DOSE-DEPENDENT ANTI-DEPRESSANT EFFECT OF CITRUS PARADISI IN REHABILITATION OF PSYCHOLOGICAL DISORDERS

With advancement in the modern world, psychological issues, including anxiety and depression has become the leading cause of mental illness. Herbal products have been explored to observe their effects in helping such conditions and citrus fruits are found to be among these helping tools. To study this, current research was designed to explore the antidepressant effects of Citrus paradisi in various doses on rodents. Swiss Albino mice were used and fresh extract of Citrus paradisi was prepared. Imipramine was used as the standard drug. Ethical approval was taken from the Board of Advanced Studies, University of Karachi. The animals were divided into controlled, standard and test groups. All the animals were recruited from the animal house of University of Karachi. Forced Swimming Test was used to analyses the efficacy. It is a standard test, which is commonly used to evaluate the behavioral response in animals using a cylinder at room temperature. After the completion of dosing and monitoring the animals for 30 days, it was observed that marked improvement in the test animal’s behavior with significant p-values indicating that the extract has improved the animal’s psychological response towards the external stress and animals showed noteworthy improvement in the mobile phase as compared to the immobile phase. It can be concluded from the experimental results that protocol established in this study can be used as an alternative for patients suffering with depression as an herbal option, keeping in mind the other co morbidities of patient and drug interactions of Citrus paradisi.

Cloning and Sequencing of Mycobacterium Tuberculosis Genes and Used As Dna Vaccines

Tuberculosis (TB) is said to be the oldest disease of history and still considered as a major problem in the world especially in developing countries like Pakistan. It needs continuous efforts to work on its prevention and cure. Mycobacterium tuberculosis (M. tb) bacterium is the etiological agent of this disease and almost one third population of world is infected with it. Almost 1.5 million deaths/year are due to active TB. Asia carries two-third of the total burden of TB, and Pakistan is at 5th position among 22 high burden countries. Efforts are made to control and prevent TB by improving diagnostic tests, effective medication, care through DOTS program and development of new vaccines. BCG vaccine is available as control measure but its efficacy is very limited in most parts of the world. There is need to develop better and state of the art vaccines like DNA vaccines. The current study was designed to produce construct of M. tb genes and later used them as DNA vaccine in mice model. The two M. tb genes SodC and Mpt51 were selected and their gene sequence was downloaded from Tuberculist database. Primers were designed manually. PCR conditions were optimized for each gene and PCR products were confirmed through restriction digestion. PCR products were cloned into pTZ57R/A vector and transformed into chemically competent E.coli cells. The clones were confirmed through restriction digestion and finally by sequence analysis. Finally the constructs were made in pND vector. The finally confirmed clones were stored at -70 C as glycerol stocks and later were used for sub-cloning purposes in mammalian expression vectors to finally develop DNA vaccines. In this regard, twenty Balb/c female mice were used having age of eight weeks each. Eight eight animals were used for inoculation with SodC-pND and Mpt51-pND respectively. Four were used as negative control (Normal Saline group). All animals were bled through tail after three weeks intervals and finally through cardiac puncture at nine weeks post inoculation. All antisera were strongly positive on dot blot and Agar Gel Immunodiffusion test. The results of this study proved that both SodC-pND and Mpt51-pND DNA vaccines are good enough to produce strong humoral response in mice model and hold a promise to be used in anti-TB vaccines in future