Identification of Genetic Variants in Exon 4 of TP53 in Lung Carcinoma and in Silico Prediction of Their Significance.

Lung cancer is a severe and the leading cause of cancer related deaths in men and women all over the world. Tumor suppressor protein (TP53) encoded by the TP53 gene which plays a pivotal role in various cellular tumor suppression processes viz cell cycle arrest and apoptosis. Henceforth, the present study was aimed to TP53 exon4 variants from lung carcinoma. Histopathologic and clinically proven 20 patients of lung cancer were enrolled in this study the average age of patients was 45 ± 8 years which categorized as early onset of lung cancer. Genomic DNA was isolated from the blood specimen of patients. Extracted DNA was subjected to PCR amplification for exon 4 of TP53 using appropriate primers and subsequently amplified products were applied to nucleotide alterations via using the DNA sanger sequencing. The genetic analysis documented five variants in exon4 of TP53 which include viz. 4 substitutions [c.215 > C at codon 72, C. 358-359AA > GG at codon 120] were highly prevalent, occurring in 63% and 25% frequency in patients. Other two variants viz. C. 358 A > C at codon 120, C. 365T > G at codon 122 were present at frequency of 15% whilst one deletion variant [152 del C] was found with 5% frequency. Furthermore, alterations on codon 72, 120,122 and 51 were characterized as possibly damaging by Poly Phen-2 and decreased stability using stability bioinformatic tool. Taken together all these findings infer that TP53 gene involved in modulation and susceptibility to lung cancer.

Atypical Case of Highly Mutated h-TERT Promoter in Germline Genome from Buccal Mucosa Cancer.

Buccal mucosa cancer has an aggressive nature as it rapidly grows and penetrates with high recurrence rate. Strikingly, carcinoma of buccal mucosa is the most common cancer of oral cavity in India. Recently, telomerase and telomere biology have been implicated in the pathogenesis and progression in various cancers via regulation of telomere maintenance by telomerase expression which is controlled by telomerase reverse transcriptase (TERT) promoter. Strikingly, h-TERT promoter mutations have been incriminated in regulation of telomerase gene expression. Here, we present a 35 years old male with intense coughing, short breathlessness and fever since 15 days, was admitted to the pulmonary unit. He was a chronic smoker and gutka user. The cytopathological analysis of gastric aspirate revealed buccal mucosa carcinoma of IV stage. We identified h-TERT promoter mutations in isolated genomic DNA from whole blood using DNA sequencer. Genetic analysis disclosed that h-TERT promoter region was highly mutated in this patient. Identified mutations include C.-248 del G, C.-272 del G, C.-279 del G, C.-331 del G, C.-349 del G, C.-351 del C, C.-360 G > A, C.-362 T > A, C.-371 del T and C.-372 del T. Further, all identified mutations were subjected to predict the pathologic functional consequences using bioinformatics tools viz TFsitescan and CiiiDER which showed either loss or gain of transcription factors binding sites in h-TERT promoter. This is a unique case in which total 9 mutations were observed in h-TERT promoter in a single case. In conclusion, all together these mutations in h-TERT promoter may alter the epigenetics and subsequently the tenacity of binding transcription factors which are of functional significance.

Identification of h-TERT Promoter Mutations in Germline DNA from North Indian Lung Carcinoma Patients.

Lung cancer is a severe and the leading cause of cancer related deaths worldwide. The recurrent h-TERT promoter mutations have been implicated in various cancer types. Thus, the present study is extended to analyze h-TERT promoter mutations from the North Indian lung carcinoma patients. Total 20 histopathologically and clinically confirmed cases of lung cancer were enrolled in this study. The genomic DNA was extracted from venous blood and subjected to amplification using appropriate h-TERT promoter primers. Amplified PCR products were subjected for DNA Sanger sequencing for the identification of novel h-TERT mutations. Further, these identified h-TERT promoter mutations were analysed for the prediction of pathophysiological consequences using bioinformatics tools such as Tfsitescan and CIIDER. The average age of patients was 45 ± 8 years which was categorized in early onset of lung cancer with predominance of male patients by 5.6 fold. Interestingly, h-TERT promoter mutations were observed highly frequent in lung cancer. Identified mutations include c. G272A, c. T122A, c. C150A, c. 123 del C, c. C123T, c. G105A, c. 107 Ins A, c. 276 del C corresponding to -168 G>A, -18 T>A, -46 C>A, -19 del C, -19 C>T, -1 G>A, -3 Ins A, -172 del C respectively from the translation start site in the promoter of the telomerase reverse transcriptase gene which are the first time reported in germline genome from lung cancer. Strikingly, c. -18 T>A [C.T122A] was found the most prevalent variant with 75% frequency. Notwithstanding, other mutations viz c. -G168A [c. G272A] and c. -1 G>A [c. G105A] were found to be at 35% and 15% frequency respectively whilst the rest of the mutations were present at 10% and 5% frequency. Additionally, bioinformatics analysis revealed that these mutations can lead to either loss or gain of various transcription factor binding sites in the h-TERT promoter region. Henceforth, these mutations may play a pivotal role in h-TERT gene expression. Taken together, these identified novel promoter mutations may alter the epigenetics and subsequently various transcription factor binding sites which are of great functional significance. Thereby, it is plausible that these germline mutations may involve either as predisposing factor or direct participation in the pathophysiology of lung cancer through entangled molecular mechanisms.

Common Allergens Prevalent in and Around Ambala, Haryana: An Intradermal Study among Patients with Asthma and Allergic Rhinitis and Atopic Dermatitis.

OBJECTIVE: Identification of allergens by intradermal test in patients with asthma, allergic rhinitis, and eczema. MATERIALS AND METHODS: Intradermal test was performed in 100 patients of Ambala over an 8-year period to identify the common allergens. A total of 197 allergens including 50 types of pollen, 19 fungi, 17 insects, 14 types of dust, 6 animal dander, 7 types of fabric and feather and 82 types of foods, dust mite, and parthenium were tested. RESULTS: In this study, the major allergens were pollen (51%) followed by foods (28.9%), insects (26.9%), fungus (12.6%), dusts (6.7%). Among pollen allergens, Brassica campestris (8%) was a major allergen followed by Ageratum conyzoides (7%) and Artemisia scoparia (6%) Cannabis sativa, Cynodon dactylon and Maerua arenaria (5%). Among the fungal group, Alternaria tenuis, Aspergillus flavus, Aspergillus fumigates, Candida albicans, Penicillium sp., Rhizopus nigricans (3%), Fusarium solani (2%) were found. In the insect group, mosquito (7%), ant (6%), grasshopper (5%), locust (male), moth, and house fly (4%) were the major allergens. Among the dust allergens, grain dust rice (3%), straw dust, house dust, and grain dust bajra (2%) were found. Among the food allergens, prawn (5%), almonds, Baker's yeast, Bengal gram (3%) and mushroom, mango ripe, rajma, cinnamon, chocolate, beans fresh, and areca nut (2%) were found. CONCLUSION: It can be said that the knowledge may help to create a mapping of allergens in this area and help to treat patients by immunotherapy or avoidance strategy.

Association of tuberculosis and diabetes Mellitus: an analysis of 1000 consecutively admitted cases in a tertiary care hospital of North India.

INTRODUCTION: The association of Tuberculosis and Diabetes Mellitus is a cause of concern for the health sector. The coexistence of these two highly prevalent diseases has made the already existing treatments very complex. This issue is of particular significance to developing countries like India that bear a significant burden of these two diseases. METHODS: Retrospective analysis of 1000 consecutively admitted patients in a tertiary care hospital were analyzed for the coexistence of Tuberculosis and Diabetes Mellitus. RESULTS: The study found that a significant proportion of diabetic patients had coexistent tuberculosis (65.5%). Rural population was predominantly affected in both the genders. The study observed that the coexistence of these two conditions increased with advanced age. CONCLUSION: The coexistence of Diabetes Mellitus with Tuberculosis needs to identified early and adequately addressed. The rural population needs to be educated about these two conditions and seek timely medical care.

Fluorescent microscopy and Ziehl-Neelsen staining of bronchoalveolar lavage, bronchial washings, bronchoscopic brushing and post bronchoscopic sputum along with cytological examination in cases of suspected tuberculosis.

OBJECTIVES: Ever since the discovery of Mycobacterium tuberculosis in 1882, many diagnostic methods have been developed. However "The gold standard" for the diagnosis of tuberculosis (TB) is still the demonstration of acid fast Bacilli (AFB) by microscopic examination of smear or bacteriological confirmation by culture method. MATERIALS AND METHODS: In suspected 75 patients with active pulmonary TB, the materials obtained bronchoscopically, were bronchoalveolar lavage (BAL), bronchial brushings, bronchial washings and post bronchoscopic sputum. Four smears were made from each of the specimen. Fluorescent Staining, Ziehl-Neelsen (ZN), Pap and May Grunwald-Giemsa (MGG) stains were carried out for cytological examination. RESULTS: Fluorescent stain yielded maximum AFB positivity in all the methods, that is 36 (48%) in post fibre-optic bronchoscopy (FOB) sputum and 19 (25.33%) by fluorescence microscopy in both bronchial brushings and bronchial washings. Maximum yield of AFB with ZN staining 12 (16%) was equal to the post FOB sputum and bronchial brushings samples. It was followed by 6 cases (8%) in BAL and 4 (5.3%) in bronchial washings. The cytological examination was suggestive of TB in only 8 (10.66%) cases in bronchial washings and 6 (8%) cases in post FOB collection. It was equal in BAL and Bronchial brushings each that is 5 (6.67%). CONCLUSION: Bronchoscopy is a useful diagnostic tool and fluorescent microscopy is more sensitive than ZN and cytology. On X-ray examination, other diseases like malignancy or fungus can also mimick TB. So apart from ZN staining or fluorescence microscopy, Pap and MGG stain will be worthwhile to identify other microorganisms.

Guidelines for Diagnosis and Management of Bronchial Asthma: Joint Recommendations of National College of Chest Physicians (India) and Indian Chest Society.

Bronchial asthma is an important public health problem in India with significant morbidity. Several international guidelines for diagnosis and management of asthma are available, however there is a need for country-specific guidelines due to vast differences in availability and affordability of health-care facilities across the globe. The Indian Chest Society (ICS) and the National College of Chest Physicians (NCCP) of India have collaborated to develop evidence-based guidelines with an aim to assist physicians at all levels of health-care in diagnosis and management of asthma in a scientific manner. Besides a systematic review of the literature, Indian studies were specifically analysed to arrive at simple and practical recommendations. The evidence is presented under these five headings: (1) definitions, epidemiology and impact, (2) diagnosis, (3) pharmacologic management of stable disease, (4) management of acute exacerbations, and (5) non-pharmacologic management and special situations. The modified grade system was used for classifying the quality of evidence as 1, 2, 3, or usual practice point (UPP). The strength of recommendation was graded as A or B depending upon the level of evidence.

Effect of health education on compliance in asthma.

Non-compliance is a major problem in the treatment of any chronic disease. Asthma is one such chronic disease where non-compliance is a major problem. Several factors for non-compliance like cost of the drugs and relief of symptoms on taking medications for a short time are common to all chronic diseases but in asthma, inhalational therapy especially in elderly because of senile changes, economic factors and comorbidities is an added factor. Health education can improve the compliance. So a study was undertaken to observe the causes of non-compliance in asthmatics, to compare these causes in elderly and young asthmatics and to see the effect of health education on non-compliance in asthmatics. One hundred patients of bronchial asthma, group A consisting 50 patients > or = 65 years old and group B 50 patients < 40 years attending tuberculosis and chest diseases hospital, Patiala were studied. Initial compliance and reasons for non-compliance, initial usage of inhalational devices and their techniques of inhalation were studied. Patients were educated on asthma and inhalational techniques.These patients were followed up at 15th day, 1st, 2nd and 3rd months for changes in compliance and lung functions. At the initial stage, non-compliance was observed in 30 patients (60%) amongst elderly and only 15 patients (30%) amongst the young. Cause for non-compliance was cost in 6, memory in 11, both cost and memory in 7 and relief of symptoms in 6 in elderly and 4, 0, 0 and 11 in young respectively. Only 23 patients (40%) amongst the elderly and 33 patients (66%) amongst the young were put on inhalational therapy. There were statistically significant differences between elderly and young with respect to all the above factors. Out of these 23 elderly, only 7 (30.4%) were confident of the technique of inhalation and demonstrated it correctly.The health education resulted in significant improvement in compliance in both the groups, but more so in the young. The compliance for medications improved from a baseline of 40% to 88% amongst elderly and from 70% to 96% amongst young. There was an improvement in the confidence and technique of inhalation from 44% to 86% amongst elderly and from 64% to 98% amongst young at follow-ups. Mean PEFR improved statistically significantly from 75.25% to 81.13% in elderly and 84.38% to 89.74% in young asthmatics. However, during the follow-up at 2nd month, it was realised that 7 patients amongst elderly and 1 amongst young could not be sustained on inhalational medications because of the cost factor, therefore they were put on only oral medications which were supplied free of cost to them and thus at followup at 3rd month, these patients were not on inhalational medications any more.

Impact of interaction of polymorphic forms of p53 codon 72 and N-acetylation gene (NAT2) on the risk of lung cancer in the North Indian population.

The interaction of genetic and environmental factors can determine individual susceptibility to various cancers. We studied the influence of NAT2 and codon 72 p53 polymorphisms on 151 patients with lung cancer and an equal number of matched population controls. Polymorphisms of NAT2 and p53 were determined by PCR-RFLP techniques. The results were analyzed using logistic regression analysis. A statistically significant relationship between NAT2*5 and NAT2*6 alleles and lung cancer risk was observed. In addition, the population with slow acetylator alleles for NAT2*5 and NAT2*6 had a significantly higher risk of lung cancer compared with rapid acetylator alleles both in smokers and nonsmokers. The combined genotype of heterozygous arginine (Arg)/proline (Pro), Pro/Pro, and slow acetylator alleles of NAT2*5 and NAT2*6 showed higher, although not significant, risk of lung cancer compared with Arg/Arg and rapid acetylator alleles of NAT2*5 and NAT2*6. In conclusion, these findings suggest that the influence of NAT2 genotype, alone or in combination with p53 genotype, may confer increased susceptibility to lung cancer.

No association of DNA ligase-I polymorphism with the risk of lung cancer in north-Indian population.

DNA ligases play an essential role in repair, replication, and recombination of DNA, and catalyzes the formation of a phosphodiester bond at a nick junction on single- and double-strand breaks. We have conducted a hospital-based case-control study to examine the role of polymorphism of DNA repair gene ligase I (LIGI) in the context of lung cancer risk for north Indian population. One hundred, fifty-one primary lung cancer cases and an equal number of matching hospital controls were collected. The LIGI polymorphism was determined by using the PCR-RFLP method. The association between polymorphisms in the LIGI gene with the risk of lung cancer was estimated by computing odds ratios (ORs) and a 95% confidence interval (CI) using a Multivariate Logistic Regression Analysis. The risk for lung cancer was not associated for individuals featuring LIGI (AC) (OR -0.8, 95% CI = 0.44-1.40) and (AA) (OR -0.8, 95% CI = 0.41-1.80) genotypes. The DNA repair gene (LIGI) may not be playing an important role in modulating the risk of lung cancer in the north Indian population.

Dengue shock syndrome in newborn: a case series.

Four cases of dengue shock syndrome were seen during an epidemic of dengue fever. Three cases recovered following appropriate management.