A novel diblock copolymer of (monomethoxy poly [ethylene glycol]-oleate) with a small hydrophobic fraction to make stable micelles/polymersomes for curcumin delivery to cancer cells

Curcumin is a potent natural anticancer agent, but its effectiveness is limited by properties such as very low solubility, high rate of degradation, and low rate of absorption of its hydrophobic molecules in vivo. To date, various nanocarriers have been used to improve the bioavailability of this hydrophobic biomaterial. This study investigates the encapsulation of curcumin in a novel nanostructure of monomethoxy poly(ethylene glycol)-oleate (mPEG-OA) and its anticancer effect. Tests were done to determine the critical micelle concentration (CMC), encapsulation efficiency, drug-loading efficiency, and cytotoxicity (against U87MG brain carcinoma cells and HFSF-PI3 cells as normal human fibroblasts) of some nanodevice preparations. The results of fluorescence microscopy and cell-cycle analyses indicated that the in vitro bioavailability of the encapsulated curcumin was significantly greater than that of free curcumin. Cytotoxicity evaluations showed that half maximal inhibitory concentrations of free curcumin and curcumin-loaded mPEG-OA for the U87MG cancer cell line were 48 μM and 24 μM, respectively. The Annexin-V-FLUOS assay was used to quantify the apoptotic effect of the prepared nanostructures. Apoptosis induction was observed in a dose-dependent manner after curcumin-loaded mPEG-OA treatments. Two common self-assembling structures, micelles and polymersomes, were observed by atomic force microscopy and dynamic light scat­tering, and the abundance of each structure was dependent on the concentration of the diblock copolymer. The mPEG-OA micelles had a very low CMC (13.24 μM or 0.03 g/L). Moreover, atomic force microscopy and dynamic light scattering showed that the curcumin-loaded mPEG-OA polymersomes had very stable structures, and at concentrations 1,000 times less than the CMC, at which the micelles disappear, polymersomes were the dominant structures in the dispersion with a reduced size distribution below 150 nm. Overall, the results from these tests revealed that this nanocarrier can be considered as an appropriate drug delivery system for delivering curcumin to cancer cells. © 2014 Erfani-Moghadam et al

Inheritance of RAPD marker in female grass carp (Ctenopharyngodon idella), male bighead carp (Hypophthalmichthys nobilis) and their F1 hybrids

Genetic variation and inheritance of randomly amplified polymorphic DNA (RAPD) markers in female grass carp (Ctenopharyngodon idella) and male bighead carp (Hypophthalmichthys nobilis) and their F1 offspring hybrid have been studied. For this purpose, genomic DNA was extracted from muscle tissue according to phenol-chloroform method, and six decamer primers were used for amplifying polymorphic DNA. Results from 77 produced bands showed that all RAPD bands pattern of parents were present in F1 offspring hybrid, which indicated the high influence and dominance of the mentioned markers. The low difference in polymorphisms in used markers between F1 offspring hybrid and parents shows that parents are heterozygous in some loci, which can cause low difference in hybrids of two distinct genera. Although our results revealed that RAPD markers had a suitable efficiency in distinguishing parents from F1 hybrids, genetic diversity of triploid and diploid F1 hybrid were not detected by introduced primers

Effects of incubation and larval rearing temperature on histological changes of muscles in Caspian trout (Salmo trutta caspius Kessler, 1877) larvae

Out of considerable environmental factors, temperature and its possible effects on life stages of Caspian trout investigated by natural incubation (8 ͦC) condition simulation comparing with cold (4 ͦC) and warm (12 ͦC) constant incubation temperatures in 3 well equipped incubators by water recycling systems. Green eggs triple treatments of wild and F1 cultured brooders were incubated. Incubation implemented in dark by using REDD water and DO–pH – temperature digital monitoring ended to yolk sac absorption and entering larval stage. Numbers and diameters of white fiber muscles measured and significant differences considered between three thermal treatments (P<0.05) in both wild and cultured stocks. The numbers of white fiber muscles in warm treatment by highest means (72.54) and lowest diameter (8.46 micrometer) compared with Cold treatment white fiber muscles diameter (20.59 micrometer) and numbers (50.72) which were the highest diameter and lowest numbers means between treatments. Hatching success stated considerable mortality for cold treatments and 8°C incubator improved the best temperature in wild treatment. Incubation temperature induced significant effect on white fiber muscles stated considerable index for flesh precursor muscles condition which is subject of natural stocks rehabilitation and domestication projects considerations

Association of fibroblast growth factor (FGF-21) as a screening biomarker for chronic progressive external ophthalmoplesia

Purpose: To investigate whether or not fibroblast growth factor (FGF-21) can be used as a screening biomarker in chronic progressive external ophthalmoplesia (CPEO) patients. Methods: FGF-21 concentration was measured in the serum of 24 patients with CEPO phenotype and 24 control samples by enzyme-linked immunosorbent assay (ELISA) and determined the deletion of mitochondrial genome by multiplex polymerase chain reaction (PCR). Results: FGF-21 concentration in 50% of CPEO patients showed notable differences from that in control subjects. FGF-21 concentration ratio in patient group, 2 disorder control groups (mitochondrial and non-mitochondrial) and normal group, respectively, was 294.87 ± 42.10 (p 50 years age group who show acute symptoms. © Pharmacotherapy Group, Faculty of Pharmacy, University of Benin, Benin City, 300001 Nigeria. All rights reserved

Dengue viruses and promising envelope protein domain III-based vaccines

Abstract Dengue viruses are emerging mosquito-borne pathogens belonging to Flaviviridae family which are transmitted to humans via the bites of infected mosquitoes Aedes aegypti and Aedes albopictus. Because of the wide distribution of these mosquito vectors, more than 2.5 billion people are approximately at risk of dengue infection. Dengue viruses cause dengue fever and severe life-threatening illnesses as well as dengue hemorrhagic fever and dengue shock syndrome. All four serotypes of dengue virus can cause dengue diseases, but the manifestations are nearly different depending on type of the virus in consequent infections. Infection by any serotype creates life-long immunity against the corresponding serotype and temporary immunity to the others. This transient immunity declines after a while (6 months to 2 years) and is not protective against other serotypes, even may enhance the severity of a secondary heterotypic infection with a different serotype through a phenomenon known as antibody-depended enhancement (ADE). Although, it can be one of the possible explanations for more severe dengue diseases in individuals infected with a different serotype after primary infection. The envelope protein (E protein) of dengue virus is responsible for a wide range of biological activities, including binding to host cell receptors and fusion to and entry into host cells. The E protein, and especially its domain III (EDIII), stimulates host immunity responses by inducing protective and neutralizing antibodies. Therefore, the dengue E protein is an important antigen for vaccine development and diagnostic purposes. Here, we have provided a comprehensive review of dengue disease, vaccine design challenges, and various approaches in dengue vaccine development with emphasizing on newly developed envelope domain III-based dengue vaccine candidates. Keywords: Dengue virus Envelope protein Chimeric vaccine Disease Immunogenicit

Molecular study of the populations of Artemia partenogenetica in Iran using PCR-RFLP Method

Considering the importance of genetic studies to manifest inter population differences in species, samples of Artemia partenogenetica were collected from seven inland lakes including Shoor and Inche-Borun lakes in Golestan Province, Hoze-Soltan and Namak lakes in Qom Province, Maharloo and Bakhteghan lakes in Fars Province and Mighan pool in Markazi Province. A total of 210 samples were subjected to DNA extraction by phenol-chloroform method. Primers were designed on a ribosomal fragment (16SrRNA) of the species' mtDNA sequence and the PCR was conducted on the samples. Digestion of the PCR product with approximately 1584bp lengths by 10 restriction endonuclease (AluI, EcoRI, Eco47I, HaeIII, HindIII, HinfI, MboI, MspI, RsaI, TaqI) showed 12 different haplotypes: 4 haplotypes in Shoor and Inche-Borun, 1 in Namak and Hoze-Soltan, 3 in Mighan pool, 1 in Bakhtegan and Maharloo and 3 in Maharloo. Haplotype diversity values within collected samples varied from zero in Hoze-Soltan, Namak and Bakhteghan samples to 0.7425 in Inche-Borun and Shoor while nucleotide diversity varied from zero in Hoze-Soltan, Namak and Bakhteghan, to 0.0077 in Mighan. The minimum nucleotide diversity among samples was zero between Hoze-Soltan vs. Namak and the maximum was 0.1700 between Inche-Borun and Shoor vs. Mighan. Nucleotide divergences among samples were least in Inche-Borun vs. Shoor (%-0.02) and most in Inche-Borun and Shoor vs. Mighan (%16.18), averaging to %3.40. The evolutionary distances between 12 haplotype showed that the maximum value belonged to Mighan haplotypes vs. Inche-Borun and Shoor haplotypes. Regarding the digestive patterns produced by each enzyme in the studied region, Eco47I is introduced as the population-specific marker of A. partenogenetica in Iran. Test of population differentiation based on haplotype frequencies were statistically significant (P≤0.001) with the exception of Hoze-Soltan vs. Namak and Inche-Borun vs. Shoor. We conclude that there are enough evidences in haplotypic level for dividing A. partenogenetica in Iran into five populations: Hoze-Soltan and Namak, Mighan, Maharloo, Bakhtegan, Incheh-Borun and Shoor

Investigation of solvent effect and NMR shielding tensors of p53 tumor-suppressor gene in drug design

The p53 tumor-suppressor gene encodes a nuclear phosphoprotein with cancer- inhibiting properties. The most probable cancerous mutations occur as point mutations in exons 5 up to 8 of p53, as a base pair substitution that encompasses CUA and GAT sequences. As DNA drug design represents a direct genetic treatment of cancer, in the research reported computational drug design was carried out to explore, at the Hartree–Fock level, effects of solvents on the thermochemical properties and nuclear magnetic resonance (NMR) shielding tensors of some atoms of CUA involved in the hydrogen-bonding network. The observed NMR shielding variations of the solutes caused by solvent change seemed significant and were attributed to solvent polarity, and solute–solvent and solvent–solute hydrogen-bonding interactions. The results provide a reliable insight into the nature of mutation processes. However, to improve our knowledge of the hydration pattern more rigorous computations of the hydrated complexes are needed

Nanodiagnostic method for colorimetric detection of Mycobacterium tuberculosis 16S rRNA

A nanodiagnostic method using nucleic acid sequence-based amplification (NASBA) and gold nanoparticle probes (AuNP probes) was developed for colorimetric detection of Mycobacterium tuberculosis. The primers targeting 16S rRNA were used for the amplification of mycobacterial RNA by the isothermal NASBA process. The amplicons were hybridized with specific gold nanoparticle probes. The RNA-DNA hybrids were colorimetrically detected by the accumulation of gold nanoparticles. Using this method, 10 CFU ml-1 of M. tuberculosis was detected within less than 1 h. Results obtained from the clinical specimens showed 94.7% and 96% sensitivity and specificity, respectively. No interference was encountered in the amplification and detection of M. tuberculosis in the presence of non-target bacteria, confirming the specificity of the method. © 2009 Humana Press Inc

Detection of icaAD gene and biofilm formation in Staphylococcus aureus isolates from wound infections

Wound infections are a common cause of staphylococcal infections. An ability of S.aureus is to adhere and form biofilm on host surfaces. Biofilm is an exopolysaccharide, a slime matrix around multiple layers of cells and is mediated by expression of the icaADBC operon. The present study evaluated the biofilm forming capacity and the presence of icaAD gene among S.aureus isolated from wound infections. Slime production assay was performed by cultivation on Congo Red Agar plate. In addition, Quantitative biofilm formation determined by microtiter plate assay PCR method used for detection of icaAD gene. Fifty strains were identified, 54 of the isolates produced black colonies on CRA plate, 52 were positive biofilm forming, and all strains carried the icaAD gene. Regarding the ability of Saureus to form biofifms helps the bacterium to survive hostile environments within the host, suggests that biofilm production is a risk factor for infection. It is important in rapid diagnosis and treatment biofilm forming strains, because biofilm formation may lead to increased antimicrobial resistance and create a significant impediment to wound healing