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Correlations between abnormalities of morphological details and DNA fragmentation in human sperm
Nguyen, Hiep Tuyet Thi,Dang, Hong Nhan Thi,Nguyen, Thai Thanh Thi,Nguyen, Trung Van,Dang, Thuan Cong,Nguyen, Quoc Huy Vu,Le, Minh Tam The Korean Society for Reproductive Medicine 2022 Clinical and Experimental Reproductive Medicine Vol.49 No.1
Objective: As the associations of sperm DNA fragmentation with morphology have not been examined in detail, this study aimed to investigate the relationship between abnormalities of morphological details and DNA integrity in human sperm. Methods: In this cross-sectional study, men from infertile couples were enrolled at Hue Center for Reproductive Endocrinology and Infertility, Vietnam. Conventional semen parameters, including morphological details, were analyzed following the World Health Organization 2010 criteria. Sperm DNA fragmentation was evaluated using a sperm chromatin dispersion assay. The relationships and correlations between semen parameters, sperm morphology, and the type of halosperm and the DNA fragmentation index (DFI) were analyzed. Results: Among 130 men in infertile couples, statistically significant differences were not found in the sperm halo type between the normal and abnormal sperm morphology groups. The percentage of round-head spermatozoa was higher in the DFI >15% group (16.98%±12.50%) than in the DFI ≤15% group (13.13% ±8.82%), higher values for amorphous heads were found in the DFI >15% group, and lower values for tapered heads were observed in the DFI ≤15% group; however, these differences were not statistically significant. Small-halo sperm and the DFI were positively correlated with round-head sperm (r=0.243, p=0.005 and r=0.197, p=0.025, respectively). Conclusion: The rate of general sperm morphological abnormalities in semen analysis was not related to sperm DNA integrity. However, round sperm heads were closely associated with sperm DNA fragmentation.
Flexible activity of glycosyltransferase UGT88D7 from Lamiales
Nguyen Huy Thuan,Le Thi Tuoi,Jae Won Park,Jae Kyung Sohng 한국당과학회 2012 한국당과학회 학술대회 Vol.2012 No.1
TDP-L-rhamnose–producing E. coli host containing Lamiales-derived UGT88D7 glycosyltransferase was employed to generate flavone rhamnosides and glucosides from apigenin, baicalein and luteolin. In comparison with the published literatures, this gene has showed markely flexible activity because it is able to transfer glucose, glucoronic acid and rhamnose moieties to aglycones. Although apigenin was manifested as the most favourite substrate baicalein and luteolin seemed to be good candidates to yield glycosylated products. The productivities of bioconversion of apigenin, baicalein and luteolin were 75.4, 36.1 and 32.4%, respectively, in the presence of methylated cyclodextrin as mediator. The formation of product was confirmed by TLC, HPLC and ESI-MS/MS measurement. The structural identification of products is being continued by NMR method. Data revealed the use of NDP-sugars flexible glycosyltransferase as an effective tool to produce various kinds of flavonoid glycosides by combinatorial biosynthesis in E. coli.
Protein engineering for glycosylation of flavonoid in Escherichia coli by in-vivo
Nguyen Huy Thuan,Ju Yong Shin,Joo-Ho Lee,Jae Kyung Sohng 한국당과학회 2013 한국당과학회 학술대회 Vol.2013 No.1
Glycodiversification and glycorandomization, invaluable tools for generating biochemical diversity, can be catalyzed by flexible glycosyltransferases (GTs), which attach various activated sugar “donors” onto different type of “acceptor” molecules. Among discovered and successfully developed protein engineering techniques, creation of hybrid protein have been evaluated as very powerful approach to produce enzyme with new activity. Theoretically, hybrid protein was built up by different fragments from various parent molecules. Particularly, engineered GTs have been used as wonderful catalysts for making new glycosylated compound, for example, the generation of new hybrid glycopeptides or glycosylation of scopoletin by GT from Arabidopsisthalianaorproducing novel urdamycin derivatives, etc. Applying protein modeling, we have constructed a hybrid protein HT1 that is composed of DnrS-derived N-terminal and DesVII-originated C-terminal. The enzyme activity was tested using various activated sugar produced Escherichia coli system (such as TDP-rhamnose, UDP-xylose and UDP-6-deoxy-allose) in combination with the transformation of phenolic substrate (flavonoid, stilbene, quinone, etc). The initial result showed that hybrid protein significant change in regio-specific activity in comparison with parent molecule (DnrSandDesVII), particularly, enabling produce flavonol and flavanone-O-rhamnoside such as quercetin, kaempferol, myricetin, andnaringenin-O-rhamnoside. The experiment of glyco-diversification is continuously carrying out to determine the level of enzyme’s promiscuity and which type of the most favorite substrate was recruited. Therefore, the chimeric protein HT1 is apromising tool to divert the natural products. Furthermore it is possible to improve or alter enzyme activity by domain swapping from far related glycosyltransferases.
Microbial production of astilbin, a bioactive rhamnosylated flavanonol, from taxifolin
Thuan, Nguyen Huy,Malla, Sailesh,Trung, Nguyen Thanh,Dhakal, Dipesh,Pokhrel, Anaya Raj,Chu, Luan Luong,Sohng, Jae Kyung Rapid Communications of Oxford Ltd in association 2017 World journal of microbiology biotechnology Vol.33 No.2
<P>Flavonoids are plant-based polyphenolic biomolecules with a wide range of biological activities. Glycosylated flavonoids have drawn special attention in the industries as it improves solubility, stability, and bioactivity. Herein, we report the production of astilbin (ATN) from taxifolin (TFN) in genetically-engineered Escherichia coli BL21(DE3). The exogenously supplied TFN was converted to ATN by 3-O-rhamnosylation utilizing the endogeneous TDP-l-rhamnose in presence of UDP-glycosyltransferase (ArGT3, Gene Bank accession number: At1g30530) from Arabidopsis thaliana. Upon improving the intracellular TDP-l-rhamnose pool by knocking out the chromosomal glucose phosphate isomerase (pgi) and d-glucose-6-phosphate dehydrogenase ( zwf) deletion along with the overexpression of rhamnose biosynthetic pathway increases the biotransformation product, ATN with total conversion of similar to 49.5 +/- 1.67% from 100 mu M of taxifolin. In addition, the cytotoxic effect of taxifolin-3-O-rhamnoside on PANC-1 and A-549 cancer cell lines was assessed for establishing ATN as potent antitumor compound.</P>
A Phosphate Starvation-Inducible Ribonuclease of Bacillus licheniformis
( Thanh Trung Nguyen ),( Minh Hung Nguyen ),( Huy Thuan Nguyen ),( Hoang Anh Nguyen ),( Thi Hoi Le ),( Thomas Schweder ),( Britta Jurgen ) 한국미생물 · 생명공학회 2016 Journal of microbiology and biotechnology Vol.26 No.8
The BLi03719 protein of Bacillus licheniformis DSM13 belongs to the most abundant extracellular proteins under phosphate starvation conditions. In this study, the function of this phosphate starvation inducible protein was determined. An amino-acid sequence analysis of the BLi03719-encoding gene showed a high similarity with genes encoding the barnase of Bacillus amyloliquefaciens FZB42 and binase-like RNase of Bacillus pumilus SARF-032. The comparison of the control strain and a BLi03719-deficient strain revealed a strongly reduced extracellular ribonuclease activity of the mutant. Furthermore, this knockout mutant exhibited delayed growth with yeast RNA as an alternative phosphate and carbon source. These results suggest that BLi03719 is an extracellular ribonuclease expressed in B. licheniformis under phosphate starvation conditions. Finally, a BLi03719 mutant showed an advantageous effect on the overexpression of the heterologous amyE gene under phosphate-limited growth conditions.
Minh Tam Le,Hiep Tuyet Thi Nguyen,Trung Van Nguyen,Thai Thanh Thi Nguyen,Hong Nhan Thi Dang,Thuan Cong Dang,Quoc Huy Vu Nguyen The Korean Society for Reproductive Medicine 2023 Clinical and Experimental Reproductive Medicine Vol.50 No.2
Objective: This study aimed to compare the efficacy of physiological intracytoplasmic sperm injection (PICSI) and intracytoplasmic sperm injection (ICSI) in terms of the fertilization rate and embryo quality using sibling oocyte cycles. Methods: This prospective, cross-sectional study collected data from 76 couples who underwent their first cycle at the Hue Center for Reproductive Endocrinology and Infertility, Vietnam, between May 2019 and November 2021. The inclusion criteria were cycles with at least eight oocytes and a sperm concentration of 5×10<sup>6</sup>/mL. Sperm parameters, sperm DNA fragmentation (SDF), fertilization, and the quality of cleavage-stage embryos on day 2 and blastocysts on day 5 were examined. Results: From 76 ICSI cycles, 1,196 metaphase II (MII) oocytes were retrieved, half of which were randomly allocated to either the PICSI (n=592) or ICSI (n=604) treatment group. The results showed no significant difference between the two groups in terms of fertilization (72.80% vs. 75.33%, p=0.32), day 2 cleavage rate (95.13% vs. 96.04%, p=0.51), blastulation rate (52.68% vs. 57.89%), and high-quality blastocyst rate (26.10% vs. 31.13%, p=0.13). However, in cases where SDF was low, 59 cycles consisting of 913 MII oocytes produced a considerably higher blastulation rate with PICSI than with ICSI (50.49% vs. 35.65%, p=0.00). There were no significant differences between the pregnancy outcomes of the PICSI and ICSI embryo groups following embryo transfer. Conclusion: Using variable sperm quality provided no benefit for PICSI versus ICSI in terms of embryo outcomes. When SDF is low, PICSI appears to be able to produce more blastocysts.