The use of the internal transcribed spacer region for phylogenetic analysis of the microsporidian parasite Enterocytozoon hepatopenaei infecting whiteleg shrimp (Penaeus vannamei) and for the development of a nested PCR as its diagnostic tool.

The increasing economic losses associated with growth retardation caused by Enterocytozoon hepatopenaei (EHP), a microsporidian parasite infecting penaeid shrimp, require effective monitoring. The internal transcribed spacer (ITS)-1 region, the non-coding region of ribosomal clusters between 18S and 5.8S rRNA genes, is widely used in phylogenetic studies due to its high variability. In this study, the ITS-1 region sequence (~600-bp) of EHP was first identified, and primers for a polymerase chain reaction (PCR) assay targeting that sequence were designed. A newly developed nested-PCR method successfully detected the EHP in various shrimp (Penaeus vannamei and P. monodon) and related samples, including water and feces collected from Indonesia, Thailand, South Korea, India, and Malaysia. The primers did not cross-react with other hosts and pathogens, and this PCR assay is more sensitive than existing PCR detection methods targeting the small subunit ribosomal RNA (SSU rRNA) and spore wall protein (SWP) genes. Phylogenetic analysis based on the ITS-1 sequences indicated that the Indonesian strain was distinct (86.2%) from other strains collected from Thailand and South Korea, and also showed the internal diversity among Thailand (N = 7, divided into four branches) and South Korean (N = 5, divided into two branches) samples. The results revealed the ability of the ITS-1 region to determine the genetic diversity of EHP from different geographical origins.

Effect of YC-1102 on the Improvement of Obesity in High-Fat Diet-Induced Obese Mice.

Obesity is one of the major risk factors for metabolic diseases worldwide. This study examined the effects of YC-1102, an extract derived from the roots of Rosa multiflora, on 3T3-L1 preadipocytes and high-fat diet (HFD)-induced obese mice. In vivo experiments involved the oral administration of YC-1102 (100, 150, and 200 mg/kg body weight) daily to mice for eight weeks. YC-1102 was found to downregulate the expressions of PPARγ and C/EBPα during adipogenesis, inhibiting adipocyte differentiation and upregulating the expression of PGC-1α for energy metabolism to enhance mitochondrial biogenesis and fatty acid oxidation. It has been shown that daily administration of YC-1102 to mice receiving a HFD prevented an increase in body weight and the accumulation of body fat. YC-1102 administration also reduced TG, TC, and LDL cholesterol levels, as well as glucose and leptin levels, and increased adiponectin levels, thus effectively inhibiting the metabolism of lipids. YC-1102-treated mice showed significant reductions in the mRNA expression of PPARγ and C/EBPα. The levels of PGC-1α involved in energy metabolism increased significantly in the YC-1102-treated mice when compared to the HFD-treated mice. According to the findings of this study, YC-1102 has a dual mechanism that reduces transcription factors that promote the differentiation of adipocytes and increases transcription factors that promote energy consumption.

Lactiplantibacillus plantarum HY7718 Improves Intestinal Integrity in a DSS-Induced Ulcerative Colitis Mouse Model by Suppressing Inflammation through Modulation of the Gut Microbiota.

Inflammatory bowel disease (IBD), a chronic condition that causes persistent inflammation in the digestive system, is closely associated with the intestinal microbiome. Here, we evaluated the effects of Lactiplantibacillus plantarum HY7718 (HY7718) on IBD symptoms in mice with dextran sulfate sodium (DSS)-induced colitis. Oral administration of HY7718 led to significant improvement in the disease activity index score and the histological index, as well as preventing weight loss, in model mice. HY7718 upregulated the expression of intestinal tight junction (TJ)-related genes and downregulated the expression of genes encoding pro-inflammatory cytokines and genes involved in the TLR/MyD88/NF-κB signaling pathway. Additionally, HY7718 reduced the blood levels of pro-inflammatory cytokines, as well as reversing DSS-induced changes to the composition of the intestinal microbiome. HY7718 also increased the percentage of beneficial bacteria (Lactiplantibacillus and Bifidobacterium), which correlated positively with the expression of intestinal TJ-related genes. Finally, HY7718 decreased the population of pathogens such as Escherichia, which correlated with IBD symptoms. The data suggest that HY7718 improves intestinal integrity in colitis model mice by regulating the expression of TJ proteins and inflammatory cytokines, as well as the composition of the intestinal microflora. Thus, L. plantarum HY7718 may be suitable as a functional supplement that improves IBD symptoms and gut health.

Exosomal miR-205-5p Improves Endometrial Receptivity by Upregulating E-Cadherin Expression through ZEB1 Inhibition.

Endometrial receptivity is a complex process that prepares the uterine endometrium for embryo implantation; insufficient endometrial receptivity is one of the causes of implantation failure. Here, we analyzed the microRNA expression profiles of exosomes derived from both receptive (RL95-2) and non-receptive (AN3-CA) endometrial epithelial cell (EEC) lines to identify exosomal miRNAs closely linked to endometrial receptivity. Among the 466 differentially expressed miRNAs, miR-205-5p was the most highly expressed in exosomes secreted from receptive RL95-2 cells. miR-205-5p, enriched at the adhesive junction, was closely related to endometrial receptivity. ZEB1, a transcriptional repressor of E-cadherin associated with endometrial receptivity, was identified as a direct target of miR-205-5p. miR-205-5p expression was significantly lower in the endometrial tissues of infertile women than in that of non-infertile women. In vivo, miR-205-5p expression was upregulated in the post-ovulatory phase, and its inhibitor reduced embryo implantation. Furthermore, administration of genetically modified exosomes overexpressing miR-205-5p mimics upregulated E-cadherin expression by targeting ZEB1 and improved spheroid attachment of non-receptive AN3-CA cells. These results suggest that the miR-205-5p/ZEB1/E-cadherin axis plays an important role in regulating endometrial receptivity. Thus, the use of exosomes harboring miR-205-5p mimics can be considered a potential therapeutic approach for improving embryo implantation.

Emerging Roles of Using Small Extracellular Vesicles as an Anti-Cancer Drug.

Small extracellular vesicles (sEVs) are emerging as a novel therapeutic strategy for cancer therapy. Tumor-cell-derived sEVs contain biomolecules that can be utilized for cancer diagnosis. sEVs can directly exert tumor-killing effects or modulate the tumor microenvironment, leading to anti-cancer effects. In this review, the application of sEVs as a diagnostic tool, drug delivery system, and active pharmaceutical ingredient for cancer therapy will be highlighted. The therapeutic efficacies of sEVs will be compared to conventional immune checkpoint inhibitors. Additionally, this review will provide strategies for sEV engineering to enhance the therapeutic efficacies of sEVs. As a bench-to-bedside application, we will discuss approaches to encourage good-manufacturing-practice-compliant industrial-scale manufacturing and purification of sEVs.

Neurophysiological insights into impaired mentalization in borderline personality disorder an electroencephalography study.

Introduction: Borderline personality disorder (BPD) is characterized by interpersonal and emotional instabilities, recurring suicidal tendencies, and feelings of emptiness. Childhood adverse event is reported in 70%-80% of cases involving BPD. Furthermore, the deficiency in mentalization capacity plays a significant role in emotion dysregulation and social interaction problems within individuals with BPD. This study explored the relationship among childhood adverse experiences, mentalization capacity, and neurophysiological activity in patients with BPD. Methods: Resting-state electroencephalography was used to identify the neural correlates associated with childhood adversity and mentalization deficits. The participants included 45 patients with BPD and 15 healthy controls. Results: The BPD group exhibited reduced alpha activity during eyes-closed rest, indicating heightened arousal even during relaxation. Correlations were found between the power spectral density (PSD) and mentalization capacity in the delta and theta ranges, suggesting an association between PSD and emotional awareness and expression. Gamma activity negatively correlated with psychic equivalence, implying a blurring of the boundaries between internal mental experiences and the external world. Conclusion: These findings offer insights into the pathophysiology of BPD, provide potential diagnostic markers, and suggest personalized treatment approaches based on mentalization traits.

Repeated ventricular bigeminy by trigeminocardiac reflex despite atropine administration during superficial upper lip surgery: A case report.

BACKGROUND: The trigeminocardiac reflex (TCR) is usually caused by an increased parasympathetic tone when pressure or traction is applied to the surrounding tissue of the trigeminal nerve. However, the inexperienced anesthesiologists may have challenges on the management of TCR patients. CASE SUMMARY: This is the case of an 18-year-old woman diagnosed with hemangioma of the upper lip. During the operation, about 1 h after surgery started, a constant 1:1 premature ventricular complex was detected, and blood pressure was decreased when approaching the deeper part with more strong traction for exposure of the part. Although the management of arrhythmias, such as lidocaine and atropine, was injected, arrhythmia induced by surgical stimulation could not be eliminated completely. As the traction repeated, bradycardia was also repeated, despite injecting additional atropine. Therefore, the anesthesiologist and the surgeon decided to perform the operation only to the extent that the vascular tissue was selectively removed only at the site without the reflex. CONCLUSION: With TCR, anesthesiologists should perform appropriate monitoring. In addition to proper drug administration, surgeons should be consulted to cope with stopping the surgery and setting the scope of the surgery even if the site is superficial.

The miR-182-5p/NDRG1 Axis Controls Endometrial Receptivity through the NF-κB/ZEB1/E-Cadherin Pathway.

Endometrial receptivity is essential for successful pregnancy, and its impairment is a major cause of embryo-implantation failure. MicroRNAs (miRNAs) that regulate epigenetic modifications have been associated with endometrial receptivity. However, the molecular mechanisms whereby miRNAs regulate endometrial receptivity remain unclear. Therefore, we investigated whether miR-182 and its potential targets influence trophoblast cell attachment. miR-182 was expressed at lower levels in the secretory phase than in the proliferative phase of endometrium tissues from fertile donors. However, miR-182 expression was upregulated during the secretory phase in infertile women. Transfecting a synthetic miR-182-5p mimic decreased spheroid attachment of human JAr choriocarcinoma cells and E-cadherin expression (which is important for endometrial receptivity). miR-182-5p also downregulated N-Myc downstream regulated 1 (NDRG1), which was studied further. NDRG1 was upregulated in the secretory phase of the endometrium tissues and induced E-cadherin expression through the nuclear factor-κΒ (NF-κΒ)/zinc finger E-box binding homeobox 1 (ZEB1) signaling pathway. NDRG1-overexpressing or -depleted cells showed altered attachment rates of JAr spheroids. Collectively, our findings indicate that miR-182-5p-mediated NDRG1 downregulation impaired embryo implantation by upregulating the NF-κΒ/ZEB1/E-cadherin pathway. Hence, miR-182-5p is a potential biomarker for negative selection in endometrial receptivity and a therapeutic target for successful embryo implantation.

Psychiatric and Psychosocial Factors of Suicide Decedents and Survivor of Suicide Loss: Psychological Autopsy Study of Incheon City in South Korea.

In South Korea, the suicide rate is more than double the OECD average, and precise identification of the cause is required for suicide prevention. Psychological autopsy is used to reveal factors related to suicidal behavior; however, such studies are lacking in Korea. This study investigated the factors related to suicide using psychological autopsies in Incheon, a major city in Korea. In total, 46 cases were investigated using the Korea-Psychological Autopsy Checklist (K-PAC), and data on mental health conditions and psychosocial factors of suicide decedents and their families were analyzed. It was estimated that 87% of individuals of suicides had a mental health condition before death, but only 15.2% continued treatment or counseling. Most individuals who died of suicide showed warning signs before death, but only 19.6% of survivors of suicide loss noticed them. Mental health concerns before and after the death of the individual were also identified in more than half of their families. To prevent suicide, intensive and continuous treatment for psychiatric conditions and prompt recognition of active response to suicide warning signs are required. Care for the mental health of family members is also important.

Detection of infectious hypodermal and hematopoietic necrosis virus and white spot syndrome virus in whiteleg shrimp (Penaeus vannamei) imported from Vietnam to South Korea.

In this study, whiteleg shrimp (Penaeus vannamei) imported from Vietnam were collected from South Korean markets, and examined for 2 viruses: infectious hypodermal and hematopoietic necrosis virus (IHHNV, recently classified as decapod penstyldensovirus-1), and white spot syndrome virus (WSSV). Among 58 samples, we detected IHHNV in 23 samples and WSSV in 2 samples, using polymerase chain reaction and sequencing analyses. This is the first report of IHHNV and WSSV detection in imported shrimp, suggesting that greater awareness and stricter quarantine policies regarding viruses infecting shrimp imported to South Korea are required.

Chemical fingerprinting of Codonopsis pilosula and simultaneous analysis of its major components by HPLC-UV.

Although Codonopsis pilosula (C. pilosula) has long been considered as an important herbal medicine, no analytical method of marker compounds for quality assessment is registered in the Korean Pharmacopoeia. We developed a simple and robust analytical method of three marker components lobetyolin (1), lobetyol (2), and tangshenoside I (3) using HPLC-UV method. We also confirmed the three marker components using UPLC-qTOF/MS method. Various extraction conditions were optimized to achieve three marker compounds with faster extraction kinetics and higher recovery. The analytical condition was then validated by determining the linearity, accuracy, precision, limit of detection, limit of quantification, recovery, repeatability, robustness, and stability. By this method, the three markers were successfully quantified in 38 commercial samples along with three related species that are sometimes used as alternatives to C. pilosula. Finally, principal component and hierarchical clustering analyses were conducted to show the practicality of the method developed for the quality evaluation of C. pilosula.

Three-dimensional volumetric gray-scale uterine cervix histogram prediction of days to delivery in full term pregnancy.

OBJECTIVE: Our aim was to figure out whether volumetric gray-scale histogram difference between anterior and posterior cervix can indicate the extent of cervical consistency. METHODS: We collected data of 95 patients who were appropriate for vaginal delivery with 36th to 37th weeks of gestational age from September 2010 to October 2011 in the Department of Obstetrics and Gynecology, Korea University Ansan Hospital. Patients were excluded who had one of the followings: Cesarean section, labor induction, premature rupture of membrane. Thirty-four patients were finally enrolled. The patients underwent evaluation of the cervix through Bishop score, cervical length, cervical volume, three-dimensional (3D) cervical volumetric gray-scale histogram. The interval days from the cervix evaluation to the delivery day were counted. We compared to 3D cervical volumetric gray-scale histogram, Bishop score, cervical length, cervical volume with interval days from the evaluation of the cervix to the delivery. RESULTS: Gray-scale histogram difference between anterior and posterior cervix was significantly correlated to days to delivery. Its correlation coefficient (R) was 0.500 (P = 0.003). The cervical length was significantly related to the days to delivery. The correlation coefficient (R) and P-value between them were 0.421 and 0.013. However, anterior lip histogram, posterior lip histogram, total cervical volume, Bishop score were not associated with days to delivery (P >0.05). CONCLUSION: By using gray-scale histogram difference between anterior and posterior cervix and cervical length correlated with the days to delivery. These methods can be utilized to better help predict a cervical consistency.

A monoclonal antibody specific to glucosyltransferase B of Streptococcus mutans GS-5 and its glucosyltransferase inhibitory efficiency.

Glucosyltransferase-B (GTFB) of Streptococcus mutans is considered a virulence factor because of its activity in the production of insoluble glucan, which is key to the bacterial attachment onto dental surfaces, leading to the formation of dental caries. Local passive immunization with monoclonal antibodies against GTFB is considered to be an effective way to prevent dental caries. Here we amplified a 1.3 kb fragment of the N-terminal half of the gtfB gene (193-1530) of S. mutans by PCR and expressed the truncated protein (GTFBN). The expressed, purified protein was used as an immunogen in BALB/c mice. We selected and established one hybridoma (HBN8) that was capable of producing anti-GTFBN using ELISA, dot blot, and Western blot analyses. The monoclonal anti-GTFBN antibody was purified by affinity chromatography and its isotype was confirmed as IgG2a. The anti-GTFBN antibody inhibited the enzymatic activity of crude glucosyltransferase of S. mutans GS-5 in a dose-dependent manner. These data suggest that the anti-GTFBN antibody could be used as a vaccine to prevent the aggregation of S. mutans on tooth surfaces, and thus prevent the formation of dental caries.

Anti-inflammatory activity of Taraxacum officinale.

Taraxacum officinale has been widely used as a folkloric medicine for the treatment of diverse diseases. The dried plant was extracted with 70% ethanol to generate its ethanol extract (TEE). For some experiments, ethyl acetate (EA), n-butanol (BuOH) and aqueous (Aq) fractions were prepared in succession from TEE. TEE showed a scavenging activity in the 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay, a diminishing effect on intracellular reactive oxygen species (ROS) level, and an anti-angiogenic activity in the chicken chorioallantoic (CAM) assay. In the carrageenan-induced air pouch model, TEE inhibited production of exudate, and significantly diminished nitric oxide (NO) and leukocyte levels in the exudate. It also possessed an inhibitory effect on acetic acid-induced vascular permeability and caused a dose-dependent inhibition on acetic acid-induced abdominal writhing in mice. Suppressive effects of TEE on the production of NO and expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) in lipopolysaccharide (LPS)-stimulated macrophages were also assessed. Among the fractions, the n-butanol fraction (BuOH) was identified to be most effective in the CAM assay. Collectively, Taraxacum officinale contains anti-angiogenic, anti-inflammatory and anti-nociceptive activities through its inhibition of NO production and COX-2 expression and/or its antioxidative activity.

Supramolecular ordering of amide dendrons in lyotropic and thermotropic conditions.

Self-assembled superstructures of amide dendrons, from first to third generation including monodendrons and covalently linked dimers, were extensively examined, and the supramolecular ordering processes in thermotropic and lyotropic conditions were compared. The superstructures as determined by X-ray diffraction and DSC revealed that the first and second generation dendrons showed nearly identical superstructures regardless of the assembly conditions. But, the third generation dendrons showed a more sensitive self-organizing behavior. The structure obtained from the gel state was lamellar with a more extended conformation, while the structure from the melt state revealed the columnar superstructures of contracted branches. The superstructure formed from the gel state also showed a structural change upon raising the temperature and assumed a structure similar to the thermotropically driven one, implying that the structure formed from the gel is thermodynamically unstable. The formation of lamellar- or cylinder-type superstructures from amide dendrons was primarily dependent on the shape of dendrons, which is associated with the branch size (generation) and the surrounding conditions.

Anti-angiogenic activity of the methanol extract and its fractions of Ulmus davidiana var. japonica.

This study aimed to elucidate anti-angiogenic activity of Ulmus davidiana var. japonica that has been widely used in folk medicine. The methanol extract (UDE) of Ulmus davidiana var. japonica concentration-dependently displayed a strong inhibition in the chick chorioallantoic membrane (CAM) angiogenesis. The n-butanol fraction of UDE and subsequent 30% MeOH subfraction were identified to be most responsible for the anti-angiogenic activity.

Anti-inflammatory and anti-angiogenic activities of Gastrodia elata Blume.

Gastrodia elata Blume rhizome has been traditionally used as a folk medicine for centuries in Oriental countries. Its ethanol extract (GEE) and subsequent fractions were used to evaluate anti-angiogenic, anti-inflammatory and related activities of Gastrodia elata. GEE potently inhibited angiogenesis in the chick chorioallantoic membrane assay, and its n-butanol fraction (BuOH) exerted the higher inhibitory effect. In a dose-dependent manner, GEE inhibited vascular permeability induced by acetic acid. GEE and its BuOH fraction exerted an inhibitory activity on exudate production, leukocyte migration and nitric oxide (NO) level in rat air-pouch model. GEE caused a dose-dependent inhibition of acetic acid-induced abdominal writhing in mice. In addition, GEE inhibited NO production and expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) upon stimulation by lipopolysaccharide (LPS) in RAW264.7 macrophages. In summary, we demonstrate some novel pharmacological activities of Gastrodia elata, such as anti-angiogenic, anti-inflammatory and analgesic activities, and in vivo and in vitro inhibitory activity on NO production.

Self-organization of amide dendrons and their dendronized macromolecules.

A polymerizable methacryl unit was introduced at the focal moiety of the amide dendrons which have amide branches and alkyl periphery. Their dendronized polymers were also prepared by the radical polymerization of the methacryl units. The self-organization characteristics of dendrons and dendronized polymers were then investigated in both the organic and aqueous phases. The amide dendrons (1M and 2M) in which the focal carboxyl group was blocked with methacryl units did not form gel in organic media such as chloroform or THF, whereas amide dendrons with a free carboxyl group at the focal point form self-organized structures. In the aqueous phase, 1M and 2M formed spherical vesicular assemblies. The dendronized polymers with first and second generation dendrons, 1P and 2P, respectively, exhibited lamellar and columnar organization in toluene. In addition to hydrogen bonding between the dendritic amide branches and van der Waals interactions between the alkyl periphery, steric confinement of dendritic side groups along the polymer backbone played a key role in the packing process of the dendronized polymers. In aqueous phase, 1P and 2P showed spherical vesicular aggregates with persistent stability in the presence of Triton X-100.