Parking was available for a fee and a limited volunteer driver program was offered to patients who could not otherwise access the hospital. The pulmonary rehabilitation program followed a standard format (Nici et al 2006), with seven weeks of twice-weekly group exercise and self-management education sessions. The exercise component was individually prescribed and consisted of 30 minutes of aerobic training (walking and exercise bike) with intensity progressed weekly, and resistance training using functional tasks such as step ups and sit to stand. Sessions were conducted in the morning. Patients were included in the study if they had a diagnosis of COPD and were aged 18 years or over. Patients were excluded if

they did not speak English and selleck LY2835219 price could not participate in an interview. Individuals who were eligible to take part were contacted by an independent investigator not involved in delivery of the clinical program who provided written information and obtained consent. Nine interview questions were developed (Box 1) and reviewed by two experts in the delivery of pulmonary rehabilitation programs. The questions allowed exploration of possible reasons for and individual experiences associated with non-attendance and non-completion. All participants who undertook the semi-structured interview were given the option of doing it at their home or over the telephone. Interviews were recorded

and took 20–40 minutes to complete. Researcher triangulation was employed, with interviews conducted by one of two researchers (AK or AH) in order to reduce the potential almost for bias (Patton 1999).

Researchers were encouraged to make observational memos for use during analysis (Boije 2010). Each interview was transcribed verbatim by a single researcher. If clarification was needed on the content or meaning of an interview the participant was contacted to review the information. Demographic information collected directly from participants and from their medical record was gender, age, body mass index (BMI), lung disease severity using the Global Initiative for Obstructive Lung Disease (GOLD) criteria (Rabe et al 2007) based on recent (within six months) spirometry, smoking status, home oxygen use, living situation, comorbidities score (Charlson et al 1987) and distance between their home and the pulmonary rehabilitation venue. 1. Who suggested that you might attend a pulmonary rehabilitation program? De-identified interview transcripts were examined independently by two researchers (AK and AH). Line-by-line iterative thematic analysis (Boyatzis 1998) of the transcribed interviews took place, where descriptive codes were devised to represent the data. Three rounds of coding were used. Open coding commenced during data collection and was used to compile a hierarchical coding scheme. Axial coding was then used to refine and delineate the relationship of themes to subthemes.

These data were corroborated by in vivo experiments using IRF3/7 double-deficient mice. Whereas c-di-GMP treatment elicited Type 1 IFN in wild-type B6 mice, IRF3/7 double-deficient

mice produced very little Type 1 IFN. In fact, while a single immunization with human serum albumin (HSA) + c-di-GMP elicited HSA-specific antibodies in B6 mice, this response was virtually undetectable in IRF3/7 double knockout mice [44]. McWhirter et al. postulated that since the transcriptional responses after c-di-GMP and cytosolic DNA are similar, this may add value to the use of c-di-GMP as a small molecule adjuvant. Since c-di-GMP is nonself and non-DNA, it is able to induce similar responses as DNA without the risk of autoimmune attack or mutagenic potential associated with DNA vaccines [44]. There is a largely unmet requirement Capmatinib for safe and effective vaccine adjuvants. In fact, only a few adjuvants have been approved for use in humans and as such the development of novel adjuvants and immunostimulatory agents to enhance the http://www.selleckchem.com/products/LY294002.html innate immunity and vaccine efficacies is a high priority. The fortuitous discovery of c-di-GMP and its ability to stimulate the

host immune response has jumpstarted research to investigate its potential adjuvanticity. The initial evidence suggesting the possibility of using c-di-GMP as a mucosal adjuvant is particularly exciting since mucosal immunization poses its own set of challenges. Nevertheless, another group of small synthetic molecules, CpG-ODNs, have generated a great deal of excitement as mucosal vaccine adjuvants and a number of vaccines containing CpG-ODN are currently in clinical trials [45]. c-di-GMP may represent another candidate with equal promise as a vaccine Fossariinae adjuvant. It has been less than 5 years since the immunostimulatory properties of c-di-GMP were first observed. During the past 5 years, few laboratories have examined

the potential for c-di-GMP as a vaccine adjuvant. However, with the promising data that have come out from these studies, interest in this bacterial signaling molecule has quickly grown. Over the next few years, more data is needed to support the protective efficacy of c-di-GMP in its capacity as a potential vaccine adjuvant and both c-di-GMP immunogenicity and adjuvanticity must be evaluated in other species. In addition, understanding the mechanism underlying c-di-GMP stimulation of the host response is an important step towards the successful application of c-di-GMP as a vaccine adjuvant. Also, although some preliminary data indicate that there is no lethal cytotoxicity in normal rat kidney cells or human neuroblastoma cells as well as no adverse toxigenic or carcinogenic effects in vitro [19] and [26], the in vivo safety profile for c-di-GMP must be assessed and there is some concern that its potent immunostimulatory properties may in fact lead to excessive tissue inflammation.

The magnitude of proliferation was similar across groups: 25 subjects had an SI > 5 and 12 subjects had an SI > 10 at any time-point compared with baseline. Proliferative responses of greatest magnitude (SI > 10) across dose groups were elicited by HBcAg. The frequency of ASCA responders was low, although there were more responders in Cohort A (seven subjects, 12%) than Cohort B (one subject, 2%). There was also a slight trend toward higher IgA and IgG levels

in Cohort A. The total number of responders (IgA plus IgG) was the highest in Cohort A 80 YU (five subjects, 8%). Generally, IgA and IgG levels were low at baseline with only six subjects showing a baseline response ≥25 U. These low levels were maintained during treatment. Seven of the eight ASCA responders were also defined as responders in the ELISpot. In addition, for 80 YU, MLN0128 cost all ASCA responders also displayed ELISpot and LPA responses. No anti-HBcAg antibodies were detected at any time during the study and no anti-HBsAg antibody levels >8.4 IU/mL

were determined. Two subjects, both in Cohort A 80 YU, had anti-HBsAg levels ≥3.5 IU/L during the study. HLA testing was performed to evaluate for any HLA restriction of immune responses to GS-4774. The most frequent HLA alleles were A*02, C*07, DQB1*03, and DRB1*04. No association was found between common HLA alleles and the IFN-γ ELISpot Dinaciclib purchase response to peptides or recombinant antigens (Supplementary Table 7). In the present study, GS-4774 was generally safe and well-tolerated. The most common adverse events were injection-site reactions. Adverse events occurred more frequently in both cohorts of the highest dose group, 80 YU, and the number of individual adverse events was higher after weekly than monthly immunization. Immunization with GS-4774 led to HBV antigen-specific and treatment-emergent T-cell responses. The majority much of subjects showed a response when assessed by at

least one of the assays. GS-4774 was immunogenic at all three doses tested and both immunization regimens, weekly and monthly dosing, induced T-cell-mediated immune responses. Immunogenicity was independent of HLA alleles. LPA responses were observed in the majority of subjects with no increase in the frequency of responders related to dose or timing of dose. LPA responses were measured using recombinant HBV proteins which preferentially utilize an MHC Class II pathway resulting in a bias toward CD4+ T-cell activation [12]. The responses, therefore, may represent early CD4+ T-cell activation with GS-4774 in these subjects. The higher magnitude LPA responses with SI > 5, breadth of proliferative responses to recombinant antigens, and timing of response emergence suggested an increase in LPA responses from 10 to 40 YU doses but not from 40 to 80 YU. IFN-γ ELISpot responses were seen in fewer subjects and at later time-points than LPA responses.

The median age and time since injury were 27 years (IQR 24 to 31) and 11 weeks (IQR 8 to 16), respectively. According to the International Standards for Classification of Spinal Cord Injury, participants were categorised as American Spinal Injury Association Impairment Scale (AIS) A (n = 29), AIS B (n = 2), or

AIS C (n = 1) with neurological and motor levels ranging from T1 to L1 (see Table 1). The groups were similar at baseline. Adherence to the study protocol was reasonable. The protocol dictated that participants receive 18 training sessions over six weeks. In reality, they received a median of 18 training sessions (IQR 12 to 18) over 6 weeks (IQR 6 to 7). There were four participants from the Sydney site who received only six (1 participant), 11 (2 participants), or 12 (1 participant) sessions due to poor compliance, and one participant from the Bangladesh Selleckchem Everolimus site who received only five sessions due to back pain. All three assessors indicated that blinding had been maintained throughout Venetoclax research buy the study. The mean between-group difference for the Maximal Lean Test was –20 mm (95% CI –64 to 24). The mean betweengroup difference for the Maximal

Sideward Reach was 5% of arm length (95% CI –3 to 13). The mean betweengroup difference for the Performance item of the COPM was 0.5 points (–0.5 to 1.5). Group data for these outcomes are presented in Table 2. Individual data are presented in Table 3 (see eAddenda for Table 3). None of these findings was statistically significant and the upper end of all 95% confidence intervals fell short of the pre-determined minimally worthwhile treatment effects. The corresponding values for the secondary outcomes are also presented in Table 2. Individual data are presented in Table 3 (see eAddenda for Table 3). The results of the exploratory perprotocol analysis of all outcomes are presented in Table 4. The only notable deleterious effect was an increase in

back pain in one participant. The median rating of inconvenience of the intervention provided by experimental participants was 9 (IQR 8 to 9) where 1 was ‘extremely inconvenient’ and 10 was ‘not at all inconvenient’. The results of this study indicate no added benefit MycoClean Mycoplasma Removal Kit from a 6-week training program specifically targeting unsupported sitting. We can be confident that within the limitation of this study, the results are conclusive because the upper end of the 95% CIs from the three primary outcomes falls short of the pre-determined minimally worthwhile treatment effects. These findings are largely consistent when data from the five non-compliant experimental participants are removed although there is less precision and certainty associated with some outcomes. Needless to say, the interpretation of the results relies on what is considered a worthwhile treatment effect.

The WG was established in December 2004, just before Merck applied for a biologics

license from the FDA for their vaccine, RotaTeq®, in April 2005. Shortly after the FDA approved the license on 3 February 2006, ACIP voted on the vaccine on 21 February 2006. On 11 August 2006 the MMWR published a statement entitled Prevention of Rotavirus Gastroenteritis among Infants and Children, which constituted formal approval of the vaccine and its inclusion in the vaccination schedule [10]. Beginning in June 2007, the WG expanded it focus to include consideration of a new rotavirus vaccine, Rotarix® (Glaxo-Smith-Kline), which was ultimately licensed by FDA in April 2008. From June 2007 until February 2009, the WG met at least once monthly, and often bi-monthly in preparation for data presentations at ACIP meetings. The WG, comprising 25 members, included CDC subject matter experts; immunization safety experts; ACIP Vandetanib cost members, ex officio members and liaison Tanespimycin molecular weight representatives, and invited academic consultants. At every ACIP meeting from June 2007 until June 2008 (four meetings), the WG presented information on efficacy and safety of Rotarix®, RotaTeq® vaccine coverage and adherence with age recommendations, draft proposed recommendations for use of Rotarix®, post-licensure safety monitoring of RotaTeq®, and final recommendations for use

of Rotarix® following licensure by FDA. The ACIP voted in June 2008 to add Rotarix® to the routine infant immunization schedule, and provided guidance on use of Rotarix® vs. RotaTeq®, since there were now two licensed vaccines on the market. The WG finalized the full ACIP statement, which was published in the MMWR in February 2009 [11]. The WG has been disbanded for now, but CDC program staff continue to monitor rotavirus vaccine coverage rates, rotavirus disease rates, vaccine coverage, and vaccine safety. The WG can be reassembled at any time, if necessary. For all newly licensed and recommended vaccines, ACIP members are briefed during meetings on changes in disease epidemiology that occur following

introduction of a vaccine, and this has been the case with rotavirus vaccines. At meetings following the 2006 and 2009 recommendations for the use of RotaTeq® and Rotarix®, ACIP members were informed also about the reduction in rotavirus disease burden in the US from 2000 through 2009—the 2007–2008 and 2008–2009 rotavirus seasons were shorter, later, and characterized by substantially fewer positive rotavirus test results reported to the national surveillance system compared to the pre-vaccine era (overall number of positive test results decreased by 64% from 2000–2006 to 2007–2008) [12] and [13]. With presentations on the surveillance and epidemiology of vaccine-preventable diseases following changes in national immunization policy, the ACIP is kept informed about the impact of vaccination on the target population.

Bra knowledge – the primary outcome – was measured using a custom-designed, 50-item, self-administered questionnaire. Details of the questions

which covered bra design, bra component parts, bra sizing, as well as correct and incorrect bra fit and bra wearing habits, can be found in Appendix 1 (see eAddenda for Appendix 1). Responses included multiple choice options, true/false, and short answers; an ‘I do not know’ response was offered for every question. Face validity was verified through focus groups. Bra fit was measured using the Bra Fit Assessment test (Choice Magazine 2005) as pass/fail. To be ranked a pass, the front band had to be in contact with the sternum; the posterior and side band had to have no flesh bulging above its superior edge (too small) and was not EGFR inhibitor to move upward if the arms were raised above the head three times (too big); the cup had to have no aspect of the breast bulging above its superior

or medial edge (too small) and no wrinkles in the cup material (too big); the straps were not to be digging into (too small) or slipping off (too big) the shoulders; and the cup underwire had to be resting on the ribs and sternum, not on any breast tissue. If one or more of these six components were ranked a ‘fail’ grade in fit, and the straps or the band could not be adjusted by the assessor to achieve correct fit, an overall ‘fail’ grade was awarded in BMS-354825 in vitro the Bra Fit Assessment test. Level of breast support was measured using the Level of Breast Support test as pass/fail. To be ranked a pass for design, the bra had to be a sports bra, or any two bra combination for any bra size, or a crop top only for cup sizes A or B. Lifespan was ranked

a fail (too old) if the material/elastic or underwire of any bra, of any design, had deteriorated. Both bra design and lifespan had to pass for an overall ranking of pass in the Level of Breast Support test. Discomfort during exercise was measured using a 10-cm visual analogue isothipendyl scale where participants were asked to rate their breast discomfort when wearing this bra during sport. Bra knowledge was calculated as the mean (SD) percentage of correct answers, while lack of bra knowledge was calculated as the mean (SD) percentage of ‘I do not know’ answers. Number of participants passing the Bra Fit Assessment and Level of Breast Support tests was reported. Analysis was by intention-to-treat, whereby all participants were analysed in the groups that they were randomised to and all available data were included in the analysis. Statistical significance was set at p < 0.05, so mean difference (95% CI) or risk difference (95% CI) between groups are presented. Four sporting academies agreed to participate. Three academies declined due to time constraints of their teams and coaches.

In case of detection of amylase, the starch agar medium plate was flooded with 1% iodine solution, to observe the zone of hydrolysis. The bacterium, 2b, found to produce maximum zone of hydrolysis around the colony on the casein agar medium and on starch agar medium was selected for further study. The isolate was maintained Selleckchem Luminespib on Horikoshi medium slants (pH 10.0) and stored at 4 °C. The morphological characteristics of the selected isolate 2b obtained

on Horikoshi’s –I (pH 10.0) agar plates were studied. The shape, size and arrangement of the cells were studied in Gram-stained preparations. Endospore staining was carried out according to the method of Schaeffer and Fulton.8 Motility of 12 and 24 h old cells was observed by phase contrast microscopy of hanging-drop preparations. Growth experiments at pH 7–11 were performed on Horikoshi I broth adjusted to various pH CP-673451 ic50 values: pH 7–9 (adjusted by adding NaHCO3) and pH 10–11 (adjusted by adding). Growth at various NaCl concentrations (2–10%) and at various temperatures (4–55 °C) was investigated in Horikoshi I broth (pH 10.0). Acid production from carbohydrates was determined by the method of using thymol blue instead of bromothymol blue at pH 10.0 9 and 10. Physiological and biochemical tests such as indole production from tryptophan, methyl-red and Voges–Proskauer

tests, Simmons’ citrate utilization test, catalase and oxidase activity, urea hydrolysis, production of H2S from cysteine, nitrate reduction to nitrite, hydrolysis of casein, gelatin and starch were examined

according to Smibert and Krieg.11 The taxonomic status of the selected bacterium 2b was identified following the criteria laid down by Bergey’s Manual of Systematic Bacteriology.12 The identification was further confirmed by Microbial Type Culture Collection Center and Gene Bank (MTCC), Institute of Microbial Technology, (IMTECH), Chandigarh, India. The 16S RNA gene sequencing of the isolate was performed by National Center for Cell Sciences (NCCS), Pune, India. The purified PCR product of 16sr RNA was sequenced using ABI Prism. The sequence obtained was BLAST searched Rutecarpine and compared with sequences of other closely related members of genus Bacillus retrieved from GenBank database. Phylogenetic tree was constructed from 16S rRNA gene sequences of members of genus Bacillus using neighbour-joining method. 13 The analysis involved 39 nucleotide sequences of genus Bacillus. The sequence so obtained was taken up for running NCBI BLAST against nonredundant nucleotide database using megablast algorithm for getting homologous sequences14 and 15. Sequences showing a relevant degree of similarity were imported into the CLUSTAL W program16 and multiple sequence alignment was performed. Phylogenetic trees were constructed by different treeing algorithms: neighbour-joining,13 maximum parsimony tree17 and maximum-likelihood18 and UPGMA method19 using MEGA5.

Pooled sera from mice immunized with two doses of 1 μg PCV7 served as the quality control. Goat anti-mouse HRP conjugate was purchased from Southern Technologies (Birmingham, AL). To measure total functional antibodies, a standard opsonophagocytic assay (OPA) described by Romero-Steiner et al. [31] and [32] was utilized. Titers were calculated as the reciprocal dilution at which ≥50% bacterial killing occurred this website in comparison

to complement control wells. To assess differences in functional activity due to species specific phagocytic cells, an alternative OPA protocol using Raw 264.7, mouse monocytes (ATCC) and guinea pig complement (MP Biomedicals, Solon, OH) was also evaluated [15], [33] and [34]. A week after administering

the last dose, mice were intranasally challenged with approximately 1 × 106 CFU of log phase S. pneumoniae serotype 4, 14, or 19A suspended in 10 μl PBS. Challenge doses were later confirmed by counting the overnight growth of a 10-fold serial diluted challenge inoculum [18]. Three to five days post-challenge, each mouse was euthanized and its nasopharyngeal (NP) cavity washed as described by Moreno et al. [26] and Wu et al. [35]. As seen in the study by Moreno et al., control mice significantly cleared pneumococci six days post intranasal challenge [26]. In this study, we found three to five days post-challenge to be the optimal time point in detecting a difference between control and immunized mice. NP washes Casein kinase 1 (100 μl) were collected, diluted with equal volume of saline, and further serially diluted, Vemurafenib in vivo 3-fold, an additional five times in a 96-well plate. Fifty microliters of each dilution was cultured on blood agar plates supplemented with 2.5 mg/L gentamicin. In preliminary studies, mice cleared serotypes 4 and 19A within 4 days and serotype 14 within 5 days post-challenge. Because of these results, NP washes were conducted 3 days post-challenge of serotype 4 or 19A and 4 days post-challenge

with serotype 14. As previously defined, carriage values are the average count of Pnc colony-forming units (cfu) collected in 50 μl of nasal wash [18]. Counts were adjusted for dilution factors prior to averaging. Antibody concentrations were calculated with a 4-parameter logistic equation (ELISA for Windows, CDC). Mean or geometric mean of OPA titers (with log-transformation) and colony counts were calculated. Significant differences, P ≤ 0.05, were determined between two groups using Mann–Whitney rank sum test or t-test, within an experiment using one way analysis of variance on ranks, and for multiple pairwise comparisons using the Student–Newman–Keuls method (SigmaStat software version 2.0; Jandel scientific, Point Richmond, CA). To examine the effect of PCV7 + PsaA co-administration on IgG antibody levels, mouse immune sera were assayed before and after challenge.

The secondary objective was met as the HI antibody responses following the second vaccine dose fulfilled the CHMP criteria in all treatment groups at Day 42 and persisted through Day 182. At Day 42, in subjects who were seronegative at baseline, the seroconversion rates were 95% for those who received a single primary dose of AS03B-adjuvanted 1.9 μg HA vaccine or the non-adjuvanted vaccine, and 100% for those who received two primary doses of the AS03B-adjuvanted 1.9 μg HA vaccine or a single Ruxolitinib solubility dmso primary dose of the 3.75 μg HA AS03A-adjuvanted vaccine.

In subjects who were seropositive at baseline, seroconversion rates ranged from 73.3% for those who received a single primary dose of AS03B-adjuvanted 1.9 μg HA vaccine to 95.5% buy Ulixertinib for those who received a single primary dose of

the 3.75 μg HA AS03A-adjuvanted vaccine (Supplementary Table 1). As observed from the HI antibody GMTs, the highest HI antibody response at Day 182 (pre-booster) was observed for children who received two primary doses of the AS03B-adjuvanted 1.9 μg HA vaccine (GMT [95% CI]: 318.4 [257.8–393.1]), followed by those who received a single primary dose of the 3.75 μg HA AS03A-adjuvanted vaccine (GMT [95% CI]: 240.2 [188.1–306.6]). The HI antibody GMTs (95% CI) in groups that received a single primary dose of AS03B-adjuvanted 1.9 μg HA vaccine or a single primary non-adjuvanted vaccine dose were 176.1 (137.1–226.0) and 177.2 (140.1–224.0). Seven days after booster vaccination (Day 189), with Day 0 as the reference point, SPR, SCR, and GMFR were ≥97.2%, ≥74.6% and ≥12.1, respectively,

in all treatment groups, meeting the CHMP criteria. Using the pre-booster time point as the reference point for computation, the SCR ranged from 10.2% in the non-adjuvanted vaccine group to 28.6% in the group receiving a single primary dose of AS03B-adjuvanted 1.9 μg HA vaccine. GMFR ranged from 1.5 in the non-adjuvanted vaccine group to 2.5 in the AS03A-adjuvanted 3.75 μg HA vaccine group (Table 3). An anamnestic response in all treatment groups was suggested based on the rapid increase in HI antibody Rebamipide GMTs (1.5–2.5-fold increase), 7 days after booster vaccination (Day 189) compared with the pre-booster time point (Day 182) (Table 2). Of all subjects included in the per protocol cohort for immunogenicity, 33 from 5 study centers had not reached seroconversion (either post-vaccination HI antibody titers against the A/California H1N1/2009 strain were <1:40 for subjects who were seronegative at baseline, or post-vaccination HI antibody titers against the A/California H1N1/2009 had increased by less than 4-fold for subjects who were seropositive at baseline) and thus were considered as non-responders to the study vaccine. Of these, 15 subjects were enrolled and vaccinated in four centers in Slovakia and 18 in the center located in Estonia. The distribution of these subjects per study group and center is presented in Supplementary Table 2.

Exclusion criteria were other neuromuscular pathology in the hand (eg, De Quervain’s tenosynovitis, trigger finger), surgical interventions on the carpometacarpal joint, a Beck Depression Inventory score of more than 4 (Wang et al 2005), a State

Trait Anxiety Inventory score of 30 or more (Antunes et al 2005), or any neurological condition in which pain perception was altered (Wajon and Ada 2005). Both interventions were applied by an experienced physiotherapist with a 4-year post-graduate certificate in manual therapy and 11 years of experience in the management of musculoskeletal pain disorders. The experimental group received a neurodynamic nerve slider technique targeted to the radial nerve over the symptomatic hand for 6 sessions over 4 weeks. The technique was applied with the patient positioned in supine and the physiotherapist seated. The technique involved alternating the following two movements: shoulder TSA HDAC depression applied simultaneously with elbow flexion and wrist extension; and shoulder elevation simultaneously with elbow Idelalisib extension, wrist flexion, and ulnar deviation. These movements were alternated at a rate of approximately 2 seconds per cycle (1 second into extension and 1 second into flexion). This technique is intended to produce a sliding movement of neural structures in relation to their adjacent

tissues. Speed and amplitude of movement were adjusted such that no pain was produced. At each session, the technique was applied 3 times for 3 min separated by 1-min rest periods. Participants in the control group received a sham dose of intermittent ultrasound therapy to the thumb region for 10 minutes for 6 sessions over 4 weeks. Further detail of each intervention is available in the primary report of this trial (Villafañe et al 2012a). Pressure pain threshold is a quantitative sensory test of

tissue sensitivity and it is defined as the minimal amount of pressure that produces pain, measured via a pressure algometer (Ylinen 2007). Pressure pain thresholds near to the pathological site are thought to represent the degree of peripheral nociception, whereas pressure pain thresholds distant to the pathology are a marker of central nervous system hyper-excitability (Kamper et al 2011). The validity Tolmetin and reproducibility of algometry has been described, with higher pressure pain thresholds indicating lower pain sensitivity (Fischer 1987). Pressure pain threshold was measured contralaterally over the lateral epicondyle, thumb carpometacarpal joint at the anatomical snuffbox, the tubercle of the scaphoid bone, and the unciform apophysis of the hamate bone. The pressure applied was increased by approximately 0.1 kg/cm2 each second until the onset of pain. Three measurements were obtained from each point and the mean was used for statistical analysis. A 1-min rest period was allowed between each measurement.