The thesis comprises two parts, A and B. Part A deals with the analysis and quantification of medicinally important essential oil, by mass spectrometry method while, the part B describes the chemical studies on cladode of Opuntia dillenii. A short summary on the biosynthesis of steroids, flavonoids, fatty acids and terpenoids are also included in the thesis. Part A This part explains the chemical constituents and their quantification of medicinally important components of extracts on Ixora fulgens, I. coccinea (yellow and orange color flowers), I. polyantha, I. chinensis, Ipomoea batata (blackie), I. batata pink frost, and Cassia fistula, by mass spectrometry method. GC and GC-MS studies were carried out on extracts of different parts especially flowers, leaves and stalks of the plants which led to the identification of 230 chemical compounds from hexane extracts, 324 chemical compounds were identified from chloroform extract of Ixora species. Moreover, 26 metabolites were characterized from methanol extracts of Ixora chinensis flowers extract. From Ipomoea species, 106 chemical compounds were identified from hexane extracts, while, 72 compounds were identified from chloroform extract of the plant. Moreover, 57 phytochemicals were identified from hexane extract, whereas, 51 metabolites were identified from chloroform extracts of C. fistula plant. Different class of compounds were identified from this extracts including hydrocarbons, long chain fatty acids and their esters, alcohols, aromatic acids and their esters, terpenes and vitamins. Antibacterial, antifungal, antioxidant, anti-inflammatory, and anti-cancer activities have also been carried out from all the above mentioned extracts. Part B Total thirteen different fractions of vaccum liquid chromatography (VLC) of antidepressant butanol phase of Opuntia dillenii were selected for chemical analysis. For this purpose GC and GC-MS studies were carried out, which led to the identification of 409 chemical compounds.
آج کا طالب علم غیر ذمہ دارہے نحمدہ ونصلی علی رسولہ الکریم امّا بعد فاعوذ بااللہ من الشیطن الرجیم بسم اللہ الرحمن الرحیم معزز اسا تذہ کرام اور میرے ہم مکتب ساتھیو!السلام علیکم ! آج مجھے جس موضوع پر گفتگو کرنی ہے وہ ہے:’’آج کا طالب علم غیر ذمہ دار ہے‘‘ جنابِ صدر! آج کا طالب علم واقعی غیر ذمہ دار ہے، اس کی ہمہ وقت الیکٹرانک میڈیا کے ساتھ نشست ، پورا وقت غیر ضروری پروگرام کی سماعت، جملہ اوقات ضرور یہ کے ضیاع میں دلچسپی، یہ تمام امور اسی بات کے غماض ہیں کہ اس دور میں علم کے طالب غیر ذمہ دار ہیں۔ جنابِ والا! تعلیم کے حصول میں چستی ،لگن اور دلچسپی انتہائی اہمیت کی حامل ہیں ، چاک وچوبند طالب علم حصول علم میں کوئی دقیقہ فروگذاشت نہیں کرتا۔ تساہل، غفلت، سستی اور کاہلی کے چیتھڑوں میں ملبوس نونہال کسی میدان میں بھی کار ہائے نمایاں سرانجام نہیں دے سکتا اور یہی خصلت ِقبیحہ اسے غیر ذمہ دار بناتی ہے۔ صدرِ ذی وقار! آج کا طالب علم غیر ذمہ دار کیوں ہے، اس لیے کہ اسے وقت کی قدرنہیں ہے، اپنے عظیم لمحات زیست وہ لہو د لعب میں گزار دیتا ہے۔ وقت کا ضیاع اور اس عظیم نعمت کی بے قدری اس کی فطرت ثانیہ بن چکی ہے۔ وقت کی قدرنہ کرنے والانو نہال کبھی شجر سایہ دار نہیں بن سکتا اور ایسی چیز اس کے جسم و جان سے ذمہ داری کی قوت لایموت کوختم کردیتی ہے۔ محترم سامعین! جدید سائنسی ایجاد موبائل کے غیر ضروری استعمال نے اس سے صفت ذمہ داری چھین لی ہے اور وہی ہمہ وقت اس ایجاد سے وابستہ رہنے کے باعث دیگر ضروری امور کی انجام دہی سے قاصر رہتا ہے، نیز اس میں مشغولیت کی بدولت اپنے وقت...
The growing number of diabetes coupled with the harsh side effects of some synthetic drugs has led to the increasing search for alternative medicine which are relatively cheap with minimum side effects. Objective: To investigate the anti-diabetic effect of ethanolic extract of Carica papaya and Psidiumguajava extracts separately and in combination, and the effect of treatment on hepato-renal performance. Methods: Papaya and guava leaves were macerated in 98% ethanol for extraction. Results: The result showed that ethanolic leaf extract of P. Guajava and C. Papaya significantly (P< 0.05) reduced blood glucose level, Total Cholesterol (TC), triglycerides (TG), low density lipoprotein (LDL) and significant increase in high density lipoprotein (HDL) as compared to diabetic control group. Serum activities of liver enzymes such as aspartate aminotransferase (AST), alkaline phosphatase (ALP) and alanine aminotransferase (ALT) was slightly elevated that indicate hepatotoxic nature of plant extracts. The level of gamma-glutamyl transferase (GGT), bilirubin and HbA1C level were restored to normal range. A slight reduction in body weight (P>0.05) of treated groups was observed. Serum urea and creatinine concentration was restored to normal level in treated rats with regeneration of kidney function. Conclusions: The study concluded that individual and combined plant extract show anti-hyperglycemic effect but the combined remedy at this dose show hepatotoxicity
Molecular Genetic and Epigenetic Characterization of Tumor Suppressor Rbl2/p130 in Human Breast Cancer Tumor suppressor retinoblastoma like protein-2 (Rbl2/p130) regulate G1/S transition through its binding with basal transcription factors (E2F4 and E2F5). Activities of Rbl2 protein are regulated through phosphorylation and acetylation in a cell cycle dependent manner. Deregulated expression of Rbl2 and E2F4/5 may lead to neoplastic alterations. The involvement of Rbl2 and E2F4/5 genes in the development and progression of breast cancer among local Pakistani population was investigated. Various genetic and epigenetic mechanisms were probed to explore their potential involvement in gene expressions and tumor growth formation. Rbl2/p130 gene was comprehensively screened for mutations in all exons through single strand conformational polymorphism (SSCP) analysis followed by direct sequencing of suspected samples. Total 200 breast cancer women with mean age of ~45 years were participated in this study. Out of which 126 were at stage I (SI), 40 at stage II (SII) and 34 at stage III (SIII) of breast cancer. Moreover, 138 patients were from invasive ductal carcinoma (IDC), 48 from Invasive lobular carcinoma (ILC) and 14 from ductal carcinoma in situ (DCIS). Among these, 79 mutations in blood and 87 mutations in tissues were detected. Out of these, 12 mutations were previously reported whereas the remaining were found novel. Rbl2 gene was found heavily mutated in exons 19-22 encompassing pocket and carboxyl terminal domain of protein. Among total 28 mutations in exon-21, an A>G transition in codon 1083 that codes for an acetylatable lysine present in the bipartite nuclear localization signals (NLS) 1081PSKRLR1086, was observed. This lysine was mutated to an arginine in 102 (51%) blood and 118 (59%) tissue samples, which suggests that impairment in acetylation status of Rbl2 proteins as well as alterations in nuclear localization might be a key event in tumor initiation and progression. Rbl2/p130 gene was found down regulated while E2F4 and E2F5 genes were upregulated in all study cohorts devised on the basis of age, disease severity and tissue origin. Deregulated expression of these genes was investigated in connection to xi their promoter methylation status using methylation specific PCR (MSP) of bisulphite converted genomic DNA. Rbl2 promoter was found hyper-methylated while promoters of E2F4/5 genes were hypo-methylated in patient‟s samples, suggesting a potential role of promoter methylation in breast carcinogenesis. The CpG positions (-1, +3, +15 and +75) in Rbl2 promoter were specifically found methylated, however positions (-8, +9, +21, +28, +47 and +52) were un-methylated. Rbl2 promoter methylation was found positively associated with various disease stages. Dynamic positioning of nucleosomes is pivotal in determining levels of gene expression especially on or around transcription start site (TSS) of a gene. Nucleosome position around TSS of Rbl2/p130 was determined using micrococcal nuclease (MNAse) digestion assay and ChIP-PCR. Region between -145 -to- +140 around TSS was scanned for 6 positions (P1=-137 -to- +69; P2=-90 -to- +69; P3=-33 -to- +140; RX=-137 -to- +7; F1=-54 -to- +99; F2=-11 -to- +140). About 66% breast tumors and 26.6% ANCT samples were positive for P1. The difference was found statistically significant (p < 0.0001) with an odd ratio (OD) of 9.143, suggesting that nucleosome formation in this region is ~9 times more probable in tumor samples, whereas P2, P3, RX, F1 and F2 were not that suitable for nucleosome positioning. These results indicate that nucleosomes are present slightly downstream of TSS under normal physiological circumstances, that upon breast carcinogenesis slides 55 bases upstream of the TSS, aligning position +1 at the center of nucleosome, hence hindering access to the transcriptional machinery. Breast tumor samples were found methylated at H3K4 to a lesser extent and these modifications were not found associated with Rbl2 proteins, although in precipitates where Rbl2 protein as enriched small fractions of K4 methylation were observed. Conversely, H3K9 tri-methylation as well as Suv39 H2 methyltransferase were found positively associated with Rbl2 proteins, which suggests a putative role of Rbl2 proteins in creating a pre-apoptotic environment in tumor tissues. Our results, highlights a cumulative impact of alterations in genetic and epigenetic mechanisms that culminate in neoplastic transformation. It is obvious from these findings that a cell cycle regulator like Rbl2 can act as a transducer molecule that provide platform for various other molecules to manipulate physiological environment of the cell. With this, we hypothesize that activities of Rbl2/p130 proteins are not limited to tumor suppressor only, rather it has the potential to regulate other physiological and biochemical activities of the cell.