrodes, 40 seafarers (57.1%) preferred the one-piece one for cardiopulmonary resuscitation because the feedback on the depth and frequency of thorax compressions was regarded as helpful. Germany is the first flag state that legally requires merchant seagoing ships to carry an AED. Thus, it is of interest to the community of scientists and health care providers in maritime medicine to get information from the German experience. Cell press Our results demonstrate that 81.7% of the nautical officers delivered an effective defibrillation shock without training in the handling of AEDs. After resuscitation training, all ship officers shocked effectively and none of the participants touched the manikin during shocking. Our results in nautical officers are comparable with other study populations. In a recent study of 236 laypersons, 85.6% were able to deliver a shock by a mean time to shock of 77.5 seconds. After minimal training, 92.8% were able to deliver a shock. The time to shock decreased to 55.0 seconds after training.

9%) children. Table 2 shows descriptive statistics of the response at the three tests at baseline, and 15 min after the mask had been placed and the inhalation started. No statistically significant difference in reaction time (P = 0.17) was found at baseline between the two sessions (23). N2O/O2 inhalation significantly increased the reaction time with 183 ms (P < 0.001), whereas no effect was found 10 and 30 min after the mask had been

removed. (Table 3) Baseline values for tooth-pulp pain sensitivity were not statistically significantly different between the two sessions (Table 4). At the test 15 min after the mask had been placed and inhalation started, the average value was 92.7 μA during inhalation of N2O/O2 and 54.0 μA during inhalation of atmospheric. This represents a statistically highly significant reduction in tooth-pulp pain sensitivity of 38.7 μA (P < 0.001) (Table 4). Vadimezan in vitro After adjustment for increase in reaction time,

however, this effect could not be demonstrated. No effect was found 10 and 30 min after the mask was removed. Baseline pressure pain thresholds of the masseter muscle did also not show any difference between the two sessions. At the test 15 min after the mask had been placed and inhalation started, the average value was 312.5 kPa during inhalation of N2O/O2ir and 231.7 kPa during inhalation of atmospheric air. This represents a statistically buy Fulvestrant highly significant increase of 80.8 kPa (P < 0.001)

(Table 4). This effect was reduced to 47.8 kPa, but still statistically significant (P < 0.005) after adjustment for increase in reaction time. In contrast to the findings for tooth-pulp pain sensitivity, an effect on pressure-induced muscle pain could still be seen 10 min after the mask had been removed (P = 0.03), even after adjustment for increase in reaction time (P = 0.04). No effect was found 30 min after the mask Thalidomide had been removed. The VAS score for overall discomfort from the two experimental pain tests was almost identical (N2O/O2 inhalation sessions: average: 1.23; SD: 0.19; atmospheric air sessions: average: 1.18, S.D.: 0.18), and the difference is not statistically significant. This finding was not influenced by adjustment for increase in reaction time. The present study has not been able to show any analgesic effect on tooth-pulp sensitivity, after the increase in reaction time caused by the drug has been taken into account. In contrast, an analgesic effect on pressure-induced muscle pain was found, also after adjustment for the increases in reaction time. We opted to assess both tooth-pulp pain sensitivity and jaw muscle pain sensitivity because different oro-facial tissues may have different sensitivity to painful stimuli and different responses to analgesic interventions. Furthermore, both odontogenic types of pain and musculoskeletal pains are frequently encountered in children.

9%) children. Table 2 shows descriptive statistics of the response at the three tests at baseline, and 15 min after the mask had been placed and the inhalation started. No statistically significant difference in reaction time (P = 0.17) was found at baseline between the two sessions (23). N2O/O2 inhalation significantly increased the reaction time with 183 ms (P < 0.001), whereas no effect was found 10 and 30 min after the mask had been

removed. (Table 3) Baseline values for tooth-pulp pain sensitivity were not statistically significantly different between the two sessions (Table 4). At the test 15 min after the mask had been placed and inhalation started, the average value was 92.7 μA during inhalation of N2O/O2 and 54.0 μA during inhalation of atmospheric. This represents a statistically highly significant reduction in tooth-pulp pain sensitivity of 38.7 μA (P < 0.001) (Table 4). PFT�� After adjustment for increase in reaction time,

however, this effect could not be demonstrated. No effect was found 10 and 30 min after the mask was removed. Baseline pressure pain thresholds of the masseter muscle did also not show any difference between the two sessions. At the test 15 min after the mask had been placed and inhalation started, the average value was 312.5 kPa during inhalation of N2O/O2ir and 231.7 kPa during inhalation of atmospheric air. This represents a statistically check details highly significant increase of 80.8 kPa (P < 0.001)

(Table 4). This effect was reduced to 47.8 kPa, but still statistically significant (P < 0.005) after adjustment for increase in reaction time. In contrast to the findings for tooth-pulp pain sensitivity, an effect on pressure-induced muscle pain could still be seen 10 min after the mask had been removed (P = 0.03), even after adjustment for increase in reaction time (P = 0.04). No effect was found 30 min after the mask find more had been removed. The VAS score for overall discomfort from the two experimental pain tests was almost identical (N2O/O2 inhalation sessions: average: 1.23; SD: 0.19; atmospheric air sessions: average: 1.18, S.D.: 0.18), and the difference is not statistically significant. This finding was not influenced by adjustment for increase in reaction time. The present study has not been able to show any analgesic effect on tooth-pulp sensitivity, after the increase in reaction time caused by the drug has been taken into account. In contrast, an analgesic effect on pressure-induced muscle pain was found, also after adjustment for the increases in reaction time. We opted to assess both tooth-pulp pain sensitivity and jaw muscle pain sensitivity because different oro-facial tissues may have different sensitivity to painful stimuli and different responses to analgesic interventions. Furthermore, both odontogenic types of pain and musculoskeletal pains are frequently encountered in children.