The Inhibition Activity of Natural Methoxyflavonoid from Inula britannica on Soluble Epoxide Hydrolase and NO Production in RAW264.7 Cells.

Soluble epoxide hydrolase (sEH) is an enzyme targeted for the treatment of inflammation and cardiovascular diseases. Activated inflammatory cells produce nitric oxide (NO), which induces oxidative stress and exacerbates inflammation. We identify an inhibitor able to suppress sEH and thus NO production. Five flavonoids 1-5 isolated from Inula britannica flowers were evaluated for their abilities to inhibit sEH with IC50 values of 12.1 ± 0.1 to 62.8 ± 1.8 µM and for their effects on enzyme kinetics. A simulation study using computational chemistry was conducted as well. Furthermore, five inhibitors (1-5) were confirmed to suppress NO levels at 10 µM. The results showed that flavonoids 1-5 exhibited inhibitory activity in all tests, with compound 3 exhibiting the most significant efficacy. Thus, in the development of anti-inflammatory inhibitors, compound 3 is a promising natural candidate.

Engineering homoeologs provide a fine scale for quantitative traits in polyploid.

Numerous staple crops exhibit polyploidy and are difficult to genetically modify. However, recent advances in genome sequencing and editing have enabled polyploid genome engineering. The hexaploid black nightshade species Solanum nigrum has immense potential as a beneficial food supplement. We assembled its genome at the scaffold level. After functional annotations, we identified homoeologous gene sets, with similar sequence and expression profiles, based on comparative analyses of orthologous genes with close diploid relatives Solanum americanum and S. lycopersicum. Using CRISPR-Cas9-mediated mutagenesis, we generated various mutation combinations in homoeologous genes. Multiple mutants showed quantitative phenotypic changes based on the genotype, resulting in a broad-spectrum effect on the quantitative traits of hexaploid S. nigrum. Furthermore, we successfully improved the fruit productivity of Boranong, an orphan cultivar of S. nigrum suggesting that engineering homoeologous genes could be useful for agricultural improvement of polyploid crops.

Effects of light wavelength exposure during in vitro blastocyst production on preimplantation development of mouse embryos.

CONTEXT: Despite the absence of light within the body, the application of microscopy during stages of in vitro embryo production has led to the discovery of light irradiation effects on embryo preimplantation development. AIMS: To determine the optimal light irradiation wavelengths at various embryo stages for improving the preimplantation development of mouse embryos and the quality (total cell number) of blastocysts. METHOD: All in vitro procedures of zygote or 2-cell embryo manipulation, embryo monitoring, and culture medium exchange were conducted under visible (390-750nm), blue (445-500nm), green (500-575nm), yellow (575-585nm), or red (620-750nm) light irradiation wavelength. KEY RESULTS: We found that blue, green, and yellow light irradiation during in vitro blastocyst production from zygotes significantly improved blastocyst production and quality, compared to visible and red light irradiation. However, 2-cell embryos exposed to yellow light during in vitro blastocyst production produced significantly more high-quality blastocysts than did 2-cell embryos exposed to visible, blue, green, or red light. After exposure to blue and green - but not yellow - light during in vitro zygote manipulation, yellow light irradiation during embryo monitoring and culture medium exchange triggered significant retardation of preimplantation development. CONCLUSION: These results demonstrate that yellow light irradiation during in vitro blastocyst production, regardless of embryo stage, improves preimplantation development of mouse embryos. IMPLICATIONS: The present study will contribute to produce greater high-quality blastocysts and reduce experimental errors generated by light exposure during mouse embryo-related studies.

Simple oral exercise with chewing gum for improving oral function in older adults.

BACKGROUND: As general and oral health are closely interrelated, promoting oral health may extend a healthy life expectancy. AIMS: To evaluate the long-term effects of simple oral exercise (SOE) and chewing gum exercise on mastication, salivation, and swallowing function in adults aged ≥ 65 years. METHODS: Ninety-six participants were assigned to control, SOE, and GOE (chewing gum exercise with SOE) groups. The SOE comprised exercises to improve mastication, salivation, and swallowing function. Control group participants performed no exercises. The intervention period was 8 weeks, followed by a 3-week maintenance period. The Mixing Ability Index (MAI), occlusal force, unstimulated saliva, and repetitive saliva swallowing test were evaluated at baseline and 2, 5, 8, and 11 weeks later. Self-reported discomfort was re-evaluated after 8 weeks. RESULTS: After 8 weeks, mean MAI differences from baseline significantly increased in both groups; the increase in the GOE group was largest and four times higher than in the control group. Mean differences of occlusal force from baseline increased by 56 N (SOE group) and 60 N (GOE group). The increase of salivation was greater in the SOE (3.6-fold) and GOE (2.2-fold) groups than in the control group. Furthermore, 27% and 18% of SOE and GOE group participants, respectively, were re-categorized as having good swallowing function. Participants reported less discomfort as oral functions improved. DISCUSSION: These findings may facilitate the development of clinical practice guidelines for optimal oral care in older adults. CONCLUSION: While both SOE and GOE may improve oral function in older adults, GOE is recommended for those with impaired mastication. TRIAL REGISTRATION: KCT0003305, retrospectively registered 31/10/2018.

Dog cloning from post-mortem tissue frozen without cryoprotectant.

Successful reproductive cloning depends on obtaining intact donor nuclei from viable cells, ideally isolated by tissue biopsy of a living donor. However, owners and veterinarians often freeze deceased animals, which eventually causes damage to cellular micro-organelles due to the formation of intracellular water crystals. In the present study, we have reported the production of viable cloned puppies using donor nuclei of cells obtained from frozen carcasses. Five cases of deceased and frozen canine specimens were presented to be cloned. Skin fibroblast cell lines were successfully established for four specimens. Significant longer time was needed for the cell growth from frozen tissues (4 days) to reach 80% confluency compared to fresh tissue and frozen tissues frozen for 1- or 2-days. Similarly, SA-ßgal positive cells (death cells) were significantly higher in frozen cells for 2- or 4- days compared to samples from fresh or frozen (1 day) sources. The cloning efficiency (CE) and the pregnancy rates (PR) of frozen cells were lower than those obtained from fresh or living donors (CE 2.4 ± 1.8% vs. 0.6 ± 0.3%, PR 21.7 ± 16.1% vs. 7.7 ± 5.3% for fresh vs. frozen, respectively). Here we demonstrate is the possibility to produce healthy offspring from cell lines obtained from frozen tissue collected post-mortem.

Antimicrobial activity of Curcuma xanthorrhiza nanoemulsions on Streptococcus mutans biofilms.

In this study, an optimal nanoemulsion formulation for Curcuma xanthorrhiza oil (Xan) was investigated using different sonication times. The antimicrobial effects of the nanoemulsion, the original emulsion, distilled water (DW), and Listerine, on Streptococcus mutans biofilms were compared. The optimum ultrasonic time, determined in terms of droplet size and stability, was found to be 10 min. Cell viability was the lowest on exposure to the nanoemulsion, and significantly different compared with exposure to DW or Listerine. The emulsion's effect was similar to that of the nanoemulsion, but was non-uniform with a high interquartile range. Confocal microscope analysis revealed that the live/dead cell ratio in the nanoemulsion was 50% and 40% less than those in DW and Listerine, respectively. Biofilm treated with the nanoemulsion was thinner than biofilms exposed to the other treatments. Xan nanoemulsions exhibited stable and strong antimicrobial effects due to nano-sized particles, highlighting their potential use in oral health treatment.

Unilateral Mastication Evaluated Using Asymmetric Functional Tooth Units as a Risk Indicator for Hearing Loss.

BACKGROUND: Some previous studies reported hearing ability can be reduced by impaired masticatory ability, but there has been little evidence reported of an association between hearing loss and unilateral mastication. Therefore, this study aimed to investigate the relationship between unilateral mastication (UM), estimated from individual functional tooth units (FTUs), and hearing loss in a representative sample of Korean adults. METHODS: The analyzed data were obtained from 1,773 adults aged 40-89 years who participated in Korean national survey. Hearing loss was defined as a pure-tone average of >25 dB at frequencies of 0.5, 1, 2, and 4 kHz in either ear. In each subject, UM was calculated as the difference in the sums of the FTU scores, which is an index of posterior tooth occlusion, on the two sides of the oral cavity. The scores were used to classify the UM into low, moderate, and high. The adjusted odds ratios (aORs) and their 95% confidence intervals (CIs) were calculated in multivariable logistic regression analyses. RESULTS: When controlling for sociodemographic factors, the aOR for hearing loss was 3.12 (95% CI, 1.21-8.03) for high UM relative to low UM. This association remained in a fully-adjusted model containing factors related to noise exposure (aOR 2.88; 95% CI, 1.12-7.46). CONCLUSION: Adults with high UM as measured using FTUs showed a higher occurrence of hearing loss than those with low UM.

Application of powdered activated carbon coating to fabrics in a hybrid filter to enhance mercury removal.

Elemental mercury (Hg0) is predominant constituent of flue gas emitted from coal-fired power plants. Adsorption has been considered the best available technology for removal of Hg0 from flue gas. However, adsorbent injection increases the amount of ash generated. In the present study, powdered activated carbon (PAC) was coated on polytetrafluoroethylene/glass fiber filters to increase Hg0 removal while concurrently reducing the amount of ash generated. The optimal PAC coating rate was determined in laboratory experiments to ensure better Hg0 removal with low pressure drop. When PAC of particle size less than 45 µm was used, and the areal density was 50 g/m2, the pressure drop remained under 30 Pa while the Hg0 removal efficiency increased to 15.8% from 4.3%. The Hg0 removal efficiency also increased with decrease in filtration velocity. The optimal PAC coating rate was applied on a hybrid filter (HF), which was combined with a bag filter and an electrostatic precipitator in a single chamber. Originally designed to remove fine particulates matter, it was retrofitted to the flue gas control device for simultaneous Hg0 removal. By employing the PAC coating, the Hg removal efficiency of the HF increased to 79.79% from 66.35%. Also, a temporary reduction in Hg removal was seen but this was resolved following a cleaning cycle in which the dust layer was removed.

Facile Determination of Sodium Ion and Osmolarity in Artificial Tears by Sequential DNAzymes.

Despite high relevance of tear osmolarity and eye abnormality, numerous methods for detecting tear osmolarity rely upon expensive osmometers. We report a reliable method for simply determining sodium ion-based osmolarity in artificial tears using sequential DNAzymes. When sodium ion-specific DNAzyme and peroxidase-like DNAzyme were used as a sensing and detecting probe, respectively, the concentration of Na⁺ in artificial tears could be measured by absorbance or fluorescence intensity, which was highly correlated with osmolarity over the diagnostic range (R² > 0.98). Our approach is useful for studying eye diseases in relation to osmolarity.

Clinical Assessment of a New Caries Activity Test Using Dental Plaque Acidogenicity in Children under Three Years of Age.

OBJECTIVES: The aim of this study was to assess the validity of a new caries activity test that uses dental plaque acidogenicity in children with deciduous dentition. STUDY DESIGN: Ninety-two children under the age of three years old underwent clinical examination using the dft index and examinations with two caries activity tests. Plaque samples for the new Cariview(®) test and the saliva sample for the conventional Dentocult SM(®) test were collected, incubated, and scored according to each manufacturers' instruction. The data were analysed using ANOVA and Spearman correlation analyses to evaluate the relationships between the test results and the caries experience. RESULTS: The mean dft index of all of the subjects was 4.73, and 17.4% of the subjects were caries-free. The levels of caries risk based on the new Cariview test score significantly increased with the caries experience (p < 0.01). The test results revealed a stronger correlation with caries indices (dft and dt index) than the conventional SM colony counting method (r = 0.43, r = 0.39, p < 0.01). CONCLUSIONS: The new caries activity test to analyse the acidogenic potential of whole microorganisms from dental plaques can be used to evaluate caries risk in children with deciduous teeth.

Development of a high-yield technique to isolate spermatogonial stem cells from porcine testes.

PURPOSE: To date, the methods available for isolating spermatogonial stem cells (SSCs) from porcine testicular cells have a low efficiency of cell separating. Therefore, we tried to develop a novel isolation technique with a high-yield cell separating ability to isolate SSCs from porcine testes. METHODS: We confirmed the presence of SSCs by measuring alkaline phosphatase (AP) activity and SSC-specific gene expression in neonatal porcine testis-derived testicular cells. Subsequently, the isolation of SSCs from testicular cells was performed using different techniques as follows: differential plating (DP), double DP, Petri dish plating post-DP, magnetic-activated cell sorting (MACS), and MACS post-DP. Positive AP staining was used to assess and compare the isolation efficiency of each method. RESULTS: Petri dish plating post-DP resulted in the highest isolation efficiency. The putative SSCs isolated using this method was then further characterized by analyzing the expression of SSC-specific genes and -related proteins, and germ cell-specific genes. OCT4, NANOG, EPCAM, THY1, and UCHL1 were expressed transcriptionally, and OCT4, NANOG, SOX2, TRA-1-60, TRA-1-81, and PLZF were expressed translationally in 86 % of the isolated SSCs. In contrast, no difference was observed in the percentage of cells expressing luteinizing hormone receptor (LHR), a Leydig cell-specific protein, or GATA4, a Sertoli cell-specific protein, between SSCs and negative control cells. In addition, transcriptional expression of VASA, a primordial germ cell-specific marker, and DAZL, a premeiotic germ cell-specific marker, wasn't and was detected, respectively. CONCLUSIONS: We successfully developed a novel high-yield technique to isolate SSCs from porcine testes to facilitate future porcine SSC-related research.

Thin Free Perforator Flap as a Viable Option for Forefoot Reconstruction: Comprehensive Evaluation of Outcomes.

BACKGROUND: Reconstruction of forefoot defects often necessitates free tissue transfer due to the limited availability of local tissue. Microsurgical reconstruction of the forefoot remains challenging as it requires durable yet thin, soft tissue coverage for functional restoration. This study aimed to evaluate the efficacy of free thin perforator flaps for forefoot reconstruction, with a focus on functional outcomes. METHODS: Patients who underwent microsurgical forefoot reconstruction between March 2020 and September 2023 were reviewed. Development of postoperative complications and functional outcomes were evaluated. RESULTS: In total 53 patients were analyzed, with a mean age of 62.7 years. The most common etiology of defects was oncologic surgery, followed by chronic wound, with a majority involving the plantar side. Super-thin anterolateral thigh flap and superficial circumflex iliac artery perforator flap were predominantly used. The median thickness of the flap was 4.0 mm. Digital vessels were the most frequently used as recipients. All but one flap survived, resulting in a success rate of 98.1 percent. Postoperative flap-site complications developed in 20 patients, the majority of which were resolved with conservative treatment. The median hospital stay was 8.0 days, and the mean time for initiating weight-bearing ambulation was 12.4 days. In postoperative foot function index questionnaire survey, the overall score remained 9.41, showing minimal impairment, and did not differ according to defect size and locations. Secondary debulking operation was necessitated in seven patients. CONCLUSION: Use of thin free perforator flaps might provide reliable outcomes with rapid recovery and excellent postoperative function in the forefoot reconstruction.

Distinguishing discolored caries lesions using biofluorescence and dental bleaching: an in vitro simulation model study: Discolored caries lesion using biofluorescence and dental bleaching.

BACKGROUND: Distinguishing between discoloration caused by caries and organic stains is challenging for dentists in clinical settings. Biofluorescence (BF)-bleaching assesses caries lesions by evaluating BF changes after removing organic stains through dental bleaching, leaving cariogenic discoloration. This study aimed to apply BF-bleaching to a simulation model mimicking cariogenic discoloration and compare the BF color changes between organic staining and cariogenic discoloration. METHODS: Thirty artificial caries lesions in bovine incisors were equally divided into three groups: non-stained (NS), organic-stained (OS), and cariogenic-stained (CS) groups. The specimens were treated with bleaching agent, then BF color of each specimen was evaluated using red BF intensity (ΔR), BF hue angle (h°), and hyperspectral BF spectrum. RESULTS: The ΔR of CS was approximately 2.74 and 1.73 times higher than that of OS, at baseline and after bleaching for 20 minutes, respectively. After 20 min of bleaching, the h° of CS increased by approximately 8.1° compared to the baseline, while maintaining the red BF hue range (345‒15°). In contrast, the BF hue of OS shifted from orange (15‒45°) to yellow (45‒75°) simultaneously, and the h° change was approximately 21.9°. Both CS and OS exhibited first emission peaks near 515 nm, and CS showed second peaks in the red range (620‒780 nm). After bleaching, the first peaks were restored to the sound enamel direction (peak at 486 nm), whereas the second peak of red BF in CS was maintained. CONCLUSION: Applying BF-bleaching to discolored caries lesions allowed differentiation between cariogenic discoloration and organic staining based on BF color changes.

Detection of pit and fissure sealant microleakage using quantitative light-induced fluorescence technology: an in vitro study.

This in vitro study aimed to evaluate the feasibility of quantitative light-induced fluorescence (QLF) technology for detecting the presence and severity of microleakage of pit and fissure sealants. The areas of interest (AOIs) were 160 pits and fissures of 40 extracted permanent teeth. Fluorescent images were acquired using a QLF device, and the maximum fluorescence loss ΔFmax of each AOI was analyzed. After staining and cross-sectioning of the teeth, histological dye penetration was scored on a scale of 0 to 3. The relationship between ΔFmax and microleakage depth was analyzed, and the areas under the curve (AUCs) were calculated. The │ΔFmax│ increased as microleakage depth increased. The ΔFmax values of microleakage areas showed a strong significant correlation with the histological scores of dye penetration (r = - 0.72, P = 0.001). AUC analysis showed a high diagnostic accuracy for microleakage depth (AUC = 0.83-0.91). The highest AUC of 0.91 was found when differentiating the outer half microleakage of the sealant (histological score 0 vs. 1-3). QLF technology is effective in assessing the presence and severity of microleakage, suggesting its potential for noninvasive detection and monitoring of sealant microleakage in clinical settings.

Association of cranial base suture/synchondrosis fusion with severity of increased intracranial pressure in Crouzon syndrome.

This study investigated how the fusion states of the cranial base is related to the degree of increased intracranial pressure (ICP) in patients with Crouzon syndrome. This retrospective cohort study enrolled patients who were diagnosed with Crouzon syndrome between May 2007 and April 2022. We categorized the patients into three groups: A, B, and C, according to the severity of increased ICP and the number of cranial vault remodeling procedures for corrective operation. The preoperative fusion states of the cranial base sutures/synchondroses were examined using facial bone computed tomography and compared between groups. Overall, 22 patients were included in Groups A, B, and C, including 8, 7, and 7 patients, respectively. The preoperative average grades of the total cranial base suture/synchondrosis fusion appeared to significantly increase with severity, except for the frontoethmoidal suture, which showed the opposite tendency. In the subgroup analysis, frontosphenoidal, sphenoparietal, sphenosquamosal, parietomastoid, and occipitomastoid suture and petro-occipital synchondrosis were associated with earlier fusion in the more severe group. Premature closure of the cranial base sutures/synchodroses seems to be associated with increased ICP severity in patients with Crouzon syndrome. Precise evaluation of minor sutures/synchondroses at the first visit might help build subsequent operative plans and predict disease prognosis.

Coix Sprouts Affect Triglyceride Metabolism in Huh7 Cells and High-Fat Diet-Induced Obese Mice.

Lipolysis is the hydrolysis of triglycerides (TGs), commonly known as fats. Intracellular lipolysis of TG is associated with adipose triglyceride lipase (ATGL), which provides fatty acids during times of metabolic need. The aim of this study was to determine whether Coix lacryma-jobi L. var. ma-yuen Stapf (Coix) sprouts (CS) can alleviate obesity through lipolysis. Overall, we investigated the potential of CS under in vitro and in vivo conditions and confirmed the underlying mechanisms. Huh7 cells were exposed to free fatty acids (FFAs), and C57BL/6J mice were fed a 60% high-fat diet. When FFA were introduced into Huh7 cells, the intracellular TG levels increased within the Huh7 cells. However, CS treatment significantly reduced intracellular TG levels. Furthermore, CS decreased the expression of Pparγ and Srebp1c mRNA and downregulated the mutant Pnpla3 (I148M) mRNA. Notably, CS significantly upregulated ATGL expression. CS treatment at a dose of 200 mg/kg/day resulted in a significant and dose-dependent decrease in body weight gain and epididymal adipose tissue weight. Specifically, the group treated with CS (200 mg/kg/day) exhibited a significant modulation of serum lipid biomarkers. In addition, CS ameliorated histological alterations in both the liver and adipose tissues. In summary, CS efficiently inhibited lipid accumulation through the activation of the lipolytic enzyme ATGL coupled with the suppression of enzymes involved in TG synthesis. Consequently, CS show promise as a potential anti-obesity agent.

Anti-biofilm activity of a novel nanoemulsion containing Curcuma xanthorrhiza oil.

OBJECTIVES: We aimed to solubilize Curcuma xanthorrhiza oil (CXO) using nanoemulsification and evaluate its inhibitory effects against biofilm formation. METHODS: The components of CXO were evaluated through high-performance liquid chromatography (HPLC) analysis. Healthy human saliva was inoculated onto hydroxyapatite discs to form microcosm biofilms for four days and treated six times with each antimicrobial agent: distilled water (DW), CXO emulsion (EM), CXO nanoemulsion (NE), and positive controls (Listerine and chlorhexidine). Biofilm fluorescence imaging was performed using quantitative light-induced fluorescence, and cell viability and dry-weight measurements were obtained. We compared the bacterial cell and extracellular polysaccharide (EPS) biovolume and thickness using confocal laser scanning microscopy (CLSM). RESULTS: HPLC analysis revealed that CXO was composed of approximately 47% xanthorrhizol. Compared with DW, NE exhibited significantly lower red fluorescence intensity and area (42% and 37%, p < 0.001 and p < 0.001, respectively), and reduced total and aciduric bacterial cell viability (7.3% and 3.9%, p < 0.001, p = 0.01, respectively). Furthermore, the bacterial cell and EPS biovolume and thickness in NE decreased by 40-80% compared to DW, similar to chlorhexidine. Conversely, EM showed a significant difference only in cell viability against total bacteria when compared with DW (p = 0.003), with EPS biovolume and thickness exhibiting higher values than DW. CONCLUSIONS: Nanoemulsification successfully solubilized CXO and demonstrated superior anti-biofilm effects compared to the emulsion form. CLINICAL SIGNIFICANCE: These findings suggest the potential use of NE as a novel antimicrobial agent for preventing oral diseases.

Real-time optical detection of endodontic infection using bacterial autofluorescence.

OBJECTIVES: For successful root canal treatment (RCT), it is essential to objectively assess the presence and activity of bacteria in the root canal system. However, current methods rely on subjective observations of root canal exudates. This study aimed to confirm whether real-time optical detection using bacterial autofluorescence can evaluate endodontic infection status by assessing the red fluorescence (RF) detected from root canal exudates. METHODS: During RCT, endodontic paper points were used to collect root canal exudates scored using conventional organoleptic tests to assess the severity of root canal infections. RF on the paper points was assessed using quantitative light-induced fluorescence (QLF) technology. RF intensity and area from the paper points were quantified, and their correlations with infection severity were assessed using their organoleptic scores. The oral microbiome composition of RF samples was compared with non-red fluorescent (non-RF) samples. RESULTS: The RF detection rate was nil and >98% in the non-infectious and severe groups. The RF intensity and area significantly increased with infection severity (p<0.001) and showed strong correlations with organoleptic scores (r=0.72, 0.82, respectively). The diagnostic accuracy for detecting root canal infection using RF intensity was good to excellent (AUC = 0.81-0.95) and increased with infection severity. The microbial diversity of the RF samples was significantly lower than that of the non-RF samples. Gram-negative anaerobic bacteria such as Prevotella and Porphyromonas were more predominant in RF samples. CONCLUSIONS: Optical detection using bacterial autofluorescence can objectively evaluate endodontic infection status in real-time by assessing the RF of endodontic root canal exudates. CLINICAL SIGNIFICANCE: This real-time optical technology can be utilised to detect endodontic bacterial infection without conventional incubation, allowing clinicians to determine the endpoint of chemomechanical debridement and increase the positive outcomes of RCTs.

Effects of cumulus cells on the in vitro cytoplasmic maturation of immature oocytes in pigs.

The exposure of cumulus cells to nuclear matured oocytes can be regulated through the forced delay of nuclear maturation or the alteration of in vitro maturation (IVM) time in cumulus-oocyte complexes (COCs). However, to date, no evidence has been presented for the enhancement of cytoplasmic maturation by them, indicating irrelevance of cumulus cells in cytoplasmic maturation. Therefore, in order to identify the requirement of cumulus cells in achieving the cytoplasmic maturation of immature oocytes, this study investigated the effects of cumulus cells on the in vitro cytoplasmic maturation of oocytes within COCs derived from porcine medium antral follicles (MAFs) post-the completion of nuclear maturation. For these, with IVM of COCs for 44 h (control), cumulus cell-free oocytes with completed nuclear maturation were in-vitro-matured additionally for 0, 6, or 12 h, and then a variety of factors representing the cytoplasmic maturation of oocytes were analyzed and compared. As the results, the IVM of COCs for 32 h showed complete nuclear maturation and incomplete cytoplasmic maturation. Moreover, after the removal of cumulus cells from COCs with the completion of nuclear maturation, IVM for an additional 6 or 12 h resulted in significant increases in the size of the perivitelline space, the proportion of oocytes with a normal intracellular mitochondrial distribution and a normal round first polar body, and the preimplantation development into the 2-cell and blastocyst stages after parthenogenetic activation. Simultaneously, they showed significant reduction in the level of intracellular reactive oxygen species and no significant differences in the total number of blastocysts. Furthermore, oocytes obtained by this approach did not significantly differ from control oocytes produced by IVM of COCs for 44 h. Our results demonstrate that the cumulus cells enclosing COCs derived from porcine MAFs are not essential for the completion of cytoplasmic maturation after complete nuclear maturation by COCs.

Comparison of Quality, Antioxidant Capacity, and Anti-Inflammatory Activity of Adlay [Coix lacryma-jobi L. var. ma-yuen (Rom. Caill.) Stapf.] Sprout at Several Harvest Time.

Recently, there has been a growing interest in the consumption of plant-based foods such as vegetables and grains for the purpose of disease prevention and treatment. Adlay seeds contain physiologically active substances, including coixol, coixenolide, and lactams. In this study, adlay sprouts were cultivated and harvested at various time points, specifically at 3, 5, 7, 9, and 11 days after sowing. The antioxidant activity of the extracts was evaluated using assays such as DPPH radical scavenging, ABTS radical scavenging, reducing power, and total polyphenol contents. The toxicity of the extracts was assessed using cell culture and the WST-1 assay. The aboveground components of the sprouts demonstrated a significant increase in length, ranging from 2.75 cm to 21.87 cm, weight, ranging from 0.05 g to 0.32 g, and biomass, ranging from 161.4 g to 1319.1 g, as the number of days after sowing advanced, reaching its peak coixol content of 39.38 mg/g on the third day after sowing. Notably, the antioxidant enzyme activity was highest between the third and fifth days after sowing. Regarding anti-inflammatory activity, the inhibition of cyclooxygenase 2 (COX-2) expression was most prominent in samples harvested from the ninth to eleventh days after sowing, corresponding to the later stage of growth. While the overall production mass increased with the number of days after sowing, considering factors such as yield increase index per unit area, turnover rate, and antioxidant activity, harvesting at the early growth stage, specifically between the fifth and seventh days after sowing, was found to be economically advantageous. Thus, the quality, antioxidant capacity, and anti-inflammatory activity of adlay sprouts varied depending on the harvest time, highlighting the importance of determining the appropriate harvest time based on the production objectives. This study demonstrates the changes in the growth and quality of adlay sprouts in relation to the harvest time, emphasizing the potential for developing a market for adlay sprouts as a new food product.