• Title/Summary/Keyword: Molecular approach

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Rapid Detection of Streptococcus mutans Using an Integrated Microfluidic System with Loop-Mediated Isothermal Amplification

  • Jingfu Wang;Jingyi Wang;Xin Chang;Jin Shang;Yuehui Wang;Qin Ma;Liangliang Shen
    • Journal of Microbiology and Biotechnology
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    • v.33 no.8
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    • pp.1101-1110
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    • 2023
  • Streptococcus mutans is the primary causative agent of caries, which is one of the most common human diseases. Thus, rapid and early detection of cariogenic bacteria is critical for its prevention. This study investigated the combination of loop-mediated isothermal amplification (LAMP) and microfluid technology to quantitatively detect S. mutans. A low-cost, rapid microfluidic chip using LAMP technology was developed to amplify and detect bacteria at 2.2-2.2 × 106 colony-forming units (CFU)/ml and its detection limits were compared to those of standard polymerase chain reaction. A visualization system was established to quantitatively determine the experimental results, and a functional relationship between the bacterial concentration and quantitative results was established. The detection limit of S. mutans using this microfluidic chip was 2.2 CFU/ml, which was lower than that of the standard approach. After quantification, the experimental results showed a good linear relationship with the concentration of S. mutans, thereby confirming the effectiveness and accuracy of the custom-made integrated LAMP microfluidic system for the detection of S. mutans. The microfluidic system described herein may represent a promising simple detection method for the specific and rapid testing of individuals at risk of caries.

Genome-wide association studies to identify quantitative trait loci and positional candidate genes affecting meat quality-related traits in pigs

  • Jae-Bong Lee;Ji-Hoon Lim;Hee-Bok Park
    • Journal of Animal Science and Technology
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    • v.65 no.6
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    • pp.1194-1204
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    • 2023
  • Meat quality comprises a set of key traits such as pH, meat color, water-holding capacity, tenderness and marbling. These traits are complex because they are affected by multiple genetic and environmental factors. The aim of this study was to investigate the molecular genetic basis underlying nine meat quality-related traits in a Yorkshire pig population using a genome-wide association study (GWAS) and subsequent biological pathway analysis. In total, 45,926 single nucleotide polymorphism (SNP) markers from 543 pigs were selected for the GWAS after quality control. Data were analyzed using a genome-wide efficient mixed model association (GEMMA) method. This linear mixed model-based approach identified two quantitative trait loci (QTLs) for meat color (b*) on chromosome 2 (SSC2) and one QTL for shear force on chromosome 8 (SSC8). These QTLs acted additively on the two phenotypes and explained 3.92%-4.57% of the phenotypic variance of the traits of interest. The genes encoding HAUS8 on SSC2 and an lncRNA on SSC8 were identified as positional candidate genes for these QTLs. The results of the biological pathway analysis revealed that positional candidate genes for meat color (b*) were enriched in pathways related to muscle development, muscle growth, intramuscular adipocyte differentiation, and lipid accumulation in muscle, whereas positional candidate genes for shear force were overrepresented in pathways related to cell growth, cell differentiation, and fatty acids synthesis. Further verification of these identified SNPs and genes in other independent populations could provide valuable information for understanding the variations in pork quality-related traits.

Colloidal Optics and Photonics: Photonic Crystals, Plasmonics, and Metamaterials

  • Jaewon Lee;Seungwoo Lee
    • Current Optics and Photonics
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    • v.7 no.6
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    • pp.608-637
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    • 2023
  • The initial motivation in colloid science and engineering was driven by the fact that colloids can serve as excellent models to study atomic and molecular behavior at the mesoscale or microscale. The thermal behaviors of actual atoms and molecules are similar to those of colloids at the mesoscale or microscale, with the primary distinction being the slower dynamics of the latter. While atoms and molecules are challenging to observe directly in situ, colloidal motions can be easily monitored in situ using simple and versatile optical microscopic imaging. This foundational approach in colloid research persisted until the 1980s, and began to be extensively implemented in optics and photonics research in the 1990s. This shift in research direction was brought by an interplay of several factors. In 1987, Yablonovitch and John modernized the concept of photonic crystals (initially conceptualized by Lord Rayleigh in 1887). Around this time, mesoscale dielectric colloids, which were predominantly in a suspended state, began to be self-assembled into three-dimensional (3D) crystals. For photonic crystals operating at optical frequencies (visible to near-infrared), mesoscale crystal units are needed. At that time, no manufacturing process could achieve this, except through colloidal self-assembly. This convergence of the thirst for advances in optics and photonics and the interest in the expanding field of colloids led to a significant shift in the research paradigm of colloids. Initially limited to polymers and ceramics, colloidal elements subsequently expanded to include semiconductors, metals, and DNA after the year 2000. As a result, the application of colloids extended beyond dielectric-based photonic crystals to encompass plasmonics, metamaterials, and metasurfaces, shaping the present field of colloidal optics and photonics. In this review we aim to introduce the research trajectory of colloidal optics and photonics over the past three decades; To elucidate the utility of colloids in photonic crystals, plasmonics, and metamaterials; And to present the challenges that must be overcome and potential research prospects for the future.

A genome-wide approach to the systematic and comprehensive analysis of LIM gene family in sorghum (Sorghum bicolor L.)

  • Md. Abdur Rauf Sarkar;Salim Sarkar;Md Shohel Ul Islam;Fatema Tuz Zohra;Shaikh Mizanur Rahman
    • Genomics & Informatics
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    • v.21 no.3
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    • pp.36.1-36.19
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    • 2023
  • The LIM domain-containing proteins are dominantly found in plants and play a significant role in various biological processes such as gene transcription as well as actin cytoskeletal organization. Nevertheless, genome-wide identification as well as functional analysis of the LIM gene family have not yet been reported in the economically important plant sorghum (Sorghum bicolor L.). Therefore, we conducted an in silico identification and characterization of LIM genes in S. bicolor genome using integrated bioinformatics approaches. Based on phylogenetic tree analysis and conserved domain, we identified five LIM genes in S. bicolor (SbLIM) genome corresponding to Arabidopsis LIM (AtLIM) genes. The conserved domain, motif as well as gene structure analyses of the SbLIM gene family showed the similarity within the SbLIM and AtLIM members. The gene ontology (GO) enrichment study revealed that the candidate LIM genes are directly involved in cytoskeletal organization and various other important biological as well as molecular pathways. Some important families of regulating transcription factors such as ERF, MYB, WRKY, NAC, bZIP, C2H2, Dof, and G2-like were detected by analyzing their interaction network with identified SbLIM genes. The cis-acting regulatory elements related to predicted SbLIM genes were identified as responsive to light, hormones, stress, and other functions. The present study will provide valuable useful information about LIM genes in sorghum which would pave the way for the future study of functional pathways of candidate SbLIM genes as well as their regulatory factors in wet-lab experiments.

Copper Oxide Spike Grids for Enhanced Solution Transfer in Cryogenic Electron Microscopy

  • Dukwon Lee;Hansol, Lee;Jinwook Lee;Soung-Hun Roh;Nam-Chul Ha
    • Molecules and Cells
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    • v.46 no.9
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    • pp.538-544
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    • 2023
  • The formation of uniform vitreous ice is a crucial step in the preparation of samples for cryogenic electron microscopy (cryo-EM). Despite the rapid technological progress in EM, controlling the thickness of vitreous ice on sample grids with reproducibility remains a major obstacle to obtaining high-quality data in cryo-EM imaging. The commonly employed classical blotting process faces the problem of excess water that cannot be absorbed by the filter paper, resulting in the formation of thick and heterogeneous ice. In this study, we propose a novel approach that combines the recently developed nanowire self-wicking technique with the classical blotting method to effectively control the thickness and homogeneity of vitrified ice. With simple procedures, we generated a copper oxide spike (COS) grid by inducing COSs on commercially available copper grids, which can effectively remove excess water during the blotting procedure without damaging the holey carbon membrane. The ice thickness could be controlled with good reproducibility compared to non-oxidized grids. Incorporated into other EM techniques, our new modification method is an effective option for obtaining high-quality data during cryo-EM imaging.

Identification of Cell Type-Specific Effects of DNMT3A Mutations on Relapse in Acute Myeloid Leukemia

  • Seo-Gyeong Bae;Hyeoung-Joon Kim;Mi Yeon Kim;Dennis Dong Hwan Kim;So-I Shin;Jae-Sook Ahn;Jihwan Park
    • Molecules and Cells
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    • v.46 no.10
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    • pp.611-626
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    • 2023
  • Acute myeloid leukemia (AML) is a heterogeneous disease caused by distinctive mutations in individual patients; therefore, each patient may display different cell-type compositions. Although most patients with AML achieve complete remission (CR) through intensive chemotherapy, the likelihood of relapse remains high. Several studies have attempted to characterize the genetic and cellular heterogeneity of AML; however, our understanding of the cellular heterogeneity of AML remains limited. In this study, we performed single-cell RNA sequencing (scRNAseq) of bone marrow-derived mononuclear cells obtained from same patients at different AML stages (diagnosis, CR, and relapse). We found that hematopoietic stem cells (HSCs) at diagnosis were abnormal compared to normal HSCs. By improving the detection of the DNMT3A R882 mutation with targeted scRNAseq, we identified that DNMT3A-mutant cells that mainly remained were granulocyte-monocyte progenitors (GMPs) or lymphoid-primed multipotential progenitors (LMPPs) from CR to relapse and that DNMT3A-mutant cells have gene signatures related to AML and leukemic cells. Copy number variation analysis at the single-cell level indicated that the cell type that possesses DNMT3A mutations is an important factor in AML relapse and that GMP and LMPP cells can affect relapse in patients with AML. This study advances our understanding of the role of DNMT3A in AML relapse and our approach can be applied to predict treatment outcomes.

Evaluation of genetic differentiation and search for candidate genes for reproductive traits in pigs

  • Elena Romanets;Siroj Bakoev;Timofey Romanets;Maria Kolosova;Anatoly Kolosov;Faridun Bakoev;Olga Tretiakova;Alexander Usatov;Lyubov Getmantseva
    • Animal Bioscience
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    • v.37 no.5
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    • pp.832-838
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    • 2024
  • Objective: The use of molecular genetic methods in pig breeding can significantly increase the efficiency of breeding and breeding work. We applied the Fst (fixsacion index) method, the main focus of the work was on the search for common options related to the number of born piglets and the weight of born piglets, since today the urgent task is to prevent a decrease in the weight of piglets at birth while maintaining high fertility of sows. Methods: One approach is to scan the genome, followed by an assessment of Fst and identification of selectively selected regions. We chose Large White sows (n = 237) with the same conditions of keeping and feeding. The data were collected from the sows across three farrowing. For genotyping, we used GeneSeek GGP Porcine HD Genomic Profiler v1, which included 68,516 single nucleotide polymorphisms evenly distributed with an average spacing of 25 kb (Illumina Inc, San Diego, CA, USA). Results: Based on the results of the Fst analysis, 724 variants representing selection signals for the signs BALWT, BALWT1, NBA, and TNB (weight of piglets born alive, average weight of the 1st piglets born alive, total number born alive, total number born). At the same time, 18 common variants have been identified that are potential markers for both the number of piglets at birth and the weight of piglets at birth, which is extremely important for breeding work to improve reproductive characteristics in sows. Conclusion: Our work resulted in identification of variants associated with the reproductive characteristics of pigs. Moreover, we identified, variants which are potential markers for both the number of piglets at birth and the weight of piglets at birth, which is extremely important for breeding work to improve reproductive performance in sows.

Targeting cell surface glucose-regulated protein 94 in gastric cancer with an anti-GRP94 human monoclonal antibody

  • Hyun Jung Kim;Yea Bin Cho;Kyun Heo;Ji Woong Kim;Ha Gyeong Shin;Eun-bi Lee;Seong-Min Park;Jong Bae Park;Sukmook Lee
    • BMB Reports
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    • v.57 no.4
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    • pp.188-193
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    • 2024
  • Gastric cancer (GC), a leading cause of cancer-related mortality, remains a significant challenge despite recent therapeutic advancements. In this study, we explore the potential of targeting cell surface glucose-regulated protein 94 (GRP94) with antibodies as a novel therapeutic approach for GC. Our comprehensive analysis of GRP94 expression across various cancer types, with a specific focus on GC, revealed a substantial overexpression of GRP94, highlighting its potential as a promising target. Through in vitro and in vivo efficacy assessments, as well as toxicological analyses, we found that K101.1, a fully human monoclonal antibody designed to specifically target cell surface GRP94, effectively inhibits GC growth and angiogenesis without causing in vivo toxicity. Furthermore, our findings indicate that K101.1 promotes the internalization and concurrent downregulation of cell surface GRP94 on GC cells. In conclusion, our study suggests that cell surface GRP94 may be a potential therapeutic target in GC, and that antibody-based targeting of cell surface GRP94 may be an effective strategy for inhibiting GRP94-mediated GC growth and angiogenesis.

Research progress on hydrogel-based drug therapy in melanoma immunotherapy

  • Wei He;Yanqin Zhang;Yi Qu;Mengmeng Liu;Guodong Li;Luxiang Pan;Xinyao Xu;Gege Shi;Qiang Hao;Fen Liu;Yuan Gao
    • BMB Reports
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    • v.57 no.2
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    • pp.71-78
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    • 2024
  • Melanoma is one of the most aggressive skin tumors, and conventional treatment modalities are not effective in treating advanced melanoma. Although immunotherapy is an effective treatment for melanoma, it has disadvantages, such as a poor response rate and serious systemic immune-related toxic side effects. The main solution to this problem is the use of biological materials such as hydrogels to reduce these side effects and amplify the immune killing effect against tumor cells. Hydrogels have great advantages as local slow-release drug carriers, including the ability to deliver antitumor drugs directly to the tumor site, enhance the local drug concentration in tumor tissue, reduce systemic drug distribution and exhibit good degradability. Despite these advantages, there has been limited research on the application of hydrogels in melanoma treatment. Therefore, this article provides a comprehensive review of the potential application of hydrogels in melanoma immunotherapy. Hydrogels can serve as carriers for sustained drug delivery, enabling the targeted and localized delivery of drugs with minimal systemic side effects. This approach has the potential to improve the efficacy of immunotherapy for melanoma. Thus, the use of hydrogels as drug delivery vehicles for melanoma immunotherapy has great potential and warrants further exploration.

Twindemic Threats of Weeds Coinfected with Tomato Yellow Leaf Curl Virus and Tomato Spotted Wilt Virus as Viral Reservoirs in Tomato Greenhouses

  • Nattanong Bupi;Thuy Thi Bich Vo;Muhammad Amir Qureshi;Marjia Tabassum;Hyo-jin Im;Young-Jae Chung;Jae-Gee Ryu;Chang-seok Kim;Sukchan Lee
    • The Plant Pathology Journal
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    • v.40 no.3
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    • pp.310-321
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    • 2024
  • Tomato yellow leaf curl virus (TYLCV) and tomato spotted wilt virus (TSWV) are well-known examples of the begomovirus and orthotospovirus genera, respectively. These viruses cause significant economic damage to tomato crops worldwide. Weeds play an important role in the ongoing presence and spread of several plant viruses, such as TYLCV and TSWV, and are recognized as reservoirs for these infections. This work applies a comprehensive approach, encompassing field surveys and molecular techniques, to acquire an in-depth understanding of the interactions between viruses and their weed hosts. A total of 60 tomato samples exhibiting typical symptoms of TYLCV and TSWV were collected from a tomato greenhouse farm in Nonsan, South Korea. In addition, 130 samples of 16 different weed species in the immediate surroundings of the greenhouse were collected for viral detection. PCR and reverse transcription-PCR methodologies and specific primers for TYLCV and TSWV were used, which showed that 15 tomato samples were coinfected by both viruses. Interestingly, both viruses were also detected in perennial weeds, such as Rumex crispus, which highlights their function as viral reservoirs. Our study provides significant insights into the co-occurrence of TYLCV and TSWV in weed reservoirs, and their subsequent transmission under tomato greenhouse conditions. This project builds long-term strategies for integrated pest management to prevent and manage simultaneous virus outbreaks, known as twindemics, in agricultural systems.