Enhanced expression of Organic Cation Transporters in bronchial epithelial cell layers following insults associated with asthma – impact on salbutamol transport

Increasing evidence suggests Organic Cation Transporters (OCT) might facilitate the absorption of inhaled bronchodilators, including salbutamol, across the lung epithelium. This is essentially scarred and inflamed in asthma. Accordingly, the impact of epithelial insults relevant to asthma on OCT expression and salbutamol transport was evaluated in air-liquid interfaced layers of the human broncho-epithelial cell line Calu-3. These were physically injured and allowed to recover for 48 h or exposed to the pro-inflammatory stimulant lipopolysaccharide (LPS) for 48 h and the aeroallergen house dust mite (HDM) for 8 h twice over 48 h. Increases in transporter expression were measured following each treatment, with the protein levels of the OCTN2 subtype consistently raised by at least 50%. Interestingly, OCT upregulation upon LPS and HDM challenges were dependent on an inflammatory event occurring in the cell layers. Salbutamol permeability was higher in LPS exposed layers than in their untreated counterparts and in both cases, was sensitive to the OCT inhibitor tetraethylammonium. This study is the first to show epithelial injury, inflammation and allergen abuse upregulate OCT in bronchial epithelial cells, which might have an impact on the absorption of their substrates in diseased lungs

Anti-IL5 therapy for asthma and beyond

Airway inflammation is considered to be the primary component contributing to the heterogeneity and severity of airway disorders. Therapeutic efficacies of diverse novel biologics targeting the inflammatory pathways are under investigation. One such target is IL-5, a type-1 cytokine that is central to the initiation and sustenance of eosinophilic airway inflammation. Over the past decade, anti-IL5 molecules have been documented to have mixed therapeutic benefits in asthmatics. Post hoc analyses of the trials reiterate the importance of identifying the IL-5-responsive patient endotypes. In fact, the currently available anti-IL5 treatments are being considered beyond asthma management; especially in clinical complications with an underlying eosinophilic pathobiology such as hypereosinophilic syndrome (HES) and eosinophilic granulomatosis and polyangitis (EGPA). In addition, closer analyses of the available data indicate alternative mechanisms of tissue eosinophilia that remain uncurbed with the current dosage and delivery platform of the anti-IL5 molecules. Keywords: Eosinophil, IL-5, Eosinophilic asthma, Hypereosinophilic syndrome (HES), Churg-strauss syndrome, Chronic bronchitis, Eosinophilic granulamatosis and polyangitis (EGPA), Chronic obstructive pulmonary disorder (COPD), Mepolizumab, Reslizumab, Benralizuma

A prospective randomised comparative study of unilateral paravertebral block with conventional spinal anesthesia for inguinal hernia repair

Background: Inguinal hernia repair can be performed under satisfactory anaesthetic conditions using general, regional and peripheral nerve block anaesthesia. Unilateral spinal anaesthesia provides optimal anaesthesia, with stable haemodynamics and minimal adverse events .The paravertebral block being segmental in nature can be expected to produce some advantages and may be a viable technique. Objective: Primary objective of the study was to compare the block characteristics-time required for performing the block, time to surgical anesthesia, time to ambulation, time to first analgesic, adverse events between the two groups. Secondary objective is to compare the post operative analgesia between the two groups. Methodology: About 60 consenting male patients posted for inguinal hernia repair were randomized into two groups to receive either paravertebral block (Group P, n=30) at T10 with 15 ml of 0.5% bupivacaine and at L1 with 5ml of 0.5% bupivacaine or spinal anesthesia (Group S) with 12.5 mg of 0.5% hyperbaric bupivacaine and primary outcome secondary outcome were noted. Results: Time to perform the block and time to reach surgical anesthesia were significantly higher in the patients of group P as compared to group S (p<0.001). Time to ambulation was significantly shorter in group P than compared to group S (p<0.001). Haemodynamic parameters mean arterial pressure and heart rate were found to be more stable in group P than group S (p<0.05). Minimal adverse events were noted in both the group and it was statistically not significant. Conclusion: It can be concluded that both spinal anesthesia and paravertebral block can be used for patients undergoing inguinal hernia repair. Spinal anesthesia provides adequate analgesia and motor blockade and also less time to perform block and to reach surgical anesthesia. On the other hand paravertebral block provides good haemodynamic stability as well as less time to ambulation, minimal adverse events, however the expertise related to perform, procedure related time and prolonged onset of effect are the main concerns

CT and Functional MRI to Evaluate Airway Mucus in Severe Asthma

BACKGROUND: Intraluminal contributor(s) to airflow obstruction in severe asthma are patient-specific and must be evaluated to personalize treatment. The occurrence and functional consequence of airway mucus in the presence or absence of airway eosinophils remain undetermined. OBJECTIVE: The objective of this study was to understand the functional consequence of airway mucus in the presence or absence of eosinophils and to identify biomarkers of mucus-related airflow obstruction. METHODS: Mucus plugs were quantified on CT scans, and their contribution to ventilation heterogeneity (using MRI ventilation defect percent [VDP]) was evaluated in 27 patients with severe asthma. Patients were dichotomized based on sputum eosinophilia such that the relationship between mucus, eosinophilia, and ventilation heterogeneity could be investigated. Fractional exhaled nitric oxide (Feno) and related cytokines in sputum were measured. RESULTS: Mucus plugging was present in 100% of asthma patients with sputum eosinophils and 36% of those without sputum eosinophils (P = .0006) and was correlated with MRI VDP prebronchodilator (r = 0.68; P = .0001) and postbronchodilator (r = 0.72; P \u3c .0001). In a multivariable regression, both mucus and eosinophils contributed to the prediction of postbronchodilator MRI VDP (R CONCLUSIONS: Both airway eosinophils and mucus can contribute to ventilation heterogeneity in patients with severe asthma. Patients in whom mucus is the dominant cause of airway obstruction have evidence of an upregulated IL-4/IL-13 pathway that could be identified according to increased Feno level

A sputum bioassay for airway eosinophilia using an eosinophil peroxidase aptamer

Abstract Eosinophils are granulocytes that play a significant role in the pathogenesis of asthma and other airway diseases. Directing patient treatment based on the level of eosinophilia has been shown to be extremely effective in reducing exacerbations and therefore has tremendous potential as a routine clinical test. Herein, we describe the in vitro selection and optimization of DNA aptamers that bind to eosinophil peroxidase (EPX), a protein biomarker unique to eosinophils. Fifteen rounds of magnetic bead aptamer selection were performed prior to high throughput DNA sequencing. The top 10 aptamer candidates were assessed for EPX binding using a mobility shift assay. This process identified a lead aptamer candidate termed EAP1-05 with low nanomolar affinity and high specificity for EPX over other common sputum proteins. This aptamer sequence was further optimized through truncation and used to develop an easy-to-use colourimetric pull-down assay that can detect EPX over a concentration range from 1 – 100 nM in processed sputum. Forty-six clinical samples were processed using a new sputum dispersal method, appropriate for a rapid assessment assay, that avoids centrifugation and lengthy processing times. The assay showed 89% sensitivity and 96% specificity to detect eosinophilia (compared to gold standard sputum cytometry), with results being produced in under an hour. This assay could allow for an easy assessment of eosinophil activity in the airway to guide anti-inflammatory therapy for several airway diseases

Omalizumab in patients with severe asthma and persistent sputum eosinophilia

Omalizumab, a recombinant humanized monoclonal antibody targeting the IgE molecule, is the first biologic approved for moderate-to-severe allergic asthmatics, who remain uncontrolled despite high dose inhaled corticosteroid and bronchodilators. Steroid-sparing effect of omalizumab has not been demonstrated in asthmatics with persistent airway eosinophilia in a randomised controlled trial till date. From this double-blind, placebo-controlled, multi-centred, randomized parallel group design, we report that omalizumab is possibly inadequate to control sputum eosinophilia, and therefore may not have a steroid-sparing effect, especially in those maintained on oral corticosteroids daily. This needs to be confirmed or refuted in a larger trial, which may be a challenge with respect to recruitment, since there are currently three additional biologics available to prescribe. Trial registration Clinicaltrials.gov, NCT02049294, Registered 30th January 2014, https://clinicaltrials.gov/ct2/show/NCT02049294Medicine, Faculty ofNon UBCMedicine, Department ofReviewedFacult