Muhammad -
The Recipient of The Qur’an
Muhammad was the only child of his parents – Sayyed ‘Abd-Allah bin ‘Abd Al-Muttalib Al- Hashemi and Sayyedah Aminah bint Wahb Al-Zuhriyya. He was born in Makkah, a commercial metropolis and a pilgrims’ hub in the Arabian Peninsula. His father passed away before his birth, and the mother when he was six. A shepherd in early life and a successful merchant in adulthood, he married Sayyedah Khadijah bint Khuwaylid at the age of 25. The happy couple became the parents of two sons and four daughters.
Muhammad ibn Abd-Allah was reputed to be a friendly and trustworthy gentleman with an impeccable character. He was admired for his generosity as well as his wisdom. The mutually hostile factions often called upon him to act as an arbiter. At home, he was known for his caring and helpful attitudes.
As he was attaining maturity, he was getting deeply disturbed by the social and moral environment around him. He found the idolatry and fetishism unholy and social injustices distressing. He increasingly began to search for a response to his own agony at the injustice and chaos around him by resorting to meditation and seclusion. He would go away from home for several days in a row and stay in a small cave on Mount al-Noor, it is a cave hiding in shyness and seclusion with a difficult access, named Hira.
One midnight, during the last week of the month of Ramadan that the Cave lightened up. It witnessed the presence of rows over rows of angels led by the Arch Angel, Gabriel. Muhammad got bewildered. It brought him awe, fear and foreboding. It was an experience unique of its kind; unique...
Peace initiatives during the regime of the Prophet attain to how understand to studied be to needs (صلى الله عليه وسلم) Muhammad and maintain peace in a diverse society. The measures for peace, taken by the Prophet (ﷺ) can be divided into two types: The Internal Steps and The External Steps. One of the fundamental objectives of his prophetic annunciation was ‘purification and refinement’, which was the core of his internal measures for peace. These measures produced moral values such as love and harmony, reconciliation and sacrifice among them. These personality traits were then translated into the establishment of equitable justice system and reciprocally, a just system leads to shape behaviors contributing to peace and harmony. For steps external the on focused also (صلى الله عليه وسلم) Prophet The peace. These external measures include the treaty of al-Madīnah, the agreement with the Christian delegation of Najrān, the treat of Ḥudaybiyah and the announcement of the amnesty at the conquest of Mekkah. Thus, the study of the Prophet’s (ﷺ) life unfolds that if we want global peace, we have to follow the principles, practiced. (صلى الله عليه وسلم) Prophet the by down laid and
Cancer is one of the major causes accounting for death and illness both in developed and underdeveloped countries. The second largest reason of death in Pakistan is cancer. For the treatment of cancer, a range of biomedical and non-biomedical therapeutic options are being used. However the overall recovery and survival is low. Thus signifying the development of new therapeutic agents and strategies to cure cancer and associated malignancies. Interleukin 24 (IL-24) with its unique proapoptotic and cancer cell selective properties is a potential biopharmaceutical candidate among the recently developed family of protein therapeutics. Since its discovery IL-24 has been prime focus for its recombinant production in various forms to improve biological activity and explore the mechanism of apoptosis induction. The present study reports the recombinant production of IL-24 in biologically active form, using bacterial expression system, its structural and functional characterization for subsequent use in therapeutics as well as in the studies to decipher the mechanisms of unique cancer-specific growth suppressing properties of IL-24. The present study also reports to achieve partial soluble expression of IL-24 as SUMO-IL-24 in E. coli. This study also reports recombinant production of IL-24 with native N-terminus, and recombinant production of fusion protein RGD-IL-24. The cDNA encoding IL-24 was synthesized from the total RNA extract of peripheral blood mononuclear cells using gene specific primers by reverse transcription polymerase chain reaction (RT-PCR). The IL-24 cDNA was cloned in pET-22b(+) expression vector between NdeI and BamHI restriction sites to generate pET-IL-24 recombinant plasmid. To express IL-24 as fusion protein SUMO-IL-24, the IL-24 cDNA was cloned in pE-SUMOstar expression vector between BsaI and XbaI restriction sites to generate pSUMO-IL-24 recombinant plasmid. To express IL-24 as RGD-IL-24 fusion protein a v modified IL-24 cDNA with coding sequence of RGD at 5ʹ end was cloned in pET-22b(+) expression vector between NdeI and XhoI restriction sites to generate pET-RGD-IL-24 recombinant plasmid. The recombinant plasmids pET-IL-24, pET-RGD-IL-24 and pSUMO-IL-24 were used to transform competent E. coli DH5α and positive transformants were selected by ampicillin/kanamycin resistance and further screened by colony PCR. Presence of IL-24 inserts in recombinant plasmids pET-IL-24, pET-RGD-IL-24 and pSUMO-IL-24 was confirmed by restriction analysis. Recombinant plasmids were analyzed for correct orientation and in frame integration of IL-24 insert by DNA sequencing with T7 promoter and T7 terminator primers. Subsequently the recombinant plasmids pET-IL-24, pET-RGD-IL-24 and pSUMO-IL-24 were used to transform competent E. coli BL21 CodonPlus (DE3) RIPL. The expression of recombinant proteins SUMO-IL-24, M-IL-24 and RGD-IL-24 was obtained by induction with 0.5 mM IPTG at an OD600 of 0.5 for three hours in LB broth supplemented with ampicillin/kanamycin. The expression of recombinant proteins was >30% of the total proteins in E. coli cellular extract. The growth of E. coli transformed with recombinant plasmid and expression of recombinant proteins were optimized with respect to media, duration of cultivation, inducer and temperature. M-IL-24 and RGD-IL-24 obtained in the form of insoluble inclusion bodies were solubilized in non ionic detergent NLS. Solubilized M-IL-24 and RGD-IL-24 were refolded and further purified by using anion exchange chromatography on FPLC up to >95 % purity. The fusion protein SUMO-IL-24 was purified using IMAC. SUMOstar protease was used to cleave the fusion protein SUMO-IL-24 into SUMO and IL-24 fragments. Cleaved IL-24 has native N-terminus starting with glutamine (Q) as in human hence it is referred as N-IL-24. N-IL-24 was purified using IMAC to >95 %purity. Refolded and purified recombinant proteins were characterized for secondary structure contents using circular dichroism spectroscopy. Molecular weights of the recombinant vi proteins were characterized by MALDI-TOF MS. Structural modeling of recombinant proteins was performed using in silico computational analysis. Initially biological activity of M-IL-24 was assessed on human cervical cancer cell line HeLa and human colon colorectal carcinoma cell line HCT116. Cytotoxicity of M-IL-24 was further measured on HeLa using neutral red assay and fluorescence activated cell sorting. Cytotoxic effects of N-IL-24 and RGD-IL-24 were initially observed on human cerebrum glioma cell line SF767 and human cervical cancer cell line HeLa. Biological activities of recombinant N-IL-24 and RGD-IL-24 were assessed in detail on HeLa using neutral red assay, FDAPI staining and crystal violet cell adhesion assay. Polyclonal antibodies against IL-24 were raised in Swiss mice and were characterized in post-immune sera using alkaline phosphatase conjugated rabbit anti-mouse IgG antibodies by immunodot blot, ELISA and western blot analysis. Native N-terminus is very important feature of recombinant therapeutic proteins in cancer treatment and the results of present study reveal that N-IL-24 was successfully expressed and purified in biologically active 3-D conformation with native N-terminus. Secondly N-IL-24 recombinant protein showed maximum apoptotic properties among three recombinant forms of IL-24. Decrease in cell survival rate of HeLa, cerebrum glioma cells (SF767) and HCT116 proves N-IL-24 a successful pharmaceutical candidate in future for therapy of various cancers in human. In addition the model mice injected with M-IL-24 subcutaneously retained good health during immunization period. This also proves that recombinant proteins N-IL-24 and M-IL-24 were successfully sufficient purified and safe for bio pharmaceutical applications.