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.

A study to estimate prevalence and risk factors of Obstructive Sleep Apnoea Syndrome in a semi-urban Indian population.

Obstructive Sleep Apnea Syndrome (OSAS) has been recognised as a major cause of morbidity and mortality in developing countries like India. There is still a paucity of Indian studies regarding the prevalence of OSAS. The current single centre prospective cross-sectional study was undertaken to know prevalence estimates for key symptoms and features that can indicate the presence of OSAS in an Indian population. A survey was conducted on subjects with age groups ≥ 25 years at King George's Medical University, Lucknow, Uttar Pradesh, India from August 2009 to July 2011. Data was recorded during the interview on the basis of Berlin Questionnaire (BQ). Risk factors for OSAS were also evaluated. Risk group categorization for OSAS was done with the help of a questionnaire and overnight polysomnography was performed in each group to measure apnea and hypopnea index (AHI). Out of 1816 subjects, 1512 (response rate 83.3%) finally participated in the survey with mean age 42.6±11.2 years, males 67.9% and females 32.1%. Of them 6.2% were found to be at high-risk OSAS; 12.2% were obese (Body Mass Index ≥30 kg/m2) and 33.5% of the obese population were at high-risk OSAS. Among high-risk patients with OSAS, 62.4% had hypertension. Statistically significant and independent risk factors found for OSAS were obesity, large neck size, alcoholism and use of sedatives/tranquillizers. High-risk category predicted an AHI ≥5 with a sensitivity of 86.3% (95% CI 73.1-93.8), specificity of 93.1% (95% CI 89.1-95.7), positive and negative predictive values of 70.9% (95% CI 57.9-81.4) and 97.2% (95% CI 94.1-98.8) respectively. It can be concluded that BQ questionnaire can still be used as a pre-assessment tool for predicting persons at risk for OSAS in clinical practice. Further studies on estimation of prevalence of OSAS by applying BQ are warranted in near future from other regions of India.

Coccidioidomycosis where you least expect it.

An elderly man without history of travel presented with complaints of intermittent fever for 2 months, cough with scanty expectoration for 15 days and history of weight loss of 5 kg in 1 year. The chest X-ray and CT scan of the thorax showed dispersed centrilobular nodules and patchy subpleural consolidation in both lungs with mediastinal lymphadenopathy. He underwent bronchoscopy and bronchoalveolar lavage culture grew Pseudomonas aeruginosa He was prescribed antibiotics based on culture sensitivity; however, patient continued to have symptoms. All relevant blood investigations were within normal limits. He underwent CT-guided biopsy of the right lung lesion during which clearing of the radio-opacities present in the initial CT scan and appearance of fresh lesions in different locations were observed. Migratory shadows were suspected. Fine-needle aspiration cytology showed features suggestive of coccidioidomycosis for which antifungals were started. After 1 month, he improved symptomatically and chest X-ray showed clearance of shadows.

Pitfalls of AHI system of severity grading in obstructive sleep apnoea.

Obstructive sleep apnoea (OSA) is the major underlying co-morbidity in many of the non-communicable diseases (NCD) due to obesity as a common risk factor. Incidence and prevalence of OSA is on the constant rise ever since this entity came to forefront three decades ago. Precise treatment of underlying OSA is extremely important in major NCDs like diabetes mellitus, hypertension, endocrine disorders and vascular diseases. OSA is subcategorized in to mild, moderate and severe based of apnoea-hypopnea index (AHI). Based on the severity grading, treatment of OSA ranges from life style modifications to oral appliances, continuous positive airway pressure (CPAP) and surgeries. AHI system of severity grading in OSA has several inherent shortcomings and using AHI system for severity grading as the holy grail is likely to be counter-productive. AHI system equates apnoea and hypopnea as equal events, whereas physiological effects vary significantly. AHI system does not account duration of apnoea or body position during apnoeic events. We discuss at length the pitfalls of AHI system of severity grading in OSA.

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.

Recurrent pleural effusion secondary to endometriosis: a rare malady.

Thoracic endometriosis is a rare entity, wherein thoracic endometrial tissue deposition occurs. Recurrent pleural effusion is one of its manifestations. Diagnosis and treatment are often challenging, owing to the rarity of the condition and recurrences. We present an interesting case of a young woman of childbearing age who presented with recurrent haemorrhagic pleural effusion. She was referred to our centre after her initial video-assisted thoracoscopic biopsy was inconclusive. She was re-evaluated, and video-assisted thoracoscopic biopsy confirmed thoracic endometriosis. She underwent pleurodesis two times and currently on hormonal treatment, maintaining well.

Cardiac tamponade and massive pleural effusion in a young COVID-19-positive adult.

COVID-19 has a broad spectrum of cardiac manifestations, and cardiac tamponade leading to cardiogenic shock is a rare presentation. A 30-year-old man with a history of COVID-19-positive, reverse transcription polymerase chain reaction (RT-PCR) done 1 week ago and who was home-quarantined, came to the emergency department with palpitations, breathlessness and orthopnoea. His ECG showed sinus tachycardia with low-voltage complexes, chest X-ray showed cardiomegaly and left pleural effusion and two-dimensional echocardiography showed large pericardial effusion with features suggestive of cardiac tamponade. He was taken up for emergency pericardiocentesis which showed haemorrhagic pericardial fluid. Intercostal drainage insertion was done for left-sided large pleural effusion. After ruling out all the other causes for haemorrhagic pericardial effusion, the patient was started on colchicine, steroids, ibuprofen and antibiotics to which he responded. Both pericardial and pleural effusions resolved completely on follow-up.

Comparative evaluation of serum cotinine levels in chronic periodontitis and chronic obstructive pulmonary disease.

CONTEXT: Periodontitis and chronic obstructive pulmonary disease (COPD) are chronic progressive inflammatory conditions. Smoking has been associated with both chronic periodontitis and COPD. Hence, the present study was designed to correlate serum levels of cotinine with the severity of periodontal disease with or without COPD. SETTINGS AND DESIGN: A total of eighty patients, twenty healthy individuals, twenty patients with chronic generalized periodontitis without smoking and without COPD, twenty patients who are smokers with chronic periodontitis without COPD and twenty patients who are smokers with chronic periodontitis and COPD in the age range of 43-65 years were selected for the study. SUBJECTS AND METHODS: Serum cotinine level assessment, smoking history, and periodontal examination were done in all the patients and the data obtained were statistically analyzed. RESULTS: The mean serum cotinine level was highest in smokers with chronic periodontitis and COPD (93.642 ± 14.727) and it differed significantly between the four groups (P < 0.001). There is a significant positive correlation between the number of cigarettes and serum cotinine levels in both groups involving smoking. There was no significant correlation between serum cotinine level and clinical attachment loss in chronic periodontitis smokers with or without COPD. CONCLUSIONS: The result of this study indicates that increased smoking with COPD causes a higher chance of progression of periodontal destruction but it is not statistically significant. Furthermore, this study indicates that the assessment of serum cotinine levels is a reliable method to evaluate smoking exposure.

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.

Frequency of adverse events observed with second-line drugs among patients treated for multidrug-resistant tuberculosis.

BACKGROUND: Multidrug-resistant tuberculosis (MDR-TB) is considered to be a worldwide problem with notoriously difficult and challenging treatment. Adverse events associated with second-line drugs (SLDs) can have severe impact on efficient management. OBJECTIVE: To know the frequency of adverse events due to SLDs in patients of MDR-TB. DESIGN: A prospective cohort analysis of 98 MDR-TB patients enrolled between June 2009 to February 2010 was conducted in Department of Pulmonary Medicine, King George Medical University, Lucknow, India. All the patients were provided standardized regimen. Adverse events associated with treatment were recognized primarily by clinical evidence and/or laboratory investigations that were advised at baseline and whenever clinically indicated during course of treatment. Adverse events were considered major if required permanent discontinuation or substitution of drugs. RESULTS: 119 adverse events were reported in 46 (46.9%) patients. The grouped adverse events were most commonly gastrointestinal that was observed with a frequency of 48 (40.3%) followed by ototoxicity in 28 (23.6%), and neurological in 21 (17.6%). 17 (17.4%) patients had major adverse events requiring permanent discontinuation or substitution of drugs that included deafness and tinnitus in 5 (5.1%) followed by psychosis in 4 (4.1%). None of the patients stopped complete regimen due to adverse events. The treatment success rate was observed to be 71 (72.4%). CONCLUSIONS: MDR-TB can be cured successfully with appropriate combination of drugs if adverse events associated with them can be managed aggressively and timely. Newer and less toxic drugs are urgently needed to treat MDR-TB patients.

Concomittant pulmonary tuberculosis and borderline leprosy with type-II lepra reaction in single patient.

The concommitant occurrence of both tuberculosis and leprosy in a single individual are not an uncommon clinical condition but is being reported infrequently in literature. We report a case of leprosy, diagnosed previously and also diagnosed as pulmonary tuberculosis.