[Show abstract][Hide abstract] ABSTRACT: Background:
Survivin expression is often associated with aggressive tumor behavior and therapy resistance. In this study, we investigated the effect of survivin knockdown by survivin-siRNA lentiviral vector (Svv-Lent) on the response of HNSCC to chemo-radiotherapy, tumor growth and metastasis.
Four human HNSCC (OSC19, Cal27, Cal33 and FaDu) and one normal HOK cell lines were included in the study, and survivin knockdown was achieved with Svv-Lent treatment. Cell proliferation and apoptosis were measured by MTT and TUNEL assay, respectively. Transwell assays were performed to measure in vitro cell migration and matrigel invasion. Xenograft tumors were developed in nude mice by injecting Cal27 cells subcutaneously and following tail-vein injection of lung and liver metastasis.
Knockdown of survivin significantly suppressed HNSCC cell proliferation and induced apoptosis in vitro. Survivin inhibition could also significantly reduce in vitro cell migration and matrigel invasion that might be due to inactivation of matrix metalloproteinases. In vivo studies showed significant repression of Cal27 xenograft tumor growth and tissue metastasis leading to improvement in mice survival in the Svv-Lent treated group compared to controls. Our data indicated that survivin expression in HNSCC cells contributed to chemo-radioresistance, and its down-regulation increased anti-cancer effects of paclitaxel, cisplatin and radiation.
Our findings suggest that sustained survivin expression facilitates HNSCC tumor growth and confers resistance to chemo-radiotherapy. Svv-Lent therapy may be able to enhance the cytotoxic effect of commonly used anticancer drugs such as cisplatin and paclitaxel, and radiotherapy that could provide a promising strategy for the effective control of resistant head and neck cancer.
Full-text · Article · Dec 2015 · Radiotherapy and Oncology
[Show abstract][Hide abstract] ABSTRACT: Oral cancer is one of the most commonly found cancers in many South-Asian underdeveloped countries, especially among men in comparison to women. When considering the mortality rate among all types of existing cancers, in India oral cancer is the primary reason for death in men. Some of the major reasons contributing to the high mortality rate are late diagnosis, lack of treatment options and high prevalence of tobacco consumption. Oral cancer is often diagnosed at a stage when cancer cells have already become aggressive and become resistant to standard therapeutic options. Progression, apoptosis, angiogenesis, metastasis and invasion behold great capability to treat and detect cancer at the molecular level. Dysregulation of apoptosis is one of the most common molecular events known to be associated with the development of oral cancer. In this review, we discuss key apoptotic markers which can be used as prognostic and/or therapeutic targets in oral cancer.
[Show abstract][Hide abstract] ABSTRACT: Rapid and accurate diagnosis is important for preventing transmission of Mycobacterium tuberculosis. Currently available tuberculosis (TB) diagnostic methods lack desired sensitivity and specificity, and require sophisticated equipment and skilled workforce including weeks' long duration to yield results. In this study, extracellular proteins or secretory protein antigens of M. tuberculosis H37Rv have been isolated using ion exchange chromatography, immunocharacterized and exploited for the development of efficient enzyme-linked immunosorbent assay (ELISA) for diagnosis of active TB with enhanced specificity and sensitivity. Apparent molecular masses for purified proteins were found to be 6, 27, 30, 38 and 64 kDa. Out of five purified proteins, one (64 kDa) was found to be novel. Of the five proteins, four (6, 27, 30 and 38 kDa) were found significant to be used in the development of ELISA for pulmonary and extra-pulmonary TB. The immune responses of serum samples of TB patients and other healthy subjects against the above-mentioned antigens' cocktail were evaluated. Critical parameters of newly developed ELISA were optimized and it was observed that the cocktail antigens have a greater specificity (98.06 %) and sensitivity (98.67 %) as compared to other commercially available diagnostic tests. The present findings suggest that the developed ELISA is an effective tool for routine screening and early-stage diagnosis of TB.
[Show abstract][Hide abstract] ABSTRACT: Metabolic syndrome (MS) is a cluster of metabolic abnormalities and serves as a precursor of cardiovascular disorders (CVD). The objective of the study was to determine the prevalence of MS and its association with Diabetes mellitus (DM) in an urban setup of Raipur city of Chattishgarh State. A cross sectional study was conducted on 400 randomly selected subjects (men 186, women 214 of age group 10-80 years) in the study region. Anthropometric variables, blood pressure, and lipid profile were monitored besides blood glucose in the study population. The incidence of MS and DM were worked out as per the criteria of National Cholesterol Education Programme-ATPIII (NCEPATPIII) and World Health Organization.The incidence of MS was found to be 15% and 5% by ATPIII and WHO criteria respectively. According to NCEP-ATPIII criterion, women recorded significantly high incidence of MS (65%) as compared to males (35%). About 28% of subjects with MS exhibited type II DM (males 16.6% and females 11.6%). On the other hand, the incidence of type II DM was found to be 6.7% in non-metabolic syndrome group of subjects as per NCEP-ATPIII criterion. By WHO criteria, 65% of the subjects with MS exhibited type II DM (males 35% and females 30%) while the incidence is 6.9% in non-metabolic syndrome group of subjects. Subjects in age group of 41-65 years showed the highest incidence of MS irrespective of the criteria employed. The incidence of MS differed significantly by the two criteria employed and it was more by NCEP-ATP-III criteria. The prevalence of type II DM was significantly high in subjects with metabolic syndrome and it was more in males of age group of 41-65 yrs.
Full-text · Article · Jul 2013 · Indian Journal of Public Health Research and Development
[Show abstract][Hide abstract] ABSTRACT: Acute myocardial infarction (AMI) is the leading cause of death worldwide, with early diagnosis still being difficult. Promising new cardiac biomarkers such as troponins and creatine kinase (CK) isoforms are being studied and integrated into clinical practice for early diagnosis of AMI. The cardiac-specific troponins I and T (cTnI and cTnT) have good sensitivity and specificity as indicators of myocardial necrosis and are superior to CK and its MB isoenzyme (CK-MB) in this regard. Besides being potential biologic markers, cardiac troponins also provide significant prognostic information. The introduction of novel high-sensitivity troponin assays has enabled more sensitive and timely diagnosis or exclusion of acute coronary syndromes. This review summarizes the available information on the potential of troponins and other cardiac markers in early diagnosis and prognosis of AMI, and provides perspectives on future diagnostic approaches to AMI.
[Show abstract][Hide abstract] ABSTRACT: HIV (human immunodeficiency virus) infection has now become endemic worldwide and AIDS ranks fourth among the world's top killers of mankind. A rapid and accurate HIV testing assay is pre-requisites for practical applicability of diagnostic tests. The aim of present study was to design peptides cocktail as an antigen and development of ELISA test for HIV-1/2 antibody detection, with enhanced sensitivity and specificity. A novel peptide stretch V3-I, covering immunodominant epitope corresponding to V3 hypervariable loop of gp120 antigens of selected Indian isolates, has been studied and incorporated in an antigenic cocktail of gp36, gp41, and rp24 of HIV-1/2. Peptides from these antigens were chemically synthesized and an additional cysteine residue was added at both amino- and carboxyl- terminal sequences of each peptide in order to form inter and intramolecular disulfide bond for the folding of peptides. This generated conformational epitopes with increased oligomericity and stability of peptide sequences; and attachment of antigen to the solid support of ELISA plates. The use of antigenic cocktail of folded peptides and recombinant p24 enhanced sensitivity and specificity of ELISA test. Evaluation of the test using 1123 serum samples in comparison with Boston Biomedical Incorporation (BBI) panels showed 100% sensitivity and 99.3% specificity with no cross reactivity tribulation. In conclusion, "HIV Screen test" detects HIV 1/2 antibodies with a high degree of sensitivity and specificity and could be a promising tool for seroscreening of blood during transfusion, counseling and diagnosis of HIV-1/2.
Full-text · Article · Oct 2012 · Journal of immunological methods
[Show abstract][Hide abstract] ABSTRACT: Survivin is known to be highly-expressed in various carcinomas; and is associated with their biologically aggressive characteristics and drug resistance. We have previously reported survivin as an important anti-apototic protein involved in head and neck squamous cell carcinoma (HNSCC) tumorigenesis and providing resistance to conventional cancer therapies. The purpose of present study was to investigate the potential of survivin as a therapeutic target in HNSCC. This study was designed to explore the effect(s) of survivin-siRNA therapy on the apoptosis in HNSCC cells, and its influence on cisplatin-sensitivity. Lentivirus vector was developed to deliver survivin specific siRNA into cancer cells. The data indicates that silencing of survivin-mediated by Lentivirus-siRNA therapy effectively suppressed cancer cell proliferation and induced caspase-dependent apoptosis in HNSCC cells. The study also shows that the response of HNSCC cells to cisplatin drug treatment at clinically relevant level was limited. We observed survivin to be a key factor involved in this cisplatin-resistance mechanism, and down-regulation of survivin significantly increases sensitivity of cancer cells to cisplatin-mediated apoptosis. Thus, this combination therapy acts as a multimodality regimen with significant potential to improve clinical outcomes in head and neck cancers.
Full-text · Article · Sep 2012 · Current Gene Therapy
[Show abstract][Hide abstract] ABSTRACT: Lactoglobulin (β-LG) genetic polymorphisms are important and well known due to their effects on quantitative traits and technological properties of milk. At the DNA level, polymerase chain reaction (PCR)-single-strand conformation polymorphism (SSCP) allows for the detection of unknown polymorphisms at β-LG loci. Here we describe the usefulness of the PCR-SSCP technique for β-LG typing. In the present study, we amplified and sequenced the part of promoter region and all the seven exons containing the entire coding and untranslated region for the β-lactoglobulin gene in best dairy goat breeds of India namely: Jamunapari and Jakhrana. Nine polymorphisms were detected, one in the l promoter region, four in the introns and four in the exons of the β-lactoglobulin gene. All polymorphisms were single nucleotide substitutions. The polymorphisms in the coding region did not produce any amino acid change. Key words: β-Lactoglobulin gene, dairy goats, polymorphism, single nucleotide polymorphism (SNP), single strand conformational polymorphism (SSCP).
[Show abstract][Hide abstract] ABSTRACT: Cyclooxygenase-2 (Cox-2) is an inducible enzyme involved in the conversion of arachidonic acid to prostaglandin and other eicosanoids. Molecular pathology studies have revealed that Cox-2 is over-expressed in cancer and stroma cells during tumor progression, and anti-cancer chemo-radiotherapies induce expression of Cox-2 in cancer cells. Elevated tumor Cox-2 is associated with increased angiogenesis, tumor invasion and promotion of tumor cell resistance to apoptosis. Several experimental and clinical studies have established potent anti-cancer activity of NSAID (Non-steroidal anti-inflammatory drugs) and other Cox-2 inhibitors such as celecoxib. Much attention is being focused on Cox-2 inhibitors as beneficial target for cancer chemotherapy. The mode of action of Cox-2 and its inhibitors remains unclear. Further clinical application needs to be investigated for comprehending Cox-2 biological functions and establishing it as an effective target in cancer therapy.
Full-text · Article · Mar 2011 · Current drug targets
[Show abstract][Hide abstract] ABSTRACT: The present study deals with the evaluation of the efficacy of oxaliplatin and paclitaxel combination as a potential strategy in controlling HNSCC cell proliferation and the assessment of correlation between occurrence of apoptosis and changes in expression of survivin (IAP). The panel cell lines included two HNSCC cell lines (Cal27 and NT8e) and one normal cell line (293) with differential level of survivin expression in accordance with chemosensitivity. The cytotoxicity and effect of drugs on apoptosis was determined, separately and in combination. Combined treatment of cells with paclitaxel and oxaliplatin resulted in significantly higher cytotoxicity as compared to individual single drug treatment. Cytotoxicity was prominent in paclitaxel to oxaliplatin (pacl-oxal) sequence treatment with an approximate two-fold increase in apoptosis as compared to oxaliplatin to paclitaxel (oxal-pacl) sequence treatment. Paclitaxel treatment also caused increased survivin expression showing reduced apoptosis at low concentration. Oxaliplatin, when combined with paclitaxel, decreased the survivin level with increased cell death. Inhibition of survivin by a small interfering RNA (siRNA) method also increased the sensitivity of the cancer cell lines to paclitaxel whereas over-expression of survivin in the transfected 293-cell line provided resistance. In conclusion, the interaction between drugs was synergistic and schedule-dependent. Survivin played a critical role in paclitaxel resistance through the suppression of apoptosis, and a significant induction of apoptosis was observed when oxaliplatin was combined with paclitaxel at least in part by the down-regulation of survivin.
Full-text · Article · Nov 2010 · Current cancer drug targets
[Show abstract][Hide abstract] ABSTRACT: Oxaliplatin is integrated in treatment strategies against a variety of cancers including radiation protocols. Herein, as a new strategy we tested feasibility and rationale of oxaliplatin in combination with radiation to control proliferation of head and neck squamous cell carcinoma (HNSCC) cells and discussed survivin-related signaling and apoptosis induction.
Cytotoxicity and apoptosis induced by radiation and/or oxaliplatin were examined in relation to survivin status using two HNSCC cell lines viz., Cal27 and NT8e, and one normal 293-cell line. Survivin gene knockdown by siRNA was also tested in relevance to oxaliplatin-mediated radiosensitization effects.
Survivin plays a critical role in mediating radiation-resistance in part through suppression of apoptosis via a caspase-dependent mechanism. Oxaliplatin treatment significantly decreased expression of survivin in cancer cells within 24-72 h. Apoptotic cells and caspase-3 activity were increased parallely with decrease in cell viability, if irradiated during this sensitive period. The cytotoxicity of oxaliplatin and radiation combination was greater than additive. Survivin gene knockdown experiments have demonstrated the role of survivin in radiosensitization of cancer cells mediated by oxaliplatin.
Higher expression of survivin is a critical factor for radioresistance in HNSCC cell lines. Pre-treatment of cancer cells with oxaliplatin significantly increased the radiosensitivity through induction of apoptosis by potently inhibiting survivin.
Full-text · Article · Aug 2010 · Radiotherapy and Oncology
[Show abstract][Hide abstract] ABSTRACT: Mycobacterium tuberculosis is one of the most dangerous human pathogens
evolved on the Planet that poses big challenge to the scientists working on diagnostics and therapeutics. There is an urgent requirement for the development of a novel candidate as an immunoprophylactic agent (vaccine). The complete genome sequencing of M. tuberculosis
H37 Rv had paved way for the identification of candidate antigens of prophylactic significance. Numerous proteins especially membrane proteins acting as immunogens or subunit vaccines are profiled and investigated for immunoprophylactic activity. The protein of 71 kDa, which elicits the response of both B cell/T cell has been used for the development of candidate vaccine. However the use of proteinaceious antigens in
immunoprophylaxis is not encouraging for various reasons. The lipid based immunogenic antigens find to be more promising in this regard, since lipids escape the coventional antigen processing and are routed through CD1 system by a different pathway. The lipid antigens are endocytosed through a pathway involving protease resistant proteins such as saposin.
Saposin molecules help CD1 molecules to present the antigen to the T Cell and NKT cells. The NKT cells play a larger regulatory role and have immunomodulatory properties and hence NKT cells are attractive targets for the development of immunotherapies. Another advantage of lipid Ag induced immune response is its specificity, which is attributed by δ,γ region of the variable chain of the TCR. This region of TCR are not diverse in it function. Hence the rate of elimination of mycobacterium will be rapid if the lipid Ag is challenged. The other advantages of priming with lipid moieties include, i. rate of mutation of CD1 molecules is not rapid, limited polymorphism of CD1 etc. The use of liposome technology in enhancing
the immune response is under active invesitgation. Mycobacterial mannophosphoinositides (PIMs) encapsulated in liposomes made of egg phosphatidylcholine (EPC) and cholesterol (CH) (2:1.5 molar ratio) were able to induce humoral and delayed type hypersensitivity (DTH, foot-pad swelling reaction) responses in mice without the help of any carrier protein.
Animals immunized with this liposome glycophospholipid antigen demonstrated enhanced percent survival on intravenous challenge with virulent M. tuberculosis. The paper deals with the immunoprophylactic studies carried out with M. tuberculosis derived lipid based antigens.
[Show abstract][Hide abstract] ABSTRACT: Tuberculosis is the second most infectious disease and is the leading cause of death in the tropical World. The Mycobacterium tuberculosis infects about 1.7 billion people every year and is responsible for more than 2 million deaths globally each year. Overall, one-third of the world’s population is currently infected with the M. tb bacillus without demonstrable symptoms.
A delayed or missed diagnosis of active infection and prevalence of drug resistant strains of M. tuberculosis are major causes of transmission and mortality. With the threat of such an epidemic looming, and despite an enormous amount of research since the time of Koch, we still have no
simple, rapid, sensitive, and specific test to detect and differentiate most or all patients with active tuberculosis from those with quiescent lesions, previous vaccination, or other diseases, or even from those who are completely healthy. Tuberculosis co-infected with HIV has further
complicated the diagnosis since. HIV-positive patients with active TB are more likely to have negative sputum smears and atypical chest X-rays than their HIV-negative counterparts. Timely and accurate identification of subjects infected with M. tuberculosis and rapid laboratory confirmation of tuberculosis are two effective public health measures that can be taken to combat the tuberculosis epidemic across the globe. The focus of this paper is to review the diagnostic potential of proteinaceous and non proteinaceous antigens in various diagnostic assays and promises of liposome based antibody or antigen detection methods for diagnosis of tuberculosis
[Show abstract][Hide abstract] ABSTRACT: Survivin, an inhibitor of apoptosis, is overexpressed in cancer. It has been implicated in both prevention of apoptosis and cell cycle regulation. We investigated the distribution of antiapoptotic protein survivin in 29 oral squamous cell carcinoma (OSCC) and 16 oral premalignant lesions. It has been suggested that wild-type p53 represses survivin expression. Therefore, we investigated the status of p53 in relation to survivin to determine the potential involvement in oral tumorigenesis.
Oral cancer tissues were freshly obtained at the time of surgery and classified as per general rules of head and neck cancer (TNM classification). Immunohistochemistry and reverse transcriptase-polymerase chain reaction were conducted to study the expression of survivin and p53. The Fisher's exact test was employed to determine the association of survivin and p53 with clinicopathologic parameters of the subjects being studied.
Positive staining for survivin was found in 72% OSCC and 44% oral premalignant lesions with no immunoreactions in the corresponding normal tissues. For p53, 59% OSCC, 38% premalignant lesions, and 14% normal tissues were positive. Importantly, about half of the p53-positive OSCC and premalignant tissues also showed survivin positivity (28% OSCC and 18% premalignant lesions). Further, it is observed that the number of survivin positive cells was significantly higher in the p53-positive group. Survivin is expressed in a varying proportion of cells, and in majority of patients it was localized in cytoplasm, whereas p53 is strictly restricted to the nucleus. The survivin expression levels in both primary OSCC and premalignant lesions were significantly higher than in normal oral tissues (OSCC, p < .0008; premalignant lesions, p < .04). No significant correlations between survivin and p53 expression with clinicopathologic parameters were found.
Frequent overexpression of apoptosis regulators, survivin and p53, in OSCC as well as in oral premalignant lesions were found. Overexpression of these 2 markers in premalignant lesions suggest a role in early stages of oral carcinogenesis.
[Show abstract][Hide abstract] ABSTRACT: A total of 1360 subjects with clinically confirmed pulmonary and extra-pulmonary tuberculosis (TB) and other non-tuberculous conditions.
To develop a rapid, sensitive and specific diagnostic test for the detection of the glycolipid antigen of Mycobacterium tuberculosis in a variety of clinical samples.
Affinity-purified rabbit anti-glycolipid antibodies (IgG) were coupled to liposome particles (0.2-0.4 microm) in the presence of 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride and N-hydroxysuccinamide to prepare the working reagent of the TB/M card test.
Antibody-conjugated liposomes, when determined with the glycolipid antigens present in the specimens, formed a dark blue agglutination within 4 min. No clumping was observed in samples from normal healthy subjects or patients with other diseases. The test was shown to be effective in detecting glycolipid antigens of M. tuberculosis in clinical samples from patients with active TB with as low as 1 ng/ml analytical sensitivity, 97.4% clinical sensitivity and 96.9% specificity.
The TB/M card test was found to be comparatively economical (4 Indian Rupees or US$ 0.09/test), rapid (4 min) and seems fairly useful for mass testing of a variety of biological specimens (cerebrospinal, pleural and synovial fluids, serum, tissue biopsy extract) from patients with tuberculous meningitis, pulmonary TB and other extra-pulmonary TB in endemic countries.
Full-text · Article · Nov 2007 · The International Journal of Tuberculosis and Lung Disease
[Show abstract][Hide abstract] ABSTRACT: In June 1981, the Centers for Disease Control and Prevention reported the first evidence of a new disease that would later become known as Acquired Immunodeficiency Syndrome (AIDS). HIV and Aids: Basic Elements and Priorities is a concise collection of all aspects of this disease and a source of readily available knowledge. It examines all currently advocated preventive measures such as health education, condom use, safer sex practices, and treatment of sexually transmitted infections. Coverage includes: up-to-date information on multiple dimensions of the HIV/AIDS epidemic; a discussion on new anti–retroviral therapy/drugs, new drugs under clinical trials and preventive HIV vaccine; coverage of current ethical, legal and social issues related to HIV/AIDS; an evaluation of general public awareness about HIV/AIDS; a global perspective and information about HIV/AIDS. • Recommended by the World Health Organization Eastern Mediterranean Region Office (WHO-EMRO) as a basic resource for medical faculty libraries.
[Show abstract][Hide abstract] ABSTRACT: Though the management of HIV-infected individuals is somewhat similar to that of patients with other chronic ailments, HIV infection is incurable and carries the additional burden of social stigma. Therefore, health care providers should consider the psychological and emotional aspects. With a non-judgmental attitude, health care providers can create a safe environment for discussing client's lifestyle, sexual and drug-using practices, fears, and anxieties. Health providers need to update themselves about the availability and location of various resources like medical facilities, legal aid, and social support services in order to help clients who may need them. Family members and friends of HIV-infected persons need access to accurate information, social support and referral services. The providers should accept the emotions and reactions of clients when they realise that they are HIV-infected. In countries where it is legally obligatory for doctors to report HIV infections and/or AIDS cases, the reporting should be done with due respect to privacy concerns. Patient support organisations provide significant support which can complement medical care. Health care personnel can assist patients by providing advice on what questions to ask before they choose a particular organisation.
[Show abstract][Hide abstract] ABSTRACT: Development of a preventive vaccine offers the best prospect for containing the HIV epidemic. A realistic goal of a preventive HIV vaccine would be to reduce viral load. Various approaches to developing an effective vaccine include delivering live vector vaccines to mucosal surfaces, incorporating multiple HIV proteins into recombinant live vector or DNA vaccine preparations, injecting purified DNA encoding for HIV proteins into skin or muscle, and incorporating cytokines in vaccine preparations. Liposomes offer multiple advantages as carriers of vaccine antigens. Scientific obstacles to vaccine development include lack of a suitable animal model, antigenic diversity and hypervariability of HIV, its transmission by mucosal route and by infected host cells, resistance of wild-type virus to sero-neutralisation, integration of HIV genome into the host cell chromosomes, latency of HIV in resting memory T-cells, rapid emergence of viral escape mutants in the host, and down-regulation of major histo-compatibility Class I antigens. Programme-related obstacles in developing an AIDS vaccine include inadequate political leadership, insufficient allocation of funds for AIDS vaccine research, insufficient coordination, lengthy approval process and delays in starting trials in developing countries, and insufficient standardisation of assays and reagents. Recruitment of volunteers is difficult in populations exposed to stigma, discrimination, rumours, misunderstandings, and media opinion. Due to decreasing HIV incidence rates, vaccine efficacy trials will have to be conducted at multiple centres or countries in order to achieve the necessary sample size. In human trials of potential AIDS vaccines, counselling on safer sexual practices is essential. Phase I trials of TBC-M4 Modified Vaccine Ankara HIV-1 multigenic subtype C vaccine and tgAACo9 began in December 2005 in India. The Aventis-Pasteur live recombinant vaccine is in Phase III trial in Thailand. At present, there is no effective vaccine that can prevent HIV infection in uninfected individuals or halt the progress of the disease in HIV-infected persons.