Search

The Pacific oyster, Crassostrea gigas, is the most important and valuable commercial fishery species in Korea. Its farming started 20 years ago and is still rapid expansion in Korea. In this study, to maintain the genetic diversity of this valuable marine resource, possible genetic similarity and differences between the wild population and hatchery population in Tongyeong, Korea were accessed using multiplex assays with nine highly polymorphic microsatellite loci. A total of 250 different alleles were found over all loci. Despite a long history of hatchery practices, very high levels of polymorphism (mean alleles = 22.89 and mean heterozygosity = 0.92) were detected between the two populations. No statistically significant reductions were found in heterozygosity or allelic diversity in the hatchery population compared with the wild population. However, significant genetic heterogeneity was found between two populations. These results provide no evidence to show that hatchery practice of Pacific oyster in Korea has significantly affected the genetic variability of the hatchery stock. Although further studies are needed for comprehensive determinations of the hatchery and wild populations with increased number of Pacific oyster sample collections, information on the genetic variation and differentiation obtained in this study can be applied for genetic monitoring of aquaculture stocks, genetic improvement by selective breeding and designing of more efficient conservation management guidelines for these valuable genetic materials. [ABSTRACT FROM AUTHOR]

Red sea cucumber Stichopus japonicus is the most important and valuable commercial sea cucumber species in Korea. Its farming and stock enhancement started in the early 2000s and is still rapid expansion in Korea. Therefore, the analyses of genetic status of wild and hatchery populations are necessary to maintain the genetic diversity of this valuable marine resource. In this study, possible genetic similarity and differences between the wild population and hatchery population in Jeju, Korea were accessed using multiplex assays with eight highly polymorphic microsatellite loci. High levels of polymorphism were observed between the two populations. A total of 93 different alleles were found. Although a considerable loss of unique alleles and relatively high inbreeding coefficient value were observed in the hatchery samples, no statistically significant reductions were found in heterozygosity or allelic diversity in the hatchery population, compared with the wild population. However, significant genetic heterogeneity was found between two populations. These results suggest that genetic drift has probably promoted differentiation between populations, and stocking intensity in wild populations may correlate with loss of genetic integrity. Therefore, the sustainable exploitation plans of the fishery resource should be developed by applying basic genetic principles combined with molecular monitoring. This genetic baseline information of Korean red sea cucumber has important implications for designing of genetically sustainable restocking programs and more efficient conservation management guidelines for these valuable genetic materials. [ABSTRACT FROM AUTHOR]

We developed a novel one-pot synthetic strategy for preparing monodisperse polyethylene glycol diacrylate (PEGDA) microdroplets via organoclay-assisted interfacial polymerization approach for Escherichia coli encapsulation. Based on the mechanism of spontaneous and rapid polymerization of PEGDA precursor solution with Mg-organoclay, the prepared PEGDA microdroplets have uniform size and fine round shape, with size range of 74-118 µm. The size of microdroplets can be controlled through the changing continuous phase flow rate. Organoclay-assisted polymerization method provides a unique environment to produce non-toxic ways of fabricating microorganism encapsulated microdroplets and to prohibit microdroplets merge during the processes. Furthermore, we successfully carried out to entrap E. coli inside of the PEGDA microdroplets. E. coli expressing a green fluorescent protein shows a good viability inside the PEGDA microdroplets. The in situ microfluidic synthetic method provides a novel approach for the preparation of monodisperse PEGDA microdroplets via a one-pot route. Biotechnol. Bioeng. 2012;109: 289-294. © 2011 Wiley Periodicals, Inc. [ABSTRACT FROM AUTHOR]

The article reports on the study on the genetic characterization of four East Asian giant scallop populations using microsatellite markers in Korea, China, Russia and Japan. It states that knowledge of scallop population genetics is essential for the adequate management and the improvement of genetic resources of cultivated stocks. The study found high levels of polymorphism in the East Asian giant scallop, which permitted a fine-scale analysis of population relationships. Also, the study revealed that microsatellite markers are a powerful approach by which to monitor genetic diversity in the said population.

Summer mortality, caused by thermal conditions, is the biggest threat to abalone aquaculture production industries. Various measures have been taken to mitigate this issue by adjusting the environment; however, the cellular processes of Pacific abalone (Haliotis discus hannai) have been overlooked due to the paucity of genetic information. The draft genome of H. discus hannai has recently been reported, prompting exploration of the genes responsible for thermal regulation in Pacific abalone. In this study, 413 proteins were systematically annotated as members of the heat shock protein (HSP) super families, and among them 26 HSP genes from four Pacific abalone tissues (hemocytes, gill, mantle, and muscle) were differentially expressed under cold and heat stress conditions. The co-expression network revealed that HSP expression patterns were tissue-specific and similar to those of other shellfish inhabiting intertidal zones. Finally, representative HSPs were selected at random and their expression patterns were identified by RNA sequencing and validated by qRT-PCR to assess expression significance. The HSPs expressed in hemocytes were highly similar in both analyses, suggesting that hemocytes could be more reliable samples for validating thermal condition markers compared to other tissues. [ABSTRACT FROM AUTHOR]

We characterized the total mitochondrial genome of Australian spiny lobster,Panulirus cygnus(George, 1962), which is found along the western coast of Australia. Total mitochondrial genome length ofP. cygnuswas 15 724 bp, in which 13 proteins, 2 ribosomal RNAs, 22 transfer RNAs, and a putative control region were encoded. Nine and four protein-coding genes are encoded on the H-strand and on the L-strand, respectively. According to the phylogenetic analysis,P. cygnuswas most closely related toPanulirus japonicusamong the compared six species belonging to Palinuridae. Although overall gene organization was the same, the putative control region (betweenSrRNAgene andtRNAIlel) is least similar to one another among mitochondrial genomes from the compared six species belonging to Palinuridae [ABSTRACT FROM AUTHOR]

Mitogenome of femaleRuditapes philippinarumorganism was sequenced, and genomic variation and phylogeny were examined in this study. Length of the mitogenome was 22 089 bp showing 94.28% of sequence identity with previously reported sequence. Total 707 single nucleotide polymorphisms, SNPs, were detected and 50 residues were non-synonymous SNPs among the 202 SNPs in protein-coding genes. Deleted genomic fragments with of 265 bp and 322 bp were observed in non-coding regions,ND2toND4LandND4LtotRNAIle, respectively. Phylogenic analysis confirmed that used organisms were femaleR. philippinarum, and the species has closer evolutionary distance with genusPaphiarather than genusMeretrix. Our finding will be help to set an insight for population and evolutionary genomics of Veneroida clams as well as application to marine industry. [ABSTRACT FROM AUTHOR]

Abstract Anti-lipopolysaccharide factors (ALFs) are a representative host defense protein in crustaceans. In this study, we successfully developed two novel antimicrobial peptides (AMPs), named crab-ALF2A and crab-ALF6A, which contain changes to the amino acid sequences of the lipopolysaccharide binding domain and signal peptide, respectively, of the ALF of the swimming crab Portunus trituberculatus. The crab-ALF2A peptide showed potent antimicrobial activity against the Gram-positive bacteria Bacillus cereus , Staphylococcus aureus , and Streptococcus iniae (minimal effective concentration [MEC] 1.51–1.93 μg/mL) and the Gram-negative bacteria Pseudomonas aeruginosa and Escherichia coli (MEC 1.87–1.98 μg/mL), with maximal bactericidal activity at a peptide concentration of 5 μg/mL. The crab-ALF6A peptide also showed potent antimicrobial activity against B. cereus , S. aureus , and S. iniae (MEC 1.49–2.3 μg/mL) and P. aeruginosa and E. coli (MEC 1.72–1.19 μg/mL) at a peptide concentration of 5 μg/mL. Notably, the crab-ALF2A and crab-ALF6A peptides exhibited strong activity against Candida albicans (MECs of 2.11 and 1.95 μg/mL, respectively). These activities were stable following heat treatment. Moreover, the effect of crab-ALF2A and crab-ALF6A peptide treatment on microbe cell morphology was confirmed by scanning electron microscopy. Membrane disruption and damage, and the leakage of cytoplasmic content were clearly observed. A downsizing peptide approach illustrated that the hexapeptide ALF6A8 (RVLLRL) was the shortest peptide showing significant antimicrobial activity. Our approach allows for the generation of novel antimicrobial peptides in a cost effective manner as potential next-generation antibiotics. Highlights • Novel antimicrobial peptides were designed based on the anti-LPS factor of Portunus tribuberculatus. • Membrane disruption and damage of microbes by AMPs were all clearly observed by scanning electron microscopy. • The hexapeptide (ALF6A8) was the shortest analogous form showing significant antimicrobial activity. [ABSTRACT FROM AUTHOR]

Abstract An antimicrobial peptide with 55 amino acid residues was purified by C 18 reversed-phase high-performance liquid chromatography (HPLC) from foot extract of the hard-shelled mussel, Mytilus coruscus. This peptide showed strong antimicrobial activity against Gram-positive and Gram-negative bacteria, as well as fungi. The purified peptide was determined to have a molecular mass of 6202 Da by matrix-assisted laser desorption/ionization time-of-flight mass spectrophotometry (MALDI-TOF/MS). The identified 20-amino acid sequence of the purified peak by Edman degradation shared 100% identity with the N-terminal regions of mytichitin-1, mytichitin-2, mytichitin-3, mytichitin-4, mytichitin-5, and chitinase-like protein-1, and so was named mytichitin-CBD. The cDNA of mytichitin-CBD was cloned and sequenced by rapid amplification of cDNA ends (RACE). The mRNA transcripts were mainly detected in foot tissue, and they were up-regulated and peaked at 4 h after bacterial infection. We constructed and expressed recombinant mytichitin-CBD protein which displayed antimicrobial activity against Gram-negative bacteria Gram-positive bacteria and the fungus as well as anti-parasitic activity against scuticociliates. The results of this study demonstrate that the peptide isolated from M. coruscus is related to the innate immune system of this marine invertebrate and is a possible alternative to antibiotics. Highlights • Antimicrobial effect of mytichin-chitin binding domain (mytichitin-CBD) was identified from foot extract of the Korean hard-shelled mussel, Mytilus coruscus. • 6.2 kDa of the mytichitin-CBD peptide containing 55 amino acid residues exhibited antibacterial and anti-parasite activity. • Expression levels and patterns were dependent on the tissue type and phase of infection. • Mytichitin-CBD peptide might cooperate in the innate immune defense system of Korean hard-shelled mussel, Mytilus coruscus. [ABSTRACT FROM AUTHOR]

Antimicrobial peptides (AMPs) process strong antimicrobial activity and play a major role in protect host against pathogens. Piscidins, one of main AMPs of fish, are considered as crucial effectors of innate immune response as well as modulators of adaptive immune response. In this study, we discovered two rock bream piscidin paralogs (RP6 and RP7) from leucocytes of the rock bream and investigated their gene expression patterns and antimicrobial and cytotoxic activities. Their open reading frames (ORF) consisted of 240 and 249 base pairs (bp), encoding 80 and 83 amino acid residues, respectively, and both conserved AMP 12 domain. The predicted tertiary structure of the AMP 12 domain of RP6 and RP7 is an amphipathic alpha-helix structure. RP6 and RP7 mRNAs are ubiquitously expressed in healthy fish, the most expression of both in the PBLs. RP6 expression was more induced after pathogen infection relative to that of RP7, while RP7 exhibited stronger antimicrobial and haemolytic activity compared with RP6. Ultimately, the powerful antibacterial effect of RP7 would allow a small amount of RP7 to protect the host from pathogens and RP6 may have other immune response function in rock bream in addition to direct pathogen killing, suggesting that RP6 and RP7 have important function in rock bream immune system. Also, the studies on these RP6 and RP7 will be useful as important data on the substitution of antimicrobials in aquaculture of rock bream. [ABSTRACT FROM AUTHOR]

Disturbance in the balance between pro-oxidants and anti-oxidants result oxidative stress in aerobic organisms. However, oxidative stress can be inhibited by enzymatic and non-enzymatic defense mechanisms. Superoxide dismutases (SODs) are well-known scavengers of superoxide radicals, and they protect cells by detoxifying hazardous reactive oxygen species. Here, we have identified and characterized two different SODs, CuZnSOD and MnSOD, from black rockfish (RfCuZnSOD and RfMnSOD, respectively). In silico analysis revealed the well-conserved molecular structures comprising all essential properties of CuZnSOD and MnSOD. Phylogenetic analysis revealed that both RfCuZnSOD and RfMnSOD cladded with their fish counterparts. The recombinant RfSOD proteins demonstrated their potential superoxide scavenging abilities through a xanthine oxidase assay. The optimum temperature and pH conditions for both rRfSODs were 25 °C and pH 8, respectively. Moreover, the potential peroxidation function of rRfCuZnSOD was observed in the presence of HCO 3 − . The highest peroxidation activity was observed at 100 μg/mL of rRfCuZnSOD using the MTT cell viability assay and flow cytometry. The analogous tissue-specific expression profile indicated ubiquitous expression of both RfCuZnSOD and RfMnSOD in selected tissues of healthy juvenile rockfish. An immune challenge experiment illustrated the altered expression profiles of both RfCuZnSOD and RfMnSOD against lipopolysaccharide, Streptococcus iniae , and polyinosinic-polycytidylic acid (poly I:C). Collectively, these results strengthen the general understanding of the structural and functional characteristics of SODs within the host defense system. [ABSTRACT FROM AUTHOR]

The tripartite motif-containing (TRIM) proteins are involved in a wide range of cellular processes, and the role of TRIM1 in immunity has been explored. However, fundamental studies on fish TRIM1 are lacking. In this study, we cloned and characterized TRIM1 cDNA from the Korean rose bitterling, Rhodeus uyekii (RuTRIM1). Two RuTRIM1 isoforms (RuTRIM1-X1 and RuTRIM1-X2) were identified. The coding sequence (CDS) of RuTRIM1-X1 comprised 2157 bp encoding a 718-aa protein, and the CDS of RuTRIM1-X2 comprised 1929 bp encoding a 642-aa protein. Both RuTRIM1 isoforms contained a RING finger domain, B-box 1, B-box 2, coiled-coil domain, COS box, FN3 motif, and PRY/SPRY domain. The deduced RuTRIM1-X1 and RuTRIM1-X2 proteins had high amino acid identity (76.27–98.89%) with orthologs from various other species, and a phylogenetic tree was constructed. RuTRIM1-X1 and RuTRIM1-X2 mRNA were expressed in all tissues examined, with the highest expression levels detected in the hepatopancreas. During early development, RuTRIM1-X1 and RuTRIM1-X2 mRNA levels changed differently from the gastrula period to the first feeding stage. An in vivo ubiquitination assay showed that RuTRIM1 exhibited RING-dependent E3 ubiquitin ligase activity, mainly by comparing RuTRIM1-X2 to RuTRIM1-X1. The subcellular localization of the two RuTRIM1 protein isoforms was characterized, revealing that they formed aggregates in cytoplasmic bodies in Raw264.7 cells. Interferon-γ/lipopolysaccharide-induced nuclear factor-κB signaling was negatively regulated by RuTRIM1-X1 and RuTRIM1-X2, and the negative effect was reversed in RING deletion mutants. To our knowledge, this is the first study to characterize fish TRIM1, which may play a role in the inflammatory response. [ABSTRACT FROM AUTHOR]

We isolated and characterised a cDNA encoding the lysosomal protective protein (serine protease) cathepsin A (CTSA) from rock bream ( Oplegnathus fasciatus ). The full-length rock bream CTSA (RbCTSA) cDNA (1814 bp) contains an open reading frame of 1419 bp that encodes 472 amino acids. Alignment of multiple CTSA protein sequences revealed that the active site serine and histidine residues were well-conserved among the other CTSA sequences. RbCTSA is highly expressed in the peripheral blood leukocytes, kidney, spleen, liver, intestine, gill, heart, brain, stomach, and eye. RbCTSA expression was also examined in several tissues, including whole kidneys and spleens, under bacterial and viral challenge. In general, all of the examined tissues that were infected with Edwardsiella tarda , Streptococcus iniae , or red sea bream iridovirus (RSIV) exhibited significant upregulation of RbCTSA expression compared to the controls. Our results reveal that RbCTSA may be involved in the immune responses of rock bream. [ABSTRACT FROM AUTHOR]

Abstract: Recently, marine environmental pollution has become more and more serious due to oil spills or the collapse of nuclear power plant resulting in a massive release of hazardous materials to the ocean. These harmful radioactive materials are associated with the random mutation of genes leading to deformity of individuals. So far there has been no effective system for monitoring this marine pollution. Therefore, rapid detection of DNA mutations has become increasingly important in a variety of fields including medical diagnostics and food safety. Herein, we report a rapid, sensitive, qualitative and quantitative approach for DNA detection based on the magnetophoretic assay. The Au-magnetic microparticles (MMPs) nanocomplex was used to generate a color signal and uncombined Au NPs contributed an optical signal that could be directly quantified by UV–Vis absorption spectroscopy of the supernatant after the sample was placed in a magnetic field for 2min. The linear relationship between signal intensity and the target DNA concentrations (0.1nM–1.0μM) was investigated and a limit of quantitation of 0.1nM was achieved using this method. [Copyright &y& Elsevier]

Microbial inactivation and pesticide removal by remote exposure of atmospheric air plasma were investigated in confined environments, including an airtight box and commercial refrigerator. The relative sterilization ratios of remote plasma exposure in an airtight box were found to be affected by the distance from the plasma generator, the volume of box and the time of irradiation; however, over 99% saturation was obtained within only 120 s in all experiments. The sterilization of microorganisms and the removal of pesticide in a refrigerator with a volume of 292 l were also successfully achieved, resulting in over 99% inactivation or decontamination in a few minutes. Considering the reported results by direct plasma exposure and circulation, it can be concluded that the confined environment enhances the efficient irradiation of plasma by eliminating air flow. This system can be applied to the storage to keep agricultural products freshly and exclusion of harmful materials on the products. [Copyright &y& Elsevier]

Abstract: Delicate thermodynamic control at confined space has been of importance in recent nano/micro-plasmonic sensing to be more accurate and to use lower quantity of specimens than conventional devices. In our study, experimental and computational study on thermal behavior of surface plasmon resonance (SPR) was carried out using various sizes of Au nanoparticles (NPs; 4, 18, and 57 nm) and thermo-reversible polymer, poly(N-isopropylacrylamide) (PNIPAm) on SPR chip. It was interesting that SPR angle shift on temperature was dependent on NP sizes, where the smaller size the better sensitive. It is probable that both plasmonic interaction and polymer dynamicity were collaboratively affected and presumably the latter dominated in the thermal monitoring of SPR signals. The simulations clearly indicated that the interaction between the Au NPs and polymers was strengthened by increasing temperature and NP sizes, resulting in decreasing sensitivity. These results are supportive to develop further accurate devices using plasmonics and polymer dynamics, such as plasmonic sensing devices, thermo-actuators, hybrid organic solar cell, and flexible display. [Copyright &y& Elsevier]

Abstract: Quantitative detection of biomolecules (e.g. DNA and proteins) has become increasingly important in a variety of fields, including medical diagnostics, food safety, and anti-bioterrorism. We report a simple, sensitive, and inexpensive quantitative approach for DNA detection based on the optical properties of gold nanoparticles and gold-coated magnetic nanoparticle hybrids. We employed a simple one-step reaction to synthesize gold-coated iron oxide nanoparticles (Fe3O4–Au NP). Fe3O4 nanoparticles were used as the central core for preparation of Fe3O4–Au in an aqueous state without precipitation and aggregation of nanoparticles. Citrate-coated Fe3O4 nanoparticles were prepared and subsequently coated with Au layers through reduction of HAuCl4 by citrate on the surface Fe3O4. The resulting Fe3O4–Au nanoparticles showed good paramagnetic properties and were coated with thin layers of gold atoms (∼10nm) having an average diameter of ca. 20nm. These magnetic nanoparticles were well-dispersed in water and stable at physiological pH without precipitation. Nucleic acid-functionalized Fe3O4–Au nanoparticles were then hybridized with complementary target DNA molecules. This resulted in nucleic acid-functionalized gold nanoparticles. To achieve high sensitivity, Fe3O4–Au nanoparticles were employed for the collection of gold nanoprobes that hybridized with complementary target DNA molecules. The Au–Fe3O4–Au nanocomplex remains in solution at a concentration proportional to the concentration of the target DNA and its optical properties allow it to be easily quantified using UV/Vis absorption spectroscopy. The limit of detection for this method was as low as 0.1fM. This selectable detection system is easy to operate, inexpensive, and versatile, making it a candidate for future use in predicting the seafood safety risks through detection of different DNA sequences and any related genetic mutations. [Copyright &y& Elsevier]

Abstract: An active immobilization method utilizing the metal-binding property was developed and examined for its ability to facilitate the biosensing of avian influenza virus. The special biosensing performance with optical plasmonic analysis, including surface plasmon resonance (SPR) was evaluated on gold substrate and also by SPR imaging (SPRi) and localized SPR (LSPR) system where antigen–antibody interaction occurs. A complete optical analytical system was developed by integrating microarray and fabricating nanoparticles onto a single glass chip, thus allowing specific and sensitive diagnosis with subsequent binding. Reaction condition for the maximum reactivity was optimized by SPR analysis and more sensitive interaction was performed by SPRi analysis. Furthermore, ultra-sensitive detection was successfully developed up to the target molecules of 1pgmL−1 by LSPR analysis. The advanced phase-in of enhanced plasmonic sensing system allows more efficient and sensitive detection by switching fabrication processes, which were prepared on the gold surface using the nanoparticles. This inflow contains the gold binding polypeptide (GBP)–fusion protein, which was expressed in recombinant Escherichia coli cells, was bound onto the gold substrates by means of specific interaction. The GBP–fusion method allows immobilization of proteins in bioactive forms onto the gold surface without surface modification suitable for studying antigen–antibody interaction. It was used for the detection of influenza virus, an infectious viral disease, as an example case. [Copyright &y& Elsevier]

Abstract: The antigen (Ag) CFP-10, found in tissue fluids of tuberculosis (TB) patients, is an excellent candidate for use as a sensitive TB marker for early and simplified diagnosis of TB when combined with an effective sensing method. In this study, chemical and optical optimizations were carried out using novel immuno-materials to establish a self-assembled surface plasmon resonance (SPR) optical immunosensing system for the detection of CFP-10 Ag in clinical urine samples from TB patients, which is the first clinical trial as far as we know to monitor Ag in urine. To create a simple sensing interface, a monoclonal antibody (anti-CFP-10) was immobilized directly on a gold surface, followed by blocking with cystamine. The orientation and accessibility of anti-CFP-10 were assessed by the selective binding of CFP-10. The results reveal a linear relationship between the SPR angle shift and the acid-fast bacilli stain stage (0–4) of TB patients. Significant differences were seen between urine samples from TB patients and controls. This method, using urine samples and SPR spectroscopy, is promising and can be rapidly developed into a reliable diagnostic method to monitor TB. [Copyright &y& Elsevier]

Abstract: Information on the occurrence and distribution of polychlorinated biphenyls (PCBs), organochlorine pesticides (OCPs) and polybrominated diphenyl ethers (PBDEs) in marine mammals from Korean coastal waters is not available to date. This is the first study to investigate concentrations and accumulation of PCBs, OCPs, and PBDEs in liver and blubber of minke whales and common dolphins from Korean coastal waters. The concentrations of organochlorines and PBDEs in blubber were several times higher than in liver. The concentrations of PCBs and OCPs were generally low, but the concentrations of PBDEs were similar to those reported for cetaceans elsewhere. The accumulation profiles of OCPs differed between liver and blubber, while the profiles of PCBs and PBDEs were relatively similar in both tissues. The concentrations of PCBs, DDTs, CHLs and PBDEs in common dolphins were significantly higher than those in minke whales, while there were no inter-species differences for HCHs and HCB. The predominant OCP, PCB and PBDE congeners in cetaceans were p,p′-DDE, PCB 153 and BDE 47, respectively. The concentrations of PCBs and DDTs in Korean cetaceans were close to the threshold for adverse health effects. [Copyright &y& Elsevier]

Abstract: Despite several studies that report accumulation of polychlorinated biphenyls (PCBs) and DDT in marine mammals worldwide, very few have examined polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs) and dioxin-like PCBs. In particular, no earlier studies have reported concentrations and accumulation profiles of persistent organic pollutants (POPs) in marine mammals from Korea. In this study, concentrations and accumulation features of PCDD/Fs and dioxin-like PCBs were measured in liver and blubber of minke whales and long-beaked common dolphins collected from Korean coastal waters. The concentrations of PCDFs and dioxin-like PCBs in blubber were 3–10 times higher than the concentrations measured in liver, but PCDDs were higher in liver than blubber. Total toxic equivalent (TEQ) concentrations in blubber of minke whales and common dolphins from Korean coastal waters were similar to, or higher than those reported for cetaceans and seals from other countries. Homologue and congener profiles of PCDD/Fs were different between livers and blubbers, while the profiles of dioxin-like PCBs were similar between the tissues. Concentrations of PCDD/Fs and dioxin-like PCBs in liver and blubber of dolphins were significantly higher than those measured in whales, due to differences in habitat and diet. The relative contribution of individual chemical groups to total TEQs was different between the two cetacean species, suggesting different exposures and metabolic activity. The TEQ levels in minke whales did not exceed the threshold level suggested for immunomodulation, while TEQs in all dolphin samples exceeded the suggested threshold level, implying potential adverse health effects from exposure to PCDD/Fs and PCBs. [Copyright &y& Elsevier]

An amendment to this paper has been published and can be accessed via a link at the top of the paper. [ABSTRACT FROM AUTHOR]

Myostatin (MSTN), a negative regulator of skeletal muscle development, is the major gene of interest for improving animal breeding productivity, including for aquaculture. We introduced a CRISPR/Cas9 system to disrupt Paralichthys olivaceus MSTN (PoMSTN) using microinjection. Cas9 mRNA and sgRNAs targeting the first exon of the PoMSTN gene were co-injected into embryos of P. olivaceus. On-target mutations on the PoMSTN locus were generated in somatic tissues with 75.6% efficiency, and founders harboring germline mutations were produced in the F0 generation. In the F1 generation, restriction fragment length polymorphism analysis was applied to select biallelic mutants, and heterozygous biallelic mutants with PoMSTN disruption were obtained. PoMSTN-disrupted P. olivaceus exhibited greater body thickness and increased condition factor, indicating the enhancement of muscle mass with muscle hyperplasia. Expression of PoMSTN mRNA and protein was significantly reduced in the muscle of PoMSTN-disrupted P. olivaceus , and the mRNA expression of major myogenic regulatory factors (myogenin, MyoD, and Myf5) was differentially affected in PoMSTN-disrupted mutants. These results suggest that the establishment of a CRISPR/Cas9 system in P. olivaceus provides powerful tools for related genetic studies and breeding. • A CRISPR/Cas9 system was introduced to disrupt Paralichthys olivaceus MSTN (PoMSTN). • PoMSTN−/− were obtained which exhibited greater body thickness and increased condition factor. • Expression of PoMSTN was reduced in the muscle of PoMSTN−/−. • The establishment of a CRISPR/Cas9 system in P. olivaceus provides powerful tools for genetic studies and breeding. [ABSTRACT FROM AUTHOR]

Red sea bream, a popular fish resource in Korea and Japan, is being bred in fish farms of the two countries. It is hypothesized that the genomes of red sea bream are influenced by decades of artificial selection. This study investigates the impact of artificial selection on genomes of red sea bream. Whole genome sequencing was conducted for 40 samples of red sea bream either from Ehime, Nagasaki and Tongyeong fish farms or from the wild. Population stratification based on whole genome data was investigated and the genomic regions of fish farm populations under selection were identified using XP-EHH and relative nucleotide diversity. Gene ontology analysis revealed that different functions were enriched in different fish farms. In conclusion, this study highlights the difference between independently cultured red sea bream populations by showing that influence of artificial selection acted upon completely different genes related to different functions including metabolic and developmental processes. [ABSTRACT FROM AUTHOR]

Brain-machine Interface (BMI) is a system that translates neuronal data into an output variable to control external devices such as a robotic arm. A robotic arm can be used as an assistive living device for individuals with tetraplegia. To reflect users' needs in the development process of the BMI robotic arm, our team followed an interactive approach to system development, human-centered design, and Human Activity Assistive Technology model. This study aims to explore the perspectives of people with tetraplegia about activities they want to participate in, their opinions, and the usability of the BMI robotic arm. Eight people with tetraplegia participated in a focus group interview in a semistructured interview format. A general inductive analysis method was used to analyze the qualitative data. The 3 overarching themes that emerged from this analysis were: 1) activities, 2) acceptance, and 3) usability. Activities that the users wanted to do using the robotic arm were categorized into the following 5 activity domains: activities of daily living (ADL), instrumental ADL, health management, education, and leisure. Participants provided their opinions on the needs and acceptance of the BMI technology. Participants answered usability and expected standards of the BMI robotic arm within 7 categories such as accuracy, setup, cost, etc. Participants with tetraplegia have a strong interest in the robotic arm and BMI technology to restore their mobility and independence. Creating BMI features appropriate to users' needs, such as safety and high accuracy, will be the key to acceptance. These findings from the perspectives of potential users should be taken into account when developing the BMI robotic arm., Competing Interests: All authors report having no financial, personal, or organizational conflict of interest with the work in this manuscript., (Copyright © 2022 the Author(s). Published by Wolters Kluwer Health, Inc.)

Objectives: This study aims to develop a community care model in traditional Korean medicine (TKM) by developing a community care participation model for the health of the elderly and deriving tasks to implement it., Methods: This study implemented a group interview with experts. A fact-finding survey was conducted targeting 16 local governments that are implementing a leading project to identify the status of TKM service provision and welfare service linkage in all regions. An expert group interview (FGI) targeted public and private sector experts for each job role, the former represented by those in charge of the central government's health care policy and administrative delivery system, and the latter by professors majoring in social welfare, professors majoring in health, and local TKM societies. After forming the expert groups, three expert group interviews were conducted., Results: Through collective interviews with experts, a model for providing TKM and welfare services in community integrated care was derived by dividing it into local and central government levels. The strategies and tasks for promoting TKM-oriented health welfare services were derived from 3 strategies, 8 tasks, and 20 detailed tasks., Conclusion: The core direction of the TKM health care model is the region-centered provision of TKM and welfare services. To this end, policy support for the use and linkage of health care service resources is required at the central government level, and linkage and provision of health welfare services centered on TKM are necessary through linkage and convergence between service subjects and between government health care projects., Competing Interests: CONFLICT OF INTEREST The authors declare that there are no conflicts of interest regarding the publication of this paper., (© 2022 Korean Pharmacopuncture Institute.)

Objectives: The aim of this study was to examine the status of community care services regarding traditional Korean medicine (TKM) for older adults and raise awareness on current opinions and services of TKM institutions., Methods: The National Development Institute of Korean Medicine conducted a survey of 16 local governments by sending official letters through an electronic document system from October 2020 to November 2020. The survey items included basic demographic information and information about TKM service., Results: Eleven (68.8%) of the 16 local governments provided TKM home care services. A total of 136 TKM clinics provided home care services for 598 older adults with musculoskeletal disorders. The number of TKM services provided in five or more local governments were cupping 11 (100.0%), acupuncture 11 (100.0%), education and consulting 10 (90.9%), and moxibustion 9 (81.8%). Moreover, pain (recorded on visual analogue scale) and quality of life significantly improved following TKM services ( p < 0.001)., Conclusions: Covered under medical policy, TKM homecare services could function as a viable alternative for continued medical care disrupted during the coronavirus disease 19 pandemic. In addition, standardisation and legalisation of these services could ensure and improve their efficiency.

Semisulcospira gottschei is an Asian endemic species inhabiting Korea and China. However, genetic structure analysis of the resource management of this species has not been performed. To investigate the genetic diversity among populations, microsatellites can be used to determine the geographic origins of marine and freshwater species. This study investigated the genetic structures of the Korean and Chinese populations of S. gottschei based on mitochondrial DNA (mtDNA) Cytochrome oxidase subunit I (COI) and polymorphic microsatellite loci developed from Semisulcospira coreana. Analysis of the mtDNA COI sequence revealed 43 haplotypes, which indicated no gene flow between the Korean and Chinese populations. To further elucidate the genetic structures of the Korean and Chinese populations, the population genetics of S. gottschei were analyzed using nine microsatellite markers. The genetic diversity analysis showed an average of 5.25 alleles per locus, with an average allelic richness of 4.02. Excessive homozygosity was found at all loci, which was expected to be due to the presence of null alleles at all loci. Populations of S. gottschei formed two separate clusters according to pairwise F ST and AMOVA. Also, the UPGMA tree, PCA, STRUCTURE, and GeneClass indicated separation of the 11 populations into two clusters: Korea and China. These results have potential use in the management, restoration, and distinction of the origin country of populations.

Background: An external herbal dispensary (EHD) is a type of pharmacy that provides various types of personalized herbal medicines (PHMs) to other traditional Korean medicine (TKM) institutions. Such dispensaries were legalized by the Ministry of Health and Welfare (MoHW) in 2008 in South Korea. The purpose of this study is to understand the current status of the EHD facilities and their quality controls and compare them with the good manufacture practice (GMP) guidelines to contribute to the establishment of the safety and quality control criteria for PHMs., Methods: We contacted 107 EHD representatives or people in charge of the preparation of PHMs (TKM pharmacists) and invited them to complete a survey questionnaire; of the total, 81 responded. The survey questionnaire was developed in 3 stages: drafting, revision by external experts, and final editing. It consisted of 20 questions covering 3 sections: basic characteristics of EHDs, facility, and quality control. The survey was hosted online from December 2017 to January 2018 as guided by the MoHW., Results: The completion rate was 75.7% (n = 81). In terms of facilities, the five facilities (water supply, manufacture, pest control, hygiene management and warehousing) that corresponded to the legal requirements of EHD were mostly equipped, but the types of facilities and equipment differed. Two facilities (sterilization and cross-contamination that were not legally required for EHD were found to have mostly pharmacopuncture-EHD (P-EHD), but hardly any herbal medicine-EHD (H-EHD). In our findings regarding quality control of non-medicinal herbs, sensory evaluation that included checks for foreign bodies and deterioration were conducted. In terms of the quality control of herbal medicines, residual pesticides and heavy metals tests were performed and for pharmacopuncture, pH, salinity, sterility, and endotoxin tests along with gross examination were performed. In the end, we found that 6 of the 38 standard items as required by the Korea GMP were suitable., Conclusions: In this study, detailed information for each existing EHD law was determined through a nationwide questionnaire. Moreover, the basis for its reflection in additional legal standards should be introduced so that safe herbal medicine can be prepared in EHDs.

The pinnipeds, which comprise seals, sea lions, and walruses, are a remarkable group of marine animals with unique adaptations to semi-aquatic life. However, their genomes are poorly characterized. In this study, we sequenced and characterized the genomes of three pinnipeds (Phoca largha, Callorhinus ursinus, and Eumetopias jubatus), focusing on site-wise sequence changes. We detected rapidly evolving genes in pinniped lineages and substitutions unique to pinnipeds associated with amphibious sound perception. Phenotypic convergence-related sequence convergences are not common in marine mammals. For example, FASN, KCNA5, and IL17RA contain substitutions specific to pinnipeds, yet are potential candidates of phenotypic convergence (blubber, response to hypoxia, and immunity to pathogens) in all marine mammals. The outcomes of this study will provide insight into targets for future studies of convergent evolution or gene function.

To explore the spatio-temporal dynamics of endangered fin whales (Balaenoptera physalus) within the baleen whale (Mysticeti) lineages, we analyzed 148 published mitochondrial genome sequences of baleen whales. We used a Bayesian coalescent approach as well as Bayesian inferences and maximum likelihood methods. The results showed that the fin whales had a single maternal origin, and that there is a significant correlation between geographic location and evolution of global fin whales. The most recent common female ancestor of this species lived approximately 9.88 million years ago (Mya). Here, North Pacific fin whales first appeared about 7.48 Mya, followed by a subsequent divergence in Southern Hemisphere approximately 6.63 Mya and North Atlantic about 4.42 Mya. Relatively recently, approximately 1.76 and 1.42 Mya, there were two additional occurrences of North Pacific populations; one originated from the Southern Hemisphere and the other from an uncertain location. The evolutionary rate of this species was 1.002 × 10 -3 substitutions/site/My. Our Bayesian skyline plot illustrates that the fin whale population has the rapid expansion event since ~ 2.5 Mya, during the Quaternary glaciation stage. Additionally, this study indicates that the fin whale has a sister group relationship with humpback whale (Meganoptera novaeangliae) within the baleen whale lineages. Of the 16 genomic regions, NADH5 showed the most powerful signal for baleen whale phylogenetics. Interestingly, fin whales have 16 species-specific amino acid residues in eight mitochondrial genes: NADH2, COX2, COX3, ATPase6, ATPase8, NADH4, NADH5, and Cytb.

Neural progenitor cells generate various types of neurons and glia in a tightly regulated manner. During primary neurogenesis, retinoic acid (RA) acts earlier than Notch signaling and regulates differentiation and proliferation by upregulating proneural and neurogenic genes in the neural plate. However, the relationship between Notch signaling and the retinoid pathway during late neurogenesis remains unclear. We investigated the role of Mindbomb (Mib)-mediated Notch signaling in the differentiation of neural progenitors during late neurogenesis by overexpressing Mib and administering RA to Tg[hsp70-Mib:EGFP]. The majority of cells in the p3 domain differentiated into GABAergic Kolmer-Agduhr (KA) cells in Tg[hsp70-mib:EGFP] embryos heat-shocked during late neurogenesis, whereas these phenotypes were suppressed by exogenous RA. Our observations suggest that Mib-mediated Notch signaling plays a critical role in the temporal differentiation of neural progenitors, and that the generation of late-born KA″ cells is regulated by the interplay between Mib and RA., (Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.)

Thioredoxin is a multifunctional antioxidant enzyme that belongs to the reductase family. In this study, we cloned and characterized thioredoxin 1 cDNA from the Korean rose bitterling Rhodeus uyekii (RuTrx). The full-length RuTrx cDNA consists of 674 bp with a 324 nt open reading frame (ORF) encoding a 107 aa protein. The deduced RuTrx amino acid sequence indicated a characteristic redox active site, (31)WCGPC(35). Pairwise alignment revealed RuTrx amino acid identity (55.1%-83.2%) with orthologs from various species of mammalia, amphibia, fish and bird. Phylogenetic analysis was conducted to determine the evolutionary position of RuTrx. Expression analysis showed that RuTrx transcripts were present in all of the tissues examined, and was high in the hepatopancreas of R. uyekii. During early development, the expression of RuTrx transcripts was increased. Recombinant RuTrx protein (rRuTrx) was tested for its capacity to serve as an antioxidant enzyme using a metal-catalyzed oxidation (MCO) system. The ability of rRuTrx to protect against supercoiled DNA cleavage due to oxidative nicking increased in a dose-dependent manner. In Raw264.7 cells, Dihydroethidium (DHE) staining for ROS production indicated the antioxidant activity of rRuTrx. Together, these findings suggest that RuTrx may play a role in maintaining the redox state balance in Korean rose bitterling R. uyekii.

The fish Vitellogenin (Vg) gene has been applied as a biomarker for exposure to estrogenic compounds in the aquatic environment. In this study, we cloned and characterized Vg cDNA from the Korean rose bitterling Rhodeus uyekii (Ru-Vg). The Ru-Vg cDNA encodes a 1424-amino-acid polypeptide that belongs to the VgAo1 family and contains a putative signal peptide, lipovitellin I, phosvitin, and lipovitellin II, but does not contain the vWFD domain or the C-terminal peptide. The deduced Ru-Vg protein has high amino acid identity (73.97%-32.17%) with fish Vg proteins. Pairwise alignment and phylogenetic analysis revealed that Ru-Vg is most closely related to Acheilognathus yamatsutae Vg. Ru-Vg transcripts were detected using quantitative polymerase chain reaction in all tissues tested, with the highest level of expression observed in the ovary. Ru-Vg mRNA was upregulated in R. uyekii hepatopancreas cells in response to treatment with 17β-estradiol (E2) or 17α-ethinylestradiol (EE2). Luciferase reporter expression, driven by the 5'-regulatory region of the Ru-Vg gene spanning from -1020 bp to the start codon was induced by the estrogen receptor and was synergistically activated by treatment with E2 or EE2. These results suggest that R. uyekii and the Ru-Vg gene may be useful as biomarkers for exposure to E2 or EE2.

In the present study, a rapid, sensitive and quantitative detection of influenza A virus targeting hemagglutinin (HA) was developed using hybrid structure of quantum dots (QDs) and nanoporous gold leaf (NPGL). NPGL film was prepared by dealloying bimetallic film where its surface morphology and roughness were fairly controlled. Anti-influenza A virus HA antibody (ab66189) was bound with NPGL and amine (-NH2) terminated QDs. These biofunctionalized NPGL and QDs formed a complex with the influenza virus A/Beijing/262/95 (H1N1) and the photoluminescence (PL) intensities of QDs were linearly correlated with the concentrations of the virus up to 1ng/mL while no PL was observed in the absence of the virus, or in bovine serum albumin (BSA, 1µg/mL) alone. In addition, it was demonstrated that this assay detected successfully influenza virus A/Yokohama/110/2009 (H3N2) that is isolated from a clinical sample, at a concentration of ca. 50 plaque forming units (PFU)/mL. This detection limit is 2-order more sensitive than a commercially available rapid influenza diagnostic test. From these results, the proposed assay may offer a new strategy to monitor influenza virus for public health., (Copyright © 2014 Elsevier B.V. All rights reserved.)

For achieve sensitivity in lab-on-a-chip electrochemical detection, more reliable probing methods are required, especially for repeated measurements. Spring-probes are a promising candidate method which can replace needle-like probes and alligator clips that usually produce scratches on the surface of gold electrodes due to the strong physical contacts needed for electrochemical measurements. The superior reliability of amperometric measurements by a spring-probe system was compared with results by conventional probing methods. We demonstrated that a universal spring-probe system would be potentially suitable to achieve high performance in lab-on-a-chip devices using electrochemical detection.

Major habitats for the snow crab Chionoecetes opilio are mostly found within the northwest Atlantic and North Pacific Oceans. However, the East Sea populations of C. opilio, along with its relative the red snow crab (C. japonicas), are two of the most important commercial crustacean species for fisheries on the east coast of the Korean Peninsula. The East Sea populations of C. opilio are facing declining resources due to overfishing and global climate change. Thus, an analysis of population structure is necessary for future management. Five Korean and one Russian group of C. opilio were analyzed using nine microsatellite markers that were recently developed using next-generation sequencing. No linkage disequilibrium was found between any pair of loci, indicating that the markers were independent. The number of alleles per locus varied from 4 to 18 with a mean of 12, and allelic richness per locus ranged from 4.0 to 17.1 across all populations with a mean of 9.7. The Hardy-Weinberg equilibrium test revealed significant deviation in three out of nine loci in some populations after sequential Bonferroni correction and all of them had higher expected heterozygosity than observed heterozygosity. Null alleles were presumed in four loci, which explained the homozygosity in three loci. The pairwise fixation index (F ST ) values among the five Korean snow crab populations did not differ significantly, but all of the pairwise F ST values between each of the Korean snow crab populations and the Russian snow crab population differed significantly. An UPGMA dendrogram revealed clear separation of the Russian snow crab population from the Korean snow crab populations. Assignment tests based on the allele distribution discriminated between Korean and Russian origins with 93 % accuracy. Therefore, the snow crab populations around the Korean Peninsula need to be managed separately from the populations in Bering Sea in global scale resource management. Also, this information can be used for identification of snow crab origin which is problematic in worldwide crab trade.

Noble metal nanostructure allows us to tune optical and electrical properties, which has high utility for real-world application. We studied surface plasmon-induced emission of semiconductor quantum dots (QDs) on engineered metallic nanostructures. Highly passive organic ZnS-capped CdSe QDs were spin-coated on poly-(methyl methacrylate)-covered Ag films, which brought QDs near the metallic surface. We obtained the enhanced electromagnetic field and reduced fluorescence lifetimes from CdSe/ZnS QDs due to the strong coupling of emitter wave function with the Ag plasmon resonance. Observed changes include a six-fold increase in the fluorescence intensity and striking reduction in fluorescence lifetimes of CdSe/ZnS QDs on rough Ag nanoneedle compared to the case of smooth surfaces. The advantages of using those nanocomposites are expected for high-efficiency light-emitting diodes, platform fabrication of biological and environmental monitoring, and high-contrast imaging.

Of the more than 300 octopus species, Octopus minor is one of the most popular and economically important species in Eastern Asia, including Korea, along with O. vulgaris, O. ocellatus, and O. aegina. We developed 19 microsatellite markers from Octopus minor and eight polymorphic markers were developed to analyze the genetic diversity and relationships among four octopus populations from Korea and three from China. The number of alleles per locus varied from 10 to 49, and allelic richness per locus ranged from 2 to 16.4 across all populations. The average allele number among the populations was 11.1, with a minimum of 8.3 and a maximum of 13.6. The mean allelic richness was 8.7 in all populations. The Hardy-Weinberg equilibrium (HWE) test revealed significant deviation in 19 of the 56 single-locus sites, and null alleles were presumed in five of eight loci. The pairwise F ( ST ) values between populations from Korea and China differed significantly in all pairwise comparisons. The genetic distances between the China and Korea samples ranged from 0.161 to 0.454. The genetic distances among the populations from Korea ranged from 0.033 to 0.090, with an average of 0.062; those among populations from China ranged from 0.191 to 0.316, with an average of 0.254. The populations from Korea and China formed clearly separated into clusters via an unweighted pair group method with arithmetic mean dendrogram. Furthermore, a population from muddy flats on the western coast of the Korean Peninsula and one from a rocky area on Jeju Island formed clearly separated subclusters. An assignment test based on the allele distribution discriminated between the Korean and Chinese origins with 96.9 % accuracy.

Pen-2 is a key regulator of the γ-secretase complex, which is involved in the production of the amyloid β (Aβ)-42 peptides, which ultimately lead to Alzheimer's disease (AD). While Pen-2 has been studied in vitro, Pen-2 function in vivo in the brains of transgenic (Tg) mice overexpressing human Pen-2 (hPen-2) protein has not been studied. This study aimed to determine whether Pen-2 overexpression could regulate the AD-like phenotypes in Tg mice. NSE/hPen-2 Tg mice were produced by the microinjection of the NSE/hPen-2 gene into the pronucleus of fertilized eggs. The expression of the hPen-2 gene under the control of the NSE promoter was successfully detected only in the brain and kidney tissue of NSE/hPen-2 Tg mice. Also, 12-month-old NSE/hPen-2 Tg mice displayed behavioral dysfunction in the water maze test, motor activity and feeding behavior dysfunction in food intake/water intake/motor activity monitoring system. In addition, tissue samples displayed dense staining with antibody to the Aβ-42 peptide. Furthermore, NSE/hPen-2 Tg mice exhibiting feeding behavior dysfunction were significantly more apt to display symptoms related to diabetes and obesity. These results suggest that Pen-2 overexpression in NSE/hPen-2 Tg mice may induce all the AD-like phenotypes, including behavioral deficits, motor activity and feeding behavior dysfunction, Aβ-42 peptide deposition and chronic disease induction.

Color variation is a well-known feature of sea cucumbers (Apostichopus japonicus), which are classified into three groups based on their colors of red, green and black. It is also one of the most important traits related to how they taste, and it thereby affects their market price. Attempts were made to identify single-nucleotide polymorphisms (SNPs) and to analyze differences associated with SNP genotypes between green and red color variants using HSP70 as the target gene. The HSP70 gene, which is found universally in organisms from bacteria to humans, is one of the most evolutionarily conserved genes and the most widely studied biomarker of stress response. DNA fragments of 1074 bp covering a partial sequence of the sea cucumber HSP70 gene, were amplified from both red and green variants, and subsequently analyzed for the presence of SNPs. Twenty-seven polymorphic sites in total, including heterozygous sites, were observed. Of these, six sites were found to be significantly different SNP genotypes between green and red variants. Furthermore, PCR with an internal primer designed to include an allele-specific SNP at the 3' end (site 443) showed differentiation between the two variants, 100% and 4.2% amplification in green and red variants, respectively. The validated SNPs may serve as informative genetic markers that can be used to distinguish variants at the early developmental stage, prior to color differentiation., (Copyright © 2011. Published by Elsevier Ltd.)

A localized surface plasmon resonance (LSPR)-based optical biosensor in connection with a multispot copper-capped nanoparticle array (MC-NPA) chip was proposed and developed. The copper (Cu) films, used as a shell, formed a "cap-like" layer on the top of the silica nanoparticles, used as a core, in an orderly fashion, to form the surface called a "Cu-capped nanoparticle array chip". The plasmonic properties of this nanostructure type were initially investigated while controlling the shell thickness of the deposited Cu. Also, we quantified the sensitivity of MC-NPA chip to changes in bulk refractive index (RI). As a result of its LSPR properties, the MC-NPA chip displayed a sensitivity of 67.8 nm per RI unit, and the wavelength shift of the LSPR spectrum peak was sensitive to the RI of the surrounding bulk medium, such as the biomolecular layers. Using MC-NPA chips, multiplex sensing of target DNAs from reference bacteria and clinical samples was possible in a quantitative manner with a detection limit of 10 fM (50 zmol). The optical biosensor developed in this study represents a unique approach to performing LSPR that utilizes a simple and cost-effective optical setup with disposable chips.

Superparamagnetic iron oxide nanoparticles (SPIONs) have been widely utilized for the diagnosis and therapy of specific diseases, as magnetic resonance imaging (MRI) contrast agents and drug-delivery carriers, due to their easy transportation to targeted areas by an external magnetic field. For such biomedical applications, SPIONs must have multifunctional characteristics, including optimized size and modified surface. However, the biofunctionality and biocompatibility of SPIONs with various surface functional groups of different sizes have yet to be elucidated clearly. Therefore, it is important to carefully monitor the cytotoxicity and genotoxicity of SPIONs that are surfaced-modified with various functional groups of different sizes. In this study, we evaluated SPIONs with diameters of approximately 10 nm and 100~150 nm, containing different surface functional groups. SPIONs were covered with -O⁻ groups, so-called bare SPIONs. Following this, they were modified with three different functional groups--hydroxyl (-OH), carboxylic (-COOH), and amine (-NH₂) groups--by coating their surfaces with tetraethyl orthosilicate (TEOS), (3-aminopropyl)trimethoxysilane (APTMS), TEOS-APTMS, or citrate, which imparted different surface charges and sizes to the particles. The effects of SPIONs coated with these functional groups on mitochondrial activity, intracellular accumulation of reactive oxygen species, membrane integrity, and DNA stability in L-929 fibroblasts were determined by water-soluble tetrazolium, 2',7'-dichlorodihydrofluorescein, lactate dehydrogenase, and comet assays, respectively. Our toxicological observations suggest that the functional groups and sizes of SPIONs are critical determinants of cellular responses, degrees of cytotoxicity and genotoxicity, and potential mechanisms of toxicity. Nanoparticles with various surface modifications and of different sizes induced slight, but possibly meaningful, changes in cell cytotoxicity and genotoxicity, which would be significantly valuable in further studies of bioconjugation and cell interaction for drug delivery, cell culture, and cancer-targeting applications.