HPV16-E7 Protein T Cell Epitope Prediction and Global Therapeutic Peptide Vaccine Design Based on Human Leukocyte Antigen Frequency: An In-Silico Study

International Journal of Peptide Research and Therapeutics - Cervical cancer is the second most common leading cause of women's death due to cancer worldwide, about 528,000 patients’...     

Tank bioleaching of copper from combined flotation concentrate and smelter dust

Bioleaching studies were conducted to evaluate the extraction of copper from combined flotation concentrate and smelter dust using two different dust-addition strategies, non-fractional (simultaneously with concentrate) and fractional (step-wise dust addition) in stirred tank reactors. The highest copper extraction obtained with the fractional dust addition was up to 94% while extraction using the non-fractional method leveled off at approximately 44%. Thus, an increased copper recovery of about 50% was achieved with the fractional dust-addition method. It seems that the main reason for increased copper recovery using fractional dust addition could be the control and prevention of chalcopyrite passivation (according to SEM/EDS, XRD, and quantitative mineralogy data). This leads to its improved dissolution.     

Targeting a newly established spontaneous feline fibrosarcoma cell line by gene transfer

Fibrosarcoma is a deadly disease in cats and is significantly more often located at classical vaccine injections sites. More rare forms of spontaneous non-vaccination site (NSV) fibrosarcomas have been described and have been found associated to genetic alterations. Purpose of this study was to compare the efficacy of adenoviral gene transfer in NVS fibrosarcoma. We isolated and characterized a NVS fibrosarcoma cell line (Cocca-6A) from a spontaneous fibrosarcoma that occurred in a domestic calico cat. The feline cells were karyotyped and their chromosome number was counted using a Giemsa staining. Adenoviral gene transfer was verified by western blot analysis. Flow cytometry assay and Annexin-V were used to study cell-cycle changes and cell death of transduced cells. Cocca-6A fibrosarcoma cells were morphologically and cytogenetically characterized. Giemsa block staining of metaphase spreads of the Cocca-6A cells showed deletion of one of the E1 chromosomes, where feline p53 maps. Semi-quantitative PCR demonstrated reduction of p53 genomic DNA in the Cocca-6A cells. Adenoviral gene transfer determined a remarkable effect on the viability and growth of the Cocca-6A cells following single transduction with adenoviruses carrying Mda-7/IL-24 or IFN-γ or various combination of RB/p105, Ras-DN, IFN-γ, and Mda-7 gene transfer. Therapy for feline fibrosarcomas is often insufficient for long lasting tumor eradication. More gene transfer studies should be conducted in order to understand if these viral vectors could be applicable regardless the origin (spontaneous vs. vaccine induced) of feline fibrosarcomas.     

<Emphasis Type="Italic">Salicola mahdashtensis</Emphasis> sp. nov., an extremely halophilic bacterium isolated from Mahdasht saline spring in Iran

A novel extremely halophilic bacterium, designated strain R1^T, was isolated from saline spring bed soil collected from Mahdasht in Alborz province, Iran. Strain R1^T is gram negative, motile, reddish orange pigmented, rod shape, does not form endospores, facultative anaerobe required at least 17.5% salt and 20% total salinity for growth. Optimal growth was occurred in 20% salt and growth observed in salt more than 30%. pH ranges between 5.5 to 8.5, optimum pH for growth is 6.5. Cellular fatty acids are C_10:0, C_12:0, C_12:03-OH, C_16:0 Nalcohol, C_16:0, C_18:3ω6c (6, 9, 12) and C_18:1ω9c. Phylogenetic analysis of 16S rRNA gene sequence showed a close proximity to Salicola salis (99.2%) and Salicola marasensis (99.0%) in the Gammaproteobacteria. The G + C content of type strain was 61.3 mol %. DNA?DNA hybridization indicated that the level of relatedness to Salicola salis was 65.1% and that to Salicola marasensis was 63.5%. Further differences were apparent in antibiotic resistance, oxidase activity, nitrate reduction, hydrolysis of pectin and growth on citrate medium, utilize glucose and lactose. Based on the phenotypic and genotypic data presented, strain R1^T should be the type strain of a new species of genus Salicola which has been named Salicola mahdashtensis. The type strain is R1^T (KCTC 32441, MTCC 11814).     

Thermal coefficients of the methyl groups within ubiquitin

Physiological processes such as protein folding and molecular recognition are intricately linked to their dynamic signature, which is reflected in their thermal coefficient. In addition, the local conformational entropy is directly related to the degrees of freedom, which each residue possesses within its conformational space. Therefore, the temperature dependence of the local conformational entropy may provide insight into understanding how local dynamics may affect the stability of proteins. Here, we analyze the temperature dependence of internal methyl group dynamics derived from the cross-correlated relaxation between dipolar couplings of two CH bonds within ubiquitin. Spanning a temperature range from 275 to 308 K, internal methyl group dynamics tend to increase with increasing temperature, which translates to a general increase in local conformational entropy. With this data measured over multiple temperatures, the thermal coefficient of the methyl group order parameter, the characteristic thermal coefficient, and the local heat capacity were obtained. By analyzing the distribution of methyl group thermal coefficients within ubiquitin, we found that the N-terminal region has relatively high thermostability. These results indicate that methyl groups contribute quite appreciably to the total heat capacity of ubiquitin through the regulation of local conformational entropy.     

New procedure for epidermal cell isolation using kiwi fruit actinidin,and improved culture of melanocytes in the presence of leukaemia inhibitory factor and forskolin

Objectives

Conventional isolation of epidermis from the dermis and disruption of epidermal sheets to liberate the cells, are performed using proteolytic enzymes such as thermolysin or collagenase. Selective population expansion of melanocytes is achieved by suppressing proliferation of keratinocytes and fibroblasts in epidermal cell suspensions, using phorbol esters and cholera toxin. Here, we introduce a new procedure for isolation of epidermal cells, using proteolytic activity of kiwi fruit actinidin, and also an improved growth medium for melanocytes in the presence of leukaemia inhibitory factor (LIF) and forskolin.

Materials and methods

Dermo‐epidermal separation and epidermal sheet cell dispersion were performed using actinidin compared to conventional proteases including collagenase, thermolysin or trypsin. Thereafter, melanocyte culture was performed in two common media and one modified medium to discover optimization for these cells.

Results

We found that dermo‐epidermal separation and epidermal sheet cell dispersion using kiwi fruit actinidin were considerably better than previously used methods, both from the aspect of less fibroblast and keratinocyte contamination, and of more viable native cells. Also, melanocytes proliferated better in phorbol ester‐ and cholera toxin‐free proliferation medium supplemented with LIF and forskolin.

Conclusion

Less contamination and higher numbers of viable cells were actinidin preferential for separation of epidermis and isolation of epidermal cells. Supplementation of LIF and forskolin to new medium increased proliferation potential of melanocytes in comparison to exogenous mitogens.

     

Oxidative aromatization of Hantzsch 1,4-dihydropyridines in the presence of mixed-addenda vanadomolybdophosphate heteropolyacid, H6PMo9V3O40

A variety of Hantzsch 1,4-dihydropyridines were oxidized to the corresponding pyridines in high yields in the presence of H(6)PMo(9)V(3)O(40), a Keggin type heteropolyacid, in refluxing acetic acid. The heteropolyacid was found to be reusable.     

Simple and Effective Purification of Recombinant Peptide Drug,hPTH (1-34), Expressed in E. coli Host

International Journal of Peptide Research and Therapeutics - Several human diseases arise from abnormality in functional proteins production in different tissues. Recombinant protein drugs produced...