Characteristic alterations of gut microbiota in uncontrolled gout

ul-Haq Asad,Lee Kyung-Ann,Seo Hoonhee,Kim Sukyung,Jo Sujin,고경민,Moon Su-Jin,Kim Yun Sung,Choi Jung Ran,Song Ho-Yeon,Kim Hyun-Sook 한국미생물학회 2022 The journal of microbiology Vol.60 No.12

Microbiome research has been on the rise recently for a more in-depth understanding of gout. Meanwhile, there is a need to understand the gut microbiome related to uric acid-lowering drug resistance. In this study, 16S rRNA gene-based microbiota analysis was performed for a total of 65 stool samples from 17 healthy controls and 48 febuxostat-treated gout patients (including 28 controlled subjects with decreased uric acid levels and 20 uncontrolled subjects with non-reduced uric acid levels). Alpha diversity of bacterial community decreased in the healthy control, controlled, and uncontrolled groups. In the case of beta diversity, the bacterial community was significantly different among groups (healthy control, controlled, and uncontrolled groups). Taxonomic biomarker analysis revealed the increased population of g-Bifidobacterium in healthy controls and g-Prevotella in uncontrolled patients. PCR further confirmed this result at the species level. Additionally, functional metagenomics predictions led to the exploration of various functional biomarkers, including purine metabolism. The results of this study can serve as a basis for developing potential new strategies for diagnosing and treating gout from microbiome prospects.

Isolation of MLL1 Inhibitory RNA Aptamers

Asad Ul-Haq,Ming Li Jin,정광원,김환묵,전광훈 한국응용약물학회 2019 Biomolecules & Therapeutics(구 응용약물학회지) Vol.27 No.2

Mixed lineage leukemia proteins (MLL) are the key histone lysine methyltransferases that regulate expression of diverse genes. Aberrant activation of MLL promotes leukemia as well as solid tumors in humans, highlighting the urgent need for the development of an MLL inhibitor. We screened and isolated MLL1-binding ssRNAs using SELEX (Systemic Evolution of Ligands by Exponential enrichment) technology. When sequences in sub-libraries were obtained using next-generation sequencing (NGS), the most enriched aptamers—APT1 and APT2—represented about 30% and 26% of sub-library populations, respectively. Motif analysis of the top 50 sequences provided a highly conserved sequence: 5΄-A[A/C][C/G][G/U][U/A]ACAGAGGG[U/A]GG[A/C] GAGUGGGU-3΄. APT1, APT2, and APT5 embracing this motif generated secondary structures with similar topological characteristics. We found that APT1 and APT2 have a good binding activity and the analysis using mutated aptamer variants showed that the site information in the central region was critical for binding. In vitro enzyme activity assay showed that APT1 and APT2 had MLL1 inhibitory activity. Three-dimensional structure prediction of APT1-MLL1 complex indicates multiple weak interactions formed between MLL1 SET domain and APT1. Our study confirmed that NGS-assisted SELEX is an efficient tool for aptamer screening and that aptamers could be useful in diagnosis and treatment of MLL1-mediated diseases.

Bioloistic-mediated Transformation of Cotton (Gossypium hirsutum L.): Embryogenic Calli as Explant

Haq Ikram-ul,Asad Shaheen,Zafar Yusuf The Korean Society of Plant Biotechnology 2005 Plant molecular biology and biotechnology research Vol.7 No.4

Genetic transformation was carried out by using biolistic gun method. The hypocotyl derived embryogenic calli (explants) of cotton (Gossypium hirsutum L.) cv. Cocker-312 were transformed with a recombinant pGreen II plasmid, in which both, bar (selection marker) and GUS (${\beta}$-glucuronidase) reporter genes were incorporated. Explants were arranged on osmoticum-containing medium (0.5M mannitol) 4 hours prior to and 16 hours after bombardment that was resulted into an increase about >80% for GUS stable expression. 3 days after bombardment, GUS assay was performed, which exhibited, $18.36{\pm}1.00$ calli showed blue spots. The transformed embryogenic calli were cultured on selection medium (@ 6 mg/L basta) for 3 months. The putative transgenic plants were developed via selective somatic embryogenesis (@1.50 mg/L basta); maximum $27.58{\pm}1.25$ somatic embryos were obtained while $17.47{\pm}1.00$ embryos developed into plantlets (@ 0.75mg/L basta). In five independent experiments, up to 7.24% transformation efficiency was recorded. The presence of the transgenes was analyzed by using PCR and southern hybridization analysis. The transgenic plants were developed with in 6-7 months, but mostly transformants were abnormal in morphology.

Human Organic Anion Transporting Polypeptide 1B3 Applied as an MRI-Based Reporter Gene

Baek Song-Ee,Ul-Haq Asad,Kim Dae Hee,Choi Hyoung Wook,김명진,최혜진,Kim Honsoul 대한영상의학회 2020 Korean Journal of Radiology Vol.21 No.6

Objective: Recent innovations in biology are boosting gene and cell therapy, but monitoring the response to these treatments is difficult. The purpose of this study was to find an MRI-reporter gene that can be used to monitor gene or cell therapy and that can be delivered without a viral vector, as viral vector delivery methods can result in long-term complications. Materials and Methods: CMV promoter-human organic anion transporting polypeptide 1B3 (CMV-hOATP1B3) cDNA or CMV-blank DNA (control) was transfected into HEK293 cells using Lipofectamine. OATP1B3 expression was confirmed by western blotting and confocal microscopy. In vitro cell phantoms were made using transfected HEK293 cells cultured in various concentrations of gadoxetic acid for 24 hours, and images of the phantoms were made with a 9.4T micro-MRI. In vivo xenograft tumors were made by implanting HEK293 cells transfected with CMV-hOATP1B3 (n = 4) or CMV-blank (n = 4) in 8-week-old male nude mice, and MRI was performed before and after intravenous injection of gadoxetic acid (1.2 μL/g). Results: Western blot and confocal microscopy after immunofluorescence staining revealed that only CMV-hOATP1B3-transfected HEK293 cells produced abundant OATP1B3, which localized at the cell membrane. OATP1B3 expression levels remained high through the 25th subculture cycle, but decreased substantially by the 50th subculture cycle. MRI of cell phantoms showed that only the CMV-hOATP1B3-transfected cells produced a significant contrast enhancement effect. In vivo MRI of xenograft tumors revealed that only CMV-hOATP1B3-transfected HEK293 tumors demonstrated a T1 contrast effect, which lasted for at least 5 hours. Conclusion: The human endogenous OATP1B3 gene can be non-virally delivered into cells to induce transient OATP1B3 expression, leading to gadoxetic acid-mediated enhancement on MRI. These results indicate that hOATP1B3 can serve as an MRI-reporter gene while minimizing the risk of long-term complications.

7Effects on Thermal and Ablative Properties of Phenolic Resin (Novolac) Blended Acrylonitrile Butadiene Rubber

Rashid Nawaz,Naghmana Rashid,Zulfiqar Ali,Asad U. Khan,M. Shahid Nazir,Noaman Ul-Haq 한국섬유공학회 2018 Fibers and polymers Vol.19 No.6

In this work we investigated the ablative response and thermal properties of phenolic resin (PR) blended acrylonitrile butadiene rubber (NBR) composites. PR was added to NBR in the proportion of 0, 5, 10, 20, 30, 40 and 50 phr by means of two-roll laboratory mill. PR remarkably improved ablation resistance and thermal properties of NBR/PR composite. The linear and mass ablation rates reduced to 21.3 % and 26.1 % respectively. The char content deposition increased from 0.19 to 26.8 %. Char layer produced by PR, obviously reduced the erosion rate of the NBR/PR composite relative to neat NBR (without PR). Detailed morphological studies of the composite and post-test (ablation) microstructure of char revealed that higher loading of PR in the rubber composite produced dense char layer firmly intact to the substrate. Furthermore, thermal stability of the composite improved by 22-23 ºC, however, thermal conductivity of the composite slightly increased by 0.115 W/mK for 50 Phr of PR loading as compared to the neat.