Increased expression of YAP1 in prostate cancer correlates with extraprostatic extension

Objective:Yes associated protein 1(YAP1)is a member of the Hippo pathway,acting as a transcriptional coactivator.To elucidate the role of YAP1 and phosphorylated(p)YAP1 in prostate cancer(PCa)tumorigenesis,we investigated their expression in clinical samples of PCa and cell lines.Methods:Fifty-four tumor,adjacent nontumor,and prostate intraepithelial neoplasia(PIN)tissues from patients with PCa after radical prostatectomy were selected from a retrospective cohort and studied using immunohistochemistry(IHC).Protein and m RNA expression levels of YAP1 were evaluated by Western blot analysis and quantitative real-time reverse transcription PCR,respectively,in cancer cell lines.Publicly available gene expression datasets were downloaded to analyze YAP1 m RNA and protein levels in PCa tissue samples.Results:IHC analysis of PCa tissues revealed that YAP1 staining intensities were moderate to weak in the nucleus and cytoplasm of tumor cells,whereas adjacent normal epithelia showed strong staining.We observed that benign prostates were characterized by higher expression levels of both nuclear(P=0.004)and cytosolic(P=0.005)YAP1.p YAP1 staining was weak in the cytoplasm and absent in the nucleus of all the tissues investigated.YAP1 expression was an indicator of extraprostatic extension(EPE).The level of YAP1 was negatively correlated with the level of the androgen receptor(AR)in The Cancer Genome Atlas dataset and Western blot analysis of cell lines.Conclusions:Our study suggested that YAP1 expression is heterogeneous in PCa tissue samples;therefore,YAP1 might play different roles in different aspects of PCa progression.This might involve AR–YAP1 interplay in PCa.

Two novel variants in CYP1B1 gene: a major contributor of autosomal recessive primary congenital glaucoma with allelic heterogeneity in Pakistani patients

AIM: To find the CYP1 B1 mutations associated with primary congenital glaucoma(PCG) in Pakistani consanguineous pedigrees. METHODS: After getting informed consent, 11 consanguineous pedigrees belonging to different ethnic groups were enrolled. Detailed medical history was recorded and pedigrees were drawn. The standard ophthalmological examination was done to characterize the phenotype. Genomic DNA was extracted from 10 mL whole blood and coding exons and exon intron boundaries of CYP1 B1 gene were directly sequenced. Bioinformatics tools were used to model the mutant protein and predict the effect of novel variants on protein structure and function. RESULTS: Sequencing analysis revealed 5 different CYP1 B1 variants in 7 families(7/11; 64%), including two novel variants. A common mutation, p.R390 H was found in four families, whereas p.P437 L was found once in a family. Two novel variants, a homozygous non sense variant p.L13* and a compound heterozygous variant, p.P350 T along with p.V364 M were segregating with PCGin two families. All the patients had the variable onset and severity of the disease. The success rate of early clinical interventions was observed dependent on mutation types and position. Two different haplotypes were associated with frequently found mutation, p.R390 H. CONCLUSION: Identification of novel CYP1 B1 variants reassert the genetic heterogeneity of Pakistani PCG patients. The patients with missense mutations show severe phenotypic presentations and poor vision after surgical interventions as compare to patients with null variants. This may help to better understand the role of CYP1 B1 mutations in the development of PCG and its course of pathogenicity.

Effect of Putrescine on Low-Temperature Acclimation in Chlamydomonas reinhardtii

Putrescine is reported to be necessary for cold acclimation under low-temperature stress.In this study,the effect of low-temperature on some physiological and biochemical parameters has been investigated using the green algae Chlamydomonas reinhardtii.The lipid peroxidation rate,amount of Rubisco protein,activities of antioxidant enzymes and gene expression of polyamine biosynthesis(odc2,and spd1),heat shock proteins(hsp70c,hsp90a,and hsp90c),and PSII repair mechanisms(psba,rep27,and tba1)were determined to understand the low-temperature response.Exogenous putrescine application significantly increased Rubisco protein concentration and catalase enzyme activities under low-temperature stress.Moreover,real-time RT-PCR results and gene expression analysis showed that polyamine metabolism induced gene expression at low-temperatures in the first 24 h.In the same way,the gene expression of heat shock proteins(hsp70c,hsp90a,and hsp90c)decreased under low-temperature treatment for 72 h;however,application of putrescine enhanced the gene expression in the first 24 h.The results obtained indicated that molecular response in the first 24 h could be important for cold acclimation.The psba and tba1 expressions were reduced under low-temperatures depending on the exposure time.In contrast,the exogenous putrescine enhanced the expression level of the psba response to low-temperature at 24 and 72 h.The results obtained in this study indicate that putrescine could play a role in the PS II repair mechanisms under low-temperature stress.

Evaluation of Transgenic <i>Nicotiana tabacum</i>with <i>deh</i>E Gene Using Transposon Based IRAP Markers

In the present study, five genetically modified herbicide tolerant Nicotiana tabacum cv. TAPM24 plants with a constructed vector pCAMBIA1301a carrying dehalogenase E (dehE) gene were compared with three non-transgenic controls using Tto1 retrotransposon specific IRAP markers. dehE gene was originally characterized in Rhizobium sp. and it produced an enzyme which degraded the Dalapon herbicide. IRAP protocol was applied on transgenic and non-transgenic plants to investigate the retrotransposon based genetic variation which may appear during transformation. Polymorphism rates were calculated as 0%-20% from IRAP-PCR products among all plant samples. These results show that transformation of tobacco plant with the dehE gene may cause Tto1 retrotransposon alterations appearing as different band profiles. The findings are expected to contribute to genetic engineering studies to obtain better results and also to understand how transposons contribute to features such as transgenesis. In our knowledge, this is one of the first experimental data of transgenic N. tabacum engineered with dehE gene originated Rhizobium sp. in terms of retrotranposon based variation.

The Application of Ultraviolet Spectrophotometry(UV)on Some Water Mite Species(Acari,Hydrachnidia)

In this study an ultra violet analysis, which is a spectrophotometric method, was applied for the first time as a structural analysis of the group. The following species were collected from Karamik Lake in Afyon Province, Turkey between April and August, 2015, Hydryphantes flexiosus, Hydrodroma despiciens, Georgella helvatica, Eylais infundibulifera, Hygrobates nigromacutlatus and Torrenticola brevirostris. These species were separately analyzed by Ultraviolet Spectrophotometer (US) and the results assessed using graphical readings. The affiliations between the species were read from the graphical curves of each species and then compared in terms of the curves.

Assessment of the Phylogenetic Affiliation Levels of Water Mite(Acari,Hydrachnidia)Species with the Elemental Analysis Method

10 different species of water mites which are Georgella helvetica, Eylais extendens, Hydrachna globose, Hydrachna prosifera, Hydrachna skorikowi, Hydrodroma despiciens, Hydryphantes dispar, Limnesia fulgida, Eylais setosa, Hydryphantes flexuosus were used in this study. The total masses of these species were measured as mg with the use of an elemental analyzer to calculate the percentage of the organic components of their structures. The achieved values were assessed separately for each species and element with the interpolation method. Out of these organic elements, the amount of C with an approximately value of 50% was the highest for all species while the amounts of S which was approximately 1% was determined as the lowest for almost all species. The observed values were discussed in terms of the systematic of water mites.

Improving mechanical and antibacterial properties of PMMA via polyblend electrospinning with silk fibroin and polyethyleneimine towards dental applications

Poly(methylmethacrylate)(PMMA)is a widely used material in dental applications,particularly as denture resins.Due to thermally unstable and wet oral cavity,the implanted PMMA based resins occasionally deform and grow bacterial biofilms at the interface between oral cavity and the biomaterial.Several strategies attempted earlier to improve the bacterial resistance and mechanical performance of PMMA.Poly(ethyleneimine)(PEI)is a hyperbranched cationic polymer shown earlier to improve antibacterial activity of resins but do not improve mechanical properties of the resins alone,while silk fibroin(SF)is a natural biopolymer with unique material properties.In this study,we combined SF and PEI towards development of antibacterial and mechanically superior PMMA based materials towards overcoming its drawbacks.Using polyblend electrospinning to combine SF,PEI and PMMA,we successfully developed intrinsically antibacterial and mechanically reinforced nanofiber mats.We propose that the resulting nanofiber mats have the potential to be incorporated into PMMA based denture resin materials to overcome the problems of patients and improve their quality of life.