Evidence of Tandem Repeat and Extra Thiol-groups Resulted in the Polymeric Formation of Bovine Haptoglobin: A Unique Structure of Hp 2-2 Phenotype

Lai, Yi An,Lai, I Hsiang,Tseng, Chi Feng,Lee, James,Mao, Simon J.T. Korean Society for Biochemistry and Molecular Biol 2007 Journal of biochemistry and molecular biology Vol.40 No.6

Human plasma Hp is classified as 1-1, 2-1, and 2-2. They are inherited from two alleles Hp 1 and Hp 2, but there is only Hp 1 in almost all the animal species. Hp 2-2 molecule is extremely large and heterogeneous associated with the development of inflammatory-related diseases. In this study, we expressed entire bovine Hp in E. coli as a $\alpha\beta$ linear form. Interestingly, the antibodies prepared against this form could recognize the subunit of native Hp. In stead of a complicated column method, the antibody was able to isolate bovine Hp via immunoaffinity and gelfiltration columns. The isolated Hp is polymeric containing two major molecular forms (660 and 730 kDa). Their size and hemoglobin binding complex are significantly larger than that of human Hp 2-2. The amino-acid sequence deducted from the nucleotide sequence is similar to human Hp 2 containing a tandem repeat over the $\alpha$ chain. Thus, the Hp 2 allele is not unique in human. We also found that there is one additional -SH group (Cys-97) in bovine $\alpha$ chain with a total of 8 -SH groups, which may be responsible for the overall polymeric structure that is markedly different from human Hp 2-2. The significance of the finding and its relationship to structural evolution are also discussed.

Unveiling the Complex World of Extracellular Vesicles: Novel Characterization Techniques and Manufacturing Considerations

James J. Lai,John J. Hill,Casey Y. Huang,Gino C. Lee,Karol W. Mai,Maggie Y. Shen,Simon K. Wang 전남대학교 의과학연구소 2024 전남의대학술지 Vol.60 No.1

Extracellular vesicles (EVs) function as potent mediators of intercellular communication for many in vivo processes, contributing to both health and disease related conditions. Given their biological origins and diverse functionality from correspondingly unique “cargo” compositions, both endogenous and modified EVs are garnering attention as promising therapeutic modalities and vehicles for targeted therapeutic delivery applications. Their diversity in composition, however, has revealed a significant need for more comprehensive analytical-based characterization methods, and manufacturing processes that are consistent and scalable. In this review, we explore the dynamic landscape of EV research and development efforts, ranging from novel isolation approaches, to their analytical assessment through novel characterization techniques, and to their production by industrial-scale manufacturing process considerations. Expanding the horizon of these topics to EVs for in-human applications, we underscore the need for stringent development and adherence to Good Manufacturing Practice (GMP) guidelines. Wherein, the intricate interplay of raw materials, production in bioreactors, and isolation practices, along with analytical assessments compliant with the Minimal Information for Studies of Extracellular Vesicles (MISEV) guidelines, in conjunction with reference standard materials, collectively pave the way for standardized and consistent GMP production processes.

Evidence of Tandem Repeat and Extra Thiol-groups Resulted in the Polymeric Formation of Bovine Haptoglobin: A Unique Structure of Hp 2-2 Phenotype

( Yi An Lai ),( I Hsiang Lai ),( Chi Feng Tseng ),( James Lee ),( Simon J. T. Mao ) 생화학분자생물학회 2007 BMB Reports Vol.40 No.6

Recent Advances in Biosensors for Nucleic Acid and Exosome Detection

Zirui Fu,Yicheng Lu,James J. Lai 전남대학교 의과학연구소 2019 전남의대학술지 Vol.55 No.2

Biosensors are analytical devices for biomolecule detection that compromise three essential components: recognition moiety, transducer, and signal processor. The sensor converts biomolecule recognition to detectable signals, which has been applied in diverse fields such as clinical monitoring, in vitro diagnostics, food industry etc. Based on signal transduction mechanisms, biosensors can be categorized into three major types: optical biosensors, electrochemical biosensors, and mass-based biosensors. Recently, the need for faster, more sensitive detection of biomolecules has compeled researchers to develop various sensing techniques. In this review, the basic structure and sensing principles of biosensors are introduced. Additionally, the review discusses multiple recent works about nucleic acid and exosome sensing.

Microbiome diversity in carriers of fluoroquinolone resistant Escherichia coli

Michael A. Liss,Robin J. Leach,Elizabeth Rourke,Allison Sherrill,Teresa Johnson-Pais,Zhao Lai,Joseph Basler,James R. White,Jan E. Patterson 대한비뇨의학회 2019 Investigative and Clinical Urology Vol.60 No.2

Purpose: Fluoroquinolone-resistant (FQR) Escherichia coli causes transrectal prostate biopsy infections. In order to reduce colonization of these bacteria in carriers, we would like to understand the surrounding microbiome to determine targets for decolonization. Materials and Methods: We perform an observational study to investigate the microbiome differences in men with and without FQR organisms found on rectal culture. A rectal swab with two culturettes was performed on men before an upcoming prostate biopsy procedure as standard of care to perform “targeted prophylaxis.” Detection of FQR was performed by the standard microbiology lab inoculates the swab onto MacConkey agar containing ciprofloxacin. The extra swab was sent for 16S rRNA amplicon sequencing (MiSeq paired-end) using the V1V2 primer. Alpha and beta-diversity analysis were performed using QIIME. We used PERMANOVA to evaluate the statistical significance of beta-diversity distances within and between groups of interest. Results: We collected 116 rectal swab samples before biopsy for 16S rRNA amplicon sequencing. We identified 18 isolates (15.5%, 18/116) that were positive and had relative reduced diversity profiles (p<0.05). Enterobacteriaceae were significantly over-represented in the FQR subjects (adjusted p=0.03). Conclusions: Microbiome analysis determined that men colonized with FQR bacteria have less diverse bacterial communities (dysbiosis), higher levels of Enterobacteriaceae and reduced levels of Prevotella disiens. These results may have implications in pre/probiotic intervention studies.

Assessment of forest fire impacts on carbonaceous aerosols using complementary molecular marker receptor models at two urban locations in California's San Joaquin Valley

Bae, Min-Suk,Skiles, Matthew J.,Lai, Alexandra M.,Olson, Michael R.,de Foy, Benjamin,Schauer, James J. Elsevier 2019 Environmental pollution Vol.246 No.-

<P><B>Abstract</B></P> <P>Two hundred sixty-three fine particulate matter (PM<SUB>2.5</SUB>) samples were collected over fourteen months in Fresno and Bakersfield, California. Samples were analyzed for organic carbon (OC), elemental carbon (EC), water soluble organic carbon (WSOC), and 160 organic molecular markers. Chemical Mass Balance (CMB) and Positive Matrix Factorization (PMF) source apportionment models were applied to the results in order to understand monthly and seasonal source contributions to PM<SUB>2.5</SUB> OC. Similar source categories were found from the results of the CMB and PMF models to PM<SUB>2.5</SUB> OC across the sites. Six source categories with reasonably stable profiles, including biomass burning, mobile, food cooking, two different secondary organic aerosols (SOAs) (i.e., winter and summer), and forest fires were investigated. Both the CMB and the PMF models showed a strong seasonality in contributions of some sources, as well as dependence on wind transport for both sites. The overall relative source contributions to OC were 24% CMB wood smoke, 19% CMB mobile sources, 5% PMF food cooking, 2% CMB vegetative detritus, 17% PMF SOA summer, 22% PMF SOA winter, and 12% PMF forest fire. Back-trajectories using the Weather Research and Forecasting model combined with the FLEXible PARTicle dispersion model (WRF-FLEXPART) were used to further characterize wind transport. Clustering of the trajectories revealed dominant wind patterns associated with varying concentrations of the different source categories. The Comprehensive Air Quality Model with eXtensions (CAMx) was used to simulate aerosol transport from forest fires and thus confirm the impacts of individual fires, such as the Rough Fire, at the measurement sites.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Comparison of Molecular marker CMB and PMF models to OC. </LI> <LI> Assessment of Forest Fire impacts on OC using PMF. </LI> <LI> Forest fire confirmation from CAMx simulations based on FLAMBE emissions. </LI> </UL> </P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>