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Home > Bioassy Guided Isolation and Characterization of Secondary Metabolites Rumex Hastatus D. Don and Their Anticancer Activities

Bioassy Guided Isolation and Characterization of Secondary Metabolites Rumex Hastatus D. Don and Their Anticancer Activities

Thesis Info

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Author

Sajjad. Ahmad

Program

PhD

Institute

University of Malakand

City

Malakand

Province

KPK

Country

Pakistan

Thesis Completing Year

2017

Thesis Completion Status

Completed

Subject

Pharmacology

Language

English

Link

http://prr.hec.gov.pk/jspui/bitstream/123456789/9366/1/Sajjad%20Ahmad-Pharmacy.doc

Added

2021-02-17 19:49:13

Modified

2024-03-24 20:25:49

ARI ID

1676725581719

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The inquisitive nature of human compelled the nature to bestow myriads of resources and commodities to man. Since yonks ago, the human have been fully dependent on the earth’s ecosystem. Shortly, among the most productive natural sources, the extraordinary role of plants is prettily inevitable. Besides providing the food, shelter and living, the plants also provide health ameliorating entities known as drugs or medicine. The drugs from the plant sources are literally known as herbal medicine. Based on the very importance of herbal medicine, one of the scantly explored herb i.e., Rumex hastatus, which belongs to family Polygonaceae has been investigated over here in the terms of phytochemistry and anticancer aspects. As a lot of knowledge has been transferred from the previous era to the modern world via ethnobiology. So the R. hastatus has also been explored based on the ethnomedicine (traditional medicine) to authenticate its purported traditional uses especially for anticancer purposes. In the current study, different samples of R. hastatus have been investigated i.e., the crude sample, the solvent fractions of crude sample, the essential oil and the pure compounds. The crude sample was collected via maceration process while the isolation of essential oil was carried out following hydrodistillation via Clevenger apparatus. The fractions of different solvents have been obtained using successive solvent extraction procedure via separating funnel. Similarly, the compounds were obtained via bioassay guided isolation procedure through gravity columns with the help of thin layer chromatography. The compounds obtained were purified with pen/pencil column or small diameter gravity columns. The pure compounds obtained were identified with various techniques i.e., Mass spectroscopy, carbon NMR and proton NMR. Four compounds i.e., SA-1, SA-2, SA-3 and SA-4 were properly identified and structurally elucidated. Three compounds were in sufficient quantities, which were subjected to various anticancer assays. The anticancer assays were based on mechanistic approach. The assays performed were MTT assay, Comet assay, CAM assay, antitumor assay, anti-mutagenic assay, apoptosis assay, antithrombin assay, intercalation assay and antioxidant assay. Before these assays the phytochemical screening was carried out for the detection of different types of secondary metabolites in the samples. The screening of samples was also performed to figure out the presence of heavy metals and their amounts were figured out. The samples were also tested for acute toxicity via mice animal model. The investigations revealed that two compounds SA-1 and SA-2 were highly active against cancer cells. The SA-3 was found to be the least active compound against cancer. If we go to the detail of the test compounds, the SA-1 was found to have 46% antiangiogenic, 85% antimutagenic, 75% antioxidant, 61% antithrombin, 41% antitumor, 50% apoptosis, 42% necrosis, 67% cytotoxicity by MTT, 118/200 comet score and significant intercalation to DNA at the highest test concentration. The SA-2 has revealed 41% antiangiogenic, 70% antimutagenic, 55% antioxidant, 53% antithrombin, 37% antitumor, 61% apoptosis, 22% necrosis, 53% cytotoxicity my MTT, 91/200 comet score and moderate intercalation to DNA. The acute toxicity assay produce no visible anomaly, toxicity or lethality, while the heavy metals analysis revealed the presence of heavy metals within the permissible limits as per WHO guidelines. Based on the results of anticancer activities with various mechanisms, it may be deduced that the SA-1 was quite active against cancer and mechanistically we can say that SA-1 is strongly antimutagenic i.e., it quenches the the mutagens or carcinogens. Secondly we can see that the SA-1 has a significant antioxidant potential so it also inhibit the free radicals. Thirdly, it has also been demonstrated to cause cytotoxicity significantly. Similarly, In regards to the possible mechanistism, the SA-1 have the ability to bind with the genetic material, to quench the mutagens, to inhibit the free radicals, to inhibit the thrombin, to cause cytoxicity, to cause programmed cell death and necrosis, to break down the DNA. The SA-1 also moderately inhibit the neovascularization i.e., new blood vessels formation and inhibit the tumor. So the SA-1 has been found to possess highest potential for quenching the mutagens and inhibit the mutagenesis while the lowest potential has been found against tumorigenesis. Similarly, the SA-2 has also been demonstrated with moderate anticancer potential with the highest potential against mutagenesis and lowest potential to induce necrosis. Both of the compounds SA-1 and SA-2 have been verified in the current investigational studies to combat cancer especially the cervical and colon cancer with no toxicity profile. So further work should be performed by the dosage form designers for these compounds to hit the market.
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فاخر ہریانوی

فاخرہریانوی (۱۹۰۱ء۔۱۹۷۷ء) کا اصل نام دین محمد تھا اور تخلص فاخرؔ تھا۔ فاخرؔ ہریانہ ضلع ہوشیا ر پور میں پیدا ہوئے۔ پنجاب یونیورسٹی سے ۱۹۳۱ء میں بی او ایل کیا۔ پھر پنجاب ایجوکیشن ڈیپارٹمنٹ سے ۱۹۳۵ء میں سینئر اینگلو ورینکولر کا سرٹیفکیٹ لیا۔ تعلیم کے بعد اردو مرکز لاہور میں ملازم ہو گئے۔ جگر مراد آبادی ،اصغر گونڈوی او ر یاس یگانہ چنگیزی بھی ان دنوں اس مرکز سے منسلک تھے۔ اصغر کے چلے جانے کے بعد فاخر کو اس ادارے کا ناظم بنا دیا گیا۔ فاخر نے کچھ عرصہ پنجاب لیجسلیٹو کونسل میں مترجم کی حیثیت سے بھی کام کیا۔ ۱۹۲۹ء میں فاخر شعبہ تعلیم میں چلے گئے۔ ملازمت کے سلسلے میں وہ بہت سے علاقوں میں رہے۔ اور آخر کار پسرور میں مستقل سکونت اختیار کی۔(۴۳۲)

’’موجِ صبا‘ فاخرؔ کا واحد شائع شدہ شعری مجموعہ ہے۔ جو فروری ۱۹۶۶ء میں ایوانِ ادب لاہور سے شائع ہوا۔ اس مجموعے کا دیباچہ پروفیسر حمید احمد خان نے لکھا ہے ۔ا س کی ترتیب میں ضیاء محمد ضیاء اور طاہر شادانی کی تلاش اور تفتیش شامل ہے۔ مرتبین نے اسے سات حصوں ،حمدیہ ،جذبات و افکار،رومان ،دیہاتی نغمے ،یادِ رفتگاں ،سیاسیات اور متفرقات میں تقسیم کیا ہے۔ یہ شعری مجموعہ ۲۴۰ صفحات پر مشتمل ہے۔ اس کی تفریظ ڈاکٹر وزیر آغا نے لکھی ہے۔ اشکِ عمل ان کا غیر مطبوعہ مجموعہ ہے۔فاخر نے سب سے پہلے مسدس حالی کی بحر میں اشکِ عمل قلمبند کروائی ہے۔ اول حصہ حمد باری تعالیٰ اور دوسرا حصہ حضورؐ کی زندگی سے متعلق اہم واقعات پر مشتمل ہے۔ ان میں تبلیغ اسلام فتح مکہ اور جنگ احد بالخصوص قابل ذکر ہیں- فاخرؔ نے قرآن مجید کا منظوم ترجمہ بھی کرنا شروع کیا لیکن ادھورا چھوڑ دیا۔ پھر ایام پیری میں دوبارہ اس کا م کا عزم کیا ۔قرآن مجید کی آیاتِ کریمہ کو بغیر قافیہ ردیف...

سیرت طیبہ کی تشریعیاتی حیثیت کا تحقیقی و تاریخی مطالعہ A Research review of the legislative Status of the Sirah of Holy Prophet ﷺ

The deeds and actions of the Holy Prophet ﷺ is a practical interpretation of the Holy Qur'an. Allah Almighty has stated his Shari'ah in short but comprehensive words that “take what the Holy Prophet ﷺ has given you and which you forbid”. Therefore, his command is the command of Allah Almighty and not yours. That is why the Holy Qur'an has declared his obedience to be the obedience of Allah Almighty. And from the instructions of the selected scholars of the ummah, it is clear that the legislative status of The Sirah is authenticated. Allah Almighty has given him a special position of Shariah. As for the legislative status of Sira-e-Taiba, both the special and the general legislation are specific or special to him. Special legislation etc i.e. you have legislated for a specific person at certain times like accepting the condition of a person that he will convert to Islam if he prays only two prayers. The other person cannot join. The general legislation in which you have enacted all kinds of legislation for the common ummah is included. The center and axis of Islamic law is your caste. You have complete control over the status and sanctity, likes and dislikes as you like, for whom you can make Shariah whenever you want. It is as if you are a follower of the Shari'ah. He also has full authority in the brief description of the Qur'an, in the adherence to the Absolute, and the explanation of similarities. You are authorized to legislate as a Shari'ah and Shari'ah in all areas of worship, affairs, debates, and crimes. Therefore, in this article, the legislative status of His Sira-e-Taiba has been explained in the light of Qur'an and Hadith, Sahabah and Tabi'een, and the commandments of Imams and jurists.

Genetic Polymorphism of Milk Protein in Local Goat Breeds

The genes encoding milk proteins possess polymorphic forms which greatly influence the composition of raw milk and dairy products manufactured. The major milk proteins directly influencing the milk properties are whey proteins and caseins. The β-LG protein from whey fraction and κ-Casein from the casein complex are the most commonly studied milk proteins in different dairy animals worldwide. Several genetics variants of both proteins have been found associated with milk yield, total milk protein and fat contents as well as manufacturing properties of milk. Polymorphisms in β-LG and κ-casein gene CSN3 have been extensively studied in cattle; however, not much work has been reported in goats Present study was designed to identify polymorphisms in β-LG and CSN3 in Beetal and DDP goats of Pakistan and associations of identified variants with milk components were also explored. For genetic analysis, DNA was extracted from blood and quantified spectrophotometrically. The polymorphisms in β-LG were explored using PCR-RFLPs while CSN3 exon IV was sequenced and genotypes determined based on Prinzenberg et al., 2005 nomenclature. Total milk proteins and whey protein contents were estimated spectrophotometrically and casein protein contents determined. The total milk fat contents were estimated by Gerber’s method. The β-LG protein isoforms were also identified in both goat breeds through SDS-PAGE. Our results showed that β-LG AA genotype is most prevalent in both goats. The β-LG AA genotyped animals had higher total protein contents in milk while high fat contents were found in the milk of animals with BB genotype. Three β-LG protein isoforms with genotypes; AA, AB and BB were identified. The frequency of A allele protein variant was significantly higher in both goat breeds xvii while heterozygous AB genotype was the most prevalent one. However β-LG protein isoforms lacked any association with milk yield and composition. At CNS3 locus, Beetal carries A genotype whereas, DDP carried D genotype. The haplotypes revealed five polymorphisms, one silent c.245T>C whereas others were missense mutations; c.247A>G, c.309G>A, c.471G>A and c.591T>C. The CNS3 genotype A in corresponds to protein variant A and D to B. The variant protein B, prevalent in DDP goats, is found to be more economically beneficent in terms of cheese production. Daily milk yield and total milk fat contents were significantly elevated in Beetal goat. In conclusion, polymorphism exists in the β-LG and CNS3 genes as well as in β-LG protein in Beetal and DDP goats of Pakistan. At β-LG locus, A allele is dominant in both goats whereas at CNS3 locus A allele is dominant in Beetal and D in DDP. The genetic findings could be employed in future to explore suitable candidates for quality dairy products manufacture.