Incidence of HPV Infection in High and Low Grade Lesions of Cervix

INTRODUCTION: Carcinoma of the uterine cervix is the most common cancer in South Indian women and occupies the top rank among cancers in women in most developing countries, constituting 34% of all women’s cancers. To an estimated annual global incidence of 500,000 cervical cancers, India contributes 100,000, ie. 1/5 of the world burden.23 The magnitude of the problem is thus more than evident. The world pattern of cervical cancer, together with the age adjusted rate and ranking, clearly indicate that cervical cancer is predominantly a problem of poorer socio-economic societies.1 On the other hand, uterine cervical cancer is a favourable site for an effective control program. It is easily accessible and there is usually a long latent period of intraepithelial neoplasia which is easily recognizable by the Pap smear. Furthermore, treatment at this stage is very effective. The burden of cervical cancer in India, taken in the context of the additional problems of advanced disease at presentation, the country’s limited resources and health infrastructure, and the paucity of trained personnel emphasize the urgent need for a control program. Cancer of the cervix has been the most important problem in women in India over the past two decades. AIM OF THE STUDY: To determine the HPV status in high and low grade cervical lesions in women of reproductive age group . Setting: It is a prospective study conducted in the Institute of Social Obstetrics (ISO) & Govt. Kasturba Gandhi Hospital for Women and Children, Triplicane. Chennai attached to the Madras medical College, Chennai. Sample Size: IT is conducted in a sample of 50 women with high risk cervical lesions during the period 2008-2009. Around 350 women in the reproductive age group are screened in the colposcopy clinic and among them 50 are selected for HPV testing. MATERIALS AND METHODS : THE HC2 High-Risk HPV DNA Test using capture 2 technology is a nucleic acid hybridization assay with signal amplification that utilizes micro plate chemiluminescent detection. It is a solution phase hybridization assay which results in HPV DNA signal amplification. All my tests are done in the Lab of RENU DIAGNOSTICS. The kit is an in vitro nucleic Acid Hybridization Microplate Assay with Amplification using Microplate chemiluminescene for the Qualitative Detection of 13 High-Risk types of human papillomavirus (HPV) DNA in cervical Specimens. It is a DNA PAP Cervical Sampler. The Manufacturers are DIEGENE, 1201, Clopper road, Gatheisberg, USA. A R Med Limited. Samples are collected by cervical brushings in liquid media and subjected to the test. The assay uses RNA probes that react with 13 DNA targets. These RNA-DNA hybrids are captured by monoclonal antibodies bound to the well plate and detected with anti hybrid antibodies by chemiluminesence. Step 1: Denature DNA [~1 hour]To initiate the lab process (labeling tubes for identification, etc.). 1. Cervical specimen is added to sample tube. 2. Denaturation agent is added to tube. Step 2: Mixing of probes and hybridization [~1.5 hours] 1. Probe B cocktail for the high-risk HPV types is prepared and Added to the Tubes. (Probe B is specific for oncogenic HPV types.) 2. The tubes are placed in a water bath at 65°C for 30 minutes. 3. The samples are washed several times using a standard laboratory reagent. Step 3: Hybrid capture [~1.5 hours] 1. The processed samples are transferred to a microtiter plate provided in the kit. 2. It is placed on a mechanical shaker for 30 minutes (allowing the RNADNA hybrids created in Step 2 to be “captured” by antibodies attached to the walls of the microtiter well). Step 4: Detection for labeling [~1 hour] 1. Additional antibodies tagged with alkaline phosphate, are added which bind with the captured materials from Step 3. When the alkaline phosphatase separates from the antibody complex, light is produced. Step 5: Detection, validation, and interpretation [~1 hour] 2. The light released during Step 4 is measured using a luminometer with integrated computer software. Any sample that emits light as bright as or brighter than the light released by a positive control is considered a positive signal for HPV. Inclusion Criteria: Women in the age group 25-35 yrs, Women who are leading an active sexual life, Women who have HSIL and HSIL in colposcopic findings. Exclusion Criteria: Post menopausal women, Who are not leading an active sexual life, Invasive cancer cervix, Normal colposcopic findings, Women with gross active PID or cervicitis. SUMMARY:\ud Although oncogenic HPV infection has been established as a causative factor of the precursors of cancer cervix as well as their progression to higher grade and eventually to malignancy, there are some other predisposing factors which play a substantive role in the causation and progression of these lesions. This study has tried to delineate these risk factors in cases of cervical dysplasia. This study is a prospective study conducted in a sample of 50 women in the reproductive age group . Women with LSIL AND HSIL Lesions are selected ,cervical samples are taken from them and couriered to Renu Diagnostics lab. HPV infection is detected by HC 2 technique. It is an In vitro nucleic Acid Hybridization Microplate Assay with Amplification using Microplate chemiluminescene for the Qualitative Detection of 13 High-Risk types of human papillomavirus (HPV) DNA in cervical Specimen’s. The results are coded in RLU. HPV infection showed higher incidence among 28 to 30 years of age, among low SES and among illiterates. 100% of HSIL showed HPV infection and 60% of LSIL showed positive infection. HPV infection shows higher incidence among low social status where the genital hygiene is poor and among illiterate women who have a very poor knowledge about sources of prevention. Women of low SES also get married soon and have decreased birth spacing which increases the incidence. Following these women 73% of these lesions were inflammatory ,treated with antibiotics and followed up with colposcopy every 6 months. 9 cases found to be moderate dysplasia and treated by LLETZ. 10 cases found to be mild dysplasia, Cryotherapy was done in these women. 1 lesion turned out to be CIN3 and proceeded with TAH. CONCLUSION: Accumulated evidence based on etiologic associations and the differential world pattern points to cervical cancer being a preventable disease. Sexual hygiene and the use of barrier contraception (condom) may largely achieve this objective but there is a need for long-term education and acceptance. Improvements in socio-economic standards would automatically reduce morbidity and mortality but this again is a long-term process. Primary prevention, then, involves the education of a large segment of the population, especially the high risk groups, about sexual hygiene, barrier contraception and control of HPV infection. Secondary prevention assumes vital importance in the context of the hurdles in implementing primary prevention methods. In a large country such India with a large, growing population and limited resources, population screening by Pap smear is neither pragmatic nor cost-effective. It is thus essential that we evolve our own strategies. Women with an abnormal cervix should have additional studies including: Pap smear, biopsy, colposcopy and HPV DNA testing wherever possible

A REVIEW ON MEDICAL ADVANTAGES AND CHEMICAL CONSTITUENTS OF CARICA PAPAYA LINN.

Papaya is an evergreen blossoming plant having rich characteristic thing; it is recognized to offer different therapeutic purposes of premium. The normal item is sound and delicious. The entire plant parts such as root, bark, peel, seeds, blooms, leaves, and squash are known to have helpful properties. It is utilized for the treatment of a various infections such as dengue fever, warts, corns, sinuses, skin inflammation, against diabetic, glandular tumors, blood pressure, digestive disorders, constipation, antibacterial, antifertility, anti-HIV, expel worms, invigorate regenerative organs and many, as needs be it can be seen as a nutraceutical. The present review focuses on salient features of nutritional composition, health benefits, medical advantages, and synthetic constituents of papaya

The human ankyrin 1 promoter insulator sustains gene expression in a β-globin lentiviral vector in hematopoietic stem cells.

Lentiviral vectors designed for the treatment of the hemoglobinopathies require the inclusion of regulatory and strong enhancer elements to achieve sufficient expression of the β-globin transgene. Despite the inclusion of these elements, the efficacy of these vectors may be limited by transgene silencing due to the genomic environment surrounding the integration site. Barrier insulators can be used to give more consistent expression and resist silencing even with lower vector copies. Here, the barrier activity of an insulator element from the human ankyrin-1 gene was analyzed in a lentiviral vector carrying an antisickling human β-globin gene. Inclusion of a single copy of the Ankyrin insulator did not affect viral titer, and improved the consistency of expression from the vector in murine erythroleukemia cells. The presence of the Ankyrin insulator element did not change transgene expression in human hematopoietic cells in short-term erythroid culture or in vivo in primary murine transplants. However, analysis in secondary recipients showed that the lentiviral vector with the Ankyrin element preserved transgene expression, whereas expression from the vector lacking the Ankyrin insulator decreased in secondary recipients. These studies demonstrate that the Ankyrin insulator may improve long-term β-globin expression in hematopoietic stem cells for gene therapy of hemoglobinopathies

Developing Online Tutors and Mentors in Sri Lanka through a Community Building Model: Predictors of Satisfaction

This paper discusses the results of a tutor mentor development program that utilized a community building model to train online tutors and mentors in higher education institutions and professional organizations in Sri Lanka. Based on WisCom; an instructional design model for developing online wisdom communities, this tutor mentor development program which utilized a blended format of face-to-face and online activities in MOODLE, attempted to build a learning community between trainees, both academics and professionals who represented diverse disciplines and organizations. A regression model examined predictors of learner satisfaction, using four independent variables: Community Building, Interaction, Course Design, and Learner Support. Interaction emerged as a strong predictor of Learner Satisfaction explaining 50.2% of the variance in Learner Satisfaction. This finding shows the importance of designing interactive learning activities to support learning online, and contradicts the general belief that Sri Lankan participants would be less likely to interact online because they come from a traditional education system that encourages passivity and reception of ideas from a more learned teacher. Qualitative analysis showed evidence of several types of learning online as a result of collaborative group interaction, as well as issues that contributed to non-participation. Factors that motivated participants to stay engaged in learning could be classified into three categories: (1) general enjoyment, interest and motivation; (2) collaborative learning and community building; and (3) knowledge building. These results suggest that the online learning design based on WisCom led to learner satisfaction and supported interaction and collaborative learning in the Sri Lankan socio-cultural context

Creating New β-Globin-Expressing Lentiviral Vectors by High-Resolution Mapping of Locus Control Region Enhancer Sequences.

Hematopoietic stem cell gene therapy is a promising approach for treating disorders of the hematopoietic system. Identifying combinations of cis-regulatory elements that do not impede packaging or transduction efficiency when included in lentiviral vectors has proven challenging. In this study, we deploy LV-MPRA (lentiviral vector-based, massively parallel reporter assay), an approach that simultaneously analyzes thousands of synthetic DNA fragments in parallel to identify sequence-intrinsic and lineage-specific enhancer function at near-base-pair resolution. We demonstrate the power of LV-MPRA in elucidating the boundaries of previously unknown intrinsic enhancer sequences of the human β-globin locus control region. Our approach facilitated the rapid assembly of novel therapeutic βAS3-globin lentiviral vectors harboring strong lineage-specific recombinant control elements capable of correcting a mouse model of sickle cell disease. LV-MPRA can be used to map any genomic locus for enhancer activity and facilitates the rapid development of therapeutic vectors for treating disorders of the hematopoietic system or other specific tissues and cell types

Glaucoma-associated long-term mortality in a rural cohort from India: the Andhra Pradesh Eye Disease Study.

AIM: To evaluate glaucoma-associated mortality in a rural cohort in India. METHODS: The study cohort comprised individuals aged 40 years and above who took part in the Andhra Pradesh Eye Disease Study (APEDS1) during 1996-2000. All participants underwent detailed comprehensive eye examination. Glaucoma was defined using International Society of Geographic and Epidemiologic Ophthalmology criteria. This cohort was followed up after a decade (June 2009 to January 2010; APEDS2). Mortality HR analysis for ocular risk factors was performed using Cox proportional hazards regression after adjusting for sociodemographic, lifestyle and clinical variables. RESULTS: In APEDS1, 2790 individuals aged more than or equal to 40 years were examined. 47.4% were male. Forty-five participants had primary open angle glaucoma (POAG) and 66 had primary angle closure disease (PACD). Ten years later, 1879 (67.3%) were available, 739 (26.5%) had died and 172 (6.2%) had migrated; whereas 22 of the 45 (48.8%) with POAG and 22 of the 66 (33.3%) with PACD had died. In univariate analysis, a higher mortality was associated with POAG (HR 1.9; 95% CI 1.23 to 2.94), pseudoexfoliation (HR 2.79; 95% CI 2.0 to 3.89), myopia (HR 1.78; 95% CI 1.54 to 2.06) and unit increase in cup:disc ratio (HR 4.49; 95% CI 2.64 to 7.64). In multivariable analysis, only cup:disc ratio remained independently associated with mortality (HR 2.5; 95% CI 1.3 to 5.1). The association remained significant when other ocular parameters were included in the model (HR 2.1; 95% CI 1.03 to 4.2). CONCLUSIONS: This is the first longitudinal study to assess the association of glaucoma and mortality in a rural longitudinal cohort in India. Increased cup:disc ratio could be a potential marker for ageing and would need further validation