Assessment of left ventricular function in subjects with chronic obstructive pulmonary disease.

Chronic obstructive pulmonary disease (COPD) is the world's third leading cause of death, posing a significant public health challenge. Heart failure is an important milestone in the natural history of this disease's ever-decreasing course. Cor pulmonale alone is sometimes insufficient to explain the scenario, as many heart failures are refractory even after standard treatment. A possible explanation is involvement of the left ventricle (LV). However, there is currently no consensus on a therapeutic approach to LV dysfunction in COPD. Some previous studies were conducted on COPD patients regardless of the presence or absence of other factors influencing LV function. This study attempted to rule out all known confounding factors/co-morbidities that influence LV function. We discovered that LV diastolic dysfunction was common in subjects at all stages of COPD. We believe that all COPD patients, regardless of stage, should have screening echocardiography.

Detection of clinically-relevant EGFR variations in de novo small cell lung carcinoma by droplet digital PCR.

Targeted therapy that utilizes tyrosine kinase inhibitors (TKIs), specific to epidermal growth factor receptors (EGFR) has changed the landscape of treatment of non-small cell lung cancer (NSCLC). The success or failure of this approach depends on presence of certain variations in the tyrosine kinase domain of EGFR gene. Generally, patients diagnosed with Small cell lung cancer (SCLC) are considered ineligible for TKI therapy owing to the absence of EGFR variations. . However, there is evidence of these variations being detected in SCLCs, both in de-novo and in transformed SCLCs (TKI-treated adenocarcinomas). Despite the presence of clinically-relevant EGFR variations in SCLCs, the response to TKIs has been inconsistent.  Liquid biopsy is a well-established approach in lung cancer management with proven diagnostic, prognostic and predictive applications. It relies on detection of circulating tumor-derived nucleic acids present in plasma of the patient. In this study, a liquid biopsy approach was utilized to screen 118 consecutive lung cancer patients for four clinically-relevant variations in EGFR gene, which included three activating/sensitizing variations (Ex18 G719S, Ex19del E746-A750 and Ex21 L858R) and one acquired/resistance (Ex20 T790M, de novo) variation by droplet digital PCR, the most advanced third generation PCR technique. As expected, clinically-relevant EGFR variations were found in majority of the non-small cell lung cancer cases. However, among the handful of small cell lung cancer samples screened, sensitizing variations (Ex18 G719S and Ex21 L858R) were seen in almost all of them. Interestingly, Ex20 T790M variation was not detected in any of the cases screened.  The results of our study indicate that EGFR variations are present in SCLCs and highly sensitive liquid biopsy techniques like ddPCR can be effectively utilized for this purpose of screening EGFR variations in such samples.

A study of Spirometric parameters in non asthmatic allergic rhinitis.

INTRODUCTION: Allergic rhinitis (AR) is a common IgE -mediated inflammatory condition characterised by sneezing, nasal congestion, itching and rhinorrhoea. Nasal allergy is a strong risk factor for the onset of asthma in adults. Bronchial hyper-responsiveness (BHR) is a distinct feature of pathophysiology in asthma. Spirometric parameters like Forced Expiratory Volume in first second [FEV1] and Forced Expiratory Flow [FEF 25-75%] are known to be impaired in patients with allergic rhinitis. We studied these parameters in subjects of AR who have never experienced any chest symptoms. It is well known that, subjects with allergic rhinitis are at greater risk of developing overt bronchial asthma in future. METHODS: All patients presented with symptoms of allergic rhinitis without history of bronchial asthma were included. Patients those who were clinically diagnosed with allergic rhinitis were evaluated with absolute eosinophilic count, serum IgE and Spirometric assessment. In spirometry, post bronchodilator FEV1 reversibility and post bronchodilator FEF 25-75% values were used to assess lower airway abnormalities. RESULTS: Among 61 subjects, 32 were males and 29 were females. The maximum numbers [28] of patients were in 21-35 age group. Absolute eosinophil count was elevated in 38% of patients. 33% of patients showed elevated IgE values above 1000. 43% of patients showed FEV1 reversibility which is a sign of Bronchial hyperreactivity. 5% of patients showed impaired post bronchodilator FEF 25-75% which indicates presence of small airway disease. There was significant correlation between FEV1 reversibility and elevated IgE. CONCLUSION: Impaired spirometric parameters indicate coexistence of bronchial impairment and hence predisposition to progression from allergic rhinitis alone to overt asthma in future. Thus careful evaluation of lower airway has to be done to rule out coexisting subclinical asthma.

Oral Candidal Load and Oral Health Status in Chronic Obstructive Pulmonary Disease (COPD) Patients: A Case-Cohort Study.

OBJECTIVE: The objective of this study was to determine the candidal load of the patients with Chronic Obstructive Pulmonary Disease (COPD) and evaluate the oral health status of subjects with COPD. Material and Methods. N = 112 COPD subjects and N = 100 control subjects were included in the study. The selection of COPD cases was confirmed based on the set criteria from the American College of Physicians. The oral health status was assessed as per WHO criteria to determine the score of decayed, missing, and filled teeth (DMFT), significant caries index (SiC), community periodontal index and treatment needs (CPITN), and oral hygiene index-simplified (OHI-S). Gram staining was performed to identify Candida using the whole saliva. Quantitative evaluation of the candidal load was carried out using Sabouraud Dextrose Agar (SDA). Chrome agar was used to differentiate between the commensal carriages. A statistical analysis paired t-test and 95% confidence interval (CI) for proportions was carried out using STATA software. RESULTS: Candidal growth was found in 21.42% (n = 24) of COPD cases and 1.1% (n = 11) of control cases (p < 0.05) (95% CI 0.45, 0.59). The DMFT score was 8.26 in COPD subjects and 4.6 in controls, the SiC score was 16.42 in COPD subjects and 10.25 in controls, and the CPITN score for both COPD and control cases was score 2. CONCLUSION: In conclusion, there was a higher candidal load among subjects suffering from COPD. Theophylline medication can be a risk factor for increased candidal load in COPD patients.

Coinfection of SARS-CoV-2 and MTB: how not to miss the wood for the trees.

Scarce data exist about the coinfection of SARS-CoV-2 and Mycobacterium tuberculosis (MTB). A young woman who was undergoing treatment for multiple sclerosis was brought to our hospital with a COVID-19 positive status. On further evaluation, her chest X-ray showed right upper and mid-zone opacity, which lead to the suspicion of MTB. Her sputum came positive for acid-fast bacilli (AFB) staining and cartridge-based nucleic acid amplification test (CBNAAT) confirmed it, and rifampicin resistance was not detected. She was started on an antitubercular regimen. She was discharged, and by the end of the intensive phase of treatment, her symptoms subsided, but her sputum CBNAAT still showed the presence of TB bacillus.

Quantitative analysis of cell-free DNA by droplet digital PCR reveals the presence of EGFR mutations in non-malignant lung pathologies.

Tyrosine kinase inhibitors (TKIs) targeting epidermal growth factor receptor (EGFR) are effectively used in treatment of non-small cell lung cancer (NSCLC). Mutation profile of tyrosine kinase domain of EGFR determines the eligibility of the patients for tyrosine kinase inhibitor (TKI) therapy. Liquid biopsy, which relies on circulating tumor-derived nucleic acids, has emerged as an effective tool in lung cancer management with proven diagnostic, prognostic and predictive applications. We screened 100 subjects, suspected to have lung malignancy, for four hotspot mutations including three activating (G719S, Ex19del E746-A750 and L858R) and one acquired (T790M, de novo) in EGFR gene by droplet digital PCR (ddPCR). While 97 subjects were subsequently confirmed to have lung malignancy based on histo/cytopathological studies, three cases turned out to be non-malignant lung pathologies that were completely cured by antibiotic therapy. Intriguingly, ddPCR revealed the presence of EGFR mutations in these non-malignant subjects. Two cases showed the presence of G719S and T790M mutations respectively and another had compound mutations (T790M and L858R). The detection of EGFR mutations in non-malignant pulmonary conditions opens up a new area of research.

Non-invasive detection of EGFR mutations by cell-free loop-mediated isothermal amplification (CF-LAMP).

Targeting epidermal growth factor receptor (EGFR) through tyrosine kinase inhibitors (TKI) is a successful therapeutic strategy in non-small cell lung cancer. However, the response to TKI therapy depends on specific activating and acquired mutations in the tyrosine kinase domain of the EGFR gene. Therefore, confirming the EGFR status of patients is crucial, not only for determining the eligibility, but also for monitoring the emergence of mutations in patients under TKI therapy. In this study, our aim was to develop a cost effective, yet sensitive, technique that allows the detection of therapeutically-relevant EGFR hotspot mutations at isothermal conditions in a non-invasive manner. Previously, we developed an allele-specific loop-mediated isothermal amplification (AS-LAMP) assay for screening germline and somatic de novo T790M EGFR mutation in lung cancer patients. In this study, we used cell free DNA as a template in AS-LAMP assay (CF-LAMP) for non-invasive detection of two hotspot EGFR mutations (T790M, and L858R) and compared its efficiency with ultrasensitive droplet digital PCR (ddPCR) assay. The results of CF-LAMP assay were consistent with those obtained in ddPCR assay, indicating the robustness of the method. CF-LAMP may serve as a valuable and cost-effective alternative for liquid biopsy techniques used in molecular diagnosis of non-small cell lung cancer.

Detection of epidermal growth factor receptor T790M mutation by allele-specific loop mediated isothermal amplification.

INTRODUCTION: Targeted therapy using specific inhibitors against tyrosine kinases (TKs) is a paradigm in non-small-cell lung cancer management. However, the success of TK inhibitor (TKI) therapy depends on certain activating or acquired mutations, which render sensitivity or resistance to TKIs in the patients. The acquisition of epidermal growth factor receptor (EGFR) T790M point mutation is the most common mechanism of resistance to TKI in non-small cell lung cancer. A number of molecular strategies are now available for molecular testing of non-small cell lung cancers. However, almost all of them are cost-intensive and laborious and require high-end advanced equipment. Thus, assays that are rapid, simple, and cost-effective, yet sensitive, are most ideal in clinical settings for screening such therapeutically relevant mutations. MATERIALS AND METHODS: Allele-specific loop-mediated isothermal amplification assay (AS-LAMP), which is a variant of the original LAMP assay, is a promising diagnostic technique for screening single-nucleotide polymorphisms. Using commercially available plasmid constructs as template DNA, AS-LAMP assay for EGFR T790M mutation was optimized with six different sets of reaction mixture containing varying concentrations of buffer and primers. The results of AS-LAMP assay were further validated by ultrasensitive droplet digital polymerase chain reaction. RESULTS: Only one of the six sets of reaction mixture could accurately distinguish between wild type and mutated DNA, indicating that the primers and buffer are the two most critical components that determine the accuracy of AS-LAMP. The optimized AS-LAMP assay was further used to screen germ line and somatic T790M mutations in non-small cell lung cancer using blood and tissue samples collected from patients. CONCLUSION: Development of an accurate and rapid diagnostic assay that can detect resistant mutations without the need for sequencing is highly useful for clinicians in deciding on the eligibility of patients for TKI therapy. Considering its several inherent advantages, AS-LAMP assay could become an effective molecular tool for screening baseline or acquired EGFR T790M mutations in non-small cell lung cancer patients.