Synthetic ShK-like Peptide from the Jellyfish Nemopilema nomurai Has Human Voltage-Gated Potassium-Channel-Blocking Activity.

We identified a new human voltage-gated potassium channel blocker, NnK-1, in the jellyfish Nemopilema nomurai based on its genomic information. The gene sequence encoding NnK-1 contains 5408 base pairs, with five introns and six exons. The coding sequence of the NnK-1 precursor is 894 nucleotides long and encodes 297 amino acids containing five presumptive ShK-like peptides. An electrophysiological assay demonstrated that the fifth peptide, NnK-1, which was chemically synthesized, is an effective blocker of hKv1.3, hKv1.4, and hKv1.5. Multiple-sequence alignment with cnidarian Shk-like peptides, which have Kv1.3-blocking activity, revealed that three residues (3Asp, 25Lys, and 34Thr) of NnK-1, together with six cysteine residues, were conserved. Therefore, we hypothesized that these three residues are crucial for the binding of the toxin to voltage-gated potassium channels. This notion was confirmed by an electrophysiological assay with a synthetic peptide (NnK-1 mu) where these three peptides were substituted with 3Glu, 25Arg, and 34Met. In conclusion, we successfully identified and characterized a new voltage-gated potassium channel blocker in jellyfish that interacts with three different voltage-gated potassium channels. A peptide that interacts with multiple voltage-gated potassium channels has many therapeutic applications in various physiological and pathophysiological contexts.

Capillary gingival hemangioma in a cat.

A 12-year-old spayed female American short-haired cat presented with a palatal gingival mass located between the right maxillary third incisor and the canine teeth. The mass was dark red and had a narrow attachment to the gingival margin of the canine tooth. The mass was completely removed by marginal excision and the histopathological diagnosis was a capillary hemangioma. The mass did not relapse until 1 year later; however, the tooth was extracted because of cervical resorption of the right maxillary canine immediately adjacent to the mass resection site. This report presents a rare case of the gingival hemangioma in a cat and the possibility of a causal relationship between the occurrence of external cervical tooth resorption and hemangioma resection.

Development of Reproducible and Scalable Culture Conditions for In Vitro Maintenance of Pig Embryonic Stem Cells Using the Sandoz Inbred Swiss Mouse Thioguanine-Resistant Ouabain-Resistant Cell Line as a Feeder Layer.

Feeder cells play a crucial role in maintaining the pluripotency of embryonic stem cells (ESCs) by secreting various extrinsic regulators, such as extracellular matrix (ECM) proteins and growth factors. Although primary mouse embryonic fibroblasts (MEFs) are the most widely used feeder cell type for the culture of ESCs, they have inevitable disadvantages such as batch-to-batch variation and labor-intensive isolation processes. Here, we revealed that the Sandoz inbred Swiss Mouse (SIM) thioguanine-resistant ouabain-resistant (STO) cell line, an immortalized cell line established from mouse SIM embryonic fibroblasts, can be used as a feeder layer for in vitro culture of authentic pig ESCs instead of primary MEFs. First, the expression of genes encoding ECM proteins and growth factors was analyzed to compare their secretory functions as feeder cells. Quantitative real-time polymerase chain reaction (qPCR) showed that the gene expression of these pluripotency-associated factors was downregulated in STO cells compared to primary MEFs of similar density. Therefore, subsequent optimization of the culture conditions was attempted using higher STO cell densities. Notably, pig ESCs cultured on STO cell density of 3 × (187,500 cells/cm2) exhibited the most similar pluripotent state to pig ESCs cultured on primary MEF density of 1 × (62,500 cells/cm2), as determined by alkaline phosphatase staining, qPCR, and immunocytochemistry. In addition, pig ESCs cultured on STO cell density of 3 × formed complex teratoma containing multiple types of tissues derived from all three germ layers. Our culture conditions using optimal STO cell density can be applied to fields requiring reproducible and scalable production of pig ESCs, such as preclinical research and cellular agriculture.

High-Resolution Tactile-Sensation Diagnostic Imaging System for Thyroid Cancer.

In this study, we propose the direct diagnosis of thyroid cancer using a small probe. The probe can easily check the abnormalities of existing thyroid tissue without relying on experts, which reduces the cost of examining thyroid tissue and enables the initial self-examination of thyroid cancer with high accuracy. A multi-layer silicon-structured probe module is used to photograph light scattered by elastic changes in thyroid tissue under pressure to obtain a tactile image of the thyroid gland. In the thyroid tissue under pressure, light scatters to the outside depending on the presence of malignant and positive properties. A simple and easy-to-use tactile-sensation imaging system is developed by documenting the characteristics of the organization of tissues by using non-invasive technology for analyzing tactile images and judging the properties of abnormal tissues.

Primary well differentiated hepatic liposarcoma in a meerkat (Suricata suricatta).

A 9-year-old female meerkat (Suricata suricatta) succumbed to progressive abdominal distension, anorexia, and depression. Necropsy revealed an extensively distended abdomen with ascites and markedly enlarged liver. The liver had multiple yellowish masses and displaced the thoracic cavity and abdominal organs. There was no evidence of metastatic lesions based on the gross and microscopic findings. Histologically, the liver mass was composed of locally invasive well-differentiated neoplastic adipocytes with Oil Red O-positive lipid vacuoles. Immunohistochemistry revealed positive immunoreactivity to vimentin, S-100 and negative to pancytokeratin, desmin, smooth muscle actin (SMA), ionized calcium-binding adapter molecule 1 (IBA-1). Thus, the primary well differentiated hepatic liposarcoma was diagnosed based on gross, histological and immunohistochemistry results.

The Role of Fucoxanthin as a Potent Nrf2 Activator via Akt/GSK-3ß/Fyn Axis against Amyloid-ß Peptide-Induced Oxidative Damage.

Increasing evidence is suggesting that amyloid-ß peptide (Aß), a characteristic of Alzheimer's disease (AD), induces oxidative stress and mitochondrial dysfunction, leading to neuronal death. This study aimed to demonstrate the antioxidant and anti-apoptotic effects of fucoxanthin, a major marine carotenoid found in brown algae, against neuronal injury caused by Aß. Non-toxic dose range of fucoxanthin (0.1-5 µM) were selected for the neuroprotective study against Aß25-35. The PC12 cells were pretreated with different concentrations of fucoxanthin for 1 h before being exposed to 10 µM Aß25-35 for another 24 h. The present results showed that fucoxanthin inhibited Aß25-35-induced cell death by recovering cell cycle arrest and decreasing intracellular reactive oxygen species (ROS) level. The compound enhanced mitochondrial recovery and regulated apoptosis related proteins including B-cell lymphoma 2 (Bcl-2) and Bcl-2-associated X protein (Bax) from Aß25-35-induced oxidative stress. Concomitantly, fucoxanthin increased the expression of nuclear factor E2-related factor 2 (Nrf2) and its downstream phase II detoxifying enzymes including NADPH: quinone oxidoreductase-1 (NQO-1), glutamate cysteine ligase modifier subunit (GCLm), and thioredoxin reductase 1 (TrxR1), whereas it decreased the expression of cytoplasmic Kelch-like ECH-associated protein 1 (Keap1). Moreover, pretreatment of fucoxanthin reduced Fyn phosphorylation via protein kinase B (Akt)-mediated inhibition of glycogen synthase kinase-3ß (GSK-3ß), which increased the nuclear localization of Nrf2, suggesting that the compound enhanced Nrf2 expression by the activation of upstream kinase as well as the dissociation of the Nrf2-Keap1 complex. Further validation with a specific phosphatidylinositol 3-kinase (PI3K) inhibitor LY294002 demonstrated that the fucoxanthin-mediated Nrf2 antioxidant defense system was directly associated with the Akt/GSK-3ß/Fyn signaling pathway. In silico simulation revealed that the oxygen groups of fucoxanthin participated in potent interactions with target markers in the Nrf2 signaling pathway, which may affect the biological activity of target markers. Taken together, the present results demonstrated that the preventive role of fucoxanthin on Aß-stimulated oxidative injury and apoptosis via Akt/GSK-3ß/Fyn signaling pathway. This study would provide a useful approach for potential intervention for AD prevention.

Splenic epidermoid cyst in a dog.

Splenic epithelial cysts are rare in humans and have not been reported in animals, to our knowledge. During a routine medical examination of a 12-y-old castrated male Maltese dog, a splenic mass was found and subsequently removed via splenectomy. Histologically, a well-defined multilocular cyst in the spleen was lined mostly by simple cuboidal, multifocally by stratified cuboidal, or occasionally by stratified squamous epithelium. Immunohistochemically, the lining cells were positive for cytokeratin and negative for vimentin, CD31, and Wilms tumor protein 1. The case was diagnosed as a primary splenic epidermoid cyst.

Consistency of Product Quality for SB5, an Adalimumab Biosimilar.

BACKGROUND: Biologics, regardless of whether they are biosimilars or reference products, are inherently variable due to their size, complexity, and the manufacturing process involved to produce them. Since a drift or evolution of quality attributes of a biologic may impact its clinical safety or efficacy, it is critical for the manufacturer to carefully control the manufacturing process and monitor the quality attributes of a biologic. OBJECTIVE: The aim of this study was to demonstrate that the quality profile of the SB5 drug product has been consistent over its production history from 2013 to 2022. SB5 is a biosimilar referencing adalimumab (Humira, trademark of AbbVie Biotechnology Ltd) and SB5 has been approved by 14 regulatory authorities including the European Commission in August 2017 (brand name Imraldi™) and the US Food and Drug Administration in July 2019 (brand name Hadlima™). METHODS: A total of 93 SB5 drug product batches manufactured between 2013 and 2022 were analyzed for a series of release parameters to evaluate the consistency in their critical quality attributes including purity, charge variants, and functional activities (TNF-α binding activity and TNF-α neutralizing potency). RESULTS: The purity, charge variants, and functional activities of all batches were consistent over time and within the stringent acceptance criteria defined by regulatory agencies to ensure the safety and efficacy of SB5. CONCLUSION: The data presented in this study provide evidence that the quality of SB5 has remained consistent and tightly controlled even through process changes such as manufacturing site transfers and change in formulation.

Staged melanoma excision requires larger margins for tumor clearance and results in low rates of recurrence.

There is controversy regarding the optimal surgical modality and ideal recommended margins for treating melanoma in situ (MIS) and invasive melanoma (IM). Although wide local excision is recommended, staged excision offers excellent margin control and low recurrence rates. In this manuscript, we reviewed a 10-year experience of staged excisions for the treatment of MIS and IM. A retrospective review was performed of 130 MIS and 32 IM cases treated with staged excision from April 2012 to April 2022. Staged excision was performed on the head and neck in 102 (79%) MIS and 23 (72%) IM cases. Approximately 10% of cases required surgical margins above the current recommendations (11 (9%) MIS and 6 (19%) IM). Twenty-three (19%) MIS and 7 (22%) IM cases required more than one excision to obtain clearance. Recurrence rates among MIS and IM were 0.0% and 0.6%, respectively. Upstaging occurred in 5 (4%) MIS and 7 (22%) IM cases. Complex repairs were performed on 82 (63%) MIS and 17 (53%) IM cases. Our findings revealed that staged excision provides effective margin control and low recurrence rates. Approximately 10% of patients required margins greater than the current recommendations, leading to larger defects and more complex repairs.

Applying Occam's Razor and Descending the Reconstructive Ladder: The Modified Cheek Advancement Flap for Reconstruction of Nasal Defects.

Nasal reconstruction has important functional and cosmetic considerations, as proper repair of nasal defects is necessary to maintain function of the nasal airway and to recreate the normal appearance of this central facial structure. Cheek advancement flaps provide matched, mobile, and highly vascularized tissue for the reconstruction of nasal defects, allowing for the concealment of incisions within natural creases in a one-stage approach. However, cheek advancement flaps are often underutilized for nasal reconstruction because of their difficulty in restoring nasal contour. We describe reconstruction of 19 nasal dorsal and sidewall defects 0.8 to 3 cm in size. We incorporated a periosteal anchoring suture to maintain/restore nasal contour and additionally removed a half standing cone inferior to the defect to prevent encroachment of the nasal ala or alar crease. All patients were evaluated at least 3 months postoperatively. In all patients, we were able to restore concavity of the nasofacial sulcus, preserve the biconvex nasal tips, prevent alar flaring and retraction, and conserve the alar groove. All patients had excellent functional and cosmetic outcomes. We believe this modified cheek advancement flap provides functionally and aesthetically superior results and can be considered as a first-line approach for repair of nasal dorsal and sidewall defects in subselected patients.

Monitoring Cellular Immune Responses after Consumption of Selected Probiotics in Immunocompromised Mice.

Probiotics are currently considered as one of tools to modulate immune responses under specific clinical conditions. The purpose of this study was to evaluate whether oral administration of three different probiotics (Lactiplantibacillus plantarum CJLP243, CJW55-10, and CJLP475) could evoke a cell-mediated immunity in immunodeficient mice. Before conducting in vivo experiments, we examined the in vitro potency of these probiotics for macrophage activation. After co-culture with these probiotics, bone marrow derived macrophages (BMDMs) produced significant amounts of proinflammatory cytokines including interleukin-6 (IL-6), IL-12, and tumor necrosis factor-α (TNF-α). Levels of inducible nitric oxide synthase (inos) and co-stimulatory molecules (CD80 and CD86) were also upregulated in BMDMs after treatment with some of these probiotics. To establish an immunocompromised animal model, we intraperitoneally injected mice with cyclophosphamide on day 0 and again on day 2. Starting day 3, we orally administered probiotics every day for the last 15 d. After sacrificing experimental mice on day 18, splenocytes were isolated and co-cultured with these probiotics for 3 d to measure levels of several cytokines and immune cell proliferation. Results clearly indicated that the consumption of all three probiotic strains promoted secretion of interferon-γ (IFN-γ), IL-1ß, IL-6, IL-12, and TNF-α. NK cell cytotoxicity and proliferation of immune cells were also increased. Taken together, our data strongly suggest that consumption of some probiotics might induce cell-mediated immune responses in immunocompromised mice.

Online information analysis on pancreatic cancer in Korea using structural topic model.

Inappropriate information on a deadly and rare disease can make people vulnerable to problematic decisions, leading to irreversible bad outcomes. This study explored online information exchanges on pancreatic cancer. We collected 35,596 questions and 83,888 answers related to pancreatic cancer from January 1, 2003 to May 31, 2020, from Naver, the most popular Korean web portal. We also collected 8495 news articles related to pancreatic cancer during the same period. The study methods employed were structural topic modeling, keyword frequency analysis, and qualitative coding of medical professionals. The number of questions and news articles increased over time. In Naver's questions, topics on symptoms and diagnostic tests regarding pancreatic cancer increased in proportion. The news topics on new technologies related to pancreatic cancer from various companies increased as well. The use of words related to back pain-which is not an important early symptom in pancreatic cancer-and biomarker tests using blood increased over time in Naver's questions. Based on 100 question samples related to symptoms and diagnostic tests and an analysis of the threaded answers' appropriateness, there was considerable misinformation and commercialized information in both categories.

Effects of extremely high concentrations of polystyrene microplastics on asexual reproduction and nematocyst discharge in the jellyfish Sanderia malayensis.

Numerous studies have assessed the detrimental effects of microplastics (MPs) on aquatic invertebrates due to their ubiquitous and persistent nature. In this study, the toxic effects of MPs were examined on the polyp and ephyrae of the marine hydrozoan Sanderia malayensis. The jellyfish were exposed to different sizes (1-6 µm) of non-functionalized polystyrene microbeads at a concentration of 1 × 104 particles mL-1. The MPs randomly attached to the external and internal parts of the jellyfish body, and the longest MP attachment was 52 days during the depuration after initial exposure (for 24 h). Consistent seventeen-day exposure to MPs significantly reduced the asexual reproduction of the S. malayensis polyps. To assess if the MPs can stimulate nematocyst discharge in polyp and ephyrae stages via direct contact, they were exposed to particle sizes up to 430 µm. None of the MPs or their aggregates, including the 430 µm particles, induced nematocyst discharge. These results suggest that prolonged exposure to relatively high MP concentrations affects the early stages of jellies and provides evidence for the no effect on nematocyst discharge.

Fluoxetine in the environment may interfere with the neurotransmission or endocrine systems of aquatic animals.

Antidepressants are extensively used to treat the symptoms of depression in humans, and the environmentally discharged drugs potentially threaten aquatic organisms. In this study, the acute toxic effects of fluoxetine (FLX) were investigated in two aquatic organisms, the freshwater polyp (Hydra magnipapillata) and Javanese medaka (Oryzias javanicus). The median lethal concentration (LC50) of FLX in H. magnipapillata was 3.678, 3.082, and 2.901 mg/L after 24, 48, and 72 h, respectively. Morphological observations of the FLX-exposed H. magnipapillata showed that 1.5 mg/L FLX induced the contraction of the tentacles and body column. The LC50 of FLX in O. javanicus was 2.046, 1.936, 1.532, and 1.237 mg/L after 24, 48, 72, and 96 h, respectively. Observation of the behavior of the FLX-exposed fish showed that FLX reduced their swimming performance at a minimum concentration of 10 µg/L. The half-maximal effective concentration (EC50) of FLX for swimming behavior in O. javanicus was 0.135, 0.108, and 0.011 mg/L after 12, 24, and 96 h, respectively. Transcriptomic analyses indicated that FLX affects various physiological and metabolic processes in both species. FLX exposure induced oxidative stress, reproductive deficiency, abnormal pattern formation, DNA damage, and neurotransmission disturbance in H. magnipapillata, whereas it adversely affected O. javanicus by inducing oxidative stress, DNA damage, endoplasmic reticulum stress, and mRNA instability. Neurotransmission-based behavioral changes and endocrine disruption were strongly suspected in the FLX-exposed fish. These results suggest that FLX affects the behavior and metabolic regulation of aquatic organisms.

Dieckol Ameliorates Aß Production via PI3K/Akt/GSK-3ß Regulated APP Processing in SweAPP N2a Cell.

The proteolytic processing of amyloid precursor protein (APP) by ß-secretase (BACE1) and γ-secretase releases amyloid-ß peptide (Aß), which deposits in amyloid plaques and contributes to the initial causative events of Alzheimer's disease (AD). In the present study, the regulatory mechanism of APP processing of three phlorotannins was elucidated in Swedish mutant APP overexpressed N2a (SweAPP N2a) cells. Among the tested compounds, dieckol exhibited the highest inhibitory effect on both intra- and extracellular Aß accumulation. In addition, dieckol regulated the APP processing enzymes, such as α-secretase (ADAM10), ß-secretase, and γ-secretase, presenilin-1 (PS1), and their proteolytic products, sAPPα and sAPPß, implying that the compound acts on both the amyloidogenic and non-amyloidogenic pathways. In addition, dieckol increased the phosphorylation of protein kinase B (Akt) at Ser473 and GSK-3ß at Ser9, suggesting dieckol induced the activation of Akt, which phosphorylated GSK-3ß. The specific phosphatidylinositol 3-kinase (PI3K) inhibitor LY294002 triggered GSK-3ß activation and Aß expression. In addition, co-treatment with LY294002 noticeably blocked the effect of dieckol on Aß production, demonstrating that dieckol promoted the PI3K/Akt signaling pathway, which in turn inactivated GSK-3ß, resulting in the reduction in Aß levels.

Risk-based prediction model for selecting eligible population for lung cancer screening among ever smokers in Korea.

BACKGROUND: This study developed a new lung cancer risk prediction model for the Korean population and evaluated the performance, compared to the previously reported risk models developed in Western countries. METHODS: Among the 6,811,893 people who received health examinations from the Korean National Health Insurance Service, 969,351 ever-smokers (40-79 years) were included. Performance of Bach, Lung Cancer Risk Models for Screening, PLCOM2012, Pittsburgh, and Liverpool Lung Project models were evaluated. The ever-smokers were divided into the training and validation datasets by random sampling. The lung cancer risk model was developed and validated in the Korean population. The efficiency of model-based selection for lung cancer screening was compared with the eligible criteria of the National Lung Screening Trial (NLST). RESULTS: The Korean lung cancer risk model showed the area under the curve and expected/observed (E/O) ratio of 0.816 and 0.983 in the training dataset and 0.816 and 0.988 in the validation dataset. The Korean lung cancer risk model included age-mean of age, square of age-mean of age, sex, square root of pack-years of smoking, years since cessation, physical activity, alcohol consumption, body mass index, and medical history of chronic pulmonary obstructive disease, emphysema, pneumoconiosis, and interstitial pulmonary disease. Compared with the NLST criteria, the Korean lung cancer risk model's cut-off criteria (>2.1%) had more improved sensitivity (61.4% vs. 44.3%) and positive predictive value (4.1% vs. 2.9%). The Korean lung cancer risk model showed better discrimination and calibration than previously developed models in Western population. CONCLUSIONS: The Korean lung cancer risk model can select eligible population for low-dose computed tomography screening among the Asian population. The efficiency of risk model-based selection for lung cancer screening is superior to that of fixed criteria-based selection.