Production and characterization of a monoclonal antibody to Francisella tularensis lipopolysaccharide

Having the capacity to detect and identify pathogens that can be employed in a bioterror attack is critical from both a public health and defence perspective. Immunodiagnostic assays are useful tools for enhancing such detection capabilities. In order to develop an immunodiagnostic assay for the detection of Francisella tularensis, a murine monoclonal antibody (MAb) was developed, using the live vaccine strain (LVS) of F. tularensis as the inoculating antigen. A single MAb, F94G2-1, which is specific for the lipopolysaccharide (LPS) of this bacterium was developed and characterized. An indirect ELISA using purified LPS was effective in determining reactivity of the MAb against its target. An immunodotblot and a manually printed antigen microarray were also tested as suitable detection methods. Both assays showed that MAb F94G2-1 has excellent specificity for F. tularensis LPS and demonstrate the utility of using the same MAb in a variety of immunodiagnostic applications.     

Pilot crossover trial of Reiki versus rest for treating cancer-related fatigue

Fatigue is an extremely common side effect experienced during cancer treatment and recovery. Limited research has investigated strategies stemming from complementary and alternative medicine to reduce cancer-related fatigue. This research examined the effects of Reiki, a type of energy touch therapy, on fatigue, pain, anxiety, and overall quality of life. This study was a counterbalanced crossover trial of 2 conditions: (1) in the Reiki condition, participants received Reiki for 5 consecutive daily sessions, followed by a 1-week washout monitoring period of no treatments, then 2 additional Reiki sessions, and finally 2 weeks of no treatments, and (2) in the rest condition, participants rested for approximately 1 hour each day for 5 consecutive days, followed by a 1-week washout monitoring period of no scheduled resting and an additional week of no treatments. In both conditions, participants completed questionnaires investigating cancer-related fatigue (Functional Assessment of Cancer Therapy Fatigue subscale [FACT-F]) and overall quality of life (Functional Assessment of Cancer Therapy, General Version [FACT-G]) before and after all Reiki or resting sessions. They also completed a visual analog scale (Edmonton Symptom Assessment System [ESAS]) assessing daily tiredness, pain, and anxiety before and after each session of Reiki or rest. Sixteen patients (13 women) participated in the trial: 8 were randomized to each order of conditions (Reiki then rest; rest then Reiki). They were screened for fatigue on the ESAS tiredness item, and those scoring greater than 3 on the 0 to 10 scale were eligible for the study. They were diagnosed with a variety of cancers, most commonly colorectal (62.5%) cancer, and had a median age of 59 years. Fatigue on the FACT-F decreased within the Reiki condition (P=.05) over the course of all 7 treatments. In addition, participants in the Reiki condition experienced significant improvements in quality of life (FACT-G) compared to those in the resting condition (P <.05). On daily assessments (ESAS) in the Reiki condition, presession 1 versus postsession 5 scores indicated significant decreases in tiredness (P <.001), pain (P <.005), and anxiety (P<.01), which were not seen in the resting condition. Future research should further investigate the impact of Reiki using more highly controlled designs that include a sham Reiki condition and larger sample sizes.     

Inhibition of pyocyanin-potentiated IL-8 release by steroids in bronchial epithelial cells

Airway epithelial cells are the first targets of environmental stimuli and local cytokines. Pyocyanin-induced synergism with interleukin (IL)-1 or tumour necrosis factor (TNF) in triggering IL-8 release has been documented previously. In this study, IL-8 mRNA and protein expression were examined in cultured human bronchial epithelial cells (BEAS-2B) stimulated with pyocyanin alone, and in combination with IL-1beta or phorbol 12,13-dibutyrate (PDBu) in the absence and presence of a group of glucocorticoids. IL-8 mRNA was measured by RT-PCR, and IL-8 protein by ELISA (cell supernatants). Pyocyanin alone produced no increase in IL-8 mRNA and release. However, pyocyanin upregulated the stimulatory effect of IL-1beta or PDBu on the release of IL-8 in a dose-dependent manner. The stimulatory effect of pyocyanin on the IL-1beta- or PDBu-stimulated IL-8 release was reduced in the presence of dexamethasone, budesonide, and fluticasone. Budesonide and fluticasone were 10-fold more potent than dexamethasone. The protein kinase C (PKC) inhibitor, Go6976, also significantly reduced the stimulatory effect of pyocyanin on IL-1beta, and PDBu increased IL-8 release. In conclusion, this study shows that PKC signal pathway seems to be involved in the pyocyanin-mediated upregulation of the IL-1beta and PDBu-induced IL-8 release in BEAS-2B cells. These findings suggest that a vicious cycle perpetuating inflammation may exist in the biologic milieu of bronchiectatic patients infected with Pseudomonas aeruginosa due to the production of pyocyanin. The priming action of pyocyanin appears to be blocked by glucocorticoids, thus providing in vitro data in support of the clinical efficacy of inhaled glucocorticoids as anti-inflammatory drugs.     

Comparison between two methods for estimating pneumothorax size from chest X-rays

AIM: To compare the estimated size of spontaneous pneumothoraces using the established Rhea inter-pleural distances method with the CT-derived Collins method. METHOD: Adult patients with spontaneous pneumothorax treated conservatively were identified from the ED databases. X-rays were reviewed independently by two researchers and measured according to the methods described by Rhea and Collins. Estimates of size derived by the two methods were compared using bias plot analysis techniques. RESULTS: A total of 156 X-rays in 57 patients were identified. A total of 82% were male with a median age of 22 years. Pneumothoraces varied in size from 4% to 88%. The average difference between methods was 4% (Collins method estimating larger size) with 95% limits of agreement -3.8% to 11.7%. Agreement was very close for small pneumothoraces but deteriorated with increasing pneumothorax size (Collins methods estimated larger pneumothorax size). CONCLUSION: The Rhea method for estimating pneumothorax size is acceptably accurate for smaller pneumothoraces but may significantly under-estimate the size of larger pneumothoraces.     

Estimating the rate of re-expansion of spontaneous pneumothorax by a formula derived from computed tomography volumetry studies

Background

Available evidence suggesting that the rate of re‐expansion of spontaneous pneumothorax is 1.25%/day is based on a small sample and mathematical modelling‐based estimates.

Aim

To estimate the rate of re‐expansion of spontaneous pneumothoraces by a formula derived from computed tomography volumetry studies.

Methods

This retrospective study included adult patients with spontaneous pneumothorax, identified from patient management databases, who were treated conservatively. Medical records were reviewed to confirm that no intervention such as aspiration or catheter drainage had occurred. Radiographs were reviewed independently by two researchers and measured according to the method described by Collins et al. Their formula was used to estimate pneumothorax size on each date. The rate of re‐expansion was defined as the change in size (%)/number of days between radiographs. Patients were excluded if they did not have at least two radiographs taken, at least 1 day apart. Data were analysed using cluster analysis by patient to minimise the effect of repeated measures from an individual patient.

Results

88 episodes were identified in 57 patients. 82% were men and the patients had a median age of 22 years. The average rate of re‐expansion was 2.2%/day (95% confidence interval 1.4% to 3.0%), but varied between −7.5% and 13.4%/day.

Conclusion

Spontaneous pneumothoraces treated conservatively re‐expand at an average rate of 2.2%/day.A few studies have explored the rate of resolution of spontaneous pneumothoraces, and all have had small sample sizes. Current “best evidence” derives from the report of Kircher and Swartel,¹ who used a mathematical approach based on the area (not volume) of the collapsed lung on a radiograph compared with the area of the hemithorax and serial radiographs to estimate that pneumothoraces re‐expanded at a rate of 1.25%/day when treated with bed rest. Recently, more sophisticated methods for estimating the volume of a pneumothorax have been developed using radiographic thoracic gas volume measurement² and helical computed tomography,³ but no studies have investigated the rate of re‐expansion of pneumothoraces using these methods.We investigated the rate of re‐expansion of spontaneous pneumothoraces in patients treated conservatively.