Streamlined Adeno-Associated Virus Production Using Suspension HEK293T Cells.

Recombinant adeno-associated viruses (rAAVs) are valuable viral vectors for in vivo gene transfer, also having significant ex vivo therapeutic potential. Continued efforts have focused on various gene therapy applications, capsid engineering, and scalable manufacturing processes. Adherent cells are commonly used for virus production in most basic science laboratories because of their efficiency and cost. Although suspension cells are easier to handle and scale up compared to adherent cells, their use in virus production is hampered by poor transfection efficiency. In this protocol, we developed a simple scalable AAV production protocol using serum-free-media-adapted HEK293T suspension cells and VirusGEN transfection reagent. The established protocol allows AAV production from transfection to quality analysis of purified AAV within two weeks. Typical vector yields for the described suspension system followed by iodixanol purification range from a total of 1 × 1013 to 1.5 × 1013 vg (vector genome) using 90 mL of cell suspension vs. 1 × 1013 to 2 × 1013 vg using a regular adherent cell protocol (10 × 15 cm dishes). Key features • Adeno-associated virus (AAV) production using serum-free-media-adapted HEK293T suspension cells. • Efficient transfection with VirusGEN. • High AAV yield from small-volume cell culture. Graphical overview.

Back pain and school bag weight - a study on Indian children and review of literature.

Back pain in children is a complex phenomenon, which has been shown to be associated with physical and psychosocial factors. None of the studies on Indian children have investigated the possible association between back pain and psychosocial factors. We decided to determine the prevalence of back pain in students and association with both physical and psychosocial factors. A cross-sectional study was conducted on 163 students from two city schools. Physical factors were evaluated using a structured questionnaire. Strengths and difficulties questionnaire was used to assess psychosocial factors. The studied sample consisted of 49.7% boys and 50.3% girls. Mean school bag weight was 6.1±2.4 kg (range: 2.2-15.6 kg). Mean weight of school bag as a percentage of body weight was 13.5±6.2%. Approximately 76% students carried more than 10% of their body weight. The prevalence of back pain (over last 1 month) was 53.9%. No associations were found between back pain and weight of school bag, weight of school bag as a percentage of body weight and physical activities. Significant associations were found between back pain and number of hours spent watching television [odds ratio (OR): 3.3, 95% confidence interval (CI): 1.5-7.5; P=0.003], history of injury to the back (OR: 4.8, 95% CI: 1.47-15.8, P=0.003), presence of a family member complaining of back pain (OR: 1.98, 95% CI: 1.25-3.14, P=0.002), conduct (P=0.042) and hyperactivity problems (P=0.045). Thus, there appears a need to consider psychosocial factors along with physical factors when evaluating a young student with back pain.

Body Image and its Relation with Body Mass Index among Indian Adolescents.

OBJECTIVE: To evaluate association of body mass index with perception and attitude towards bodyweight, shape and body image among adolescents. Methods: This cross-sectional study was done on 1811 adolescents. Attitude towards body image was assessed by using a self-administered Multidimensional Body-Self Relations Questionnaire. Perceived body shape was measured using the Stunkard scale. RESULTS: Adolescents showed significant difference (P<0.005) in perceptions and behaviors related to appearance, fitness, health, body areas and weight across various body mass index and socioeconomic categories. Girls articulated significantly higher (P<0.005) body dissatisfaction than boys. CONCLUSION: Attitudes and perceptions towards body image differ with sex, body mass index and socioeconomic class.

Public awareness and perception of clinical trials: Quantitative study in Pune.

CONTEXT: Studies have reported that clinical research has experienced tremendous growth during past few decades with many multinational pharmaceutical companies recruiting millions of Indians in clinical trials (CTs). However, there is hardly any literature that talks about the participants, their knowledge, and awareness of CTs. It is important that the general public is aware about CTs so that they can take their own informed decision to participate in CTs. AIM: To assess public awareness, perceptions, and attitudes toward CTs and their views on various methods to create awareness about CTs. MATERIALS AND METHODS: Cross sectional survey was conducted with 200 non trial participants (NTPs) and 40 trial participants (TPs). RESULTS: TPs were significantly (P < 0.0001) older than NTPs. More than 80% of both TPs and NTPs mentioned participation in CT helps advance medical science and strongly felt that there is a need to create awareness about CTs. Nearly 70% of TPs could not remember the phase of the trial while 20% did not know which type of trial they had participated. The main reason for participation in the trial was physician's advice. About 80% of both TPs and NTPs felt that participation in CT will increase with free medications and advice from friends/relatives who had good experience with trial. CONCLUSION: Results of this pilot study revealed need to create CT awareness among the general public. However, considering ethno-cultural, regional, and literacy-level differences throughout the country, a nationwide study would be appropriate to provide reliable results about awareness of CTs among Indians.

Validity and reliability of English and Marathi Oswestry Disability Index (version 2.1a) in Indian population.

STUDY DESIGN: A total of 200 patients with low back pain (LBP) completed an English and Marathi Oswestry Disability Index (ODI) questionnaires (100 each), visual analogue scale, and Roland-Morris Disability Questionnaire. OBJECTIVE: To validate the English and Marathi versions of ODI (version 2.1a). SUMMARY OF BACKGROUND DATA: Patient-orientated assessment methods are important in the evaluation of treatment outcome. The ODI is one of the condition-specific questionnaires recommended for the use of patients with LBP. METHODS: An adaptation of the ODI (version 2.1a) for Marathi language was carried out according to established guidelines. RESULTS: Average age of patients who answered the English ODI was 42 ± 15, whereas that of Marathi-speaking patients was 52 ± 15 years. About 40% were males. The Cronbach α reliability score was 0.877 for English and 0.943 for Marathi. Forty-seven and 53 of these patients were retested with English and Marathi ODI within 2 weeks (to assess test-retest reliability). The intraclass correlation coefficient (ICC) for the test-retest reliability of the questionnaire was 0.877 and 0.943 for English and Marathi respectively. The ODI scores correlated with visual analogue scale pain intensity (r = 0.67, P < 0.0001) and Roland-Morris Disability Questionnaire score (r = 0.71, P < 0.0001) for English and visual analogue scale (r = 0.325, P < 0.001) and Roland-Morris Disability Questionnaire scores (r = 0.503, P < 0.0001) for Marathi. The receiver operating characteristic curve analysis showed comparable performance in discriminating the existence of sign and symptoms (area under curve = 0.947, P < 0.0001, 95% confidence interval: 0.893-0.999 for English and Marathi (area under curve: 0.834, P < 0.0001, 95% confidence interval: 0.735-0.933) for severe compared with nonsevere LBP proving discriminative validity. Results showed that English ODI is valid and reliable. CONCLUSION: The Marathi version of Oswestry questionnaire is reliable and valid, and shows psychometric characteristics as good as the English version. It should represent a valuable tool for use in future patient-orientated outcome studies for population with LBP in India.

Public awareness of clinical trials: A qualitative pilot study in Pune.

CONTEXT: Medical expertise combined with availability of patients with varied diseases have resulted in rapid increase in number of clinical trials (CTs) recruiting millions of patients in India. Yet, few researchers have tried to understand if the public in India is aware of CTs. AIMS: To explore the awareness, perceptions of and attitude towards participating in CTs among general public in Pune. MATERIALS AND METHODS: Focus group discussions (FGDs) and interviews were conducted by contacting people in the community of various age groups and socio economic status with 7 Trial participants (TPs) and 17 Non Trial Participants (NTPs). The survey tool consisted of open-ended questions that assessed the awareness and attitudes of the individuals regarding the CTs. Interview were recorded on paper and translated from (Marathi) local language to English for analysis. Qualitative analysis was used to report the findings. RESULTS: Most participants could associate CTs with medicine or development of new medicine; however they did not have a good understanding of the manner and safeguards with which CTs are conducted. Participants were not aware about different types of CTs and phases of the CTs. CTs were felt to be of benefit to the community and advancement of science. However, due to fear of adverse effects, 80% of the respondents were not ready to participate in the CTs. CONCLUSIONS: The Indian Pharmaceutical company is the world's 3(rd) largest by volume as per Dr. Shivathanu Pillai's report 17th March 2010, in spite of that it has been noticed that the awareness about CTs is very low; therefore there is a need to create awareness about CTs which helps the participants to participate in CTs based on their own decision. These FGD findings require validation in a larger sample.