Therapeutic implication of MicroRNA-320a antagonist in attenuating blood clots formed during venous thrombosis.

Venous thrombosis (VT) is a complex multi-factorial disease and a major health concern worldwide. Its clinical implications include deep vein thrombosis (DVT) and pulmonary embolism (PE). VT pathogenesis involves intricate interplay of various coagulants and anti-coagulants. Growing evidences from epidemiological studies have shown that many non-coding microRNAs play significant regulatory role in VT pathogenesis by modulating expressions of large number of gene involved in blood coagulation. Present study aimed to investigate the effect of human micro RNA (hsa-miR)-320a antagonist on thrombus formation in VT. Surgery was performed on Sprague-Dawley (SD) rats, wherein the inferior vena cava (IVC) was ligated to introduce DVT. Animals were divided into four groups (n = 5 in each group); Sham controls (Sham), IVC ligated-DVT (DVT), IVC ligated-DVT + transfection reagent (DVT-NC) and IVC ligated-DVT + miR320a antagonist (DVT-miR-320a antagonist). IVC was dissected after 6 h and 24 h of surgery to estimate thrombus weight and coagulatory parameters such as levels of D-dimer, clotting time and bleeding time. Also, ELISA based biochemical assays were formed to assess toxicity of miRNA antagonist in animals. Our experimental analysis demonstrated that there was a marked reduction in size of thrombus in hsa-miR-320a antagonist treated animals, both at 6 h and 24 h. There was a marked reduction in D-dimer levels in hsa-miR-320a antagonist treated animals. Also, blood clotting time was delayed and bleeding time was increased significantly in hsa-miR-320a antagonist treated rats compared to the non-treated and Sham rats. There was no sign of toxicity in treated group compared to control animals. Hsa-miR-320a antagonist could be promising therapeutic target for management of VT.

Prevalence and Determinants of Sarcopenic Obesity in Older Adults: Secondary Data Analysis of the Longitudinal Ageing Study in India (LASI) Wave 1 Survey (2017-18).

INTRODUCTION: Sarcopenic obesity (SO) represents the confluence of two epidemics-an aging population and an increasing rate of obesity. The two diseases may act synergistically, and SO may significantly affect morbidity and mortality. However, the burden is not defined to drive the policy changes. Hence the present study was done to estimate the prevalence and predictors of SO in India. METHODS: We did a secondary data analysis of the 72,250 older adults who participated in the first wave of the Longitudinal Aging Study in India (2017-18). Possible sarcopenia was defined as per the guidelines by the Asian Working Group for Sarcopenia (AWGS) criteria. The modified criterion of overweight and obesity for Asian adults was used to categorize obesity. Presence of both sarcopenia and obesity depicted SO. Weighted analysis was done to estimate the prevalence of SO, and multinomial bivariate logistics regression was used to identify the predictors of SO. RESULTS: The overall prevalence of obesity, sarcopenia, and SO was 27.1%, 41.9%, and 8.7%, respectively. The mean age, weight, body mass index (BMI), and blood pressure of adults with SO were significantly higher compared to others. Higher age, urban residence, west and south regions of India, consumption of tobacco or alcohol, no physical activity, and presence of diabetes contribute to SO. CONCLUSION: The burden of SO seems to be less but amounts to a massive number in an aging country. We stress increased screening of the geriatric age group and advocate increased physical activity and dietary modifications to realize the concept of healthy aging.

NMR metabolomic and microarray-based transcriptomic data integration identifies unique molecular signatures of hypersensitivity pneumonitis.

Hypersensitivity pneumonitis (HP) is an immune-mediated granulomatous interstitial lung disease (ILD) that results from repeated inhalation of certain antigens. Despite major advances in research, pathophysiology of the disease remains poorly understood. The present study combines metabolomic and transcriptomic data to determine alterations in HP subjects as compared with healthy controls. Metabolic signatures were identified in serum, exhaled breath condensate (EBC) and bronchoalveolar lavage fluid (BALF) of HP patients using proton nuclear magnetic resonance (NMR) metabolomics. The expression of three metabolites, i.e., lactate, pyruvate, and proline, was found to be significantly altered in all three biofluids. The potential of differential diagnosis based on these three metabolites was investigated by including a group of patients with sarcoidosis, which is another type of granulomatous ILD. In addition, differentially expressed transcriptomic fingerprints in blood samples were identified by analyzing a Gene Expression Omnibus (GEO) database. The transcriptomics analysis of these microarray-based data revealed 59 genes to be significantly dysregulated in patients with HP. Over representation analysis of the metabolites and genes of interest was performed using IMPaLA (Integrated Molecular Pathway Level Analysis) version 12. Integrated analysis of serum metabolite signatures and blood gene expression suggests dysregulation of PI3K-AKT signaling and TCA cycle pathways in these patients. This preliminary study is a step towards better understanding of the pathogenesis of HP by identification of differentially expressed metabolites and transcriptomic fingerprints. These molecular signatures may be explored as diagnostic markers for differentiating HP from other lung diseases.

Global metabolome profiling of exhaled breath condensates in male smokers with asthma COPD overlap and prediction of the disease.

Asthma-chronic obstructive pulmonary disease (COPD) overlap, termed as ACO, is a complex heterogeneous disease characterised by persistent airflow limitation, which manifests features of both asthma and COPD. These patients have a worse prognosis, in terms of more frequent and severe exacerbations, more frequent symptoms, worse quality of life, increased comorbidities and a faster lung function decline. In absence of clear diagnostic or therapeutic guidelines, ACO presents as a challenge to clinicians. The present study aims to investigate whether ACO patients have a distinct exhaled breath condensate (EBC) metabolic profile in comparison to asthma and COPD. A total of 132 age and BMI matched male smokers were recruited in the exploratory phase which consisted of (i) controls = 33 (ii) asthma = 34 (iii) COPD = 30 and (iv) ACO = 35. Using nuclear magnetic resonance (NMR) metabolomics, 8 metabolites (fatty acid, propionate, isopropanol, lactate, acetone, valine, methanol and formate) were identified to be significantly dysregulated in ACO subjects when compared to both, asthma and COPD. The expression of these dysregulated metabolites were further validated in a fresh patient cohort consisting of (i) asthma = 32 (ii) COPD = 32 and (iii) ACO = 40, which exhibited a similar expression pattern. Multivariate receiver operating characteristic (ROC) curves generated using these metabolites provided a robust ACO classification model. The findings were also integrated with previously identified serum metabolites and inflammatory markers to develop a robust predictive model for differentiation of ACO. Our findings suggest that NMR metabolomics of EBC holds potential as a platform to identify robust, non-invasive biomarkers for differentiating ACO from asthma and COPD.

Proteomics in idiopathic pulmonary fibrosis: the quest for biomarkers.

Idiopathic pulmonary fibrosis (IPF) is a debilitating chronic progressive and fibrosing lung disease that culminates in the destruction of alveolar integrity and dismal prognosis. Its etiology is unknown and pathophysiology remains unclear. While great advances have been made in elucidating the pathogenesis mechanism, considerable gaps related to information on pathogenetic pathways and key protein targets involved in the clinical course of the disease exist. These issues need to be addressed for better clinical management of this highly challenging disease. Omics approach has revolutionized the entire area of disease understanding and holds promise in its translation to clinical biomarker discovery. This review outlines the contribution of proteomics towards identification of important biomarkers in IPF in terms of their clinical utility, i.e. prognosis, differential diagnosis, disease progression and treatment monitoring. The major dysregulated pathways associated with IPF are also discussed. Based on numerous proteomics studies on human and animal models, it is proposed that IPF pathogenesis involves complex interactions of several pathways such as oxidative stress, endoplasmic reticulum stress, unfolded protein response, coagulation system, inflammation, abnormal wounding, fibroblast proliferation, fibrogenesis and deposition of extracellular matrix. These pathways and their key path-changing mediators need further validation in large well-planned multi-centric trials at various geographical locations for successful development of clinical biomarkers of this confounding disease.

Metabolomic fingerprinting and systemic inflammatory profiling of asthma COPD overlap (ACO).

BACKGROUND: Asthma-COPD overlap (ACO) refers to a group of poorly studied and characterised patients reporting with disease presentations of both asthma and COPD, thereby making both diagnosis and treatment challenging for the clinicians. They exhibit a higher burden in terms of both mortality and morbidity in comparison to patients with only asthma or COPD. The pathophysiology of the disease and its existence as a unique disease entity remains unclear. The present study aims to determine whether ACO has a distinct metabolic and immunological mediator profile in comparison to asthma and COPD. METHODS: Global metabolomic profiling using two different groups of patients [discovery (D) and validation (V)] were conducted. Serum samples obtained from moderate and severe asthma [n = 34(D); n = 32(V)], moderate and severe COPD [n = 30(D); 32(V)], ACO patients [n = 35(D); 40(V)] and healthy controls [n = 33(D)] were characterized using gas chromatography mass spectrometry (GC-MS). Multiplexed analysis of 25 immunological markers (IFN-γ (interferon gamma), TNF-α (tumor necrosis factor alpha), IL-12p70 (interleukin 12p70), IL-2, IL-4, IL-5, IL-13, IL-10, IL-1α, IL-1ß, TGF-ß (transforming growth factor), IL-6, IL-17E, IL-21, IL-23, eotaxin, GM-CSF (granulocyte macrophage-colony stimulating factor), IFN-α (interferon alpha), IL-18, NGAL (neutrophil gelatinase-associated lipocalin), periostin, TSLP (thymic stromal lymphopoietin), MCP-1 (monocyte chemoattractant protein- 1), YKL-40 (chitinase 3 like 1) and IL-8) was also performed in the discovery cohort. RESULTS: Eleven metabolites [serine, threonine, ethanolamine, glucose, cholesterol, 2-palmitoylglycerol, stearic acid, lactic acid, linoleic acid, D-mannose and succinic acid] were found to be significantly altered in ACO as compared with asthma and COPD. The levels and expression trends were successfully validated in a fresh cohort of subjects. Thirteen immunological mediators including TNFα, IL-1ß, IL-17E, GM-CSF, IL-18, NGAL, IL-5, IL-10, MCP-1, YKL-40, IFN-γ, IL-6 and TGF-ß showed distinct expression patterns in ACO. These markers and metabolites exhibited significant correlation with each other and also with lung function parameters. CONCLUSIONS: The energy metabolites, cholesterol and fatty acids correlated significantly with the immunological mediators, suggesting existence of a possible link between the inflammatory status of these patients and impaired metabolism. The present findings could be possibly extended to better define the ACO diagnostic criteria, management and tailoring therapies exclusively for the disease.

Metabolomic signatures of asthma-COPD overlap (ACO) are different from asthma and COPD.

INTRODUCTION: Asthma-chronic obstructive pulmonary disease (COPD) overlap, termed as ACO, is a complex heterogeneous disease without any clear diagnostic or therapeutic guidelines. The pathophysiology of the disease, its characteristic features, and existence as a unique disease entity remains unclear. Individuals with ACO have a faster lung function decline, more frequent exacerbations, and worse quality of life than those with COPD or asthma alone. OBJECTIVES: The present study aims to determine whether ACO has a distinct metabolic profile in comparison to asthma and COPD. METHODS: Two different groups of patients were recruited as discovery (D) and validation (V) cohorts. Serum samples obtained from moderate and severe asthma patients diagnosed as per GINA guidelines [n = 34(D); n = 32(V)], moderate and severe COPD cases identified by GOLD guidelines [n = 30(D); 32(V)], ACO patients diagnosed by joint GOLD and GINA guidelines [n = 35(D); 40(V)] and healthy controls [n = 33(D)] were characterized using nuclear magnetic resonance (NMR) spectrometry. RESULTS: Multivariate and univariate analysis indicated that 12 metabolites [lipid, isoleucine, N-acetylglycoproteins (NAG), valine, glutamate, citric acid, glucose, L-leucine, lysine, asparagine, phenylalanine and histidine] were dysregulated in ACO patients when compared with both asthma and COPD. These metabolites were further validated in a fresh cohort of patients, which again exhibited a similar expression pattern. CONCLUSIONS: Our findings suggest that ACO has an enhanced energy and metabolic burden associated with it as compared to asthma and COPD. It is anticipated that our results will stimulate researchers to further explore ACO and unravel the pathophysiological complexities associated with the disease.

Long term treatment of metformin impedes development of chemoresistance by regulating cancer stem cell differentiation through taurine generation in ovarian cancer cells.

Development of resistance poses a significant challenge to effective first-line platinum based therapy for epithelial ovarian cancer patients. Cancer Stem Cells are envisaged as a critical underlying factor for therapy resistance. Thus, there is a critical need for developing approaches to diminish the enrichment of cancer stem cells and acquirement of resistance. Administration of metformin, a commonly prescribed drug against Type II diabetes exhibited promising effect in the management of ovarian cancer. However, the effect of long term administration of low dose of metformin as an adjuvant to cisplatin and paclitaxel during acquirement of chemoresistant phenotype has not been investigated so far. Using two isogenic cellular chemoresistant models (A2780 and OAW42) developed in the presence or absence of metformin, we demonstrated the ability of metformin to impede the development of resistance through increased drug sensitivity, increased proliferation, and reduced migratory abilities of the resistant cells. Metformin introduction also decreased the cancer stem cell population, expression of specific biomarkers and pluripotent genes. Further metabolic profiling of these cells using 1H-Nuclear Magnetic Resonance spectroscopy revealed significant modulation in taurine and histidine levels in resistant cells developed in the presence of metformin. Intriguingly, taurine treatment considerably reduced the cancer stem cell population and chemoresistance in resistant cells, indicating a novel role of taurine in differentiation of ovarian cancer stem cells. Altogether this is the first report on the potential role of metformin for targeting the cancer stem cell population via up regulation of taurine, leading to impediment in the acquirement of chemoresistance.

Trend and pattern of various types of cancer with special reference to gall bladder cancer in north bengal medical college, west bengal, India: a 3 years record based study.

BACKGROUND: Global burden of cancer is on rise and trends and pattern of cancers are rapidly changing different geographic and population groups. Gall bladder cancers are emerging with increasing proportion among select areas and groups and understanding these variations are important for appropriate strategies and interventions. However, absence of a well-developed universal cancer registry system in India, institution based secondary data analysis can generate useful information. The present study was conducted to determine the trend and pattern of cancer cases (with special reference to gall bladder cancer) treated in a tertiary care hospital in northern part of West Bengal. METHODS: Record based cross-sectional study was conducted in Department of Radiotherapy, North Bengal Medical College and Hospital. All newly registered cases between (2010 and 2012) were reviewed. RESULTS: A total of 2058 cancer cases were recorded during the 3-year period. Of these, major types of cancers were oro-pharynx (16.1%), breast (15.4%), cervix (13.2%), lung (12.7%), gall bladder (6.5%) stomach cancer (6.4%), etc., Increasing proportions was observed for breast and gall bladder cancers. The proportion of gallbladder cancer cases in 2010, 2011, and 2012 were 3.8%, 7.3% and 7.8%, respectively. Among 134 gall bladder cancer cases, 93.3% were females, 85.1% alcoholics, 57.4% had a history of fatty liver, 94% had adeno/adenosquamous carcinomas, and 65.7% were metastatic in nature. CONCLUSIONS: Increasing trend is observed in gall bladder cancer cases emphasizing the need for further large scale studies.

Fine needle aspiration cytology of non-hematological neoplasms in pediatric age group: Our experience.

BACKGROUND: The role of aspiration cytology has largely been ignored in pediatric population. The present study was undertaken to evaluate the role of fine needle aspiration cytology (FNAC) in non-hematological neoplasms in children in our institution, which is a rural tertiary care center. MATERIALS AND METHODS: A total of 88 cases of non-hematological pediatric mass lesions were studied in which cytopathological diagnosis could be corroborated with histopathology. RESULTS: Out of all the cases, 70 (80%) cases were benign tumors and 18 (20%) were malignant tumors. Fibroadenoma (37.9%) comprised the majority of cases in the benign category while small round cell tumors (SRCTs) (44.4%) comprised the majority of cases in the malignant category. Definite diagnosis could be offered based on the cytomorphology in 79.5% cases, while in 20.5% of cases only a broad cytological classification could be offered. Among the malignant lesions, FNAC showed 100% sensitivity while a specific diagnosis was made in 90% of cases. CONCLUSION: FNAC proved to be a rapid and fairly accurate tool in diagnosing non-hematological tumors in the pediatric age group.

Cytohistopathological correlation of a case of squamous cell carcinoma of gallbladder with lymph node metastasis.

Primary carcinomas of the gallbladder are rare malignancies and adenocarcinoma is the more common subtype. Primary squamous cell carcinoma of the gallbladder has rarely been diagnosed by aspiration cytology. Here, we present a case of a 62-year old female patient suffering from abdominal complaints who underwent ultrasound-guided fine-needle aspiration cytology and was diagnosed as keratinizing squamous cell carcinoma of gallbladder. The diagnosis was confirmed on subsequent histopathological examination that also revealed metastasis in the cystic lymph node. Histogenesis and biological behavior of squamous cell carcinoma of gallbladder remains a matter of debate but the role of aspiration cytology in diagnosing these lesions cannot be undermined.

Cytologic diagnosis of undifferentiated high grade pleomorphic sarcoma of breast presenting with brain metastasis.

Primary sarcoma of breast are rare. Diagnosis by aspiration cytology is difficult due to nonspecific cytomorphologic features. An initial presentation with neurological symptoms due to metastasis of breast sarcoma to the brain has not been previously reported. Here, we describe a case of a 60-year-old female who presented with headache, dizziness and convulsion and was subsequently diagnosed with undifferentiated high grade pleomorphic sarcoma of breast with cerebellar metastasis.

Giant adrenal myelolipoma with hemorrhage masquerading as retroperitoneal sarcoma.

Adrenal myelolipomas are functionally inactive, rare adrenal tumors which are usually small in size and are discovered incidentally. Giant symptomatic myelolipomas have rarely been reported in medical literature. Here, we describe the case of a 40-year-old female patient who presented to the surgical outpatient department with left flank pain. An ultrasonogram of the abdomen suggested a large retroperitoneal tumor which was then surgically resected. Histopathological examination of the resected specimen revealed a giant adrenal myelolipoma with intratumoral hemorrhage. The patient was relieved of symptoms and was free of any complaints in follow-up.

Oncocytic lesion of parotid gland: A dilemma for cytopathologists.

Oncocytes are epithelial cells with abundant, granular, eosinophilic cytoplasm due to presence of numerous large mitochondria of varied sizes. The presence of oncocytes in salivary glands can occur in a variety of conditions. Here, we present a rare case of a 68 year old male patient who presented with a 6 cm diameter swelling in the right parotid region. A fine needle aspiration cytology done from the lesion showed a cellular oncocytic lesion. A possibility of oncocytoma was entertained. Histopathology of the mass showed a rare entity called diffuse hyperplastic oncocytosis. Originally believed to be a metaplastic process, oncocytes can occur in various lesions ranging from hyperplastic conditions to malignant neoplasms. However, diagnosis on cytological smears can be very challenging for the cytopathologist.

Computed tomography guided fine needle aspiration cytology of mass lesions of lung: Our experience.

CONTEXT: Computerized tomography (CT) guided fine needle aspiration cytology (FNAC) of lung lesions has rapidly emerged as a less-invasive, cheap, rapid and fairly accurate diagnostic aid in lung lesions. AIMS: The purpose of this present study was to evaluate the effectiveness of CT-guided FNAC in the diagnosis of pulmonary mass lesions (both benign and malignant) and to determine the complication rate of this procedure. SETTINGS AND DESIGN: We conducted an institution-based, prospective study on 127 patients who presented with pulmonary mass lesions. MATERIALS AND METHODS: After proper consent was obtained, CT-guided transthoracic fine needle aspiration was done and their diagnoses were confirmed by appropriate methods. The results were analyzed statistically. RESULTS: Out of 127 cases selected for the study, 59.8% were males while the rest were females. Cough was the most common symptom present in 71.2% cases, followed by weight loss (62.4%). 21.2% cases were cytologically benign. Adenocarcinoma (54.2%) was the commonest malignant tumor. FNAC provided at least 96% sensitivity and 100% specificity in diagnosing lung tumors. Among the benign lesions, specific diagnoses were obtained in 48.1% cases. Thus, altogether a specific diagnosis was obtained in 109 of 127 cases, i.e. 85.8%. No major complication was noted. CONCLUSIONS: CT-guided FNAC is an extremely valuable and fairly accurate diagnostic aid of intrathoracic mass lesions, with a reasonable rate of complication.

Post-salpingectomy endometriosis: An under-recognized entity.

We report a case of a 48-year old lady, who presented with complaints of lower abdominal pain and menorrhagia for the last four months. The patient had undergone bilateral salpingectomy four years back by the Pomeroy technique. Ultrasonography revealed an ovarian cyst on the right side. A total abdominal hysterectomy with bilateral salpingo-oophorectomy was performed and the specimen was sent for histopathological examination. It revealed that the normal mucosa of the tubectomy stump was completely replaced by endometrial tissue. Tubal endometriosis remains an under-recognized entity, due to less extensive routine sampling of the fallopian tubes, and they may be also be associated with other pathologies, as was in the present case.