Thyroid-Friendly Soft Materials as 3D Cell Culture Tool for Stimulating Thyroid Cell Function.

The disruption of thyroid hormones because of chemical exposure is a significant societal problem. Chemical evaluations of environmental and human health risks are conventionally based on animal experiments. However, owing to recent breakthroughs in biotechnology, the potential toxicity of chemicals can now be evaluated using 3D cell cultures. In this study, the interactive effects of thyroid-friendly soft (TS) microspheres on thyroid cell aggregates are elucidated and their potential as a reliable toxicity assessment tool is evaluated. Using state-of-the-art characterization methods coupled with cell-based analysis and quadrupole time-of-flight mass spectrometry, it is shown that TS-microsphere-integrated thyroid cell aggregates exhibit improved thyroid function. Specifically, the responses of zebrafish embryos, which are used for thyroid toxicity analysis, and the TS-microsphere-integrated cell aggregates to methimazole (MMI), a known thyroid inhibitor, are compared. The results show that the thyroid hormone disruption response of the TS-microsphere-integrated thyroid cell aggregates to MMI is more sensitive compared with those of the zebrafish embryos and conventionally formed cell aggregates. This proof-of-concept approach can be used to control cellular function in the desired direction and hence evaluate thyroid function. Thus, the proposed TS-microsphere-integrated cell aggregates may yield new fundamental insights for advancing in vitro cell-based research.

Hydroxyapatite microbeads containing BMP-2 and quercetin fabricated via electrostatic spraying to encourage bone regeneration.

BACKGROUND: Hydroxyapatite (HAp) possesses osteoconductive properties, and its granular form can serve as an effective drug delivery vehicle for bone regeneration. Quercetin (Qct), a plant-derived bioflavonoid, is known to promote bone regeneration; however, its comparative and synergistic effects with the commonly used bone morphogenetic protein-2 (BMP-2) have not been investigated. METHODS: We examined the characteristics of newly formed HAp microbeads using an electrostatic spraying method and analyzed the in vitro release pattern and osteogenic potential of ceramic granules containing Qct, BMP-2, and both. In addition, HAp microbeads were transplanted into a rat critical-sized calvarial defect and the osteogenic capacity was assessed in vivo. RESULTS: The manufactured beads had a microscale size of less than 200 µm, a narrow size distribution, and a rough surface. The alkaline phosphatase (ALP) activity of osteoblast-like cells cultured with the BMP-2-and-Qct-loaded HAp was significantly higher than that of either Qct- or BMP-2-loaded HAp groups. The mRNA levels of osteogenic marker genes such as ALP and runt-related transcription factor 2 were found to be upregulated in the HAp/BMP-2/Qct group compared to the other groups. In micro-computed tomographic analysis, the amount of newly formed bone and bone surface area within the defect was significantly higher in the HAp/BMP-2/Qct group, followed by the HAp/BMP-2 and HAp/Qct groups, which is consistent with the histomorphometrical results. CONCLUSIONS: These results imply that electrostatic spraying can be an efficient strategy to produce homogenous ceramic granules and that the BMP-2-and-Qct-loaded HAp microbeads can serve as effective implants for bone defect healing.

Integration of Bioinspired Fibrous Strands with 3D Spheroids for Environmental Hazard Monitoring.

Numerous methods have been introduced to produce 3D cell cultures that can reduce the need for animal experimentation. This study presents a unique 3D culture platform that features bioinspired strands of electrospun nanofibers (BSeNs) and aquatic cell lines to compensate for shortcomings in the current cell spheroid generation techniques. The use of BSeNs in 3D zebrafish liver cell cultures is found to improve liver and reproductive functions through spheroid-based in vitro assays such as whole transcriptome sequencing and reproductive toxicity testing, with optimized properties exhibiting results similar to those obtained for fish embryo acute toxicity (FET, OECD TG 236) following exposure to environmental endocrine-disrupting chemicals (17ß-Estradiol (E2), 4-hydroxytamoxifen (4-HT), and bisphenol compounds (bisphenol A (BPA) and 9,9-Bis(4-hydroxyphenyl)fluorene (BPFL)). These findings indicate that the beneficial effects of bioinspired materials that closely mimic ECM environments can yield efficient zebrafish cells with intrinsic functions and xenobiotic metabolism similar to those of zebrafish embryos. As a closer analog for the in vivo conditions that are associated with exposure to potentially hazardous chemicals, the straightforward culture model introduced in this study shows promise as an alternative tool that can be used to further eco-environmental assessment.

Objective Verification of Physiologic Changes during Accommodation under Binocular, Monocular, and Pinhole Conditions.

BACKGROUND: To objectively investigate accommodative response to various refractive stimuli in subjects with normal accommodation. METHODS: This prospective, non-randomized clinical trial included 64 eyes of 32 subjects with a mean spherical equivalent -1.4 diopters (D). We evaluated changes in accommodative power, pupil diameter, astigmatic value, and axis when visual stimuli were applied to binocular, monocular (dominant eye, non-dominant eye, ipsilateral, and contralateral), and pinhole conditions. Visual stimuli were given at 0.25 D (4 m), 2 D (50 cm), 3 D (33 cm), and 4 D (25 cm) and accommodative response was evaluated using open view binocular autorefractor/keratometer. RESULTS: The accommodative response to binocular stimulus was 90.9% of the actual refractive stimulus, while that of the monocular stimulus was 84.6%. The binocular stimulus induced a smaller pupil diameter than did the monocular stimulus. There was no difference in accommodative response between the dominant eye and non-dominant eye or between ipsilateral and contralateral stimuli. As the refractive stimuli became stronger, the absolute astigmatic value increased and the direction of the astigmatism axis became more horizontal. Pinhole glasses required 10%-15% less accommodative power compared with the monocular condition. CONCLUSION: Binocular stimuli enable more precise and effective accommodation than do monocular stimuli. Accommodative response is composed of 90% true accommodation and 10% pseudo-accommodation, and the refractive stimulus in one eye affects the contralateral eye to the same extent. This should be taken into account when developing guidelines for wearing smart glasses while driving, as visual stimulation is applied to only one eye, but far distance attention is constantly needed. TRIAL REGISTRATION: ClinicalTrials.gov Identifier: NCT03557346.

Enhancing cartilage regeneration through spheroid culture and hyaluronic acid microparticles: A promising approach for tissue engineering.

Cell therapy using chondrocytes has shown promise for cartilage regeneration, but maintaining functional characteristics during in vitro culture and ensuring survival after transplantation are challenges. Three-dimensional (3D) cell culture methods, such as spheroid culture, and hydrogels can improve cell survival and functionality. In this study, a new method of culturing spheroids using hyaluronic acid (HA) microparticles was developed. The spheroids mixed with HA microparticles effectively maintained the functional characteristics of chondrocytes during in vitro culture, resulting in improved cell survival and successful cartilage formation in vivo following transplantation. This new method has the potential to improve cell therapy production for cartilage regeneration.

Phenology model development for Neodryinus typhlocybae: Evaluation of phenological synchrony with its host, Metcalfa pruinosa.

The invasive species Metcalfa pruinosa has inflicted significant economic losses in various European and Asian regions. To combat this pest, the parasitoid wasp Neodryinus typhlocybae has been effectively introduced in Europe. Despite its success, research on the field occurrence patterns of N. typhlocybae, particularly its phenology, remains scarce. This study aims to develop a degree-day model for predicting the adult emergence of N. typhlocybae from overwintering cocoons and to assess the phenological synchrony between N. typhlocybae adults and the nymphal stages of M. pruinosa in Korea. In this study, we estimated the thermal parameters of N. typhlocybae under field temperatures and six constant temperatures (13.92, 17.71, 18.53, 20.53, 22.78, and 24.03 °C) conditions. The lower developmental temperature was estimated using the values of the coefficient of variation for the cumulative degree days of emerged individual adults. The estimated lower developmental threshold temperature was 12.3 °C. With this developmental threshold, a degree-day model was developed, and this model well-predicted emergence in field conditions. By simulating this developed model with the actual occurrence of the nymphal stages of its host, M. pruinosa, adult wasp emergence was estimated to be 1.5 weeks later than the first instar nymph of the host but faster than other nymphal stages of M. pruinosa. Thus, the findings in this study would be helpful in determining the possibility of establishing N. typhlocybae and improving the management efficiency of M. pruinosa.

142Development of an alginate-gelatin bioink enhancing osteogenic differentiation by gelatin release.

Hydrogels are natural bioink options for cellular printing due to their high-water content and permeable three-dimensional (3D) polymeric structure, which are favorable for cellular anchoring and metabolic activities. To increase the functionality of hydrogels as bioinks, biomimetic components are often incorporated, such as proteins, peptides, and growth factors. In this study, we aimed to enhance the osteogenic activity of a hydrogel formulation by integrating both the release and retention of gelatin so that gelatin serves as both an indirect support for released ink component on cells nearby and a direct support for encapsulated cells inside a printed hydrogel, thereby fulfills two functions. Methacrylate-modified alginate (MA-alginate) was chosen as the matrix because it has a low cell adhesion effect due to the absence of ligands. The gelatin-containing MA-alginate hydrogel was fabricated, and gelatin was found to remain in the hydrogel for up to 21 days. The gelatin remaining in the hydrogel had positive effects on encapsulated cells, especially on cell proliferation and osteogenic differentiation. The gelatin released from the hydrogel affected the external cells, showing more favorable osteogenic behavior than the control sample. It was also found that the MA-alginate/gelatin hydrogel could be used as a bioink for printing with high cell viability. Therefore, we anticipate that the alginate-based bioink developed in this study could potentially be used to induce osteogenesis in bone tissue regeneration.

Support-less ceramic 3D printing of bioceramic structures using a hydrogel bath.

Volumetric bone tissue defects are beyond the intrinsic regenerative capacity of bone tissue. With the recent development of ceramic 3D printing, various bioceramic scaffolds that can induce bone regeneration are being actively developed. However, hierarchical bone is complex, with overhanging structures that require additional sacrificial support during ceramic 3D printing. Not only can this increase the overall process time and material consumption, but breaks and cracks may occur when sacrificial supports are removed from fabricated ceramic structures. In this study, a support-less ceramic printing (SLCP) process using a hydrogel bath was developed to facilitate the manufacture of complex bone substitutes. A hydrogel bath, consisting of pluronic P123 with temperature-sensitive properties, mechanically supported the fabricated structure when the bioceramic ink was extruded into the bath and promoted the cement reaction to cure the bioceramic. SLCP enables the fabrication of complex bone constructs with overhanging structures, such as the mandible and maxillofacial bones, with reduced overall processing time and material consumption. Scaffolds fabricated by SLCP showed more cell adhesion, higher cell growth rate, and osteogenic protein expression due to their rougher surface than conventionally printed scaffolds. Hybrid scaffolds were fabricated by SLCP to co-print cells and bioceramics, and SLCP provided a cell-friendly environment, exhibiting high cell viability. SLCP enables control of the shape of various cells, bioactive substances, and bioceramics and thus can be used as an innovative 3D bioprinting technique to manufacture complex hierarchical bone structures.

Fabrication of color-graded feldspathic dental prosthetics for aesthetic and restorative dentistry.

OBJECTIVE: Feasibility investigation of natural teeth shades replication on dental prosthetics fabricated via functionally graded additive manufacturing (FGAM) using combination of feldspathic porcelain (FP) and yttrium aluminum garnet cerium (Y3Al5O12:Ce, YAG:Ce) as a promising esthetic restoration option. METHODS: Color-graded feldspathic crown fabrication parameter through FGAM method was comprehensively examined from the slurry rheology, cure depth, debinding to sintering temperature. Effect of light absorbent also checked towards overcuring reaction during UV exposure by the shape comparison. Lastly, the flexural bending strength measured following ISO 6872:2015 to assure the applicability. Applying the studied parameter, natural teeth shades then imitated and investigated by alteration of FP and FP + 0.1 wt% YAG:Ce (Y-FP). Generated color across the structure captured through mobile camera, interpreted through the CIELAB coordinate and the gradation confirmed by the color differences (ΔE00) calculated using CIEDE2000 formula. RESULT: Parameter study indicated that 70 wt% of FP slurry with 3 wt% dispersant and 0.2 wt% light absorbent is favored. It produces excellent flowability in our FGAM system with less overcuring justified by edge margin reduction from 95.65° to 90.00° after UV exposure on rectangle shapes masking. The obtain structure also offers adequate flexural bending strength of 106.26 MPa (FP) and 101.36 MPa (Y-FP) after sintering at 780 °C. This validated the materials as class 2 dental prosthetics citing ISO 6872:2015. Color gradation was verified by the yellow b* value reduction (14.8 to -3.33) as it shifted from cervical to incisal area while ΔE00 further affirmed the differences from each segment in comparison with the FP and Y-FP. SIGNIFICANCE: Color gradation was successfully replicated by FP and YAG:Ce composition shift via FGAM technique. This result highlights the potential of FGAM as an alternative for fabricating dental prosthetics with high efficiency and improved esthetic appeal.

Enhancement of properties of a cell-laden GelMA hydrogel-based bioink via calcium phosphate phase transition.

To improve the properties of the hydrogel-based bioinks, a calcium phosphate phase transition was applied, and the products were examined. We successfully enhanced the mechanical properties of the hydrogels by adding small amounts (< 0.5 wt%) of alpha-tricalcium phosphate (α-TCP) to photo-crosslinkable gelatin methacrylate (GelMA). As a result of the hydrolyzing calcium phosphate phase transition involvingα-TCP, which proceeded for 36 h in the cell culture medium, calcium-deficient hydroxyapatite was produced. Approximately 18 times the compressive modulus was achieved for GelMA with 0.5 wt%α-TCP (20.96 kPa) compared with pure GelMA (1.18 kPa). Although cell proliferation decreased during the early stages of cultivation, both osteogenic differentiation and mineralization activities increased dramatically when the calcium phosphate phase transition was performed with 0.25 wt%α-TCP. The addition ofα-TCP improved the printability and fidelity of GelMA, as well as the structural stability and compressive modulus (approximately six times higher) after three weeks of culturing. Therefore, we anticipate that the application of calcium phosphate phase transition to hydrogels may have the potential for hard tissue regeneration.

Disruptive Effects of Two Curcuminoids (Demethoxycurcumin and Bisdemethoxycurcumin) on the Larval Development of Drosophila melanogaster.

Juvenile hormones (JHs) play a central role in insect development, reproduction, and various physiological functions. Curcuminoids generally exhibit a wide range of biological activities, such as antioxidant, anti-inflammatory, antibacterial, and insecticidal, and they exhibit insect growth inhibitory effects. However, research on insecticidal properties of curcuminoids has been limited. Moreover, to the best of our knowledge, studies on JHs of insects and curcuminoids are lacking. Therefore, this study aimed to identify the substances that act as JH disruptors (JHDs) from edible plants. Demethoxycurcumin (DMC) and bisdemethoxycurcumin (BDMC), two curcuminoids from the turmeric plant Curcuma longa L. inhibited the formation of a methoprene-tolerant (Met)-Taiman (Tai) heterodimer complex in Drosophila melanogaster, as shown through in vitro yeast two-hybrid assays. An artificial diet containing 1% (w/v) DMC or BDMC significantly reduced the number of D. melanogaster larvae in a concentration-dependent manner; larval development was disrupted, preventing the progression of larvae to pupal stages, resulting in an absence of adults. Building on the results obtained in this study on curcuminoids, researchers can use our study as a reference to develop eco-friendly pesticides.

Fabrication of multifunctional alginate microspheres containing hydroxyapatite powder for simultaneous cell and drug delivery.

Novel alginate-hydroxyapatite hybrid microspheres were developed for simultaneous delivery of drugs and cells as a multifunctional bone substitute for osteoporotic bone tissue regeneration. The microspheres were used to enhance osteogenesis and to carry and deliver quercetin, a representative phytoestrogen that controls bone tissue regeneration metabolism in osteoporosis patients, through sustained release over a long period. To overcome quercetin's hydrophobicity and low solubility in aqueous environments, we added it to the surface of hydroxyapatite (HAp) nanoparticles before mixing them with an alginate solution. The homogeneous distribution of the HAp nanoparticles in the alginate solution was essential for preventing nozzle clogging and achieving successfully fabricated hybrid microspheres. To this end, a 3D ultrasonic treatment was applied. Electrostatic microencapsulation was then used to fabricate hybrid alginate-HAp microspheres containing quercetin and cells. The microspheres were approximately 290.7 ± 42.5 µm (aspect ratio of 1). The sustained release of quercetin was confirmed during a test period of 20 weeks. The cells in the hybrid microspheres maintained good cell viability during the entire testing period, and their osteogenic differentiation behavior was boosted by the presence of HAp. Thus, osteogenic differentiation could be greatly improved by adding quercetin. These novel multi-biofunctional hybrid microspheres have great potential for the regeneration of osteoporotic bone tissue at indeterminate defect sites.

Assessment of acute and chronic toxicity of cyantraniliprole and sulfoxaflor on honey bee (Apis mellifera) larvae.

BACKGROUND: Recently, cyantraniliprole (CYA) and sulfoxaflor (SUL) have been considered as alternatives to neonicotinoid insecticides. In this study, we evaluated the acute and chronic toxicities of CYA and SUL on honey bee (Apis mellifera L.) larvae reared in vitro. RESULTS: In the acute toxicity test, the following test doses were used to determine the median lethal dose (LD50 ): CYA 0.007, 0.014, 0.028, 0.056 and 0.112 µg larva-1 ; SUL 2.5, 5, 10, 20 and 40 µg larva-1 . In the chronic toxicity test, the following test doses were used to determine the LD50 : CYA 0.00512, 0.0128, 0.032, 0.08 and 0.2 µg larva-1 ; SUL 0.0625, 0.125, 0.25, 0.5 and 1.0 µg larva-1 . The acute LD50 values of CYA and SUL were 0.047 and 11.404 µg larva-1 , respectively. Larvae acutely exposed to SUL had significantly lower body weight than controls, but those exposed to CYA showed no difference. The no observed adverse effect level (NOAEL) and LD50 values of the chronic toxicity tests for each insecticide were 0.00512 and 0.064 µg larva-1 for CYA, and 0.0625 µg larva-1 and 0.212 µg larva-1 for SUL, respectively. Larvae chronically exposed to SUL emerged as bees with deformed wings, reaching adult deformation rates of over 50%; however, CYA had no effect on adult deformation. CONCLUSION: Exposure to CYA increased larval mortality but did not cause any adult deformation, whereas SUL exposure increased pupal mortality and caused wing deformation in newly emerged bees. Our study may be useful for the assessment of pesticide toxicity by providing valuable findings on the effects of these insecticides on honey bee larvae. © 2022 Society of Chemical Industry.

Developmental Toxic Effects of Thiram on Developing Zebrafish (Danio rerio) Embryos.

Thiram, an oxidized dimer of dithiocarbamate, has fungicidal and ectoparasiticidal roles. This study aimed to determine the effects of thiram on the development of zebrafish (ZF) embryos. The developmental toxicity test was performed in accordance with the OECD 236 test guidelines, and ZF embryos were subjected to several thiram concentrations and a DMSO (0.01%) control. Subsequently, embryo mortalities and developmental anomalies were evaluated at different hours post fertilization (hpf). Thiram was highly toxic to ZF, with calculated median lethal concentrations (LC50) of thiram at 48 and 96 h as 13.10 ± 2.17 and 8.87 ± 2.09 µg/L, respectively. Thiram-treated embryos/larvae exhibited a variety of deformities, such as abnormal somites, reduced eye pigment, abnormal tail shape, yolk sac edema, hatching defects, and curved spines, with a median effective concentration (EC50) of 3.88 ± 1.23, 5.04 ± 1.82, 6.23 ± 0.92, 5.24 ± 2.22, 1.39 ± 0.25, and 2.60 ± 0.82 µg/L, respectively. Teratogenic index (TI) values ranged from 1.42 to 6.66 for the scored deformities. At 48 hpf, the average heartbeat of the control group was 177.20 ± 5.63 per minute, while the highest thiram-treated group (40 µg/L) was 99.50 ± 18.12 per minute. In addition, cardiac-related issues, such as pericardial edema and abnormal blood flow, were observed in thiram-treated ZF embryos. Overall, these findings suggest that thiram is teratogenic to ZF.

Multifunctional Calcium-Deficient Hydroxyl Apatite-Alginate Core-Shell-Structured Bone Substitutes as Cell and Drug Delivery Vehicles for Bone Tissue Regeneration.

In this work, we fabricated unique coiled-structured bioceramics contained in hydrogel beads for simultaneous drug and cell delivery using a combination of bone cement chemistry and bioprinting and characterized them. The core of the calcium-deficient hydroxyl apatite (CDHA) contains quercetin, which is a representative phytoestrogen isolated from onions and apples, to control the metabolism of bone tissue regeneration through sustained release over a long period of time. The shell consists of an alginate hydrogel that includes preosteoblast MC3T3-E1 cells. Ceramic paste and hydrogel were simultaneously extruded to fabricate core-shell beads through the inner and outer nozzles, respectively, of a concentric nozzle system based on a material-extruding-based three-dimensional (3D) printing system. The formation of beads and the coiled ceramic core is related to both alginate concentration and printing conditions. The size of the microbeads and the thickness of the coiled structure could be controlled by adjusting the nozzle conditions. The whole process was carried out at physiological conditions (37 °C) to be gentle on the cells. The alginate shell undergoes solidification by cross-linking in CaCl2 or monocalcium phosphate monohydrate (MCPM) solution, while the hardening and cementation of the α-tricalcium phosphate (α-TCP) core to CDHA are subsequently initiated by immersion in phosphate-buffered saline solution. This process replaces the typical sintering of ceramic processing to prevent damage to the hydrogel, cells, and drugs in the beads. The cell-loaded beads were then cultured in cell culture media where the cells could maintain good viability during the entire testing period, which was over 50 days. Cell growth and elongation were observed even in the alginate along the CDHA coiled structure over time. Sustained release of quercetin without any initial burst was also confirmed during a test period of 120 days. These novel structured microbeads with multibiofunctionality can be used as new bone substitutes for hard tissue regeneration in indeterminate defect sites.

Mechano-Responsive Piezoelectric Nanofiber as an On-Demand Drug Delivery Vehicle.

The control over biodistribution and pharmacokinetics is critical to enhance the efficacy and minimize the side effects of therapeutic agents. To address the need for an on-demand drug delivery system for precise control over the release time and the quantity of drugs, we exploited the mechano-responsiveness of piezoelectric poly(vinylidene fluoride-trifluroethylene) (P(VDF-TrFE)) nanofibers for drug delivery applications. The large surface area-to-volume ratio inherent to nanomaterials, together with the transformative piezoelectric properties, allowed us to use the material as an ultrasensitive and mechano-responsive drug delivery platform driven by the direct piezoelectric effect. The intrinsic negative zeta potential of the nanofibers was utilized to electrostatically load cationic drug molecules, where surface potential changes by exogenous mechanical actuation trigger the release of drug molecules. We show that the drug release kinetics of the P(VDF-TrFE) nanofibers depends on the fiber diameter, thus piezoelectric properties. We further demonstrated that the drug release quantity can be tuned by the applied pressure or dose of physiologically safe corporeal shockwaves as a mechanical stimulus in in vitro and ex vivo models. Overall, we demonstrated the utility of piezoelectric electrospun nanofibers for mechano-responsive controlled drug release.

3D Bioprinting of In Vitro Models Using Hydrogel-Based Bioinks.

Coronavirus disease 2019 (COVID-19), which has recently emerged as a global pandemic, has caused a serious economic crisis due to the social disconnection and physical distancing in human society. To rapidly respond to the emergence of new diseases, a reliable in vitro model needs to be established expeditiously for the identification of appropriate therapeutic agents. Such models can be of great help in validating the pathological behavior of pathogens and therapeutic agents. Recently, in vitro models representing human organs and tissues and biological functions have been developed based on high-precision 3D bioprinting. In this paper, we delineate an in-depth assessment of the recently developed 3D bioprinting technology and bioinks. In particular, we discuss the latest achievements and future aspects of the use of 3D bioprinting for in vitro modeling.

Selective Detection of an Infection Biomarker by an Osteo-Friend Scaffold: Development of a Multifunctional Artificial Bone Substitute.

Developments in three-dimensional (3D) printing technologies have led to many potential applications in various biomedical fields, especially artificial bone substitutes (ABSs). However, due to the characteristics of artificial materials, biocompatibility and infection remain issues. Here, multifunctional ABSs have been designed to overcome these issues by the inclusion of a biochemical modality that allows simultaneous detection of an infection biomarker by osteo-friend 3D scaffolds. The developed multifunctional scaffolds consist of calcium-deficient hydroxyapatite (CDHA), which has a similar geometric structure and chemical composition to human bone, and gold nanoparticles (Au NPs), which assists osteogenesis and modulates the fluorescence of labels in their microenvironment. The Au NPs were subsequently conjugated with fluorescent dye-labeled probe DNA, which allowed selective interaction with a specific target biomarker, and the fluorescent signal of the dye was temporally quenched by the Au NP-derived Förster resonance energy transfer (FRET). When the probe DNA unfolded to bind to the target biomarker, the fluorescence signal was recovered due to the increased distance between the dye and Au NPs. To demonstrate this sensing mechanism, a microbial oligonucleotide was selected as a target biomarker. Consequently, the multifunctional scaffold simultaneously facilitated osteogenic proliferation and the detection of the infection biomarker.

Transcriptome-Based Identification of Genes Responding to the Organophosphate Pesticide Phosmet in Danio rerio.

Organophosphate pesticides (OPPs) are one of the most widely used insecticides. OPPs exert their neurotoxic effects by inhibiting acetylcholine esterase (AChE). Most of the gross developmental abnormalities observed in OPP-treated fish, on the other hand, may not be explained solely by AChE inhibition. To understand the overall molecular mechanisms involved in OPP toxicity, we used the zebrafish (ZF) model. We exposed ZF embryos to an OPP, phosmet, for 96 h, and then analyzed developmental abnormalities and performed whole transcriptome analysis. Phenotypic abnormalities, such as bradycardia, spine curvature, and growth retardation, were observed in phosmet-treated ZF (PTZF). Whole transcriptome analysis revealed 2190 differentially expressed genes (DEGs), with 822 and 1368 significantly up-and downregulated genes, respectively. System process and sensory and visual perception were among the top biological pathways affected by phosmet toxicity. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis revealed significant enrichment of metabolic pathways, calcium signaling pathway, regulation of actin cytoskeleton, cardiac muscle contraction, drug metabolism-other enzymes, and phototransduction. Quantitative real-time PCR results of six DEGs agreed with the sequencing data expression profile trend. Our findings provide insights into the consequences of phosmet exposure in ZF, as well as an estimate of the potential risk of OPPs to off-target species.

Horizontal Honey-Bee Larvae Rearing Plates Can Increase the Deformation Rate of Newly Emerged Adult Honey Bees.

Rearing honey bee larvae in vitro is an ideal method to study honey bee larval diseases or the toxicity of pesticides on honey bee larvae under standardized conditions. However, recent studies reported that a horizontal position may cause the deformation of emerged bees. Accordingly, the purpose of this study was to evaluate the emergence and deformation rates of honey bee (Apis mellifera ligustica) larvae reared in horizontal and vertical positions. The study was conducted under the same laboratory conditions with three experimental groups, non-capped or capped horizontal plates and capped vertical plates. However, our results demonstrated that the exhibited adult deformation rates of the horizontal plates were significantly higher (27.8% and 26.1%) than those of the vertical plates (11.9%). In particular, the most common symptoms were deformed wings and an abnormal abdomen in the horizontal plates. Additionally, adults reared on horizontal plates were substantially smaller (10.88 and 10.82 mm) than those on vertical plates (11.55 mm). Considering these conclusions, we suggest that a vertical rearing method is more suitable when considering the deformation rates of the control groups to verify the sublethal effects of pesticides on honey bees.