DNA-Engineerable Ultraflat-Faceted Core-Shell Nanocuboids with Strong, Quantitative Plasmon-Enhanced Fluorescence Signals for Sensitive, Reliable MicroRNA Detection.

There has been enormous interest in understanding and utilizing plasmon-enhanced fluorescence (PEF) with metal nanostructures, but maximizing the enhancement in a reproducible, quantitative manner while reliably controlling the distance between dyes and metal particle surface for practical applications is highly challenging. Here, we designed and synthesized fluorescence-amplified nanocuboids (FANCs) with highly enhanced and controlled PEF signals, and fluorescent silica shell-coated FANCs (FS-FANCs) were then formed to fixate the dye position and increase particle stability and fluorescence signal intensity for biosensing applications. By uniformly modifying fluorescently labeled DNA on Au nanorods and forming ultraflat Ag shells on them, we were able to reliably control the distance between fluorophores and Ag surface and obtained an ∼186 fluorescence enhancement factor with these FANCs. Importantly, FS-FANCs were utilized as fluorescent nanoparticle tags for microarray-based miRNA detection, and we achieved >103-fold higher sensitivity than commercially available chemical fluorophores with 100 aM to 1 pM dynamic range.

Optokinetically Encoded Nanoprobe-Based Multiplexing Strategy for MicroRNA Profiling.

Multiplexed real-time analysis on multiple interacting molecules and particles is needed to obtain information on binding patterns between multiple ligands and receptors, specificity of bond formations, and interacting pairs in a complex medium, often found in chemical and biological systems, and difference in binding affinity and kinetics for different binding pairs in one solution. In particular, multiplexed profiling of microRNA (miRNA) in a reliable, quantitative manner is of great demand for the use of miRNA in cell biology, biosensing, and clinical diagnostic applications, and accurate diagnosis of cancers with miRNA is not possible without detecting multiple miRNA sequences in a highly specific manner. Here, we report a multiplexed molecular detection strategy with optokinetically (OK) coded nanoprobes (NPs) that show high photostability, distinct optical signals, and dynamic behaviors on a supported lipid bilayer (SLB) (OK-NLB assay). Metal NPs with three distinct dark-field light scattering signals [red (R), green (G), and blue (B)] and three different target miRNA half-complements were tethered to a two dimensionally fluidic SLB with mobile (M) or immobile (I) state. In situ single-particle monitoring and normalized RGB analysis of the optokinetically combinatorial assemblies among three M-NPs and three I-NPs with dark-field microscopy (DFM) allow for differentiating and quantifying 9 different miRNA targets in one sample. The OK-NP-based assay enables simultaneous detection of multiple miRNA targets in a highly quantitative, specific manner within 1 h and can be potentially used for diagnosis of different cancer types. We validated the OK-NLB assay with single-base mismatched experiments and HeLa cell-extracted total RNA samples by comparing the assay results to the quantitative reverse transcription polymerase chain reaction (qRT-PCR) results.

Posterior maxillary segmental osteotomy concomitant with sinus lift using a piezoelectric device.

Tooth loss evokes functional and esthetic problems, and adjacent teeth can move if they are not corrected in a timely manner, rendering the problems of restoring the dentition more complex. If the opposing tooth is consequently extruded, several treatment modalities may be adopted, such as occlusal adjustment, crown prosthesis, orthodontic treatment, and segmental osteotomy. Segmental osteotomy is the most aggressive and invasive treatment for restoring a problematic occlusal plane due to the complications, such as hemorrhage, loss of tooth vitality, or necrosis of the segment. Piezoelectric devices have recently been used in oral surgery and implant dentistry because they preserve the soft tissue. In this report, we describe a bilateral posterior segmental osteotomy procedure to correct a collapsed occlusal plane with sinus mucosa elevation, using piezoelectric devices to preserve the integrity of the sinus membrane and the posterior superior alveolar artery.

Epidemiology of Achilles Tendon Rupture in South Korea: Claims Data of the National Health Insurance Service from 2009 to 2017.

BACKGROUND: The incidence of Achilles tendon rupture and its trend has not been studied in Asia. The purpose of this nationwide study was to analyze the trend of incidence and surgical treatment of tendon ruptures in South Korea based on sex, age, and income level of patients, as well as seasonal variation. METHODS: A descriptive epidemiologic study was performed based on the data collected retrospectively from the Korea National Health Insurance Service. Data of all outpatients and inpatients were collected from approximately 52 million residents of South Korea, primarily diagnosed with Achilles tendon rupture from 2009 to 2017. RESULTS: A total of 112,350 patients had Achilles tendon rupture, of which 44,248 patients underwent surgical treatment during the study period. The overall, age-specific, and sex-specific incidence of Achilles tendon rupture and surgical treatment showed an increasing trend. Patients in the age group of 41 to 50 years showed the highest increase in incidence. Regarding season, higher incidence was reported during spring and summer, whereas the lowest incidence was found in winter. Higher income level was associated with increased incidence of the condition. CONCLUSIONS: The incidence of Achilles tendon rupture and surgical treatments increased rapidly in patients between 41 and 50 years of age. Patients in the higher income quintile groups experienced more Achilles tendon injury than those in lower income groups, and fewer ruptures were observed during winter.

Improvement of the osteogenic potential of ErhBMP-2-/EGCG-coated biphasic calcium phosphate bone substitute: in vitro and in vivo activity.

PURPOSE: The aim of this study was to evaluate the enhancement of osteogenic potential of biphasic calcium phosphate (BCP) bone substitute coated with Escherichia coli-derived recombinant human bone morphogenetic protein-2 (ErhBMP-2) and epigallocatechin-3-gallate (EGCG). METHODS: The cell viability, differentiation, and mineralization of osteoblasts was tested with ErhBMP-2-/EGCG solution. Coated BCP surfaces were also investigated. Standardized, 6-mm diameter defects were created bilaterally on the maxillary sinus of 10 male New Zealand white rabbits. After removal of the bony windows and elevation of sinus membranes, ErhBMP-2-/EGCG-coated BCP was applied on one defect in the test group. BCP was applied on the other defect to form the control group. The animals were sacrificed at 4 or 8 weeks after surgery. Histologic and histometric analyses of the augmented graft and surrounding tissue were performed. RESULTS: The 4-week and 8-week test groups showed more new bone (%) than the corresponding control groups (P<0.05). The 8-week test group showed more new bone (%) than the 4-week test group (P<0.05). CONCLUSIONS: ErhBMP-2-/EGCG-coated BCP was effective as a bone graft material, showing enhanced osteogenic potential and minimal side effects in a rabbit sinus augmentation model.

cDNA cloning and characterization of Type I procollagen alpha1 chain in the skate Raja kenojei.

A full-length cDNA of the Type I procollagen alpha1 [pro-alpha1(I)] chain (4388 bp), coding for 1463 amino acid residues in the total length, was determined by RACE PCR using a cDNA library constructed from 4-week embryo of the skate Raja kenojei. The helical region of the skate pro-alpha1(I) chain consisted of 1014 amino acid residues - the same as other fibrillar collagen alpha chains from higher vertebrates. Comparison on denaturation temperatures of Type I collagens from the skate, rainbow trout (Oncorhynchus mykiss) and rat (Rattus norvegicus) revealed that the number of Gly-Pro-Pro and Gly-Gly in the alpha1(I) chains could be directly related to the thermal stability of the helix. The expression property of the skate pro-alpha1(I) chain mRNA and phylogenetic analysis with other vertebrate pro-alpha1(I) chains suggested that skate pro-alpha1(I) chain could be a precursor form of the skate Type I collagen alpha1 chain. The present study is the first evidence for the primary structure of full-length pro-alpha1(I) chain in an elasmobranch.

Interaction of stem cells with nano hydroxyapatite-fucoidan bionanocomposites for bone tissue regeneration.

The combination of bioceramics with biopolymers are playing major role in the construction of artificial bone. Hydroxyapatite (HA) has been extensively studied as a material in bone repair and replacement in last two decades. In the present study, we have prepared the hydroxyapatite-fucoidan (HA-Fucoidan) nanocomposites by in situ chemical method and biologically characterized them for bone graft substitute. Biological results inferred that mineralization effect of HA-F nanocomposites shows significant enhancement compared to HA in adipose derived stem cell (ADSC). It may be due to the addition of fucoidan in the nanocomposites. The important gene expression such as osteocalcin, osteopontin, collagen and runx-2 were checked using ADSC with HA and HA-fucoidan nanocomposites and the results show that the enhancements were found at 7th day. Furthermore, we have performed in vivo study of HA-fucoidan nanocomposites with rabbit model and a slight amount of bone formation was observed in HA-fucoidan nanocomposites. Herewith, we suggest that HA-fucoidan nanocomposites will be good biomaterials for bone repair/replacement in future.

Dietary flounder skin improves growth performance, body composition, and stress recovery in the juvenile black rockfish (Sebastes schlegeli).

This study investigated the relationship between flounder skin meal (FSM) and vitamin C in mediating collagen biosynthesis. Based on the vitamin C requirements (150 mg/kg) of the black rockfish (mean body weight 10.05 ± 0.44 g), a vitamin C level of 400 mg/kg was selected, and 0, 5, 10, or 20% of the casein (purified proteins) in the diet was replaced with FSM. The feeding study was conducted for 8 weeks by using 4 experimental groups. The FSM supplementation resulted in improvement in growth performance, decrease of body lipids. Furthermore, it elevated the HDL-cholesterol levels and total protein content, reduced blood lipids, and led to rapid recovery in stress, which confirm the functionality of FSM with high collagen content.

Morphological Development of Larvae and Juveniles of Acanthopagrus schlegeli.

Newly hatched black porgy larvae (Acanthopagrus schlegeli) swam to the surface, with the mouth and anus still closed and were 1.90-2.11 mm (mean, 2.0 mm) in total length (TL). The larvae were 2.71-2.94 mm TL (mean, 2.82 mm) on day 2 after hatching. At this time, about two-thirds of the yolk was absorbed, the bladder and intestines had formed, and the mouth and anus were open. Total length was 4.32-4.66 mm (mean, 4.45 mm) at the post-larval stage on days 5-6 after hatching, and the yolk and oil globule were almost absorbed. The end of the notochord began to flex, and 6-8 caudal fin rays were visible. The larvae were 15.37-16.1 mm TL (mean, 15.83 mm) at the juvenile stage on days 30-32 after hatching, and the number of rays in all fins was completely revealed.

Biochemical characteristics of four marine fish skins in Korea.

In this study, we investigated the biochemical characteristics of the fish skins of four industrial species: olive flounder (Paralichthys olivaceus), black rockfish (Sebastes schlegeli), sea bass (Lateolabrax maculatus) and red sea bream (Pagrus major). There is high domestic demand in Korea for farming of these fish for human consumption. Crude protein contents in the skin of these fish ranged from 73% to 94% by dry weight; this was in part due to a high content of the structural protein, collagen. Among the four species, olive flounder had the thickest dermal and epidermal layers in the dorsal skin. This species was also associated with the highest extraction ratio of acid-soluble collagen. We also examined whether fish skin could be a cost-effective alternative to current fish meal sources. Our analysis indicates that, when supplemented with additional fish oils and essential amino acids, fish skin is a viable alternative for fish meal formulations.

Surface passivation for single-molecule protein studies.

Single-molecule fluorescence spectroscopy has proven to be instrumental in understanding a wide range of biological phenomena at the nanoscale. Important examples of what this technique can yield to biological sciences are the mechanistic insights on protein-protein and protein-nucleic acid interactions. When interactions of proteins are probed at the single-molecule level, the proteins or their substrates are often immobilized on a glass surface, which allows for a long-term observation. This immobilization scheme may introduce unwanted surface artifacts. Therefore, it is essential to passivate the glass surface to make it inert. Surface coating using polyethylene glycol (PEG) stands out for its high performance in preventing proteins from non-specifically interacting with a glass surface. However, the polymer coating procedure is difficult, due to the complication arising from a series of surface treatments and the stringent requirement that a surface needs to be free of any fluorescent molecules at the end of the procedure. Here, we provide a robust protocol with step-by-step instructions. It covers surface cleaning including piranha etching, surface functionalization with amine groups, and finally PEG coating. To obtain a high density of a PEG layer, we introduce a new strategy of treating the surface with PEG molecules over two rounds, which remarkably improves the quality of passivation. We provide representative results as well as practical advice for each critical step so that anyone can achieve the high quality surface passivation.

DNA-tailored plasmonic nanoparticles for biosensing applications.

The plasmonic properties of metallic nanoparticles (NPs) such as Au and Ag NPs and the plasmonic coupling between them are of enormous interest for their strong and controllable optical signal enhancement and manipulation capabilities. The strong optical properties of these plasmonic structures are promising for various biosensing applications, but the widespread use of these structures is limited largely due to the absence of high-yield synthetic method for targeted nanoprobes with nanometer precision and the poor understanding of the plasmonics of these structures. DNA is a promising material that can be used as both specific biorecognition and versatile synthetic template in forming and controlling plasmonic nanostructures and their aggregations. In this article, we provide an overview and perspective of recent advances in the use of DNA-tailored plasmonic nanostructures in biosensing applications.

Tuning and assembling metal nanostructures with DNA.

Here, we highlight the strategies for the synthesis and tuning of a variety of metal nanostructures and nanoassemblies with oligonucleotides and their applications. We have discussed the importance of and need for various metal nanostructures and the role and suitability of DNA in building these nanostructures and nanoassemblies. A large part of this article is devoted to the discussion of DNA-mediated synthetic methods for metal nanostructures. The synthetic strategies are categorized into two groups - strategy 1 that uses DNA as the ligand for metal nanoparticles and subsequent assembly or modification and strategy 2 that uses DNA template-directed assemblies of metal nanoparticles.

Inhibitory effects of corni fructus extract on angiogenesis and adipogenesis.

Natural products in Chonnam, Korea were screened via anti-angiogenesis experiments, and 1 candidate product was identified, Corni fructus, which exerted dose-dependent inhibitory effects against angiogenesis, adipogenesis, and cell adhesion. C. fructus extract (CFE) exhibits an angiogenesis inhibitory effect superior to that of the EGCG from green tea leaves. The expression level of angiogenesis and adipogenesis-related signal molecules in the western blotting was reduced by increasing the amount of added CFE. Moreover, a diet supplemented with CFE was deemed more effective in inducing weight loss in LB mice than a representative synthetic diet drug, orlistat, which incidently caused the side effect of denuding the mice of their hair. These results indicate that C. fructus may prove to be a useful anti-adipogenic compound, and these in vitro results may be reflected later under in vivo conditions.

Highly uniform and reproducible surface-enhanced Raman scattering from DNA-tailorable nanoparticles with 1-nm interior gap.

An ideal surface-enhanced Raman scattering (SERS) nanostructure for sensing and imaging applications should induce a high signal enhancement, generate a reproducible and uniform response, and should be easy to synthesize. Many SERS-active nanostructures have been investigated, but they suffer from poor reproducibility of the SERS-active sites, and the wide distribution of their enhancement factor values results in an unquantifiable SERS signal. Here, we show that DNA on gold nanoparticles facilitates the formation of well-defined gold nanobridged nanogap particles (Au-NNP) that generate a highly stable and reproducible SERS signal. The uniform and hollow gap (∼1 nm) between the gold core and gold shell can be precisely loaded with a quantifiable amount of Raman dyes. SERS signals generated by Au-NNPs showed a linear dependence on probe concentration (R(2) > 0.98) and were sensitive down to 10 fM concentrations. Single-particle nano-Raman mapping analysis revealed that >90% of Au-NNPs had enhancement factors greater than 1.0 × 10(8), which is sufficient for single-molecule detection, and the values were narrowly distributed between 1.0 × 10(8) and 5.0 × 10(9).

Arachidonic acid-induced expression of the organic solute and steroid transporter-beta (Ost-beta) in a cartilaginous fish cell line.

The organic solute and steroid transporter (OST/Ost) is a unique membrane transport protein heterodimer composed of subunits designated alpha and beta, that transports conjugated steroids and prostaglandin E(2) across the plasma membrane. Ost was first identified in the liver of the cartilaginous fish Leucoraja erinacea, the little skate, and subsequently was found in many other species, including humans and rodents. The present study describes the isolation of a new cell line, LEE-1, derived from an early embryo of L. erinacea, and characterizes the expression of Ost in these cells. The mRNA size and amino acid sequence of Ost-beta in LEE-1 were identical to that previously reported for Ost-beta from skate liver, and the primary structure was identical to that of the spiny dogfish shark (Squalus acanthias) with the exception of a single amino acid. Ost-beta was found both on the plasma membrane and intracellularly in LEE-1 cells, consistent with its localization in other cell types. Interestingly, arachidonic acid, the precursor to eicosanoids, strongly induced Ost-beta expression in LEE-1 cells and a lipid mixture containing arachidonic acid also induced Ost-alpha. Overall, the present study describes the isolation of a novel marine cell line, and shows that this cell line expresses relatively high levels of Ost when cultured in the presence of arachidonic acid. Although the function of this transport protein in embryo-derived cells is unknown, it may play a role in the disposition of eicosanoids or steroid-derived molecules.