Association of cytokines levels with epidermal growth factor receptor mutation in lung cancer patients

Background: Lung cancer is one of the most frequent types of cancer and the leading cause of cancer-related deaths. Epidermal growth factor receptor (EGFR) is a receptor tyrosine kinase (TK) being highly expressed in lung cancers. Activation of EGFR through oncogenic mutations leads to upregulation of gene expression that may heighten the inflammatory response in certain situations. EGFR acts as a key regulator and a cellular hub for inflammatory cytokine signaling, thereby promoting tumor cell proliferation, invasion, migration, metastases, and survival. The aim of the present study is to determine the serum cytokines levels and EGFR mutation status in lung cancer patients to investigate the association between the EGFR mutation status and cytokines levels with lung cancer patients. Materials and Methods: Blood and tissue samples of lung cancer patients were collected. The EGFR mutations of lung cancer patients were determined by the immunohistochemistry (IHC) and serum cytokines levels of lung cancer patients were determined using ELISA. Results: Statistically significant association of EGFR mutations with adenocarcinoma subtypes and non-smokers were found (P < 0.05). Lung cancer patients with EGFR mutations had significantly higher tumor necrosis factor-alpha levels when compared to lung cancer patients without EGFR mutations (P < 0.01), and EGFR mutation status was not significantly associated with interleukin-6 levels (P = 0.24). Conclusion: EGFR mutation detection by the IHC method is a potentially useful tool to guide clinicians for personalized treatment of lung cancer patients of adenocarcinoma subtype, and cytokines are good biomarkers for the diagnosis, prognosis, and prediction of treatment responses in lung cancer patients as well as act as therapeutic targets. This study will provide biomarkers for lung cancer diagnosis and treatments

Diagnostic utility of cytospin, cell block and immunocytochemistry in pleural effusion cytology

Background: With the introduction of cytospin, the sensitivity of diagnosing malignancies has increased mainly due to the increase in cellular yield. Cell block also gives the advantage of ancillary testing and allows for retrospective studies. Immunocytochemical markers are used to differentiate and subtype various malignancies in body effusions.Aim of the study was to compare the morphological features of both technique and to assess the diagnostic utility of cell block methods in the cytodiagnosis of pleural effusions.Methods: This was a Prospective observational comparative study of two cytopreparatory techniques. All samples were examined and processed by cytospin and cell block techniques. Continuous data were expressed as Mean±SD (standard deviation) while categorical data were expressed in number, percentage and compared by chi-square (χ2) test.Results: The final diagnosis of both cytospin (147 cases) and cell block (150 cases) techniques was divided into four broad categories: Inadequate, Benign, Suspicious and Malignant. The significant diagnostic cytospin (AUC=0.857, p<0.001) in discriminating positive and negative malignant cases with 75.00% sensitivity (95% CI=53.3-90.2) and 100.00% specificity (95% CI=86.7-100.0) and with 100.0% positive predictive value and 81.2% negative predictive value. In contrast, cell block also showed significant diagnostic but with higher accuracy (AUC=1.000, p<0.001) and sensitivity 100.00% (95% CI=86.7-100.0) and specificity 100.00% (95% CI=86.7-100.0) and 100.0% positive predictive value and 100.0% negative predictive value than cytospin technique.Conclusions: Cell block as a technique should be used in routine practice as it not only increases the diagnostic yield but ancillary test can also be done.

Predictors of co-morbidities in chronic obstructive pulmonary disease patients at a tertiary care centre in north India

Background: Chronic obstructive pulmonary disease (COPD) is a common preventable and treatable disease that is characterised by persistent respiratory symptoms and airflow limitation. COPD is characterised by an intense inflammatory process in the airways, parenchyma, and pulmonary vasculature. It is possible in some cases that the inflammatory process may overflow into the systemic circulation, promoting a generalised inflammatory reaction. Patient with COPD often have concomitant chronic illness (co-morbidities). The aim of this study is to know the pattern of co-morbidities in COPD patients.Methods: This study was a cross sectional observational study conducted on 172 COPD patients (IPD and OPD) diagnosed on the basis of GOLD guideline 2017. Co morbidities were diagnosed as per standard defined criteria laid down in the respective guidelines.Results: 55.3% of the patients with COPD had co morbidities. 18/88(20.5%) patients presented with multiple co-morbidities. 49/88, 55.7% COPD patients were affected with cardiac (either only cardiac or had multiple organs affected besides cardiac), the commonest co-morbidity. Amongst cardiac, hypertension and congestive heart failure (CHF) was the commonest (n=19/49, 38.8% each) followed by CAD/CSA/IWMI/IHD/AF. Others were metabolic (n=14/88, 15.9%), GERD (n=13/88, 14.8%), Depression (n=11/88, 12.5%). Less prevalent co-morbidities were Osteoporosis (n=8/88, 9.1%), Lung cancer (n=6/88, 6.8%), Bronchiectasis (n=5/88, 5.6%) and OSA (n=3/88, 3.4%).Conclusions: Urban indwelling, advancing age and duration of illness, presentation with low mood, loss of pleasure/ interest, appetite disturbances and heart burn with relief on taking proton pump inhibitor can be predictors of co-morbidities in COPD patients. Chance of finding co-morbidities may be multifactorial. Thus, it is important to look out for co morbidities in each and every COPD patients.

Prothrombotic state in HIV: A study on protein C, protein S, homocysteine and correlation with CD4 counts

Introduction: Human immunodeficiency virus (HIV) may result in variable haematological manifestations. Thrombotic events are more common among HIV-infected persons than the general population, possibly due to the increased inflammatory/hypercoagulable state and presence of concurrent comorbidities. Aims and Objectives: (1) Screen for coagulation abnormalities in HIV-infected patients. (2) Detect certain prothrombotic factors such as deficiency of protein C and protein S and elevation of homocysteine as possible precursors of coagulation defects in HIV patients. (3) Correlation of coagulation abnormalities with CD4 counts. Methods: A pilot study of 1-year duration conducted in the Department of Pathology in collaboration with ART centre, KGMU Lucknow. All diagnosed HIV-seropositive patients (n = 30) who were not taking Vitamin K, antithrombotic and antiplatelet drugs including aspirin, oral contraceptives and not having known protein C/S deficiency were included in the present study as cases. Apart from this, 30 age- and sex-matched healthy individuals were also included in the present study. Assessment of the bleeding time, prothrombin time and activated partial thromboplastin time, complete blood count was done. Protein C and S were measured by calorimetric assay. Serum homocysteine was measured by the semi-automated method. CD4 count was done by flow cytometry. Results: The findings of the present study suggest a relationship between HIV, its complications and thrombosis. The HIV-seropositive patients have reduced levels of haemoglobin, CD4 counts, platelet counts, mean platelet volume, protein C and S activity as compared to the healthy individuals. Thrombophilic abnormality in the form of hyperhomocysteinaemia is more frequent in HIV-infected patients. All these parameters have a definite correlation with CD4 count.

Viral aetiology of acute lower respiratory tract illness in hospitalised paediatric patients of a tertiary hospital: One year prospective study.

Context: Acute lower respiratory tract infections (ALRI), ranked as the second leading cause of death are the primary cause of hospitalisation in children. Viruses are the most important causative agents of ALRI. Aim: To study the viral aetiology of ALRI in children at a tertiary care hospital. Setting and Design: One year prospective observational study in a tertiary care hospital of King George’s Medical University, Lucknow. Material and Methods: Nasopharyngeal aspirate (NPA) was collected from children admitted with signs and symptoms of ALRI who were aged 0-14 years. Samples were transported to the laboratory at 4°C in viral transport media and processed for detection of respiratory syncytial virus (RSV) A and B, infl uenza virus A and B, adenovirus (ADV), human Boca virus (HBoV), human metapneumo virus (hMPV) and parainfl uenzavirus 1, 2, 3 and 4 using mono/multiplex real-time polymerase chain reaction (RT-PCR). STATA was used for statistical analysis. Results: In one year, 188 NPAs were screened for respiratory viruses, of which 45.7% tested positive. RSV was most commonly detected with 21.3% positivity followed by measles virus (8.5%), infl uenza A virus (7.4%), ADV (5.3%), infl uenza B virus (1.6%), hMPV (1.1%) and HBoV (0.5%). Month wise maximum positivity was seen in December and January. Positivity rate of RSV was highest in children aged < 1 year, which decreased with increase in age, while positive rate of infl uenza virus increased with increasing age. Conclusion: The occurrence of viral predominance in ALRI is highlighted.

Use of GenoType® MTBDRplus assay to assess drug resistance and mutation patterns of multidrug-resistant tuberculosis isolates in northern India.

Purpose: The emergence and spread of multidrug-resistant tuberculosis (MDR-TB) is a major public health problem. The diagnosis of MDR-TB is of paramount importance in establishing appropriate clinical management and infection control measures. The aim of this study was to evaluate drug resistance and mutational patterns in clinical isolates MDR-TB by GenoType® MTBDRplus assay. Material and Methods: A total of 350 non-repeated sputum specimens were collected from highly suspected drug-resistant pulmonary tuberculosis (PTB) cases; which were processed by microscopy, culture, differentiation and first line drug susceptibility testing (DST) using BacT/ALERT 3D system. Results: Among a total of 125 mycobacterium tuberculosis complex (MTBC) strains, readable results were obtained from 120 (96%) strains by GenoType® MTBDRplus assay. Only 45 MDR-TB isolates were analysed for the performance, frequency and mutational patterns by GenoType® MTBDRplus assay. The sensitivity of the GenoType® MDRTBplus assay for detecting individual resistance to rifampicin (RIF), isoniazid (INH) and multidrug resistance was found to be 95.8%, 96.3% and 97.7%, respectively. Mutation in codon S531L of the rpoB gene and codon S315T1 of katG genes were dominated in MDR-TB strains, respectively (P < 0.05). Conclusions: The GenoType® MTBDRplus assay is highly sensitive with short turnaround times and a rapid test for the detection of the most common mutations conferring resistance in MDR-TB strains that can readily be included in a routine laboratory workflow.

Changing patterns and trends of multidrug-resistant tuberculosis at referral centre in Northern India: A 4-year experience.

Purpose: India has a high burden of drug-resistant tuberculosis (TB), although there is little data on multidrug-resistant tuberculosis (MDR-TB). Although MDR-TB has existed for long time in India, very few diagnostic laboratories are well-equipped to test drug sensitivity. The objectives of this study were to determine the prevalence of MDR-TB, first-line drug resistance patterns and its changing trends in northern India in the 4 years. Materials and Methods: This was a prospective study from July 2007 to December 2010. Microscopy, culture by Bactec460 and p-nitro-α-acetylamino-β-hydroxypropiophenone (NAP) test was performed to isolate and identify Mycobacterium tuberculosis (M. tb) complex (MTBC). Drug sensitivity testing (DST) was performed by 1% proportional method (Bactec460) for four drugs: Rifampicin, isoniazid, ethambutol and streptomycin. Various clinical and demographical profiles were evaluated to analyse risk factors for development of drug resistance. Results: We found the overall prevalence rate of MDR-TB to be 38.8%, increasing from 36.4% in 2007 to 40.8% in 2010. we found that the prevalence of MDR-TB in new and previously treated cases was 29.1% and 43.3% ( P < 0.05; CI 95%). The increasing trend of MDR-TB was more likely in pulmonary TB when compared with extra-pulmonary TB ( P < 0.05; CI 95%). Conclusions: we found a high prevalence (38.8%) of MDR-TB both in new cases (29.1%) and previously treated cases (43.3%).This study strongly highlights the need to make strategies for testing, surveillance, monitoring and management of such drug-resistant cases.

Detection of 123 bp fragment of insertion element IS6110 Mycobacterium tuberculosis for diagnosis of extrapulmonary tuberculosis.

Purpose: Extrapulmonary tuberculosis (EPTB) is emerging problem in developing and developed countries. The diagnosis of EPTB in its different clinical presentations remains a true challenge. IS6110-based polymerase chain reaction (PCR) is used for rapid identification and positivity rate of the Mycobacterium tuberculosis complex in clinical isolates of different sites of EPTB. The present study was carried out to study the prevalence of M. tuberculosis complex in clinical isolates of EPTB at tertiary care centres in Lucknow. Materials and Methods: Seven hundred fifty-six specimens were collected from the suspected cases of EPTB which were processed for Mycobacteria by Ziehl Neelson (ZN) staining and BACTEC culture. All the specimens were also processed for IS6110-based PCR amplification with primers targeting 123 bp fragment of insertion element IS6110 of the M. tuberculosis complex. Results: Of these 756 specimens, 71(9.3%) were positive for acid fast bacilli (AFB) by ZN staining, 227(30.1%) were positive for mycobacteria by BACTEC culture and IS6110 PCR were positive for M. tuberculosis complex in 165 (20.7%) isolates. We found a significant difference in sensitivities of different tests (P<0.05). Conclusions: This study reveals the positivity of M. tuberculosis complex in clinical isolates of EPTB case in tertiary care hospitals in Northern India. 72.7% of M. tuberculosis complex was confirmed by IS6110-PCR in culture isolates from different sites of EPTB. The high prevalence of the M. tuberculosis complex was seen in lymph node aspirate and synovial fluid. However, utility of PCR may play a potentially significant role in strengthening the diagnosis of EPTB especially targeting IS6110.

Dermatophytes, related keratinophilic and opportunistic fungi in indoor dust of houses and hospitals.

Dermatophytes, related keratinophilic and opportunistic fungi were isolated from indoor dust samples of 46 hospitals and 47 houses in Kanpur. A total of 19 fungi represented by 11 genera were isolated by the hair-baiting technique from 230 and 235 samples from hospitals and houses respectively. The isolated fungi are Acremonium implicatum (Indian Type Culture Collection) ITCC 5266 , A. strictum (Germplasm Centre for Keratinophilic Fungi) GPCK 1137 , Aphanoascus fulvescens GPCK 1081 , Arthroderma simii GPCK 1275 , Chrysosporium queenslandicum ITCC 5270 , C. indicum ITCC 5269 , C. pannicola GPCK 1022 , C. tropicum GPCK 1269 , Ctenomyces serratus ITCC 5267 , Gymnoascus reessii ITCC 5265 , Malbranchea fulva GPCK 1075 , Malbranchea pulchella ITCC 5268 , Micosporum gypseum GPCK 1038 , Microsporum cookei GPCK 2001, M. fulvum GPCK 2002 , Paecilomyces lilacinum GPCK 1080 , Penicillium expansum GPCK 1082, Trichophyton mentagrophytes GPCK 2003 and T. terrestre GPCK 2004. In hospitals, the minimum frequency was of Ctenomyces serratus ITCC 5267 while the maximum frequency was of Arthroderma simii GPCK 1275 . In houses, Chrysosporium queenslandicum ITCC 5270 and C. tropicum GPCK 1269 were with minimum and maximum frequencies respectively. This makes the first report of these fungi with keratinolytic ability in the indoor dust of hospitals and houses.

Bilateral chylothorax in a child after mild trauma.

Chylothorax is a life-threatening form of pleural effusion with a varied aetiology. In this report, we describe the case of a child who developed bilateral chylothorax following minor trauma which responded well to conservative management.

Carbimazole induced ANCA positive vasculitis.

Anti-thyroid drugs, like carbimazole and propylthiouracil (PTU) are commonly prescribed for the treatment of hyperthyroidism. One should be aware of the side effects of antithyroid medications. Antineutrophil cytoplasmic antibody (ANCA)--associated vasculitis is a potentially life-threatening adverse effect of antithyroidmedications. We report a patient with Graves' disease who developed ANCA positive carbimazole induced vasculitis. The episode was characterized by a vasculitic skin rash associated with large joint arthritis, pyrexia and parotiditis but no renal or pulmonary involvement. He was referred to us for neurological evaluation because he had difficulty in getting up from squatting position and was suspected to have myositis. Carbimazole and methimazole have a lower incidence of reported ANCA positive side effects than PUT. To the best of our knowledge this is the first ANCA positive carbimazole induced vasculitis case reported from India.

Endobronchial metastasis of follicular thyroid carcinoma presenting as hemoptysis: a case report.

Endobronchial metastasis secondary to follicular thyroid carcinoma is extremely rare. Here, we report a case of follicular thyroid cancer in 58-year-old male who presented with hemoptysis. Computed tomography of the chest revealed multiple lung metastases. Flexible fiberoptic bronchoscopy revealed a fragile polypoid mass 5 cm distal to the vocal cords; biopsy taken from this mass revealed follicular thyroid carcinoma.

In vitro activity of certain antibiotics against dermatophytes.

The in vitro effect of antibiotics i.e. Hamycin, Griseofulvin and Mycostatin against Microsporum gypseum and M. fulvum showed that hamycin was comparatively less inhibitory to M. fulvum. Complete inhibition in mycelial growth and sporulation of these two dermatophytes was observed by Mycostatin at its 1000 micrograms/ml conc. after 8 days. M. fulvum did not sporulate at higher concentration of griseofulvin. However, griseofulvin completely inhibited the sporulation in M. gypseum at all the concentration of this drug, but mycelial growth of M. fulvum and M. gypseum were significantly inhibited. Mycostatin was found more inhibitory to these dermatophytes in comparison to hamycin and griseofulvin.