A non-invasive prenatal DNA screening test for Down Syndrome

Prenatal diagnosis still depends on invasive methods using cells contained in the amniotic fluid or villus cells (Suzumori, 1999). These procedures carry a small but important rate of miscarriage (Vyas, 1994). The discovery on the presence of foetal cells in maternal circulation was a turning point for the development of a safe, accurate and reliable prenatal diagnosis, which could be offered to as a routine test to any pregnant women regardless their age

Tamoxifen-loaded nanostructured lipid carrier as a drug delivery system: characterization, stability assessment and cytotoxicity

Cancer nanotherapeutics is beginning to overwhelm the global research and viewed to be the revolutionary treatment regime in the medical field. This investigation describes the development of a stable nanostructured lipid carrier (NLC) system as carrier for Tamoxifen (TAM). The TAM-loaded NLC (TAM-NLC) developed with 200 mg of TAM showed a spherical particle with the size of 46.6 nm, polydispersity index of 0.267, entrapment efficiency of 99.74% and with the zeta potential of -23.78 mV. Besides, the equivalent cytotoxicity of TAM and TAM-NLC to human (MCF-7) and mice (4T1) mammary breast cancer cell lines were observed. Incubating the formulation at the physiological pH resulted into reduced Ostwald ripening rate but without any significant change in the absorptivity. When coupled with the measurements of zeta potential and Ostwald ripening rate, the absorbance assay may be used to predict the long-term stability of drug-loaded nanoparticle formulations. The results of the study also suggest that TAM-NLC is a promising drug delivery system for breast cancer therapy. This is the first encouraging report on the in vitro effect of TAM-NLC against human and mouse mammary adenocarcinoma cell lines

An overview of in vitro research models for Alzheimer`s disease (AD)

Alzheimer’s disease (AD) is the most common form of age-related dementia. It is a neurodegenerative disease characterized by two aberrant features, the amyloid plaques and the neurofibrillary tangles which result in progressive memory loss and cognitive disturbances. This has led to devastating suffering to the patient, caregivers, family and economy of the country. As a result, scientists are putting efforts in understanding the mechanisms underlying the development of the disease as well as treatment for the disease. To do so, an ideal model is required that can mimic the development of AD,demonstrating the progressive degeneration of the neurons and formation of amyloid plaques and neurofibrillary tangles. In this review paper, currently available in vitro models for AD will be disc ussed, which include the cancer, primary culture and stem cell lines, highlighting on the benefits and limitations of each. More attention will be focused on the latest established disease-specific induced pluripotent stem cells (iPSCs)isolated from familial AD patients and Dow n syndrome patients. These models have their own advantages and limitations, therefore, more research needs to be done to come up with a model that is suitable not only for fundamental understanding of the disease but also for drug discovery and development

Potential anticancer effect of red spinach (Amaranthus gangeticus) extract.

The objective of this study was to determine the anti cancer effects of red spinach (Amaranthus gangeticus Linn) in vitro and in vivo. For in vitro study, microtitration cytotoxic assay was done using 3-(4,5-dimethylthiazol-2-il)-2,5- diphenil tetrazolium bromide (MTT) kit assay. Results showed that aqueous extract of A gangeticus inhibited the proliferation of liver cancer cell line (HepG2) and breast cancer cell line (MCF-7). The IC50values were 93.8 μg/ml and 98.8 μg/ml for HepG2 and MCF-7, respectively. The inhibitory effect was also observed in colon cancer cell line (Caco-2), but a lower percentage compared to HepG2 and MCF-7. For normal cell line (Chang Liver), there was no inhibitory effect. In the in vivo study, hepatocarcinogenesis was monitored in rats according to Solt and Farber (1976) without partial hepatectomy. Assay of tumour marker enzymes such as glutathione S-transferase (GST), gamma-glutamyl transpeptidase (GGT), uridyl diphosphoglucuronyl transferase (UDPGT) and alkaline phosphatase (ALP) were carried out to determine the severity of hepatocarcinogenesis. The result found that supplementation of 5%, 7.5% and 10% of A. gangeticus aqueous extract to normal rats did not show any significant difference towards normal control (P <0.05). The exposure of the rats to chemical carcinogens diethylnitrosamine (DEN) and 2-acetylaminofluorene (AAF) showed a significant increase in specific enzyme activity of GGT, GST, UDPGT and ALP compared to normal control (P <0.05). However, it was found that the supplementation of A. gangeticus aqueous extract in 5%, 7.5% and 10% to cancer-induced rats could inhibit the activity of all tumour marker enzymes especially at 10% (P 0.05). Supplementation of anti cancer drug glycyrrhizin at suggested dose (0.005%) did not show any suppressive effect towards cancer control (P <0.05). In conclusion, A. gangeticus showed anticancer potential in in vitro and in vivo studies

Estrogen receptor-α gene, codon 594 (G3242A) polymorphism among Iranian women with breast cancer: a case control study

A case-control study was conducted to establish a database of ESR1 polymorphisms in Iranian population in order to compare Western and Iranian (Middle East) distributions and to evaluate ESR1 polymorphism as an indicator of clinical outcome. The ESR1 gene was scanned in Iranian patients newly diagnosed invasive breast tumors, (150 patients) and in healthy individuals (147 healthy control individuals). PCR single-strand conformation polymorphism technology and direct sequencing was performed. The silent single nucleotide polymorphism (SNPs) was found, as reported previously in other studies, but at significantly different frequencies. The frequency of genotype 01 in codon 594 (ACG-ACA), (G3242A), exon 8 was significantly higher in breast cancer patients (48.0%) than in control individuals (1.4%; p = 0.001). The allele 1 in codon 594 was significantly more common in breast cancer patients with age at menarche </=12 (40.8%) than in those which their menstruation began at older than 12 years old (23.9%; p = 0.002). The allele 1 in codon 594 exhibited, the greater the frequency, the lesser the likelihood of LN metastasis. Present results demonstrated that this particular SNP marker may increase accuracy in predicting LN. Therefore, this SNP marker further increased predictive accuracy in Iranian population. These data suggest that ESR1 polymorphisms are correlated with various aspects of breast cancer in Iranian ESR1 genotype, as determined during pre-surgical evaluation, might represent a surrogate marker to increase predicting breast cancer in Iranian population

Evaluation of two cell culture media in culturing rat full term amniotic fluid cells

Introduction: Amniotic fluid (AF) consists of heterogenous population of cells with high diagnostic and therapeutic values. The study of rat amniotic fluid cells is very limited, despite the extensive use of this animal model in biomedical research. Primary culture of rat AF cells, especially from full term pregnancies has not been well established. Here we attempt to determine the suitable medium in culturing rat AF cells that would enhance the cell viability, growth rate and heterogeneity. Methods: The cell viability, growth rate and heterogeneity of rat AF cells were compared upon culturing the primary cells in two different media; Amniomax or RPMI. Cell viability study was carried out using trypan blue staining, while the growth rate was monitored based on the time required to passage the cells (population doubling time in hour). The heterogeneity of cells was examined based on the morphology of the cells. Statistical analysis was performed using t-test. Results: Amniomax was observed to provide a better culture condition in culturing rat AF cells as the cells are more viable, grow faster and more heterogenous as compared to the cells grown in RPMI. Conclusion: Amniomax is a more suitable medium for high quality and viability of full term rat AF cell culture, as compared to RPMI. Thus, warranting propagation of more rat AF cells for biomedical research

Preparation, antioxidant potential and angiotensin converting enzyme (ACE) inhibitory activity of gum Arabic-stabilised magnesium orotate nanoparticles

The present work investigated the antioxidant properties and antihypertensive activity of magnesium orotate (MgOr) using various established in vitro assays, such as β-carotene bleaching activity, 1,1-diphenyl-2-picrylhydrazyl (DPPH), and nitric oxide scavenging activity as well as angiotensin converting enzyme (ACE) inhibitory activity. Magnesium orotate nanoparticles (MgOrGANPs) were prepared using the gum arabic (GA) as stabiliser coatings for nanoparticles through freeze-drying method. The in vitro cytoxicity of MgOrGANPs against human breast cancer MCF7, liver cancer HepG2, and colon cancer HT29 was investigated. The nitric oxide (NO) and DPPH scavenging assays of MgOrGANPs showed a dose-dependent trend, while 500 and 200 μL/mL were significantly more effective than the other concentrations with an IC50 of 89.56 μg/mL and 63.22% DPPH scavenging capacity respectively. The exposure of human cancer cells to MgOrGANPs at 1.56 – 1,000 μg/mL using 3-)4,5-dimethylthiazol-2-yl(2,5-diphenyl tetrazolium bromide (MTT) inhibited the growth of cell lines examined in a dose-dependent manner. Hence, MgOrGANPs may have great potential to be applied for cancer treatments

Differentially expressed proteins in ER+ MCF7 and ER- MDA- MB-231 human breast cancer cells by RhoGDI-α silencing and overexpression

Background: The consequence of Rho GDP dissociation inhibitor alpha (RhoGDIα) activity on migration and invasion of estrogen receptor positive (ER+) and negative (ER-) breast cancer cells has not been studied using the proteomic approach. Changes in expression of RhoGDIα and other proteins interacting directly or indirectly with RhoGDIα in MCF7 and MDA-MB-231, with different metastatic potentials is of particular interest. Materials and Methods: ER+ MCF7 and ER- MDA-MB-231 cell lines were subjected to two-dimensional electrophoresis (2-DE) and spots of interest were identified by matrix-assisted laser desorption/ionization time of- flight/timeof- flight (MALDI-TOF/TOF) mass spectrometry (MS) analysis after downregulation of RhoGDIα using short interfering RNA (siRNA) and upregulated using GFP-tagged ORF clone of RhoGDIα. Results: The results showed a total of 35 proteins that were either up- or down-regulated in these cells. Here we identifed 9 and 15 proteins differentially expressed with silencing of RhoGDIα in MCF-7 and the MDA-MB-231 cells, respectively. In addition, 10 proteins were differentially expressed in the upregulation of RhoGDIα in MCF7, while only one protein was identified in the upregulation of RhoGDIα in MDA-MB-231. Based on the biological functions of these proteins, the results revealed that proteins involved in cell migration are more strongly altered with RhoGDI-α activity. Although several of these proteins have been previously indicated in tumorigenesis and invasiveness of breast cancer cells, some ohave not been previously reported to be involved in breast cancer migration. Hence, these proteins may serve as useful candidate biomarkers for tumorigenesis and invasiveness of breast cancer cells. Conclusions: Future studies are needed to determine the mechanisms by which these proteins regulate cell migration. The combination of RhoGDIα with other potential biomarkers may be a more promising approach in the inhibition of breast cancer cell migration