Nobiletin enhances mitochondrial function by regulating SIRT1/PGC-1α signaling in porcine oocytes during in vitro maturation.

Nobiletin is a natural flavonoid found in citrus fruits with beneficial effects, including anti-inflammatory, anti-cancer and anti-oxidation effects. The aim of this study was to investigate whether nobiletin improves mitochondrial function in porcine oocytes and examine the underlying mechanism. Oocytes enclosed by cumulus cells were cultured in TCM-199 for 44 h with 0.1% dimethyl sulfoxide (control), or supplemented with 5, 10, 25, and 50 µM of nobiletin (Nob5, Nob10, Nob25, and Nob50, respectively). Oocyte maturation rate was significantly enhanced in Nob10 (70.26 ± 0.45%) compared to the other groups (control: 60.12 ± 0.47%; Nob5: 59.44 ± 1.63%; Nob25: 63.15 ± 1.38%; Nob50: 46.57 ± 1.19%). The addition of nobiletin reduced the levels of reactive oxygen species and increased glutathione levels. Moreover, Nob10 promoted mitochondrial biogenesis by upregulating the protein levels of sirtuin 1 (SIRT1) and peroxisome proliferator-activated receptor-gamma coactivator 1α (PGC-1α). This resulted in an increase in the number of active mitochondria, mitochondrial DNA copy number, mitochondrial membrane potential, and ATP production, thereby enhancing mitochondrial function. The protein level of p53 decreased, followed by the phosphorylation of B-cell lymphoma 2, suggesting a reduction in mitochondria-mediated apoptosis in the Nob10 group. Additionally, the release of cytochrome c from the mitochondria was significantly diminished along with a decrease in the protein expression of caspase 3. Thus, nobiletin has a great potential to promote the in vitro maturation of porcine oocytes by suppressing oxidative stress and promoting mitochondrial function through the upregulation of the SIRT1/PGC-1α signaling pathway.

GRP78 acts as a cAMP/PKA signaling modulator through the MC4R pathway in porcine embryonic development.

Glucose-regulated protein 78 (GRP78) binds to and stabilizes melanocortin 4 receptor (MC4R), which activates protein kinase A (PKA) by regulating G proteins. GRP78 is primarily used as a marker for endoplasmic reticulum stress; however, its other functions have not been well studied. Therefore, in this study, we aimed to investigate the function of GRP78 during porcine embryonic development. The developmental quality of porcine embryos, expression of cell cycle proteins, and function of mitochondria were evaluated by inhibiting the function of GRP78. Porcine oocytes were activated to undergo parthenogenesis, and blastocysts were obtained after 7 days of in vitro culture. GRP78 function was inhibited by adding 20 µM HA15 to the in vitro culture medium. The inhibition in GRP78 function led to a decrease in G proteins release, which subsequently downregulated the cyclic adenosine monophosphate (cAMP)/PKA pathway. Ultimately, inhibition of GRP78 function induced the inhibition of CDK1 and cyclin B expression and disruption of the cell cycle. In addition, inhibition of GRP78 function regulated DRP1 and SIRT1 expression, resulting in mitochondrial dysfunction. This study provides new insights into the role of GRP78 in porcine embryonic development, particularly its involvement in the regulation of the MC4R pathway and downstream cAMP/PKA signaling. The results suggest that the inhibition of GRP78 function in porcine embryos by HA15 treatment may have negative effects on embryo quality and development. This study also demonstrated that GRP78 plays a crucial role in the functioning of MC4R, which releases the G protein during porcine embryonic development.

DeepHCS++: Bright-field to fluorescence microscopy image conversion using multi-task learning with adversarial losses for label-free high-content screening.

In this paper, we propose a novel microscopy image translation method for transforming a bright-field microscopy image into three different fluorescence images to observe the apoptosis, nuclei, and cytoplasm of cells, which visualize dead cells, nuclei of cells, and cytoplasm of cells, respectively. These biomarkers are commonly used in high-content drug screening to analyze drug response. The main contribution of the proposed work is the automatic generation of three fluorescence images from a conventional bright-field image; this can greatly reduce the time-consuming and laborious tissue preparation process and improve throughput of the screening process. Our proposed method uses only a single bright-field image and the corresponding fluorescence images as a set of image pairs for training an end-to-end deep convolutional neural network. By leveraging deep convolutional neural networks with a set of image pairs of bright-field and corresponding fluorescence images, our proposed method can produce synthetic fluorescence images comparable to real fluorescence microscopy images with high accuracy. Our proposed model uses multi-task learning with adversarial losses to generate more accurate and realistic microscopy images. We assess the efficacy of the proposed method using real bright-field and fluorescence microscopy image datasets from patient-driven samples of a glioblastoma, and validate the method's accuracy with various quality metrics including cell number correlation (CNC), peak signal-to-noise ratio (PSNR), structural similarity index measure (SSIM), cell viability correlation (CVC), error maps, and R2 correlation.

Effect of Nasogastric Tube on Aspiration Risk: Results from 147 Patients with Dysphagia and Literature Review.

Nasogastric tube (NGT) is a common feeding strategy for patients at risk of endotracheal aspiration with an oral diet. With NGT feeding, however, swallowing of small amounts saliva cannot be avoided. We investigated whether the aspiration rate when swallowing 1 mL of fluid increased in patients using an NGT in different dysphagia severities. One hundred forty-seven patients who had been receiving NGT feeding underwent a videofluoroscopic swallowing study (VFSS). During VFSS, subjects were offered 1 mL of fluid twice: initially, with the tube inserted (NGT-in) and, subsequently, with the tube removed (NGT-out). Aspiration depth was determined using the 8-point Penetration-Aspiration Scale (PAS) (0 points, no aspiration/penetration; 8 points, aspiration passing the vocal cords with no ejection efforts). PAS-diff was computed (PASNGT-in - PASNGT-out), and a positive PAS-diff (PAS-diff > 0) meant increased aspiration depth in the presence of NGT. After VFSS, diet recommendations were made according to dysphagia severity assessment: non-oral feeding (n = 59), diet modification (n = 74), and diet as tolerated (n = 13). Cognitive level (mini-mental state examination, MMSE) and general functional level (Modified Barthel Index, MBI) were compared between the PAS-diff > 0 and PAS-diff ≤ 0 groups. Aspiration severity did not significantly change after NGT removal (PASNGT-in, 2.45 ± 2.40; PASNGT-out, 2.57 ± 2.58; P = .50). Regardless of recommended diet, PAS-diff values were not significantly different (P = .49). MMSE and MBI were not significantly different (P = .23 and .94) between subjects with PAS-diff > 0 (n = 25) and PAS-diff ≤ 0 (n = 121). In conclusion, the risk of aspirating a small amount of fluid was not significantly different before and after NGT removal, regardless of swallowing function, cognitive level, or general functional level.

Sleep Quality and Attention May Correlate With Hand Grip Strength: FARM Study.

OBJECTIVE: To determine the socio-demographic, psychologic, hematologic, or other relevant factors associated with hand grip strength in Korean farmers. METHODS: A total of 528 healthy Korean farmers were enrolled. Hand grip strength was measured in both hands using a hydraulic dynamometer. Socio-demographic characteristics were assessed and anthropometric measurements were obtained. Psycho-cognitive measurements such as sleep quality (Pittsburgh Sleep Quality Index) and Go/No-Go test response time were conducted. In addition to physical measurements, serologic parameters including insulin-like growth factor 1 were measured. The factors associated with hand grip strength were analyzed using multiple linear regression analysis after adjusting for age, height, and weight. RESULTS: The mean hand grip strength was associated with the Pittsburgh Sleep Quality Index total score (ß=-0.12, p=0.01), the Go/No-Go test response time (ß=-0.18, p=0.001), vitamin D (ß=0.12, p=0.02), and insulin-like growth factor 1 levels (ß=0.1, p=0.045). In female farmers, hand grip strength was only associated with the Pittsburgh Sleep Quality Index total score (ß=-0.32, p<0.001). CONCLUSION: The results of this study demonstrate that hand grip strength was associated with sleep quality and attention in Korean farmers.