Expression Analysis of Sonic Hedgehog in Breast Cancer and its Relationship with Metastasis Involvement of Hedgehog pathway during embryogenesis and mammary development is well known. Salient members of this pathway include SHH, DHH, IHH, PTCH1, SMO and GLI1. Considering high prevalence and early onset of breast cancer among Pakistani population, reactivation of this developmental pathway is hypothesized. In the present study, expression profiling of Hedgehog pathway ligands (SHH, DHH, IHH), mediators (PTCH, SMO) and downstream effector (GLI1) in breast tumorigenesis was explored. The study was divided in two phases. The initial phase included screening of Hedgehog pathway molecules at mRNA and protein levels in Pakistani breast cancer patients. In the second phase, in-vitro models were generated for inhibition of SHH/GLI1 axis. Moreover, effect of SHH/GLI1 inhibition on EMT markers and metastatic cascade was investigated. In the first phase, expression of Hedgehog pathway molecules was evaluated using qRT-PCR and immunohistochemistry in tumor tissues (N=250) and adjacent noncancerous healthy tissues (N=250). SHH, DHH, PTCH1 and GLI1 were significantly overexpressed in tumors compared with respective controls. Significant correlation of SHH, DHH and GLI1 expression with advanced stage, grade, nodal involvement and distant metastasis was observed (p<0.05). Patients having high expression of SHH, DHH and GLI1 demonstrated poor overall survival in the cohort. Overexpression of SHH, DHH and GLI1 was observed to be significantly related with patients having early disease onset and pre-menopausal status. Moreover, Hedgehog pathway was frequently upregulated in luminal B or triple negative breast cancer affected patients. In addition, positive correlation was observed among aforementioned members of the pathway and Ki67 (r-value: 0.63-0.78) emphasizing their role towards disease progression. Expression of Vimentin and Snail was upregulated among SHH/GLI1 positive patients. Strong positive correlation of both Vimentin and Snail with SHH/GLI1 positive patients was observed. On the contrary, E-cadherin was downregulated in SHH/GLI1 positive patients and its correlation with SHH/GLI1 was also negative. xiii In the second phase, SHH knockout models were generated using CRISPR/CAS9 in MDA-MB-231 and MCF-7 cells. Additionally, both cancer cell lines were treated with GANT61 (GLI1 inhibitor) and its effect on cell viability and apoptosis was evaluated using CCK-8 and flow cytometry respectively. Effect of SHH knockout and GANT61 treatment was observed on EMT markers and Akt/ERK using qRT-PCR, western blot analysis and immunofluorescence staining. Invasion and migration assays were performed to investigate influence of SHH/GLI1 suppression on metastatic cascade. Treatment with GANT61 reduced cell viability (p<0.01) and induced apoptosis (p<0.01) in breast cancer cell lines. Expression of Vimentin and Snail was reduced in SHH knockout models and GANT61 treated cells. On the contrary, expression of E-cadherin was increased after inhibition of SHH or GLI1. Suppression of SHH/GLI1 axis also inhibited activation Akt/ ERK as well as reduced migration and invasion capacity in breast cancer cells. Based on these findings, SHH/GLI1 may be used as a prognostic marker in breast cancer patients. Moreover, inhibition of SHH/GLI1 axis provides a novel therapeutic target against aggressive breast cancer subtypes.
آہ! مولانا محمد عمران خان ندوی الازہری ۲۲؍ اکتوبر ۸۶ء کو یکایک خبر ملی کہ مولانا حافظ محمد عمران خان ندوی ازہری اپنے خاندان والوں کو دل فگار اور اپنے جاننے والوں کو دل گیر چھوڑ کر تہتر (۷۳) سال کی عمر میں اس عالم فانی سے عالم جادوانی کو سدھارے، اس خبر سے جسم و دماغ پر ایک بجلی گر پڑی۔ ان کی موت ایک خاص قسم کی قوت عمل، عزم، ادارہ، خوش انتظامی، خوش سلیقگی، خوش فکری، اور خوش و ضعی کی موت ہے، وہ اپنے پیچھے ایک درس چھوڑ گئے کہ عمل پیہم اور یقین محکم سے کس طرح اہم اور بڑے سے بڑا، کام انجام دیا جاسکتا ہے، بے سرو سامانی میں کیسے کیسے سازوسامان پیدا کیے جاسکتے ہیں، یا س کے عالم میں مخلص کارکنوں کی جماعت کی تنظیم کیسے کی جاسکتی، اور ناداری کے عالم میں بھی وہ کام انجام دیا جاسکتا ہے جو بڑی دولت کے ذریعہ سے نہیں کیا جاسکتا۔ وہ بھوپال کے رہنے والے تھے، ۱۹۴۷ء کے بعد جب بھوپال کا فرماں روا خاندان بھوپال چھوڑ کر دوسری جگہ پناہ گزین ہوگیا تھا تو مولانا عمران خان ندوی نے اپنی قوت ارادی کو بروئے کارلاکر بھوپال کے لوگوں کے دلوں پر اپنی فرماں روائی شروع کردی اور یہ نمونہ پیش کیا کہ تاج و تخت کے بغیر بھی اخلاص کی پاکیزگی، نیت کی طہارت اور عمل کی پختگی کے ساتھ فرماں روائی ہوسکتی ہے۔ انھوں نے تاج المساجد کی تعمیر جس طرح ازسرنو کی اور اس کی زینت و آرایش میں جس طرح اضافہ کیا، اور پھر اس کے ذریعہ سے جو دینی حمیت اور ایمانی حرارت پیدا کی، وہ ایسا کارنامہ ہے کہ دنیا کی بڑی مسجدوں کی تعمیر کی جب تاریخ لکھی جائے گی تو ان کا نام نامی بھی اس تاریخ میں ضرور لکھا جائے...
The concept of assertion has been playing a vital role in the linguistic to specify the meaning and nullify the uncertainties from the sentences. The numerous uses of assertion and its tools in Arabic language also played very significant role in interpreting the Qur’ān. Due to this reason rhetoricians and exegetists have much emphasized this phenomenon in their books and exegesis. One of the renowned books written by Abdul Qahir Jurjani is "دلائل الإعجاز" in which he mentioned its importance, principals and components in detail. The aim of this paper is to investigate the methods and stratigeis used by the great Muffasīr Abu Sa'ud, who tried to apply the rhetoric principals in his Tafsīr "إرشاد العقل السليم إلي مزايا القرآن الكريم" more emphasizing the phenomenon of assertion, its components, tools and importance in molding the concepts and meaning of the sentence.
Chickpea (Cicer arietinum L.) is the most important pulse crop among the major Rabi pulses of Pakistan. Fusarium wilt which is caused by Fusarium oxysporum f. sp. ciceris is considered to be the most devastating disease of this crop. Survey for the assessment of chickpea wilt disease was conducted in February - March 2005 and 2006 of major chickpea growing areas of Pakistan. During this survey, maximum disease prevalence (92.58 ± 3.69), incidence (82.52 ± 5.14) and severity (7.00 ± 0.48) was recorded in the districts Khushab, Bhakkar, Layyah, Mianwali and Sargodha. The lowest disease prevalence (29.70 ± 3.19), incidence (9.50 ± 1.72) and severity (2.1 ± 0.10) was recorded in districts Jafarabad, Dera Allah Yar, Attock, Peshawar, Rawalpindi / Islamabad. A strong correlation between disease incidence and climatic conditions in all districts was observed. Pathogenesis studies of these isolates were done in order to correlate epidemiological factors of chickpea wilt. Studies showed that seedling mortality was minimum at 3g inoculum concentration and maximum at 20g. Similarly, seedling mortality was maximum in sandy soil and minimum in clayey soil. Whereas, at low level of soil moisture disease severity was maximum and at high level it was minimum. The effect of disease development on the sap extract of leaves and roots showed maximum growth on root sap then leaves. In physiological studies the fungus grew best on Waksman Agar (WA) medium and least on Potato Dextrose Agar (PDA) medium. Glucose was found to be the best and mannitol least carbon source. Similarly potassium nitrate (KNO 3 ) was the best and ammonium sulphate (NH 4 ) 2 SO 4 was least nitrogen source. Growth of F. oxysporum was maximum at 30 0 C after 7 days of incubation, which reduced drastically below and above this temperature. Similarly, the fungus growth was best at pH 7; below and above this pH it was reduced. In case of pathogenic variability of 10 chickpea genotypes the most virulent isolate was Foc-15 (Bannu) and the least virulent was Foc-17 (Fateh-Jang) from 27 F. oxysporum f. sp. ciceris isolates. The remaining isolates showed intermediate response of variation in virulence. Cultivar ICCV2 (1±0.0) was found resistant and AUG-424 (9.0±0.0) susceptible xivagainst all the isolates. On morphological and cultural basis the isolate Foc-9 (90 mm) was found fast growing whereas, isolate Foc-22 (39 mm) was slow growing. The rest of isolates remained intermediate. The biggest size of the micro-conidia (3.7 x 4.5 μm) was obtained from the isolate Foc-14 and the smallest size (2.7 x 4.7 μm) from isolates Foc-2. Similarly the largest macro-conidia were obtained from the isolates Foc-6 (6.5 x 17.5 μm) and the smallest from isolates Foc-25 (7.5 x 20.1 μm). Maximum dry mycelium weight was obtained from isolate Foc-21 (1.56 g) and minimum from Foc-22 (0.31g). Sensitivity of F. oxysporum f. sp ciceris isolates to thirteen fungicides @ 100 ppm was observed. Benlate was found most effective fungicide against all the isolates in suppressing the fungal growth under test. The least effective fungicide were Captan and PCNB. The rest of fungicides showed intermediate response. In case of mycelial compatibility grouping out of 350 combinations only 91 combinations showed compatible reactions, whereas, 259 were identified as non-compatible. To identify genetic sources of resistance against wilt disease under artificial disease conditions in three consecutive years (2005, 2006 and 2007), 553 germplasm lines were screened. Disease observations were recorded from seedling to reproductive stages. At seedling stage, 233 genotypes were found resistant, 102 tolerant and 218 susceptible. Whereas, at reproductive stage 125 lines were resistant, 53 tolerant and 375 were observed susceptible. To evaluate the genetic relationship in the morphological traits and RAPD markers genetically diverse 38 chickpea genotypes were selected, which provided ample opportunity to create favorable gene combinations. 35 Random amplified polymorphic DNA (RAPD) markers were used to assess genetic diversity in comparison with agro- morphological traits. Agro-morphological traits have given significant results among genotypes. Days to 50% flowering ranged from 92-112, plant height ranged from 54.2- 87cm, Primary and secondary branches per plant varied from 1.4-6 and 4-20, pods per plant varied from 8-34, whereas grain yield per plant ranged from 3.47-10.2g. Disease was scored against the test genotypes, which, was recorded on 1-9 rating scale. The cluster analysis for the phylogenetic relationship among genotypes on the basis of agro- morphological traits revealed three distinct groups at 92% similarity level.