Naveed Ahmed Syed

Luminescent mesoporous LaVO4:Eu3+ core-shell nanoparticles: synthesis, characterization, biocompatibility and their cytotoxicity

A general and facile method was used for preparation of water-soluble silica coated LaVO4:Eu3+ core-shell nanoparticles. In the present study, we have discussed and compared the cytotoxicity characteristics of the synthesized LaVO4:Eu3+ and silica coated LaVO4:Eu3+ core-shell nanoparticles. X-ray diffraction (XRD), field-emission transmission electron microscopy (FE-TEM), energy dispersive X-ray analysis (EDX), Fourier-transform infrared spectroscopy (FTIR), UV/Vis absorption spectroscopy and photoluminescence spectroscopic techniques were employed to characterize the structure and morphology of the prepared products. To obtain high aqueous solubility, luminescent LaVO4:Eu3+ nanoparticles were encapsulated with silica groups, giving the nanoparticles a negatively charged surface at physiological pH. The results of XRD confirm the formation of a well-crystallized LaVO4:Eu3+ phase with a tetragonal zircon structure. Optical absorption spectra show that the optical properties of silica-coated LaVO4:Eu3+ core-shell nanoparticles are related to their sizes and shapes. Further, in order to assess cytotoxicity, we investigated whether the LaVO4:Eu3+ nanoparticles could alter biological samples once they enter human H522 and peripheral blood mono nuclear cells (PBMCs). An MTT assay was performed to measure the mitochondrial activity that reflects the number of viable cells. Silica coated LaVO4:Eu3+ core-shell nanoparticles exhibited no significant effect on the viability of both types of cells up to 24 h after exposure. Strikingly, no dose effects were detected, even at highest concentrations.

Differential Expression Profile and Genetic Variants of MicroRNAs Sequences in Breast Cancer Patients

The technology available for cancer diagnosis and prognosis is not yet satisfactory at the molecular level, and requires further improvements. Micro RNAs (miRNAs) have been recently reported as useful biomarkers in diseases including cancer. We performed a miRNA expression profiling study using peripheral blood from breast cancer patients to detect and identify characteristic patterns. A total of 100 breast cancer patients and 89 healthy patients were recruited for miRNA genotyping and expression profiling. We found that hs-miR-196a2 in premenopausal patients, and hs-miR-499, hs-miR-146a and hs-miR-196a2 in postmenopausal patients, may discriminate breast cancer patients from healthy individuals. In addition, we found a significant association between two microRNA polymorphisms (hs-miR-196a2 and hs-miR-499) and breast cancer risk. However, no significant association between the hs-miR-146a gene and breast cancer risk was found. In summary, the study demonstrates that peripheral blood miRNAs and their expression and genotypic profiles can be developed as biomarkers for early diagnosis and prognosis of breast cancer.

In-vitro cyto-toxicity, geno-toxicity, and bio-imaging evaluation of one-pot synthesized luminescent functionalized mesoporous SiO2@Eu(OH)(3) core-shell microspheres

UNLABELLED Luminescent functionalized mesoporous SiO2@Eu(OH)3 core-shell microspheres (LFMCSMs) were prepared by coating of europium hydroxide (Eu(OH)3) shell on mesoporous silica (SiO2) nanospheres via a facile one-pot process at low temperature. The FETEM images revealed that a well-defined luminescent europium hydroxide shell was successfully grafted on the surface of mesoporous silica nanospheres. These experimental results showed that the LFMCSM has a typical diameter of ca. 392 nm consisting of the silica core with about 230 nm in diameter and europium hydroxide shell with an average thickness of about 162 nm. LFMCSMs exhibited strong red emission peak upon irradiation with ultraviolet light, which originated from the electric-dipole transition (5)D0 → (7)F2 (614 nm) of Eu(3+) ion. The biocompatibility of the synthesized LFMCSMs was evaluated in vitro by assessing their cytotoxic and genotoxic effect on human hepatoblastoma (HepG2) cells using MTT, TUNEL, fluorescent staining, DNA ladder and Gene expression assays respectively. FROM THE CLINICAL EDITOR This paper describes the development of a one-pot synthesis of luminescent mesoporous SiO2@Eu(OH)3 core-shell microspheres and evaluates their favorable in vitro cyto-toxicity and geno-toxicity, and their applications in bio-imaging of these particles that emit bright red signal under UV exposure.

Mechanism of cadmium induced apoptosis in human peripheral blood lymphocytes: The role of p53, Fas and Caspase-3

Cadmium (Cd) is a major pollutant of environment. It can be fatal to human. In spite of bulk of research and literatures, the mechanism of a fatality against human is still not understood completely. Toxic and carcinogenic effects of Cd in rodents and humans are well known. However, effects of Cd on induction of apoptosis are still elusive. This study indicates immunosuppression and immunotoxicity due to Cd exposure. Present study was undertaken to determine the mechanism of cell death in vitro in human peripheral blood lymphocytes induced by Cd. Our findings suggest the toxicity due to Cd is attributed to programmed cell death-apoptosis. IC₅₀ was calculated at 21.74 μM. A significant increase of expression of the pro-apoptotic genep53, Fas and Caspase-3 in human lymphocytes was found. Cd induced p53-dependent apoptosis through cooperation between Bak upregulation without changing the Bcl-2 and Bax expression. Data of this study compel to speculate that apoptosis may also be attributed to CD95/Fas complex formation, and p53 direct apoptogenic potential at mitochondria. It was confirmed by the increased expression of Caspase-3. Although, this work does not address all the questions regarding the mechanism of Cd induced apoptosis, but these findings establish an important role of p53 and mitochondrial function during apoptosis in human lymphocyte. Moreover, based upon our findings, the role of Fas in Cd induced apoptosis is also undeniable. Hence further investigations are required to understand the different mechanism involved into apoptosis of lymphocytes due to Cd exposure.

Validation of the Antiproliferative Effects of Organic Extracts from the Green Husk of Juglans regia L. on PC-3 Human Prostate Cancer Cells by Assessment of Apoptosis-Related Genes

With the increased use of plant-based cancer chemotherapy, exploring the antiproliferative effects of phytochemicals for anticancer drug design has gained considerable attention worldwide. This study was undertaken to investigate the effect of walnut green husk extracts on cell proliferation and to determine the possible molecular mechanism of extract-induced cell death by quantifying the expression of Bcl-2, Bax, caspases-3, and Tp53. PC-3 human prostate cancer cells. In this study, we found that green husk extracts suppressed proliferation and induced apoptosis in a dose- and time-dependent manner by modulating expression of apoptosis-related genes. This involved DNA fragmentation (determined by TUNEL assay) and significant changes in levels of mRNA and the expression of corresponding proteins. An increase in expressions of Bax, caspase-3, and tp53 genes and their corresponding proteins was detected using real-time PCR and western blot analysis in PC-3 cells treated with the green husk organic extracts. In contrast, Bcl2 expression was downregulated after exposure to the extracts. Our data suggest the presence of bioactive compound(s) in walnut green husks that are capable of killing prostate carcinoma cells by inducing apoptosis and that the husks are a candidate source of anticancer drugs.

Identification of Functional SNPs in BARD1 Gene and In Silico Analysis of Damaging SNPs: Based on Data Procured from dbSNP Database

Background The BARD1 gene encodes for the BRCA1-associated RING domain (BARD1) protein. Germ line and somatic mutations in BARD1 are found in sporadic breast, ovarian and uterine cancers. There is a plethora of single nucleotide polymorphisms (SNPs) which may or may not be involved in the onset of female cancers. Hence, before planning a larger population study, it is advisable to sort out the possible functional SNPs. To accomplish this goal, data available in the dbSNP database and different computer programs can be used. To the best of our knowledge, until now there has been no such study on record for the BARD1 gene. Therefore, this study was undertaken to find the functional nsSNPs in BARD1. Result 2.85% of all SNPs in the dbSNP database were present in the coding regions. SIFT predicted 11 out of 50 nsSNPs as not tolerable and PolyPhen assessed 27 out of 50 nsSNPs as damaging. FastSNP revealed that the rs58253676 SNP in the 3′ UTR may have splicing regulator and enhancer functions. In the 5′ UTR, rs17489363 and rs17426219 may alter the transcriptional binding site. The intronic region SNP rs67822872 may have a medium-high risk level. The protein structures 1JM7, 3C5R and 2NTE were predicted by PDBSum and shared 100% similarity with the BARD1 amino acid sequence. Among the predicted nsSNPs, rs4986841, rs111367604, rs13389423 and rs139785364 were identified as deleterious and damaging by the SIFT and PolyPhen programs. Additionally, I-Mutant showed a decrease in stability for these nsSNPs upon mutation. Finally, the ExPASy-PROSIT program revealed that the predicted deleterious mutations are contained in the ankyrin ring and BRCT domains. Conclusion Using the available bioinformatics tools and the data present in the dbSNP database, the four nsSNPs, rs4986841, rs111367604, rs13389423 and rs139785364, were identified as deleterious, reducing the protein stability of BARD1. Hence, these SNPs can be used for the larger population-based studies of female cancers.

Fenugreek Induced Apoptosis in Breast Cancer MCF-7 Cells Mediated Independently by Fas Receptor Change

Trigonella foenum in graecum (Fenugreek) is a traditional herbal plant used to treat disorders like diabetes, high cholesterol, wounds, inflammation, gastrointestinal ailments, and it is believed to have anti-tumor properties, although the mechanisms for the activity remain to be elucidated. In this study, we prepared a methanol extract from Fenugreek whole plants and investigated the mechanism involved in its growth-inhibitory effect on MCF- 7 human breast cancer cells. Apoptosis of MCF-7 cells was evidenced by investigating trypan blue exclusion, TUNEL and Caspase 3, 8, 9, p53, FADD, Bax and Bak by real-time PCR assays inducing activities, in the presence of FME at 65 μg/mL for 24 and 48 hours. FME induced apoptosis was mediated by the death receptor pathway as demonstrated by the increased level of Fas receptor expression after FME treatment. However, such change was found to be absent in Caspase 3, 8, 9, p53, FADD, Bax and Bak, which was confirmed by a time-dependent and dose-dependent manner. In summary, these data demonstrate that at least 90% of FME induced apoptosis in breast cell is mediated by Fas receptor-independently of either FADD, Caspase 8 or 3, as well as p53 interdependently.

Lack of Association of BRCA1 and BRCA2 Variants with Breast Cancer in an Ethnic Population of Saudi Arabia, an Emerging High-Risk Area

Incidence of breast cancer shows geographical variation, even within areas of ethnic homogeneity. Saudi Arabia has witnessed an increase in occurrence of breast cancer in its unexplored ethnic populations over the past few years. We aimed at determining whether any association exists between single nucleotide polymorphisms in breast cancer associated gene 1 (BRCA1) and breast cancer associated gene 2 (BRCA2) and the risk of breast cancer. TaqMan based Real Time Polymerase chain reaction genotyping assays were used to determine the frequency of single nucleotide polymorphisms in BRCA1 (rs799917) and BRCA2 (rs144848) in a group of 100 breast cancer patients and unaffected age matched controls of Saudi Arabian origin. The present data revealed that neither BRCA1 nor the BRCA2 studied variant show any significant association with the disease. This study failed to find any role of the concerned variants in breast cancer either as risk or as prognostic factors. The small number of patients registered was one of the limitations of this study. In summary, comparison of mutation profile with other ethnic populations and regions reflected both differences and similarities indicating co-exposure to a unique set of risk factors. The differences could be due to exposure to particular environmental carcinogens; different lifestyle, reproductive pattern; dietary or cultural practices of Saudi Arabian women that need further investigations.

Delineating the anti-cytotoxic and anti-genotoxic potentials of catechin hydrate against cadmium toxicity in human peripheral blood lymphocytes

Catechins (flavan-3-ol) are a type of natural phenol and well-studied antioxidants. Catechin hydrate, also known as taxifolin; is non-mutagenic, low in toxicity compared to other immunomodulator antioxidants. We aimed to determine the potential of catechin hydrate to prevent the cyto-genotoxic effects of cadmium in lymphocytes; demonstrate the immuno-protective activity of catechin hydrate. Our previous study indicated that cadmium is apoptogenic. Lymphocytes were treated with catechin hydrate or cadmium and catechine hydrate combinations (range 0.1-100μM) to determine their effects on cell viability. Lymphocytes treated with 100μM catechin hydrate and 100μM cadmium showed cell viability 70.65±6.92% and 5.69±2.27%, respectively. In our previous study cadmium (10 and 20μM) induced apoptosis in 31.8% and 44.4% of lymphocytes, respectively. However, the percentage of apoptotic cells after treatment with the combination of cadmium and catechin hydrate was not significantly different from that of catechin hydrate (P>0.05). Only 7.3% and 10.5% of the lymphocytes were apoptotic after treatment with 10μM cadmium+10μM catechin hydrate and 20μM cadmium+20μM catechin hydrate, respectively. The anti-geno-cytotoxic and immuno-protective potential of catechin hydrate was also demonstrated by the non-significant expression of apoptosis-related genes after treatment with catechin hydrate.

Association of multiple drug resistance-1 gene polymorphism with multiple drug resistance in breast cancer patients from an ethnic Saudi Arabian population

Chemotherapy has been used widely to treat cancer, both as a systemic therapy and as a local treatment. Unfortunately, many types of cancer are still refractory to chemotherapy. The mechanisms of anticancer drug resistance have been extensively explored but have not been fully characterized. This study analyzed the occurrences of polymorphism (SNP) in the MDR1 gene in breast cancer patients and determined a possible association with chemotherapy. The study group included one hundred breast carcinoma patients who subsequently received chemotherapy (the regimen generally consisted of commonly used drugs such as cyclophosphamide, adriamycin, 5-fluorouracil, docetaxel and their combinations). Blood samples from 100 healthy individuals are used, as controls were also genotyped for the MDR1 gene. This investigation revealed a significant correlation with response to various regimens of chemotherapy showing a low response to therapy with the CT/TT genotype at (exon 12) 1236 codon (p<0.001). These findings demonstrate, for the first time, that the polymorphisms in (exon 12) 1236 codon of the MDR1 gene greatly influence the drug response in patients from the Arab population of Saudi Arabia.