Generative Adversarial Networks for Creating Synthetic Nucleic Acid Sequences of Cat Genome.

Nucleic acids are the basic units of deoxyribonucleic acid (DNA) sequencing. Every organism demonstrates different DNA sequences with specific nucleotides. It reveals the genetic information carried by a particular DNA segment. Nucleic acid sequencing expresses the evolutionary changes among organisms and revolutionizes disease diagnosis in animals. This paper proposes a generative adversarial networks (GAN) model to create synthetic nucleic acid sequences of the cat genome tuned to exhibit specific desired properties. We obtained the raw sequence data from Illumina next generation sequencing. Various data preprocessing steps were performed using Cutadapt and DADA2 tools. The processed data were fed to the GAN model that was designed following the architecture of Wasserstein GAN with gradient penalty (WGAN-GP). We introduced a predictor and an evaluator in our proposed GAN model to tune the synthetic sequences to acquire certain realistic properties. The predictor was built for extracting samples with a promoter sequence, and the evaluator was built for filtering samples that scored high for motif-matching. The filtered samples were then passed to the discriminator. We evaluated our model based on multiple metrics and demonstrated outputs for latent interpolation, latent complementation, and motif-matching. Evaluation results showed our proposed GAN model achieved 93.7% correlation with the original data and produced significant outcomes as compared to existing models for sequence generation.

Survival Benefit of Anticoagulation Therapy in End Stage Kidney Disease Patients with Atrial Fibrillation: A Single Center Retrospective Study.

Background and Objectives: Although the need for anticoagulation to prevent thromboembolism is increasing and non-vitamin K antagonist oral anticoagulants (NOACs) have been tried, there is still controversy about the efficacy of anticoagulation in patients with dialysis. Materials and Methods: We retrospectively analyzed the risk and benefit of anticoagulation in dialysis patients with atrial fibrillation (AF). We retrospectively analyzed all data of 89 patients who received dialysis therapy and were diagnosed with AF. Among them, 27 received anticoagulation (11 warfarin and 16 apixaban 2.5 mg twice a day), while 62 received no anticoagulation. Results: In multivariate Cox regression analysis, compared to no anticoagulation treatment, anticoagulation treatment was associated with a low incidence of all-cause mortality (hazard ratios (HR) 0.36; 95% confidence interval (CI) 0.15-0.88). Compared to no anticoagulation treatment, more anticoagulation treatment patients experienced severe bleeding (HR 4.67; 95% CI 1.26-17.25) and any bleeding (HR 2.79; 95% CI 1.01-7.74). Compared to no anticoagulation, warfarin treatment patients were associated with a low incidence of all-cause mortality (HR 0.26; 95% CI 0.09-0.81) and a high incidence of severe bleeding (HR 4.85; 95% CI 1.12-21.10). All-cause mortality and bleeding were not significantly different between no anticoagulation and apixaban treatment patients. Conclusions: In dialysis patients with AF, anticoagulation therapy is associated with an increased incidence of severe bleeding, but anticoagulation therapy is associated with a low incidence of all-cause mortality. Individualized anticoagulation therapy with careful bleeding monitoring is needed in dialysis patients with AF.

Active Compound of Pharbitis Semen (Pharbitis nil Seeds) Suppressed KRAS-Driven Colorectal Cancer and Restored Muscle Cell Function during Cancer Progression.

Kirsten rat sarcoma viral oncogene homolog (KRAS)-driven colorectal cancer (CRC) is notorious to target with drugs and has shown ineffective treatment response. The seeds of Pharbitis nil, also known as morning glory, have been used as traditional medicine in East Asia. We focused on whether Pharbitis nil seeds have a suppressive effect on mutated KRAS-driven CRC as well as reserving muscle cell functions during CRC progression. Seeds of Pharbitis nil (Pharbitis semen) were separated by chromatography and the active compound of Pharbitis semen (PN) was purified by HPLC. The compound PN efficiently suppressed the proliferation of mutated KRAS-driven CRC cells and their clonogenic potentials in a concentration-dependent manner. It also induced apoptosis of SW480 human colon cancer cells and cell cycle arrest at the G2/M phase. The CRC related pathways, including RAS/ERK and AKT/mTOR, were assessed and PN reduced the phosphorylation of AKT and mTOR. Furthermore, PN preserved muscle cell proliferation and myotube formation in cancer conditioned media. In summary, PN significantly suppressed mutated KRAS-driven cell growth and reserved muscle cell function. Based on the current study, PN could be considered as a promising starting point for the development of a nature-derived drug against KRAS-mutated CRC progression.

Safety and Efficacy of Rice Bran Supercritical CO2 Extract for Hair Growth in Androgenic Alopecia: A 16-Week Double-Blind Randomized Controlled Trial.

We conducted a 16-week double-blind randomized controlled single-center trial to evaluate the safety and efficacy of dermal rice bran supercritical CO2 extract (RB-SCE) in the treatment of androgenic alopecia. Fifty alopecia patients were randomly assigned to the experimental and placebo groups. The experimental group received a dermal application of 0.5% RB-SCE (8 mL/d) to the head skin for 16 weeks while the control group received a dermal application of placebo. Changes in hair count, diameter, and density were evaluated with a Folliscope(®). Patient satisfaction was evaluated via questionnaire and clinical photographs were rated by dermatologists. The results showed that RB-SCE significantly increased hair density and hair diameter in male subjects. Patient satisfaction and the evaluation of photographs by dermatologists also confirmed the effectiveness of RB-SCE in the treatment of alopecia. No adverse reactions related to RB-SCE were reported. Therefore, RB-SCE shows promise for use in functional cosmetics and pharmaceuticals.

Characteristics of esterified rice bran oil converted by enzymatic esterification.

In the present study, esterified rice bran oil (ERBO) was characterized using enzymatic esterification to improve stability, prevent acidification, enhance health-promoting biological activity and generate ω-3 PUFA-rich rice bran oil (RBO). Esterification reactions using RBO and ethanol were performed at 50°C under 200 bar with 3% lipozyme TL-IM (Thermomuces lanuginosa immobilized on silica gel) or RM-IM (Rhizomucor miehei immobilized on ion exchange resin) for 3 hr under supercritical CO2. The molar ratios of ethanol to RBO were 3, 6, 9 and 12, respectively. Total lipid contents and acid values decreased (maximum 83.75%),but γ-oryzanol content increased (maximum 41.33%) in esterified RBO (ERBO) prepared using TL-IM or RM-IM. In addition, DPPH radical scavenging activity of ERBO prepared by RM-IM atan ethanol to RBO molar ratio of 3 was 0.02 µg µl(-1), which was 63-fold higher than that of α-tocopherol (IC50 =1.25 µg µl(-1)). The anti-inflammatory effect of RM-IM 1:3 hydrolysate of RBO was verified showing its suppressive effect towards iNOS and Cox-2mRNA expression in a dose-dependent manner. Therefore, ERBO is a promising source of functional food, cosmetics and pharmaceuticals.

Genotoxicity of rice bran oil extracted by supercritical CO2 extraction.

Rice bran oil extracted by supercritical CO2 extraction (RB-SCE) reportedly exhibits pharmacological activities such as antioxidant and in vivo hair growth-inducing effects. Such activities raise the possibility of the development of novel hair growth-inducing agents using RB-SCE. The aim of this study was to investigate the potential genotoxic effects of RB-SCE in three short-term mutagenicity assays (bacterial reverse mutation assay, in vitro mammalian chromosomal aberration test, and in vivo micronucleus assay). RB-SCE showed no genotoxicity in the bacterial reverse mutation assay up to 5000 mg/plate and in the in vivo micronucleus test up to 600 mg/kg body weight. However, at 120 µg/mL with S9 mix and 200 µg/mL without S9 mix RB-SCE showed significantly different genotoxicity than the negative control in the in vitro chromosome aberration test. The induction of chromosomal aberrations under the present conditions may have no biological significance. We have herein demonstrated that RB-SCE can be regarded as a non-genotoxic material based on the available in vivo and in vitro results.

In vivo hair growth-promoting effect of rice bran extract prepared by supercritical carbon dioxide fluid.

The potential hair growth-promoting activity of rice bran supercritical CO2 extract (RB-SCE) and major components of RB-SCE, linoleic acid, policosanol, γ-oryzanol, and γ-tocotrienol, were evaluated with the histological morphology and mRNA expression levels of cell growth factors using real-time reverse transcriptase-polymerase chain reaction (PCR) in C57BL/6 mice. RB-SCE showed hair growth-promoting potential to a similar extent as 3% minoxidil, showing that the hair follicles were induced to be in the anagen stage. The numbers of the hair follicles were significantly increased. In addition, mRNA expression levels of vascular endothelial growth factor (VEGF), insulin-like growth factor-1 (IGF-1), and keratinocyte growth factor (KGF) were also significantly increased and that of transforming growth factor-ß (TGF-ß) decreased in RB-SCE-treated groups. Among the major components of RB-SCE, linoleic acid and γ-oryzanol induced the formation of hair follicles according to examination of histological morphology and mRNA expression levels of cell growth factors. In conclusion, our results demonstrate that RB-SCE, particularly linoleic acid and γ-oryzanol, promotes hair growth and suggests RB-SCE can be applied as hair loss treatment.

Lrmp/Jaw1 is expressed in sweet, bitter, and umami receptor-expressing cells.

Inositol 1,4,5-triphosphate-mediated calcium (IP3-Ca2+) signal cascade is an essential process in sweet, bitter, and umami taste signal transduction. Although the main components of this cascade have been identified, the candidate regulators of them in taste tissues are still unclear. In an effort to identify genes involved in taste signal transduction, we found that a gene encoding lymphoid-restricted membrane protein (Lrmp/Jaw1) was expressed in mouse taste tissues. Here we report that Lrmp/Jaw1 is specifically expressed in sweet, bitter, and umami taste receptor-expressing cells of mouse circumvallate, foliate, and fungiform papillae. In addition to this specific expression patterns, we found that Lrmp/Jaw1 is associated with type III IP3 receptor (IP3R3) via its coiled-coil domain in the COS7 heterologous expression system. These results raise the possibility that Lrmp/Jaw1 interacts with IP3R3 in taste cells and suggest an important role for Lrmp/Jaw1 in the IP3-Ca2+ signal cascade in sweet, bitter, and umami taste signal transduction.

Fluid overload is a determinant for cardiac structural and functional impairments in type 2 diabetes mellitus and chronic kidney disease stage 5 not undergoing dialysis.

BACKGROUND: Fluid overload is common in patients with diabetes and chronic kidney disease (DM and CKD; DMCKD) and can lead to structural and functional cardiac abnormalities including left ventricular hypertrophy (LVH) and left ventricular diastolic dysfunction (LVDD). Fluid overload represents a crucial step in the pathophysiological pathways to chronic heart failure in patients with end-stage renal disease. We evaluated the impact of fluid overload on cardiac alterations in patients with diabetes and non-dialysis-dependent CKD stage 5 (DMCKD5-ND) without intrinsic heart disease. METHODS: Bioimpedance spectroscopy, echocardiography, and N-terminal prohormone of B-type natriuretic peptide (NT-proBNP) measurement were performed in 135 consecutive patients on the same day. Patients were divided into groups by tertiles of overhydration/extracellular water (OH/ECW) per bioimpedance spectroscopy. RESULTS: Fluid balance markers including OH/ECW and NT-proBNP were significantly higher in the LVDD+LVH group. OH/ECW and its exacerbation were positively associated with the ratio between early mitral inflow and annular early diastolic velocities (E/e' ratio) and left ventricular mass index (LVMI). The prevalence of LVH progressively increased across increasing tertiles of OH/ECW. In multiple regression analyses, OH/ECW as a continuous and categorical variable was independently associated with the E/e' ratio and LVMI after adjustment for multiple confounding factors. CONCLUSIONS: Fluid overload was independently associated with LVDD and LVH in patients with DMCKD5-ND. Our study suggests that structural and functional cardiac abnormalities and volume status should be evaluated simultaneously in patients with early-stage DMCKD rather than only DMCKD5-ND, in addition to intensive blood pressure and glycemic control, regardless of evident cardiovascular disease.

Sample preparation method for plasma membrane proteome analysis.

The preparation of plasma membrane (PM) proteome samples is seriously difficult and time-consuming, owing to their profound hydrophobicity and low abundance. We have developed an efficient PM sample preparation method using Ultracentrifugation with Percoll and an aqueous two-phase extraction. The developed method was rapid (3 h) and provided high purities (26-fold of cell lysate) with a high yield (2.6% of whole cell lysate proteins). This method is especially useful for PM proteome studies using 2D gel electrophoresis.

Differentially expressed proteins of mesenchymal stem cells derived from human cord blood (hUCB) during osteogenic differentiation.

Multipotent mesenchymal stem cells (MSCs) derived from human umbilical cord blood (hUCB) represent promising candidates for the development of future cellular therapy strategies. MSCs have been found to be able to differentiate into various tissues. One of the primary limitations in our understanding of the biology of human MSCs is the absence of prospective markers required for the monitoring of lineage-specific differentiation. hUCB-derived MSCs have been found to have significantly greater osteogenic potential. In this study, we focused on proteins that were differentially expressed during osteogenic differentiation of hUCB-MSCs. And we analyzed the protein expression inherent to osteogenic differentiation by two-dimensional gel electrophoresis, ESI-Q-TOF, and Western blotting. Eleven differentially expressed spots were observed between the two groups (before and after differentiation) on the 2-DE map. These might also be proved as useful cytosolic biomarker proteins for osteogenesis, and might be employed in quality control of osteoblasts in cell-therapy applications.

Prenatally diagnosed ectrodactyly at 16 weeks' gestation by 2- and 3-dimensional ultrasonography: a case report.

OBJECTIVE: Ectrodactyly is a rare malformation with various presentations. The current report describes a case of ectrodactyly detected using 2-dimensional (2D) and 3-dimensional (3D) ultrasonography at 16 weeks' gestation. METHODS AND RESULTS: The 2D ultrasonographic findings were ectrodactyly in the right hand and monodactyly in the left hand, and these results were confirmed and further clarified using 3D imaging. The postmortem X-ray findings were consistent with the ultrasonography. CONCLUSION: We conclude that 3D ultrasonography can assist in clarifying 2D ultrasonography findings of hand malformations during the second trimester of pregnancy.

Human blood plasma preparation for two-dimensional gel electrophoresis.

Human plasma consists of mainly large proteins, which vary in terms of both composition and concentration with the physiological state of the individual. Alterations in protein concentrations reflect the current state of the individual's health and thus may be utilized as valuable biomarkers for a specific biological process or disease. Two-dimensional gel electrophoresis (2-DE) has proven to be a valuable method for the separation and comparison of complex protein mixtures, for example, from disease and healthy states, as this method provides information regarding the variation, relative quantities, and structures of the intact proteins. The procedures utilized for the preparation of samples for 2-DE are critical to the acquisition of high-quality results for the discovery of biomarkers. The objective of this study was to review the preparation methods of plasma for 2-DE, particularly those designed to improve the detection of proteins in low abundance in plasma on 2-DE. The use of anticoagulants and protease inhibitors during the collection of blood, the removal of abundant proteins using multicomponent immunodepletion system, and desalting procedure allow us to compile profiles of proteins occurring in low concentrations in the plasma and to improve the pattern generated during 2-DE.

Comparison of proteomes in various human plasma preparations by two-dimensional gel electrophoresis.

Serum or plasma can be utilized in a variety of studies targeted toward the discovery of disease biomarkers. In this study, the proteome profiles of plasma samples prepared using various anticoagulants (EDTA, heparin or citrate), were compared with those of serum using two-dimensional electrophoresis (2-DE). Proteins which evidenced different levels in the plasma and serum were screened and identified using ESI-Q-TOF MS/MS. The proteins which became detectable after the removal of fibrinogen from serum were identified as pigment epithelial differentiating factor (four spots), fetuin-like protein, and the hemopexin precursor. In particular, three proteins, pre-serum amyloid P component, plasma glutathione peroxidase precursor, and tetranectin, evidenced increased volume intensity only in the plasma samples prepared with EDTA.

Anti-obesity effects of yellow catfish protein hydrolysate on mice fed a 45% kcal high-fat diet.

Obesity contributes to the etiologies of a variety of comorbid conditions, such as type 2 diabetes, hypertension and cardiovascular disease. In the present study, the anti-obesity effects of yellow catfish protein hydrolysate (YPh) were observed in mice fed a 45% kcal high-fat diet (HFD) compared with those of mice treated with simvastatin. The HFD-fed control mice exhibited noticeable increase in body weight, and whole-body and abdominal fat densities, periovarian and abdominal wall-deposited fat pad weight, as well as in the levels of triglycerides (TG), blood total cholesterol (TC), low-density lipoprotein, alanine aminotransferase, aspartate aminotransferase, creatinine, blood urea nitrogen, and in the fecal TG and TC contents. However, they exhibited a decrease in serum high-density lipoprotein levels. In addition, an increase was detected in periovarian and dorsal abdominally deposited fat pad thickness, adipocyte hypertrophy, the number of steatohepatitis regions, hepatocyte hypertrophy and lipid droplet deposition-related renal tubular vacuolation degenerative lesions, along with increased hepatic lipid peroxidation and a deteriorated endogenous antioxidant defense system (glutathione, catalase and superoxide dismutase). However, all the above-mentioned obesity-related complications were dose-dependently and significantly inhibited after 84 days of thye consecutive oral administration of 125, 250 and 500 mg/kg YPh. In addition, YPh dose-dependently depleted the liver endogenous antioxidant defense system and inhibited hepatic lipid peroxidation. Overall, the effects of 250 mg/kg YPh on HFD-induced obesity and related complications were similar or more potent than those of 10 mg/kg simvastatin. These results indicate that YPh is a promising new potent medicinal ingredient for possible use in the treatment of obesity and related complications.

Blood glycemia-modulating effects of melanian snail protein hydrolysates in mice with type II diabetes.

Freshwater animal proteins have long been used as nutrient supplements. In this study, melanian snail (Semisulcospira libertina) protein hydrolysates (MPh) were found to exert anti-diabetic and protective effects against liver and kidney damage in mice with type II diabetes adapted to a 45% kcal high-fat diet (HFD). The hypoglycemic, hepatoprotective and nephroprotective effects of MPh were analyzed after 12 weeks of the continuous oral administration of MPh at 125, 250 and 500 mg/kg. Diabetic control mice exhibited an increase in body weight, and blood glucose and insulin levels, with a decrease in serum high-density lipoprotein (HDL) levels. In addition, an increase in the regions of steatohepatitis, hepatocyte hypertrophy, and lipid droplet deposit-related renal tubular vacuolation degenerative lesions were detected, with noticeable expansion and hyperplasia of the pancreatic islets, and an increase in glucagon- and insulin-producing cells, insulin/glucagon cell ratios in the endocrine pancreas and hepatic lipid peroxidation, as well as decreased zymogen contents. Furthermore, a deterioration of the endogenous antioxidant defense system was observed, with reduced glucose utilization related hepatic glucokinase (GK) activity and an increase in hepatic gluconeogenesis-related phosphoenolpyruvate carboxykinase (PEPCK) and glucose­6-phosphatase (G6pase) activity. However, all of these diabetic complications were significantly inhibited by oral treatment with MPh in a dose-dependent manner. In addition, the marked dose-dependent inhibition of hepatic lipid peroxidation, the depletion of the liver endogenous antioxidant defense system, and changes in hepatic glucose-regulating enzyme activities were also observed. The results of this study suggest that MPh exerts potent anti-diabetic effects, along with the amelioration of related complications in mice with type II diabetes. The overall effects of MPh at a dose of 125 mg/kg on HFD-induced diabetes and related complications were similar or more potent than those of metformin (250 mg/kg).

Pathogenic microorganisms, heavy metals, and antibiotic residues in seven Korean freshwater aquaculture species.

This survey was performed to estimate the levels of pathogenic microorganisms, antibiotic residues, and heavy metals in seven Korean freshwater aquaculture species including Anguilla japonica, Cyprinus carpio nudus, Oncorhynchus mykiss, Pseudobagrus fulvidraco, Semisulcospira coreana, Silurus asotus, and Trionyxs sinensis. None of the ten foodborne pathogens tested in this study were found in any of the species collected from any of the aquaculture farms. Furthermore, no banned chemicals or antibiotic residues were found in any of the species collected from any of the aquaculture farms, except enrofloxacin, which was below guideline limits (0.1 mg/kg). Finally, no species had lead, cadmium, total arsenic, or total mercury concentrations above the Ministry of Food and Drug Safety (MSDF) guidelines (0.5, 0.5, 0.1, and 0.5 mg/kg, respectively). These results ensure the safety of freshwater aquaculture species and will be useful for developing consumption advisories of freshwater fishes.

Oral Administration of a Hot Water Extract of the Softshell Turtle (Trionyx sinensis) Improves Exercise Performance.

Freshwater softshell turtle (Trionyx sinensis) extract has been used traditionally as a tonic soup, and to recover from physical fatigue. To support these claims, the forelimb grip strength of mice was measured after feeding a soft-shell turtle extract for 7 days. The T. sinensis extract significantly increased the grip strength to 1.25±0.07 N (P<0.01), which is 16.8% higher than the force on day 0. After exercising, the blood glucose levels in extract-fed mice were 202% higher and urea levels were 73% lower, which were both significantly different than the levels observed after control treatment. Lactate dehydrogenase was significantly higher by 314%, and glutathione peroxidase increased by 165%. In addition, the obesity markers, serum triglyceride and cholesterol, decreased to 62% and 49%, respectively, after mice were fed the extract. These data show that the T. sinensis extract provided more energy for forelimb exercise, prevented protein catabolism and muscle fatigue, and decreased the oxidative stress caused by an exhaustive workout.

Hot Water Extract of Leather Carp (Cyprinus carpio nudus) Improves Exercise Performance in Mice.

The hot water extract of leather carp (Cyprinus carpio nudus) has been used as a nourishing tonic soup and as an aid for recovery from physical fatigue. In this study, we investigated the effect of leather carp extract on exercise performance in mice. Swimming endurance and forelimb grip strength were assessed following oral administration of the extract (once per day for 7 days) at a dose of 0.5 mg/10 µL/g body weight. After 7 days, mice given the leather carp extract had significantly greater swimming endurance [105±18 s (P<0.05); 52% longer than day 0] and forelimb grip strength [1.18±0.05 Newton (P<0.01); 17% greater than day 0]. The extract increased muscle mass, but had little effect on body weight. Following the swimming exercise, blood glucose, glutathione peroxidase, and superoxide dismutase levels in extract-fed mice were significantly higher (145%, 131%, and 106%, respectively) than in the saline control group. Blood levels of high-density lipoprotein cholesterol were also significantly increased (128%) in mice given the extract compared to the controls. These results suggest that leather carp extract can improve physical exercise performance and prevent oxidative stress caused by exhaustive workouts.

Protective effects of ginseng leaf extract using enzymatic extraction against oxidative damage of UVA-irradiated human keratinocytes.

UVA is responsible for numerous biological effects on the skin, including premature aging characterized by wrinkles, leathery texture, and mottled pigmentation. The objective of this study was evaluating the protective effect of ginseng leaf extract prepared by Ultraflo L on skin from photodamage. Anti-wrinkle effect of ginseng leaf extract with or without Ultraflo L treatment were tested on human keratinocyte cells (HaCaT) irradiated with ultraviolet (UV) A. Ginseng leaves inhibited ROS generation, GHS depletion, and expression of MMP-2 and MMP-9 induced by UVA irradiation. The glutathione (GSH) content of the cells was significantly increased by over 25 µg mL(-1) of Ultraflo-treated extract (UTGL) as well as by over 100 µg mL(-1) of nonenzyme-treated extract (NEGL) compared to control. UTGL and NEGL treatments significantly decreased expression of metalloproteinase (MMP)-2 and 9 compared with control, but inhibitory effects of two groups on expression of MMPs were not significantly different. Overall, ULtraflo L-treated ginseng leaves inhibited ROS generation, GHS depletion, and expression of MMP-2 and MMP-9 in UVA photodamaged HaCat cells. From these results, enzyme-treated ginseng leaf extract has advantages over untreated ginseng leaves and have potential as a skin protective ingredient against UVA-induced photodamage.