Dexmedetomidine leads to the amelioration of stress response to surgery.

Objectives: To determine the impact of intravenous Dexmedetomidine (Dex) administered perioperatively on stress response markers in patients undergoing ENT surgeries. Methods: This randomized interventional study was conducted at POFs Hospital and NUMS affiliated WMC, Wah Cantt, in collaboration with IIMC Rawalpindi, Pakistan, from October 2021 to April 2022. One hundred patients aged between 15-60 years, after satisfying the inclusion standards were randomly divided into two groups C and D. Group-C (n=50) received normal saline in addition to the standard anesthesia protocol. The intervention Group-D (n=50) was administered 1µg/kg dexmedetomidine hydrochloride intravenously over 10 minutes just before the induction followed 0.5µg/kg/hr as maintenance dose till the end of surgery. Serum inflammatory biomarkers (interleukins-6, TNF-α and cortisol) were measured in blood samples in both groups, taken at 0 (T0), 30 minutes(T1), and two hours (T2) time intervals and analyzed by using Enzyme-linked immunosorbent assay (ELISA). Data was statistically analyzed using SPSS 23. Results: The patients receiving Dex showed marked decrease in serum levels of cortisol, TNF- α and interleukins-6, from 139.73 to10.18, 99.51 to 0.96 and 85.09 to 0.96 respectively. Comparison between C and D groups revealed significant differences (p≤0.05) in these serum biomarkers. Conclusions: In the present study, intravenous Dex suppressed the intraoperative rise in levels of cytokine secretion and enhanced smooth recovery with no incidence of nausea and vomiting. These effects could be attributed to the anti-inflammatory effects of dexmedetomidine.

Mutational and expressional association of the PIK3CA gene with the risk of breast cancer in the Pakistani population.

PI3K pathway is a very important pathway that is reported to be involved in breast cancer. Mutation of PI3K and p110 alpha-catalytic subunit of phosphatidylinositol 3-kinase (PIK3CA) is of high predictive and prognostic values in breast cancer. The purpose of the current study was to screen the hotspot mutations of the PIK3CA gene i.e. rs2677760, rs3806685, rs121913273 & rs121913279 along with expressional analysis of PI3K and PIK3CA genes in breast cancer female patients. For mutational analysis, TaqMan assay & Sanger sequencing were performed while for expressional analysis real-time PCR was carried out. Mutant allele C of rs2677760 was observed to be high in postmenopausal patients. The frequency of mutant allele G of rs3806685 was significantly high in breast cancer patients. All diseased and control samples were of wild type for the hotspot rs121913273 and rs121913279 with allele G for rs121913273 and A for rs121913279. Expression of the PI3K was high in breast cancer tissue samples as compared to the adjacent controls. While the expression of the thePIK3CA gene was significantly high in premenopausal breast cancer patients. It was concluded that the mutant allele C of rs2677760 might have some sort of association with the menopausal status and it could be used as a diagnostic marker in post-menopausal women if studied further. Mutant allele G of rs3806685 was also found to be associated with breast cancer. While multiallelic rs121913273 and rs121913279 showed a different trend for the studied population. For expressional analysis, PI3K showed over-expression in the cases while PIK3CA gene expression was observed to be significantly associated with pre-menopausal status.

Natural plant extracts mediated expression regulation of TGF-ß receptors and SMAD genes in human cancer cell lines.

BACKGROUND: Transforming growth factor beta (TGF-ß) superfamily has key role in cell proliferation which leads to tumor promoting activities at metastatic stage of cancer. Inhibition of transforming growth factor beta receptor (TGFßR) signaling pathway can provide better therapeutic strategy to control cancer. Natural products are best known for their safety, less toxic nature, antioxidant characteristics making them a promising candidate to inhibit TGFßR signaling pathway. METHODS AND RESULTS: Crude methanolic extracts (CMEs) of 16 selected plants were prepared by using maceration method and subjected to phytochemical assays for identification of major phytometabolites particularly cancer chemopreventive antioxidant constituents. Total flavonoid content of all plants CME was > 0.6 mg/ml exhibiting the Cichorium intybus contains comparatively highest amount of total flavonoid content (0.53 mg/ml). Scanvenging activity of all plants was determined having IC50 ranges between 2 and 88 (µg/ml) while Moringa oleifera revealed the maximum scavenging activity (IC50 2.03 µg/ml). Comparative cytotoxicity of plant extracts was evaluated in HUH and MCF-7 cell lines using 3-(4,5-Dimethylthiazol-2-Yl)-2,5-Diphenyltetrazolium Bromide (MTT) colorimetric assay. The nine active plant extracts i.e. Fagonia cretica, Argemone Mexicana, Rubus fruticosus, M. oleifera, Punica granatum, Cichorium intybus, Xanthium strumarium, Carissa opaca, Cyperus rotundus were identified based on their high antiproliferative activity > 50% against cancer cell lines and subjected to relative expression studies. Modulation of TGFß signaling molecules (i.e.TGFßR1, 2 & 3, SMAD3, SMAD5) and ubiquitous proteins i.e. SMURF1 and SMURF2 genetic expression by potent extracts was determined by RT-PCR using GAPDH (housekeeping gene) as gene of reference. CONCLUSIONS: This present study revealed that CME of Fagonia cretica and Argemone mexicana significantly inhibit TGF beta mediated signaling cascade by downregulating the gene expression fold change > 1 of TGFßR 1, 2 & 3 and receptor associated complex protein SMAD3 as compared to control.

Role of lidocaine and its effect on postoperative outcomes in abdominal cholecystectomy.

OBJECTIVE: To investigate the role of pre- and intra-operative lidocaine infusion on post-operative pain management. METHODS: The interventional, prospective study was conducted from September 2019 to June 2020 at the Pakistan Ordnance Factories Hospital, Wah Cantt, Pakistan, and comprised patients aged 18-60 years undergoing elective cholecystectomy who were randomised into intervention group A and control group B. Group A was given a bolus dose of lidocaine hydrochloride 2 mg/kg in addition to the standard anaesthesia protocol, while group B was given continuous intravenous infusion of 0.9% normal saline along with the standard protocol. Blood samples for interleukins 6 and 8 were taken at baseline, and then at 2, 6 and 8 hours Post-operatively. Data was analysed using SPSS 23. RESULTS: Of the 40 patients, 20(50%) were in each of the two groups. There was a marked decrease in interleukins 6 and 8 levels group A compared to group B (p<0.05). Interleukin 8 level showed a marked decline compared to that of interleukin 6 (p<0.05). CONCLUSIONS: A decrease in interleukins 6 and 8 levels highlighted the anti-inflammatory role of lidocaine and resulted in a decrease in post-operative opioid consumption.

SnO2nanorods/graphene nanoplatelets nanocomposites: towards fast removal of malachite green and pathogen control.

The world is facing alarming challenges of environmental pollution due to uncontrolled water contamination and multiple drug resistance of pathogens. However, these challenges can be addressed by using novel nanocomposites materials such as, SnO2/graphene nanopaletelets (GNPs) nanocomposites remarkably. In this work, we have prepared SnO2nanorods and SnO2/GNPs nanocomposites (GS-I and GS-II) with size of 25 ± 6 nm in length and 4 ± 2 nm in diameter. The optical bandgap energies change from 3.14 eV to 2.80 eV in SnO2and SnO2/GNPs nanocomposite. We found that SnO2/GNPs nanocomposite (GS-II) completely removes (99.11%) malachite green in 12 min, under UV light exposure, while under same conditions, SnO2nanorods removes only 37% dye. Moreover, visible light exposure resulted in 99.01% removal of malachite green in 15 min by GSII as compared to 24.7% removal by SnO2. In addition, GS-II nanocomposite inhibits 79.57% and 78.51% growth ofP. aeruginosaandS. aureusrespectively. A synchronized contribution of SnO2and GNPs makes SnO2/GNPs nanocomposites (GS-II) an innovative multifunctional material for simultaneous fast and complete removal of malachite green and inhibition of drug resistant pathogens.

Genetic predisposition of SNPs in miRNA-149 (rs2292832) and FOXE1 (rs3758249) in thyroid Cancer.

BACKGROUND: Many efforts have been made in recent years to investigate the alterations in protein-coding genes as well as non-coding RNAs that are playing an emerging role in the development and progression of cancers. These miRNAs are short non-coding functional RNAs that are involved in the regulation of transcriptome. In different studies, it was found that human miRNA-149 is an important microRNA that is functioning either as onco-miRNAs or acting as tumor suppressors, in different conditions. RATIONALE: Many of the miRNAs are regulating different SNPs of FOXE1 in different studies which are causing low-to-moderate penetrance of genes that initiates the development of thyroid cancer. The involvement of SNPs in miRNA-149 gene rs2292832 and FOXE1 rs3758249 with PTC for better disease prognosis and management was determined in this study and the relation between these SNPs at the genotypic level was also evaluated. MATERIALS AND METHODS: PTC patients with age and gender-matched controls were recruited in the present study. Blood samples were collected in EDTA vacutainer followed by DNA extraction by the organic method. Genotyping of rs2292832 and rs3758249 was done by ARMS-PCR and PCR- RFLP respectively. Statistical analyses were carried out by using SPSS software (version 20). RESULTS: The mutation T>C in miRNA-149 rs2292832 was significantly associated with thyroid cancer (p-value 0.0004, < 0.05) while rs3758249 G>C did not show significant association with the disease (p-value 0.124244, > 0.05). Moreover, no correlation of rs2292832 at the genotype level was observed with rs3758249. CONCLUSIONS: miRNA-149 gene SNP rs2292832 was observed in strong association with thyroid cancer. Lack of genetic association of rs3758249 of FOXE1 gene has been ruled for the disease. The statistically significant association of rs2292832 with thyroid cancer depicts its mechanistic involvement at the cellular level in Papillary Thyroid Carcinoma.

Potential risk assessment of miR-143 gene polymorphisms rs41291957 and rs353292 in colorectal cancer.

Colorectal cancer is a life-threatening and therapeutically challenging disease. Increasingly it is being deciphered that genetic and epigenetic mutations play a central role in cancer onset and progression. Excitingly, discovery of non-coding RNAs is considered to be a milestone in molecular oncology and emerging evidence is deepening our understanding about pivotal role of miRNAs in carcinogenesis. miR-143 has been experimentally verified to play an instrumental role as tumor suppressor. Recent studies suggest that single nucleotide polymorphisms rs41291957 and rs353292 in miR-143 may associate with the progression and or development of colorectal cancer. In present study 400 Pakistani subjects participated including 200 colorectal cancer patients and 200 age and gender matched healthy individuals. Blood samples and clinical information of the confirmed patients was collected from cancer diagnosis and treatment hospitals in Pakistan. The polymorphisms rs41291957 and rs353292 were genotyped in patients and controls by Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) and results were validated by Sanger sequencing. The association of the SNPs within the study group was analyzed by χ² test with p value < 0.05 as significant. Odds ratio was calculated with 95% confidence interval.Genetic predisposition to cancer was observed in presence of characteristic rs45291957 polymorphism. χ² test results show strongly significant association mi-RNA rs45291957 SNP with colorectal cancer p value 0.0111 (<0.05) along with the statistically significant correlation tested by odds ratio with 95% confidence interval. However, no significant correlation (p value 0.6683) could be found for the association of rs353292 with colorectal cancer in Pakistani population. The present study for the first time gave evidence of miR-143 rs41291957 involvement in colorectal cancer patients of Pakistani population. This target can be a useful molecular tool for the prognosis and treatment targets for colorectal cancer in Pakistani population. rs353292 genetic association can be explored for different cancers in Pakistan to completely rule out its role in cancer.

Superior antibacterial activity of ZnO-CuO nanocomposite synthesized by a chemical Co-precipitation approach.

Synthesis of highly efficient antibacterial agents has become highly important due to emergence of antibiotic resistance. Herein, Pristine ZnO and ZnO-CuO nanocomposite has been synthesized by simple chemical co-precipitation method and characterized by X-ray diffraction (XRD), microscopic and spectroscopic techniques. The prepared ZnO-CuO nanocomposite is composed of two dimensional nanosheets consisting of hexagonal ZnO and monoclinic CuO crystal phases present in coexistence. Moreover, a minute presence of Cu5Zn8 cubic phase has been evident in the XRD pattern of ZnO-CuO nanocomposite. Fourier Transform Infrared Spectroscopy (FTIR) spectrum of the prepared nanocomposite has revealed the presence of vibrational modes related to both Zn-O and Cu-O. Photoluminescence (PL) investigations depicted the formation of huge amounts of surface defects in ZnO-CuO nanocomposite as compared to pristine ZnO nanostructures. The prepared ZnO-CuO nanocomposite has efficiently killed Methicillin resistant Staphylococus aureus (s. aureus) bacterium by producing 24 mm of zone of inhibition (ZOI) comparing to 8 mm ZOI produced by pristine ZnO. The superior antibacterial activity of ZnO-CuO nanocomposite has been attributed to oxidative stress generated by electron transfer pathway and reactive oxygen species (ROS) generation.

Signaling cascades in thyroid cancer: Increasing the armory of archers to hit bullseye.

Thyroid cancer is a multifaceted and therapeutically challenging disease and rapidly accumulating experimentally verified findings have considerably improve our understanding of the molecular mechanisms which underlie its development. Substantial fraction of information has been added into existing landscape of molecular oncology and we have started to develop a sharper understanding of the underlying mechanisms of thyroid cancer. Wealth of information demystified different intracellular signaling cascades which are frequently deregulated in thyroid cancer. In vitro assays and xenografted mice based studies have helped us to identify drug targets and different synthetic and natural products are currently being tested to effectively treat thyroid cancer. Cabozantinib and vandetanib have been approved to treat medullary thyroid cancer (MTC) and two agents (lenvatinib and sorafenib) are also being used to treat radioactive-iodine refractory differentiated thyroid cancer. This review comprehensively summarizes most recent advancements in our knowledge related to dysregulated intracellular signaling cascades in thyroid cancer and how different proteins can be therapeutically exploited. (1) We discuss how loss of TRAIL mediated apoptosis occurred in thyroid cancer cells and how different strategies can be used to restore apoptosis in resistant cancer cells; (2) We provide detailed account of seemingly opposite roles of NOTCH signaling in thyroid cancers; (3) TGF/SMAD mediated signaling also needs detailed research because of context dependent role in thyroid cancer. Researchers have only begun to scratch the surface of how TGF signaling works in thyroid cancer and metastasis; and (4) Role of SHH signaling in thyroid cancer stem cells is also well appreciated and targeting of SHH pathway will be an important aspect in treatment of thyroid cancer. Better concepts and improved knowledge will be helpful for clinicians in getting a step closer to individualized medicine.

MicroRNA Regulation of Telomerase Reverse Transcriptase (TERT): Micro Machines Pull Strings of Papier-Mâché Puppets.

Substantial fraction of high-quality information is continuously being added into the existing pool of knowledge related to the biology of telomeres. Based on the insights gleaned from decades of research, it is clear that chromosomal stability needs a highly controlled and dynamic balance of DNA gain and loss in each terminal tract of telomeric repeats. Telomeres are formed by tandem repeats of TTAGGG sequences, which are gradually lost with each round of division of the cells. Targeted inhibition of telomerase to effectively induce apoptosis in cancer cells has attracted tremendous attention and overwhelmingly increasingly list of telomerase inhibitors truthfully advocates pharmacological significance of telomerase. Telomerase reverse transcriptase (TERT) is a multi-talented and catalytically active component of the telomerase-associated protein machinery. Different proteins of telomerase-associated machinery work in a synchronized and orchestrated manner to ensure proper maintenance of telomeric length of chromosomes. Rapidly emerging scientific findings about regulation of TERT by microRNAs has revolutionized our understanding related to the biology of telomeres and telomerase. In this review, we have comprehensively discussed how different miRNAs regulate TERT in different cancers. Use of miRNA-based therapeutics against TERT in different cancers needs detailed research in preclinical models for effective translation of laboratory findings to clinically effective therapeutics.

Molecular characterization of autosomal recessive non syndromic hearing loss in selected families from District Mardan, Pakistan.

Deafness is the most common sensory disorder, which affects 1/1000 neonates globally. Genetic factors are major contributors for hearing impairment. This study was conducted to explore the linkage of DFNB loci and their mutations with NSHL in selected Pakistani families. We included 10 families with history of deafness from district Mardan, Pakistan. Blood sample (5ml) along with personal and clinical information was collected from the available family members including both diseased and un-affected individuals. Genomic DNA was amplified using loci specific STR markers to investigate the linkage of DFNB loci. Family found linked with DFNB4 locus was screened for SLC26A4 mutations. One out of the ten explored families was found linked with DFNB4 locus which was further investigated for SLC26A4 gene mutation through direct DNA sequencing. Two novel mutations were observed in the studied family, one at splice donor site (164+2T>G) and the other at position 164+5C>G only in the affected members of the linked family. DFNB4 locus was found linked in the present study which harbors SLC26A4 gene. The novel mutation of SLC26A4 gene at the splice donor site results in skipping of the first coding exon and thus can lead to loss of expression of SLC26A4 product in the inner ear.

Population-specific genetic variation at microRNA-629-binding site in the 3'-UTR of NBS1 gene in prostate cancer patients.

OBJECTIVE: Prostate cancer is a genomically complex disease and recently emerging scientific evidence is adding new pieces of information into existing pool of knowledge of oncology. Prostate cancer cells tactfully rewire signaling cascades in presence or absence of androgen. We do not have finer proteome and genome based patient related information of our cancer patients. METHODOLOGY: In the present laboratory research, we studied 3' UTR C/T and A/G polymorphism in NBS1 gene in 100 prostate cancer patients and 100 healthy individuals without any previous clinical history, using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis. RESULTS: For rs13312986 A&#62;G genotypes, AA was 78% in patients and 80% in controls. AG was 21% in patients and 20% in controls. GG was 1% in patients and none was detected in control. Allelic frequencies for A was 0.885% in patients and 0.9% in controls. Allelic frequencies for G were 0.115% in patients and 0.1% in controls. For rs14448 T&#62;C genotypes, TC was 23% in patients and 20% in controls. TT was 77% in patients and 80% in controls. CC was not detected either in patients or controls. T allele was 0.885% in patients and 0.9% in controls. C allele was 0.115% in patients and 0.1%-+ in controls. CONCLUSIONS: Future studies must converge on deeper and detailed mechanisms, as miRNA regulation of NBS1 is incompletely studied and exploration of these connections constitutes one of the most exciting areas in clinical oncology.

Pharmacologically active flavonoids from the anticancer, antioxidant and antimicrobial extracts of Cassia angustifolia Vahl.

BACKGROUND: Cassia angustifolia Vahl. (commonly known as senna makkai or cassia senna), native to Saudi Arabia, Egypt, Yemen and also extensively cultivated in Pakistan, is a medicinal herb used traditionally to cure number of diseases like liver diseases, constipation, typhoid, cholera etc. This study was conducted to evaluate the in-vitro antimicrobial, antioxidant and anticancer assays and phytochemical constituents of aqueous and organic extracts of C. angustifolia leaves. METHODS: The antimicrobial activities of C. angustifolia aqueous and organic (methanol, ethanol, acetone, ethyl acetate) extracts were investigated by the disk diffusion method. These extracts were further evaluated for antioxidant potential by the DPPH radical scavenging assay. Anticancer activities of the extracts were determined by the MTT colorimetric assay. The total phenolic and flavonoid contents of C. angustifolia extracts were evaluated by the Folin-Ciocalteu method and aluminum chloride colorimetric assay, respectively. The structures of the bioactive compounds were elucidated by NMR and ESI-MS spectrometry. RESULTS: Bioactivity-guided screening of C. angustifolia extracts, led to the isolation and identification of three flavonoids quercimeritrin (1), scutellarein (2), and rutin (3) reported for the first time from this plant, showed significant anticancer activity against MCF-7 (IC50, 4.0 µg/µL), HeLa (IC50, 5.45 µg/µL), Hep2 (IC50, 7.28 µg/µL) and low cytotoxicity against HCEC (IC50, 21.09 µg/µL). Significant antioxidant activity was observed with IC50 2.41 µg/mL against DPPH radical. Moreover, C. angustifolia extracts have the potential to inhibit microbial growth of E. cloacae, P. aeruginosa, S. mercescens and S. typhi. CONCLUSION: C. angustifolia extracts revealed the presence of quercimeritrin (1), scutellarein (2), and rutin (3), all known to have useful bioactivities including antimicrobial, antioxidant and anticancer activities.

Frequency of CYP2D6*10 genotypes in Pakistani breast cancer patients taking adjuvant tamoxifen.

OBJECTIVE: To investigate the frequency of cytochrome P2D6*10 in breast cancer patients. METHODS: This retrospective study was conducted at the Nuclear Medicine, Oncology and Radiotherapy Institute, Islamabad, and the Combined Military Hospital, Rawalpindi, Pakistan, and comprised medical records of breast cancer patients from January 2000 to September 2013. Pre- and post-menopausal women with diagnosed breast cancer who were advised 20mg/day of tamoxifen as adjuvant therapy were included. The frequency of the cytochrome was determined using polymerase chain reaction-restriction fragment length polymorphism analysis. RESULTS: Of the 232 participants, 25(10.8%) had stage I disease, 127(54.7%) had stage II and 80(34.5%) had stage III disease. The overall mean age was 46.9±9.9 years. The allele frequency of cytochrome CYP2D6*1 was 431(93%) and that of CYP2D6*10 was 33(7 %). Pakistanis differed significantly from the Asian populations and other ethnic groups in the distribution of the allele cytochrome, but its frequency was comparable to South Indians. CONCLUSIONS: The frequency of CYP2D6*10 allele in Pakistani breast cancer women was comparable to the South Indians, but was significantly lower than other populations.

Association between breast cancer and tumour necrosis factor-related apoptosis-inducing ligand (TRAIL) gene 1595C/T SNP in a Pakistani population.

AIM OF THE STUDY: TRAIL-mediated signalling has emerged as an extensively studied biological mechanism reported to differentially induce apoptosis in cancer cells. However, overwhelmingly increasing experimentally verified data is shedding light on resistance against TRAIL-induced apoptosis in cancer cells. Moreover, genetic and epigenetic mutations also exert effects on the functionality of TRAIL and its receptors. In this study we investigated the association between breast cancer and polymorphisms in tumour necrosis factor-related apoptosis-inducing ligand (TRAIL) in a Pakistani Population. MATERIAL AND METHODS: Genotyping for TRAIL gene 1595 C/T polymorphism was done for 363 breast cancer patients and 193 age- and sex-matched healthy controls. DNA was extracted using standard organic methods. PCR-RFLP analysis was done for C/T polymorphism at position 1595 in exon 5 of the TRAIL gene using site-specific primers and restriction enzyme. The results were statistically evaluated by SPSS14. RESULTS: In this study, CC homozygotes were 46.3% in patients and 49.7% in controls, p = 0.729 with OR value 0.8705 (95% CI: 0.6137-1.2348). CT was statistically insignificant, p = 0.837 with OR value 0.9242 (95% CI: 0.6494-1.3154). However, the minor allele or risk allele genotype TT had a higher percentage among breast cancer patients (12.1%) than in the control group (6.7%). Since there was a statistically insignificant difference (p = 0.212, OR value 1.9098 with 95% CI 1.0019 to 3.6406) of TT genotype between the two groups, the contrastingly higher percentage of TT genotype in breast cancer patients seems to be a risk factor for the disease. Moreover, the frequency of minor allele T was also found to be higher in the patients (0.329) than in the controls (0.285). CONCLUSIONS: The TRAIL gene 1595 C/T SNP has a contradictory role in cancer development in different populations. In our population group although the percentage of homozygous risk allele TT was higher in patients it was statistically non-significant. The raised T allele and TT genotype in patients may suggest its association with breast cancer in the Pakistani population.

Penetrance and phenotypic effects of C/T polymorphism at 1595 position in exon 5 of the TRAIL gene among prostate cancer patients in Pakistani population.

It is becoming progressively more understandable that variations within the sequence of tumor suppressor genes and oncogenes may contribute to cancer progression. Increasingly it is being realized that cancer cells get resistant to pro-apoptotic signals and evidence has started to shed light on the fact that nucleotide polymorphisms may lead to suboptimal apoptotic capacity and therefore increased cancer risk. It has previously been shown that there is a relationship between C/T polymorphism at 1595 position in exon 5 of the TRAIL gene and cancer however rapidly accumulating data cannot be extrapolated to other populations due to intra- and inter-ethnic variability. The study is focused on the C/T polymorphism at 1595 position in exon 5 of the TRAIL gene in prostate cancer patients diagnosed in local population in Pakistan. 126 prostate cancer patients and 91 control subjects participated in this study. 5ml venous blood was taken from participants with informed consent. DNA was extracted using standard organic methods. PCR-RFLP analysis was done for C/T polymorphism at 1595 position in exon 5 of the TRAIL gene using site specific primers and restriction enzyme. The results were statistically evaluated in SPSS14. In this particular study it was found that there was no significant difference in major allele C genotype between patients and controls, p value > 0.05. Similar statistically non-significant difference was observed for T allele genotype in the patient and control groups. However the heterozygous genotype CT was significantly higher, p value 0.053 (-0.05), in prostate cancer patients as compared to controls. This is the first study providing a clue of relationship of C/T polymorphism role in prostate cancer development and progression in our population.

Influence of genetic variations in ABCB1 on the clinical efficacy of ondansetron - A pharmacogenetic analysis of Pakistani population.

OBJECTIVE: To study the association of C1236T single-nucleotide polymorphisms of ABCB1 gene with non-responsiveness to antiemetic treatment in post-operative patients. METHODS: The prospective, clinical trial was conducted at Combined Military Hospital, Rawalpindi, and the Institute of Biomedical and Genetic Engineering, Islamabad, in 2012-13, and comprised patients undergoing elective laparoscopic cholecystectomy. All patients were given 0.1 mg/kg ondansetron intravenously 30 minutes before the end of surgery and Deoxyribonucleic acid samples were obtained. The frequencies of genotypes of Single Nucleotide Polymorphism were determined by polymerase chain reaction followed by restriction fragment length polymorphism. RESULTS: Of the 426 patients, 201(47%) were responders having no nausea or vomiting, and 225(52.8%) were non-responders having nausea or vomiting. The incidence of post-operative nausea and vomiting during the first 2 hours after surgery was significantly lower in patients with 1236TT genotype than other 1236 genotypes (p<0.001). It was significantly higher in patients with CC genotype at 2 hours than other 1236 genotypes (p<0.001). CONCLUSIONS: Polymorphism of ABCB1 gene may be a good guide for predicting responsiveness for ondansetron.

Re-evaluating the FGFR4 (G388R) germline mutation in different cancers in Pakistani population.

BACKGROUND: Cancer is a life threatening complicated diseasethatarises because of wide-ranging environmental and cellular factors. These external and internal stresses disrupt the spatio-temporally controlled mechanisms of the cellular signalings. Accumulating evidence suggests that signal transductions are misrepresented in carcinogenesis and FGFR4 is reported to be involved in carcinogenesis. Although there is considerable evidence emphasizing the relationship between FGFR4 (G388R) mutation and carcinogenesis however rapidly accumulating data cannot be extrapolated to Pakistani population due to intra- and inter-ethnic variability. OBJECTIVE: The study is focused on the trans-membrane mutation G388R of FGFR4 genes among different types of cancers diagnosed in local population in Pakistan. METHODS: 103 breast cancer patients, 56 Lung cancer patients (both Small Cell and Non Small Cell) and 45 control subjects participated in this study. Sample of 5-ml venous blood was taken from participants with informed consent. DNA was extracted and PCR-RFLP analysis was done for G388R mutation in FGFR4 gene using site specific primers and restriction enzyme. The results were statistically evaluated in SPSS14. RESULTS: The genotypes of G388R in FGFR4 gene were in Hardy-Weinberg equilibrium. The percentage of normal homozygotes GG was found to be (48.5%) in breast cancer, (51.8%) in lung cancer. Glycine/arginine was (36.9%) in breast cancer, (42.9%) in lung cancer. CONCLUSION: We were unable to find a possible correlation between FGFR4 G388R mutation and different cancers in local population.

Antibacterial activity of local herbs collected from Murree (Pakistan) against multi-drug resistant Klebsiella pneumonae, E. coli and methyciline resistant Staphylococcus aureus.

Exploring healing power in plants emerged in prehistory of human civilization. Sustaining good health has been achieved over the millions of years by use of plant products in various traditional sockets. A major contribution of medicinal plants to health care systems is their limitless possession of bioactive components that stimulate explicit physiological actions. Luckily Pakistan is blessed with huge reservoir of plants with medicinal potential and some of them; we focused in this study for their medicinal importance.In this study we checked the antibacterial activity inherent in Ricinus communis, Solanum nigrum, Dodonaea viscose and Berberis lyceum extracts for multidrug resistance bacterial strains Klebsiella pneumonae, E. coli and methyciline resistant Staphylococcus aureus. MRSA showed sensitivity for Ricinus communis. Multidrug resistant Klebsiella pneumonae was sensitive with Pine roxburgii and Ricinus communis but weakly susceptible for Solanum nigrum. Multidrug resistant E. coli was resistant to all plant extracts. Treatment of severe infections caused by the bacterial strains used in this study with Ricinus communis, Pine roxburgii and Solanum nigrum can lower the undesired side effects of synthetic medicine and also reduce the economic burden.

Making personalized prostate cancer medicine a reality: challenges and opportunities in the re-establishment of gold standards.

Prostate cancer is a serious multidimensional disorder that arises because of misrepresentation of signaling cascades and acquired resistance against apoptosis. It is progressively becoming more insurmountable because of rheostat like switching of oncogenic signaling in androgen dependent and androgen depleted microenvironment. Additionally, oncogenic fusion proteins have been explored in prostate cancer tissues thus adding another layer of complexity to the targeting of protein network in cancer cell and generate hurdles in the standardization of therapy. In this Review we briefly describe identified oncogenic fusion transcripts in prostate cancer and suggest utilization of this biomarker for prostate cancer diagnosis alongwith standard PSA and immunohistochemistry analysis in Pakistan. We also provide overview of animal model studies to interpret the efficacy of vitamins.