Intelligent Bladder Volume Monitoring for Wearable Ultrasound Devices: Enhancing Accuracy Through Deep Learning-Based Coarse-to-Fine Shape Estimation.

Accurate and continuous bladder volume monitoring is crucial for managing urinary dysfunctions. Wearable ultrasound (US) devices offer a solution by enabling noninvasive and real-time monitoring. Previous studies have limitations in power consumption and computation cost or quantitative volume estimation capability. To alleviate this, we present a novel pipeline that effectively integrates conventional feature extraction and deep learning (DL) to achieve continuous quantitative bladder volume monitoring efficiently. Particularly, in the proposed pipeline, bladder shape is coarsely estimated by a simple bladder wall detection algorithm in wearable devices, and the bladder wall coordinates are wirelessly transferred to an external server. Subsequently, a roughly estimated bladder shape from the wall coordinates is refined in an external server with a diffusion-based model. With this approach, power consumption and computation costs on wearable devices remained low, while fully harnessing the potential of DL for accurate shape estimation. To evaluate the proposed pipeline, we collected a dataset of bladder US images and RF signals from 250 patients. By simulating data acquisition from wearable devices using the dataset, we replicated real-world scenarios and validated the proposed method within these scenarios. Experimental results exhibit superior improvements, including +9.32% of IoU value in 2-D segmentation and -22.06 of RMSE in bladder volume regression compared to state-of-the-art (SOTA) performance from alternative methods, emphasizing the potential of this approach in continuous bladder volume monitoring in clinical settings. Therefore, this study effectively bridges the gap between accurate bladder volume estimation and the practical deployment of wearable US devices, promising improved patient care and quality of life.

Kalkitoxin attenuates calcification of vascular smooth muscle cells via RUNX-2 signaling pathways.

BACKGROUND: Kalkitoxin (KT) is an active lipopeptide isolated from the cyanobacterium Lyngbya majuscula found in the bed of the coral reef. Although KT suppresses cell division and inflammation, KT's mechanism of action in vascular smooth muscle cells (VSMCs) is unidentified. Therefore, our main aim was to investigate the impact of KT on vascular calcification for the treatment of cardiovascular disease. OBJECTIVES: Using diverse calcification media, we studied the effect of KT on VSMC calcification and the underlying mechanism of this effect. METHODS: VSMC was isolated from the 6 weeks ICR mice. Then VSMCs were treated with different concentrations of KT to check the cell viability. Alizarin red and von Kossa staining were carried out to examine the calcium deposition on VSMC. Thoracic aorta of 6 weeks mice were taken and treated with different concentrations of KT, and H and E staining was performed. Real-time polymerase chain reaction and western blot were performed to examine KT's effect on VSMC mineralization. Calcium deposition on VSMC was examined with a calcium deposition quantification kit. RESULTS: Calcium deposition, Alizarin red, and von Kossa staining revealed that KT reduced inorganic phosphate-induced calcification phenotypes. KT also reduced Ca++-induced calcification by inhibiting genes that regulate osteoblast differentiation, such as runt-related transcription factor 2 (RUNX-2), SMAD family member 4, osterix, collagen 1α, and osteopontin. Also, KT repressed Ca2+-induced bone morphogenetic protein 2, RUNX-2, collagen 1α, osteoprotegerin, and smooth muscle actin protein expression. Likewise, Alizarin red and von Kossa staining showed that KT markedly decreased the calcification of ex vivo ring formation in the mouse thoracic aorta. CONCLUSIONS: This experiment demonstrated that KT decreases vascular calcification and may be developed as a new therapeutic treatment for vascular calcification and arteriosclerosis.

Multi-Task and Few-Shot Learning-Based Fully Automatic Deep Learning Platform for Mobile Diagnosis of Skin Diseases.

Fluorescence imaging-based diagnostic systems have been widely used to diagnose skin diseases due to their ability to provide detailed information related to the molecular composition of the skin compared to conventional RGB imaging. In addition, recent advances in smartphones have made them suitable for application in biomedical imaging, and therefore various smartphone-based optical imaging systems have been developed for mobile healthcare. However, an advanced analysis algorithm is required to improve the diagnosis of skin diseases. Various deep learning-based algorithms have recently been developed for this purpose. However, deep learning-based algorithms using only white-light reflectance RGB images have exhibited limited diagnostic performance. In this study, we developed an auxiliary deep learning network called fluorescence-aided amplifying network (FAA-Net) to diagnose skin diseases using a developed multi-modal smartphone imaging system that offers RGB and fluorescence images. FAA-Net is equipped with a meta-learning-based algorithm to solve problems that may occur due to the insufficient number of images acquired by the developed system. In addition, we devised a new attention-based module that can learn the location of skin diseases by itself and emphasize potential disease regions, and incorporated it into FAA-Net. We conducted a clinical trial in a hospital to evaluate the performance of FAA-Net and to compare various evaluation metrics of our developed model and other state-of-the-art models for the diagnosis of skin diseases using our multi-modal system. Experimental results demonstrated that our developed model exhibited an 8.61% and 9.83% improvement in mean accuracy and area under the curve in classifying skin diseases, respectively, compared with other advanced models.

Global well-posedness and pattern formations of the immune system induced by chemotaxis.

This paper studies a reaction-diffusion-advection system describing a directed movement of immune cells toward chemokines during the immune process. We investigate the global solvability of the model based on the bootstrap argument for minimal chemotaxis models. We also examine the stability of nonconstant steady states and the existence of periodic orbits from theoretical aspects of bifurcation analysis. Through numerical simulations, we observe the occurrence of steady or time-periodic pattern formations.

Smartphone-based spectral imaging otoscope: System development and preliminary study for evaluation of its potential as a mobile diagnostic tool.

We develop a novel smartphone-based spectral imaging otoscope for telemedicine and examine its capability for the mobile diagnosis of middle ear diseases. The device was applied to perform spectral imaging and analysis of an ear-mimicking phantom and a normal and abnormal tympanic membrane for evaluation of its potential for the mobile diagnosis. Spectral classified images were obtained via online spectral analysis in a remote server. The phantom experimental results showed that it allowed us to distinguish four different fluids located behind a semitransparent membrane. Also, in the spectral classified images of normal ears (n = 3) and an ear with chronic otitis media (n = 1), the normal and abnormal regions in each ear could be quantitatively distinguished with high contrast. These preliminary results thus suggested that it might have the potentials for providing quantitative information for the mobile diagnosis of various middle ear diseases.

White matter connectivity differences between treatment responders and non-responders in patients with panic disorder.

BACKGROUND: Panic disorder (PD) is a prevalent and highly disabling mental condition. However, less is known about relationships between biomarkers that may together predict a better response to pharmacological treatment. The objective of the present study was to compare the brain white matter (WM) connectivity between treatment-responsive patients with panic disorder (RPD) and non-responsive patients with panic disorder (NRPD) after 12 weeks of pharmacotherapy. METHODS: Sixty-four patients with PD were enrolled in this study (RPD, n = 37; NRPD, n = 27). All patients were examined by using magnetic resonance imaging at 3 Tesla. The Panic Disorder Severity Scale (PDSS), Albany Panic and Phobia Questionnaire (APPQ), Anxiety Sensitivity Inventory-Revised (ASI-R), Beck Anxiety Inventory (BAI), and Beck Depression Inventory (BDI) were administered at baseline of the study. Fractional anisotropy (FA) data were compared using tract-based spatial statistics (TBSS). RESULTS: TBSS results showed that the FA values of the patients with NRPD were significantly higher than of those with RPD in the WM regions such as the precentral gyrus, parahippocampal gyrus, posterior corona radiata, posterior thalamic radiation, posterior parts of the corpus callosum, and precuneus. Symptom severity scales, such as ASI-R scores, showed significant positive correlations of the FA values with the fronto-temporal WM regions in NRPD. CONCLUSIONS: These results suggest that structural changes to areas such as the fronto-limbic regions and the posterior part of default mode network, could influence medication response in PD. Further studies with a larger number of patients should be performed to confirm our findings.

Multimodal endoscopic system based on multispectral and photometric stereo imaging and analysis.

We propose a multimodal endoscopic system based on white light (WL), multispectral (MS), and photometric stereo (PS) imaging for the examination of colorectal cancer (CRC). Recently, the enhancement of the diagnostic accuracy of CRC colonoscopy has been reported; however, tumor diagnosis for a variety of lesion types remains challenging using current endoscopy. In this study, we demonstrate that our developed system can simultaneously discriminate tumor distributions and provide three-dimensional (3D) morphological information about the colon surface using the WL, MS, and PS imaging modalities. The results demonstrate that the proposed system has considerable potential for CRC diagnosis.

Smartphone-based multispectral imaging and machine-learning based analysis for discrimination between seborrheic dermatitis and psoriasis on the scalp.

For appropriate treatment, accurate discrimination between seborrheic dermatitis and psoriasis in a timely manner is crucial to avoid complications. However, when they occur on the scalp, differential diagnosis can be challenging using conventional dermascopes. Thus, we employed smartphone-based multispectral imaging and analysis to discriminate between them with high accuracy. A smartphone-based multispectral imaging system, suited for scalp disease diagnosis, was redesigned. We compared the outcomes obtained using machine learning-based and conventional spectral classification methods to achieve better discrimination. The results demonstrated that smartphone-based multispectral imaging and analysis has great potential for discriminating between these diseases.

Thymol inhibits RANKL-induced osteoclastogenesis in RAW264.7 and BMM cells and LPS-induced bone loss in mice.

Thymol was identified as one of key compounds contributing to the aroma of thyme leaves. We investigated the effects of thymol on receptor activator NF-κB ligand (RANKL)-induced osteoclastogenesis in murine macrophage RAW264.7 cells and bone marrow derived macrophage (BMMs) cells and lipopolysaccharide (LPS)-induced bone loss in vivo. Thymol markedly reduced RANKL-stimulated osteoclast formation and differentiation in RAW264.7 cells and BMMs cells without any cytotoxic effects. The in vitro and in vivo osteoclastogenesis inhibitory effect of thymol was assessed by calculating the quantity of TRAP (+) multinucleated cells and its inhibitory effects on the resorbing capacity were examined on calcium phosphate-coated plates. Moreover, the inhibitory effects of thymol resulted in a reduction of RANK, cathepsin K, matrix metalloproteinase-9 (MMP-9), dendritic cell-specific transmembrane protein (DC-STAMP), c-terminal myc kinase (C-MYC), C-terminal Src kinase (C-SRC), GRB2-associated-binding protein 2 (GAB2), microphthalmia-associated transcription factor (MITF), and carbonic anhydrase II genes. Similarly, activities of ERK, JNK and AKT and protein expressions of NFATc1, C-FOS, MMP-9 and cathepsin K were downregulated by thymol. More importantly, the application of thymol significantly reduced LPS-induced inflammatory bone loss in mice. In conclusion, these findings identified that thymol could be a useful therapeutic agent for the prevention of bone destructive diseases.

Mathematical modeling and its analysis for instability of the immune system induced by chemotaxis.

In this paper, we study how chemotaxis affects the immune system by proposing a minimal mathematical model, a reaction-diffusion-advection system, describing a cross-talk between antigens and immune cells via chemokines. We analyze the stability and instability arising in our chemotaxis model and find their conditions for different chemotactic strengths by using energy estimates, spectral analysis, and bootstrap argument. Numerical simulations are also performed to the model, by using the finite volume method in order to deal with the chemotaxis term, and the fractional step methods are used to solve the whole system. From the analytical and numerical results for our model, we explain not only the effective attraction of immune cells toward the site of infection but also hypersensitivity when chemotactic strength is greater than some threshold.

Smartphone-based multispectral imaging: system development and potential for mobile skin diagnosis.

We investigate the potential of mobile smartphone-based multispectral imaging for the quantitative diagnosis and management of skin lesions. Recently, various mobile devices such as a smartphone have emerged as healthcare tools. They have been applied for the early diagnosis of nonmalignant and malignant skin diseases. Particularly, when they are combined with an advanced optical imaging technique such as multispectral imaging and analysis, it would be beneficial for the early diagnosis of such skin diseases and for further quantitative prognosis monitoring after treatment at home. Thus, we demonstrate here the development of a smartphone-based multispectral imaging system with high portability and its potential for mobile skin diagnosis. The results suggest that smartphone-based multispectral imaging and analysis has great potential as a healthcare tool for quantitative mobile skin diagnosis.

Improvement of pregnancy rate by intrauterine administration of dexamethasone and recombinant human leukemia inhibitory factor at the time of embryo transfer in cattle.

Bovine embryos (day 5) were cultured to day 10 with or without 100 ng/mL PGF2α in medium supplemented with control; 100 nM Dex; 1,000 U/mL recombinant human leukemia inhibitory factor (rhLIF); or Dex+rhLIF. Although the rates to development to the blastocyst were not significantly different among groups, the hatching rate after additional culture with Dex +/or rhLIF was significantly higher in all supplemented groups than the control (p < 0.05). In the presence of PGF2α, the hatching rate was significantly restored in all supplemented groups relative to the group treated with only PGF2α and the control (p < 0.05). Embryo transfer (ET) was performed with blastocysts (day 7). PGF2α levels of control recipient cows were significantly higher in the circulatory blood samples collected 60 min after ET than in samples collected 60 min before ET (p < 0.005), and were decreased in cows injected with loading medium supplemented with Dex+rhLIF (p < 0.005). Pregnancy rate was significantly higher in the ET group that received supplemented embryo-loading medium than in the non-supplemented control (p < 0.05). The intrauterine administration of Dex and rhLIF at ET prevented increased PGF2α in circulatory blood and resulted in enhanced pregnancy rate.

Herbacetin inhibits RANKL-mediated osteoclastogenesis in vitro and prevents inflammatory bone loss in vivo.

Herbacetin is an active flavonol (a type of flavonoid) that has various biologic effects such as antioxidant, antitumor, and anti-inflammatory activities. However, one of its novel effects remains to be investigated, that is, the induction of osteoclastogenesis by the receptor activator of nuclear factor-κB ligand (RANKL). In this study, we examined the effects and mechanisms of action of herbacetin on osteoclastogenesis in RANKL-treated bone marrow-derived macrophages (BMMs) and murine macrophage RAW264.7 cells in vitro and on lipopolysaccharide (LPS)-induced bone destruction in vivo. Herbacetin significantly inhibited RANKL-induced osteoclast formation and differentiation in BMMs and RAW264.7 cells in a dose-dependent manner. Moreover, the suppressive effect of herbacetin resulted in a decrease in osteoclast-related genes, including RANK, tartrate-resistant acid phosphatase, cathepsin K, and matrix metalloproteinase-2 and -9 (MMP-9). Consistent with mRNA results, we confirmed that herbacetin treatment downregulated protein expression of MMP-9 and cathepsin K. Herbacetin also decreased induction of the osteoclastogenic transcription factor c-Fos and nuclear factor of activated T cells c1 (NFATc1) and blocked RANKL-mediated activation of Jun N-terminal kinase (JNK) and nuclear factor-κB. Herbacetin clearly inhibited the bone resorption activity of osteoclasts on plates coated with fluorescein-labeled calcium phosphate. More importantly, the application of herbacetin significantly reduced LPS-induced inflammatory bone loss in mice in vivo. Taken together, our results indicate that herbacetin has potential for use as a therapeutic agent in disorders associated with bone loss.

Induced Pluripotent Stem Cells as a Novel Tool in Psychiatric Research.

Reprogramming of somatic cells into induced pluripotent stem cells (iPSCs) provides a valuable opportunity to study neurodevelopmental and neurodegenerative psychiatric diseases by offering an unlimited source for patient-specific neuronal and glial cells. The present review focuses on the recent advancements in modeling psychiatric disorders such as Phelan-McDermid syndrome, Timothy syndrome, Rett syndrome, schizophrenia, bipolar disorder, and dementia. The treatment effects identified in studies on iPSCs using known therapeutic compounds are also summarized in this review. Here we discuss validation of cellular models and explore iPSCs as a novel drug screening tool. Although there are several limitations associated with the current methods used to study mental disorders, using iPSCs as a model system provides the advantage of rewinding and reviewing the development and degeneration of human neural cells.

Association of human microRNAs miR-22 and miR-491 polymorphisms with panic disorder with or without agoraphobia in a Korean population.

BACKGROUND: The possible involvement of microRNAs (miRNA) in psychiatric disorders has been recently recognized. Several miRNA polymorphisms have been found to be associated with panic disorder (PD) in European populations. However, the association of miRNA polymorphisms on PD has not been reported in Asian populations. We evaluated the effect of miR-22 and miR-491 polymorphisms on susceptibility to PD in a Korean population. METHODS: Genotyping for four polymorphic variants of the primary miRNA (pri-miRNA) regions of miR-22 (rs8076112 and rs6502892) and miR-491 (rs4977831 and rs2039391) was performed using blood samples of 341 Korean patients with PD and 229 healthy control subjects. To evaluate PD phenotypes, the Panic Disorder Severity Scale (PDSS) and Anxiety Sensitivity Inventory-Revised (ASI-R) were administered. RESULTS: Three single-nucleotide polymorphisms (SNPs) were found to be associated with PD: rs8076112 miR-22 and rs4977831 and miR-491 rs2039391. The rs8076112C/rs6502892C haplotypes of miR-22 and rs4977831G/rs2039391G and rs4977831A/rs2039391A haplotypes of miR-491 were significantly overrepresented in patients with PD than in healthy control subjects. In combination analysis, miR-22 rs8076112AC/rs6502892CC and rs8076112CC/rs6502892CC and miR-491 rs4977831AG/rs2039391AA were more frequent in patients with PD. Among the phenotype assessments, ASI-R scores were significantly associated with miR-22 rs6502892 in the subgroup with the agoraphobic phenotype. LIMITATIONS: The results should be considered preliminary due to the relatively small sample size and the selection of only four SNPs. CONCLUSIONS: This is the first report to show possible associations of miR-22 and miR-491 with genetic susceptibility to PD in a Korean population.

Herbacetin inhibits inducible nitric oxide synthase via JNK and nuclear factor-κB in LPS-stimulated RAW264.7 cells.

Herbacetin (3,4',5,7,8-pentahydroxyflavone), an active flavonol compound within flavonoid, has been shown to induce apoptosis in HepG2 cells and suppress hepatocyte growth factor-induced motility of human breast cancer MDA-MB-231 cells. However, the anti-inflammatory mechanisms of Herbacetin have not been researched. In this study, we examined the inflammatory responses stimulated by lipopolysaccharide (LPS) in RAW264.7 macrophage cells after pretreatment with different concentrations of Herbacetin. We found that Herbacetin decreased nitric oxide (NO) production in LPS-induced RAW264.7 and mouse bone marrow-derived macrophages. In addition, Herbacetin inhibited the LPS-induced expression of inducible nitric oxide synthase mRNA and protein in RAW264.7 cells. Treatment with Herbacetin decreased the release of proinflammatory cytokines, including TNF-α and IL-1ß. Moreover, Herbacetin inhibited the activity of JNK kinase and nuclear factor-κB, signaling molecules involved in NO production. Cell signaling analysis using Bay 11-7082 (an inhibitory κB kinase 2 inhibitor) and mitogen-activated protein kinase (MAPK) inhibitors (SB203580 for p38, SP600125 for JNK, and PD 98059 for ERK) suggested that LPS induced iNOS expression via activation of the JNK and NF-κB pathway, but not the p38 and ERK pathway. These findings suggest that Herbacetin exerts an anti-inflammatory effect through suppression of LPS-induced JNK and NF-κB signaling pathways and diminished production of proinflammatory cytokines and mediators.

The additional value of attenuation correction CT acquired during 18F-FDG PET/CT in differentiating mature from immature teratomas.

INTRODUCTION: Increased F-FDG uptake is often seen in soft-tissue components or in neuronal components of teratomas, which makes differentiation of mature and immature teratoma difficult using only F-FDG uptake. The distribution pattern of fat and calcification in teratomas is characteristic on CT, which can also be well seen on attenuation correction CT (AC-CT). We hypothesize that the fat and calcification distribution patterns on AC-CT taken during PET/CT will provide additional diagnostic information in differentiating between mature and immature teratomas. PATIENTS AND METHODS: This retrospective study included 34 patients (44 masses; mean age 32 ± 16.3 years, range 0.2-70 years) who underwent F-FDG PET/CT before surgical resection for teratomas. F-FDG equal to or higher than the liver was visually considered positive. AC-CT images acquired during PET/CT were reviewed for calcification and fat distribution patterns. AC-CT findings for immature teratomas were scattered fat and/or disperse coarse calcification. Pathologic results were categorized into mature and immature teratomas. SUVmax and AC-CT findings were correlated with pathologic results. RESULTS: Out of the 44 lesions, 11 teratomas were immature, with higher F-FDG uptake in these tumors (7.8 ± 4.10 vs. 2.1 ± 2.28, P < 0.001). SUVmax higher than 2.8 were 91% accurate, but fat and/or calcification patterns on AC-CT were extremely helpful in reducing false-positive findings based on F-FDG uptake alone. CONCLUSION: Characteristic fat and calcification patterns on AC-CT of PET/CT were extremely helpful in differentiating mature from immature teratomas, especially in mature teratomas with increased F-FDG uptake. This can potentially reduce unnecessary radiation exposure from additional contrast-enhanced CT.

Trichostatin A promotes the development of bovine somatic cell nuclear transfer embryos.

We studied the effects of trichostatin A (TSA), a histone deacetylase inhibitor, on the development of bovine somatic cell nuclear transfer (SCNT) embryos by investigating (1) the optimal concentration and treatment time of TSA for development of bovine SCNT embryos, (2) the status of histone acetylation in TSA-treated and control SCNT embryos and (3) the expression of histone acetylation- and deacetylation-related genes in TSA-treated and control SCNT embryos. We observed that 50 nM TSA-treatment for 20 h following fusion resulted in more efficient in vitro development of bovine SCNT embryos to the blastocyst stage. In regard to histone H4K5 acetylation, half of the control SCNT embryos faintly displayed histone H4K5 signals 30 min after electrofusion, while most of the TSA-treated SCNT embryos displayed histone H4K5 signals within 30 min after electrofusion. Furthermore, the expressions of HDAC1 and HDAC2 in the blastocysts were significantly lower (P<0.05) in the TSA-treated SCNT than in the control SCNT. However, the expression of GCN5 and HAT1 did not differ between the TSA-treated and control SCNT. In conclusion, we demonstrated that TSA-treatment after SCNT in bovine embryos can dramatically improve the practical applications of current cloning techniques.

Vitrification of immature bovine oocytes by the microdrop method.

This study was conducted to determine the optimal vitrification conditions for immature bovine oocytes using the microdrop method. In experiment 1, the optimal pre-equilibration period for microdrop vitrification was examined. The maturation rate of vitrified oocytes with a 3 min first pre-equilibration period (41.1%) was higher than that of vitrified oocytes with a 0 min first pre-equilibration period (21.4%), and the values of those with a 1 (33.9%) or 5 min (27.4%) first pre-equilibration period were intermediate. The value for a 1 min second pre-equilibration period (44.4%) was significantly higher (P<0.05) than those for a 0.5 (28.6%) and 2 min (21.4%) second pre-equilibration period. In experiment 2, the distribution of microtubules in matured oocytes was investigated. There was no difference among the first pre-equilibration times in terms of the rates of normal spindles in vitrified oocytes. However, this value was significantly higher (P<0.05) in the 1 min group (52.8%) compared with the 0.5 (16.7%) and 2 min groups (12.3%). In experiment 3, we investigated the developmental capacity of immature bovine oocytes vitrified under optimal pre-equilibration conditions (3 min and 1 min for the first and second pre-equilibrations, respectively). Although the total fertilization rates were significantly lower (P<0.05) in the vitrified oocytes (65.6%) compared with the control oocytes (92.4%), there was no difference in the rate of normal fertilization (2PN) between the vitrified (78.6%) and control (82.0%) oocytes. Cleavage and blastocyst rates were significantly lower (P<0.05) in vitrified oocytes (55.7 and 2.3%) than in control oocytes (84.4 and 34.7%). Thus, these results indicated that immature bovine oocytes can survive after microdrop vitrification and subsequently can be cultured to mature oocytes capable of undergoing fertilization in vitro and developing into blastocysts.

Development and apoptosis of pre-implantation porcine nuclear transfer embryos activated with different combination of chemicals.

Artificial activation of oocytes is a pre-requisite for successful cloning by nuclear transfer (NT). This study investigated effect of different combination of activation chemicals such as electric pulse (E), thimerosal (Thi) + dithiothreitol (DTT), 6-dimethylaminopurine (6-DMAP), or cycloheximide (CH) on the developmental ability and the frequency of apoptosis of porcine NT embryos during the culture in vitro. NT embryos activated with chemicals showed significantly higher developmental rate to blastocyst stage compared to embryos activated with E alone (21.5%-26.6% vs. 15.7%, respectively). Of chemicals, Thi + DTT supported higher development to blastocyst stage as compared to 6-DMAP or CH (26.6% vs. 21.5%-23.4%, respectively). Apoptosis of NT embryos were analyzed by using a terminal deoxynucleatidyl transferase-mediated deoxyuridine 5-triphosphate nick-end labeling (TUNEL) assay. The onset of apoptosis of embryos activated E alone was on Day 4, whereas embryos activated with chemicals showed apoptosis on Day 3 post-activation NT embryos exposed to chemicals for activation had higher frequency of apoptosis compared to that of embryos exposed to E alone from Day 3 to Day 7 during the culture. In conclusion, this study shows that chemical activation after fusion could increase not only the developmental ability of porcine NT embryos but also the mean cell number with an increased ratio of inner cell mass (ICM) to trophectoderm (TE) cells. However, the chemical activation also could increase the frequency of apoptosis and induced apoptosis earlier in porcine NT embryos.