The Identification of a Novel Spider Toxin Peptide, Lycotoxin-Pa2a, with Antibacterial and Anti-Inflammatory Activities.

With the increasing challenge of controlling infectious diseases due to the emergence of antibiotic-resistant strains, the importance of discovering new antimicrobial agents is rapidly increasing. Animal venoms contain a variety of functional peptides, making them a promising platform for pharmaceutical development. In this study, a novel toxin peptide with antibacterial and anti-inflammatory activities was discovered from the spider venom gland transcriptome by implementing computational approaches. Lycotoxin-Pa2a (Lytx-Pa2a) showed homology to known-spider toxin, where functional prediction indicated the potential of both antibacterial and anti-inflammatory peptides without hemolytic activity. The colony-forming assay and minimum inhibitory concentration test showed that Lytx-Pa2a exhibited comparable or stronger antibacterial activity against pathogenic strains than melittin. Following mechanistic studies revealed that Lytx-Pa2a disrupts both cytoplasmic and outer membranes of bacteria while simultaneously inducing the accumulation of reactive oxygen species. The peptide exerted no significant toxicity when treated to human primary cells, murine macrophages, and bovine red blood cells. Moreover, Lytx-Pa2a alleviated lipopolysaccharide-induced inflammation in mouse macrophages by suppressing the expression of inflammatory mediators. These findings not only suggested that Lytx-Pa2a with dual activity can be utilized as a new antimicrobial agent for infectious diseases but also demonstrated the implementation of in silico methods for discovering a novel functional peptide, which may enhance the future utilization of biological resources.

In Silico-Based Design of a Hybrid Peptide with Antimicrobial Activity against Multidrug-Resistant Pseudomonas aeruginosa Using a Spider Toxin Peptide.

The escalating prevalence of antibiotic-resistant bacteria poses an immediate and grave threat to public health. Antimicrobial peptides (AMPs) have gained significant attention as a promising alternative to conventional antibiotics. Animal venom comprises a diverse array of bioactive compounds, which can be a rich source for identifying new functional peptides. In this study, we identified a toxin peptide, Lycotoxin-Pa1a (Lytx-Pa1a), from the transcriptome of the Pardosa astrigera spider venom gland. To enhance its functional properties, we employed an in silico approach to design a novel hybrid peptide, KFH-Pa1a, by predicting antibacterial and cytotoxic functionalities and incorporating the amino-terminal Cu(II)- and Ni(II) (ATCUN)-binding motif. KFH-Pa1a demonstrated markedly superior antimicrobial efficacy against pathogens, including multidrug-resistant (MDR) Pseudomonas aeruginosa, compared to Lytx-Pa1a. Notably, KFH-Pa1a exerted several distinct mechanisms, including the disruption of the bacterial cytoplasmic membrane, the generation of intracellular ROS, and the cleavage and inhibition of bacterial DNA. Additionally, the hybrid peptide showed synergistic activity when combined with conventional antibiotics. Our research not only identified a novel toxin peptide from spider venom but demonstrated in silico-based design of hybrid AMP with strong antimicrobial activity that can contribute to combating MDR pathogens, broadening the utilization of biological resources by incorporating computational approaches.

Curcumin Disrupts a Positive Feedback Loop between ADMSCs and Cancer Cells in the Breast Tumor Microenvironment via the CXCL12/CXCR4 Axis.

Adipose tissue has a significant impact on breast cancer initiation and progression owing to its substantial proportion in the breast. Adipose-derived mesenchymal stem cells (ADMSCs) are major players in the breast tumor microenvironment (TME) as they interact with cancer cells. The intricate interaction between ADMSCs and cancer cells not only drives the differentiation of ADMSCs into cancer-associated fibroblasts (CAFs) but also the metastasis of cancer cells, which is attributed to the CXCL12/CXCR4 axis. We investigated the effects of curcumin, a flavonoid known for CXCL12/CXCR4 axis inhibition, on breast TME by analyzing whether it can disrupt the ADMSC-cancer positive loop. Using MCF7 breast cancer cell-derived conditioned medium (MCF7-CM), we induced ADMSC transformation and verified that curcumin diminished the phenotypic change, inhibiting CAF marker expression. Additionally, curcumin suppressed the CXCL12/CXCR4 axis and its downstream signaling both in ADMSCs and MCF7 cells. The CM from ADMSCs, whose ADMSC-to-CAF transformation was repressed by the curcumin treatment, inhibited the positive feedback loop between ADMSCs and MCF7 as well as epithelial-mesenchymal transition in MCF7. Our study showed that curcumin is a potent anti-cancer agent that can remodel the breast TME, thereby restricting the ADMSC-cancer positive feedback loop associated with the CXCL12/CXCR4 axis.

In silico identification of novel antimicrobial peptides from the venom gland transcriptome of the spider Argiope bruennichi (Scopoli, 1772).

As the emergence and prevalence of antibiotic-resistant strains have resulted in a global crisis, there is an urgent need for new antimicrobial agents. Antimicrobial peptides (AMPs) exhibit inhibitory activity against a wide spectrum of pathogens and can be utilized as an alternative to conventional antibiotics. In this study, two novel AMPs were identified from the venom transcriptome of the spider Argiope bruennichi (Scopoli, 1772) using in silico methods, and their antimicrobial activity was experimentally validated. Aranetoxin-Ab2a (AATX-Ab2a) and Aranetoxin-Ab3a (AATX-Ab3a) were identified by homology analysis and were predicted to have high levels of antimicrobial activity based on in silico analysis. Both peptides were found to have antibacterial effect against Gram-positive and -negative strains, and, in particular, showed significant inhibitory activity against multidrug-resistant Pseudomonas aeruginosa isolates. In addition, AATX-Ab2a and AATX-Ab3a inhibited animal and vegetable fungal strains, while showing low toxicity to normal human cells. The antimicrobial activity of the peptides was attributed to the increased permeability of microbial membranes. The study described the discovery of novel antibiotic candidates, AATX-Ab2a and AATX-Ab3a, using the spider venom gland transcriptome, and validated an in silico-based method for identifying functional substances from biological resources.

De Novo Design of AC-P19M, a Novel Anticancer Peptide with Apoptotic Effects on Lung Cancer Cells and Anti-Angiogenic Activity.

Despite the current developments in cancer therapeutics, efforts to excavate new anticancer agents continue rigorously due to obstacles, such as side effects and drug resistance. Anticancer peptides (ACPs) can be utilized to treat cancer because of their effectiveness on a variety of molecular targets, along with high selectivity and specificity for cancer cells. In the present study, a novel ACP was de novo designed using in silico methods, and its functionality and molecular mechanisms of action were explored. AC-P19M was discovered through functional prediction and sequence modification based on peptide sequences currently available in the database. The peptide exhibited anticancer activity against lung cancer cells, A549 and H460, by disrupting cellular membranes and inducing apoptosis while showing low toxicity towards normal and red blood cells. In addition, the peptide inhibited the migration and invasion of lung cancer cells and reversed epithelial-mesenchymal transition. Moreover, AC-P19M showed anti-angiogenic activity through the inhibition of vascular endothelial growth factor receptor 2 signaling. Our findings suggest that AC-P19M is a novel ACP that directly or indirectly targets cancer cells, demonstrating the potential development of an anticancer agent and providing insights into the discovery of functional substances based on an in silico approach.

Quercetin and Isorhamnetin Reduce Benzo[a]pyrene-Induced Genotoxicity by Inducing RAD51 Expression through Downregulation of miR-34a.

Benzo[a]pyrene (B[a]P) is metabolized in the liver into highly reactive mutagenic and genotoxic metabolites, which induce carcinogenesis. The mutagenic factors, including B[a]P-7,8-dihydrodiol-9,10-epoxide (BPDE) and reactive oxygen species, generated during B[a]P metabolism can cause DNA damage, such as BPDE-DNA adducts, 8-oxo-dG, and double-strand breaks (DSBs). In this study, we mechanistically investigated the effects of quercetin and its major metabolite isorhamnetin on the repair of B[a]P-induced DNA DSBs. Whole-transcriptome analysis showed that quercetin and isorhamnetin each modulate the expression levels of genes involved in DNA repair, especially those in homologous recombination. RAD51 was identified as a key gene whose expression level was decreased in B[a]P-treated cells and increased by quercetin or isorhamnetin treatment. Furthermore, the number of γH2AX foci induced by B[a]P was significantly decreased by quercetin or isorhamnetin, whereas RAD51 mRNA and protein levels were increased. Additionally, among the five microRNAs (miRs) known to downregulate RAD51, miR-34a level was significantly downregulated by quercetin or isorhamnetin. The protective effect of quercetin or isorhamnetin was lower in cells transfected with a miR-34a mimic than in non-transfected cells, and the B[a]P-induced DNA DSBs remained unrepaired. Our results show that quercetin and isorhamnetin each upregulates RAD51 by downregulating miR-34a and thereby suppresses B[a]P-induced DNA damage.

Immunomodulatory Effect and Bone Homeostasis Regulation in Osteoblasts Differentiated from hADMSCs via the PD-1/PD-L1 Axis.

Human mesenchymal stem cells (hMSCs) are promising candidates for stem cell therapy and are known to secrete programmed death-1 (PD-1) ligand 1 (PD-L1) regulating T cell-mediated immunosuppression. Given the limitations of current stem cell therapy approaches, improvements in immunomodulatory capacity and stem cell differentiation efficacy are needed. In this study, we propose novel strategies to overcome the challenges that remain in hMSC-mediated bone regeneration. We found that PD-1 is highly expressed in osteoblasts, and the PD-1/PD-L1 axis mediated the decreased proinflammatory cytokine expressions in differentiated osteoblasts cocultured with human adipose derived mesenchymal stem cells (hADMSCs). Moreover, the decrease was attenuated by PD-1/PD-L1 pathway inhibition. Osteogenic properties including osteogenic gene expression and calcium deposits were increased in osteoblasts cocultured with hADMSCs compared with those that were monocultured. Osteoblasts treated with PD-L1 and exosomes from hADMSCs also exhibited enhanced osteogenic properties, including calcium deposits and osteogenic gene expression. In our cocultured system that mimics the physiological conditions of the bone matrix, the PD-1/PD-L1 axis mediated the increased expression of osteogenic genes, thereby enhancing the osteogenic properties, while the calcium deposits of osteoblasts were maintained. Our results provide the therapeutic potentials and novel roles of the PD-1/PD-L1 axis in bone matrix for modulating the bone properties and immunosuppressive potentials that can aid in the prevention of bone diseases via maintaining bone homeostasis.

Erratum: A Vegetable and Fish Dietary Pattern Is Positively Associated with Skeletal Muscle Mass in Korean Men.

[This corrects the article on p. 1 in vol. 8, PMID: 30746343.].

Erratum: Total energy intake according to the level of skeletal muscle mass in Korean adults aged 30 years and older: an analysis of the Korean National Health and Nutrition Examination Surveys (KNHANES) 2008-2011.

[This corrects the article on p. 222 in vol. 12, PMID: 29854328.].

A Vegetable and Fish Dietary Pattern Is Positively Associated with Skeletal Muscle Mass in Korean Men.

Isolating the effects of a single nutrient or food in relation to health outcomes including increased skeletal muscle mass is a challenging task because dietary constituents are highly correlated and synergistic. Hence, diet pattern analysis may be used to investigate the role of certain diets in health outcomes. The present study investigated the dietary patterns and their relationship to skeletal muscle mass in Korean adults. Data were extracted from the 2008-2011 Korea National Health and Nutrition Examination Surveys. To explore the dietary patterns of the study subjects, factor analysis was performed using data obtained from a 24-hour recall. The skeletal muscle index according to dietary pattern scores was then investigated to estimate the changes in skeletal muscle mass. Three patterns were initially identified from the factor analysis. Of these vegetables and fish (VF) pattern was the primary factor with high reliability and was a common factor in sex-separated analyses. The VF pattern scores were positively associated with increased skeletal muscle mass in both men and women. Further analysis according to quartile levels of VF pattern scores showed a positive association between skeletal muscle mass and VF pattern in men but not in women. These results suggest that dietary patterns focused on vegetables and seafoods may contribute to increased skeletal muscle mass in Korean men but that sex difference should be considered in nutrition care for skeletal muscle health.

The roles of doctors, nurses, and industrial hygienists in the healthcare management services in Korea: a comparison of the opinions of specialized health management institutions and entrusted enterprises.

BACKGROUND: This study aimed to identify the difference of perception about the role of appointing health officers by comparing and analyzing the response of entrustment workplace (EW) and specialized health management institution (SI). This is considered an important aspect of an institutional assessment to improve the quality of health management services. METHODS: A survey questionnaire was mailed to 122 SIs and 319 EWs nationwide. The questionnaire survey was about the general characteristics of SIs and EWs and main occupations for each evaluation item. In total, 81 SIs (66.4%) and 30 EWs responded to the questionnaire. A logistic regression analysis was performed to compare the opinions of SI and EW. RESULTS: Based on the analysis, the items showing statistically significant differences were as follows. Doctors' main tasks survey: "Guidance on their wearing personal protective equipment (PPE)" (OR: 4.58), "Guidance of improvement of work environment (WE)" (OR: 3.33), etc.; Nurses' main tasks survey: "Guidance on their wearing PPE" (OR: 3.86), "Guidance for programs on health process in confined space (CS)" (OR: 0.36), "Guidance on the hearing conservation program (HCP)" (OR: 0.28), etc.; Industrial hygienist (IH)'s main tasks survey: "Guidance of work through inspection (WTI)" (OR: 0.15), "Guidance on the improvement of WE" (OR: 0.32), "Management confirmation of substances used by process and Material Safety Data Sheet (MSDS)" (OR: 0.08), "Guidance on posting or keeping of MSDS and warning signs" (OR: 0.03), "Prevention of dust-induced medical problems" (OR: 0.28), "Guidance for programs of health process in CS" (OR: 0.39), etc. CONCLUSIONS: It is necessary to educate the EWs to recognize the need for physicians to perform tasks, such as wearing a PPE, and instruction to improve WE. As for nurses' tasks, such as education about the CS and the noise work, educating the nurses of the SI is regarded necessary as the demand of the EWs is considered. With respect to the unique tasks of IH, such as WE management and instructions for wearing PPE, among several other tasks of IH, training should be provided for improved IH recognition.

Total energy intake according to the level of skeletal muscle mass in Korean adults aged 30 years and older: an analysis of the Korean National Health and Nutrition Examination Surveys (KNHANES) 2008-2011.

BACKGROUND/OBJECTIVES: Since gain or loss of skeletal muscle mass is a gradual event and occurs due to a combination of lifestyle factors, assessment of dietary factors related to skeletal muscle is complicated. The aim of this study was to investigate the changes in total energy intake according to the level of skeletal muscle mass. SUBJECTS/METHODS: A total of 8,165 subjects ≥ 30 years of age from the Korean National Health and Nutrition Examination Surveys (KNHANES) 2008-2011 were included in the analysis, and multivariate-adjusted regression analyses were performed to analyze the association of the quartiles of sarcopenia index (SI) with energy intake of the study population after adjusting for age and metabolic parameters. RESULTS: The increase in SI quartile was in proportion to the gradual decrease in systemic lipids and the anthropometric measurement of fat accumulation (P < 0.001). Subjects in higher SI quartiles tended to consume more total energy and energy-producing nutrients than those in lower quartiles (P < 0.001). After age, body weight, alcohol consumption, and metabolic parameters were adjusted in the analysis, total energy intake gradually increased according to the increase in SI quartile, and the association between total energy intake and SI was more pronounced in men. However, the risk (odd ratio) of having a low SI was not affected by any single macronutrient intake. CONCLUSIONS: In this study, total energy intake was positively associated with SI and relative skeletal mass in both men and women. However, no significant association or a weak association was observed between any single macronutrient intake and skeletal muscle mass. The data indicated that acquiring more energy intake within the normal range of energy consumption may help to maintain skeletal muscle mass.

Characteristics of occupational musculoskeletal disorders of five sectors in service industry between 2004 and 2013.

BACKGROUND: 'Work related musculoskeletal disorders (WRMSDs)' have been mostly reported in the manufacturing industry but recently the occurrence of industrial injuries has been constantly increasing in the service industry. This research is going to analyze the data about workers' compensation for WRMSDs in five different service sectors and identify characteristics of occupations with the highest approved occupations. METHODS: According to the data released from the Korea Worker's Compensation & Welfare Service, the overview of 12,730 cases of workers' compensation for WRMSDs in five service sectors from 2004 to 2013 is going to be analyzed and the source data is going to be classified by the Korean Standard Classification of Occupations to select the top five occupations that have the highest number of approval. RESULTS: After selecting each five occupations from the service sector that have work related musculoskeletal disorders, the result showed that the occupation with the highest number of approval in the health and social care sector were the early childhood educators, cooks in the school canteens in education services sector, garbage collectors in the sanitation and similar services sector, deliverymen in wholesale and retail, consumer goods repair and building cleaners in general management businesses such as those in building maintenance. The major event observed in the top five occupations was the overexertion and reaction as a cause of WRMSDs. The day when the WRMSDs mostly occurred was on Monday and the most likely time was 10 am. The median days away from work and lost working days are 29-90 days and 0-50 days respectively. The difference in each occupation was observed in year of service, age, and gender. CONCLUSIONS: 83.21% of the approved cases of workers' compensation for WRMSDs occurred in the top 25 occupations in all of the five service sectors, which meant that the approval of workers' compensation is concentrated in specific occupations. This research is going to suggest preventive measures for work related musculoskeletal disorders in the service industry and to help prioritize the preventive measures. TRIAL REGISTRATION: Not applicable.

Nutritional Status of Korean Children and Adolescents with Attention Deficit Hyperactivity Disorder (ADHD).

Attention deficit hyperactivity disorder (ADHD) has been associated with an elevated risk for obesity but this seems to be paradoxical to the fact that many youths with ADHD have symptoms of hyperactivity. People diagnosed with ADHD tend to have a high risk of developing undesirable diet habits and consequently have health related problems. However, less attention has been paid to obesity in ADHD while many efforts have been devoted to the prevention of childhood obesity in mentally normal people. Hence the purpose of this study was to explore the nutritional status and life habits of children and adolescents with ADHD (n = 76) based on degree of obesity by utilizing the Korean National Health and Nutrition Examination Survey (KNHANES) data from 2005-2013. As results the levels of blood pressure, total triglycerides and the fat intake relative to total energy intake in overweight ADHD group were higher than those in normal weight group. Interestingly, overweight ADHD subjects consumed significantly less amount of iron compared to normal weight ADHD subjects and the level of serum ferritin was lower in the overweight ADHD group (59.0 ng/mL) than in the normal weight ADHD group (47.9 ng/mL). After adjusting total energy intake, total vegetable consumption was 14.3% lower in overweight group compared to the consumption in normal weight group. These results indicate a plausible relationship of iron status and obesity in ADHD subjects but this relationship may not be specific to ADHD. A future study with case-control design is necessary to investigate the association of obesity, nutrient intake, and cognitive/mental status of ADHD.

Chronic central administration of Ghrelin increases bone mass through a mechanism independent of appetite regulation.

Leptin plays a critical role in the central regulation of bone mass. Ghrelin counteracts leptin. In this study, we investigated the effect of chronic intracerebroventricular administration of ghrelin on bone mass in Sprague-Dawley rats (1.5 µg/day for 21 days). Rats were divided into control, ghrelin ad libitum-fed (ghrelin ad lib-fed), and ghrelin pair-fed groups. Ghrelin intracerebroventricular infusion significantly increased body weight in ghrelin ad lib-fed rats but not in ghrelin pair-fed rats, as compared with control rats. Chronic intracerebroventricular ghrelin infusion significantly increased bone mass in the ghrelin pair-fed group compared with control as indicated by increased bone volume percentage, trabecular thickness, trabecular number and volumetric bone mineral density in tibia trabecular bone. There was no significant difference in trabecular bone mass between the control group and the ghrelin ad-lib fed group. Chronic intracerebroventricular ghrelin infusion significantly increased the mineral apposition rate in the ghrelin pair-fed group as compared with control. In conclusion, chronic central administration of ghrelin increases bone mass through a mechanism that is independent of body weight, suggesting that ghrelin may have a bone anabolic effect through the central nervous system.

Overexpression of 2-cysteine peroxiredoxin enhances tolerance to methyl viologen-mediated oxidative stress and high temperature in potato plants.

Oxidative stress is one of the major causative factors for injury to plants exposed to environmental stresses. Plants have developed diverse defense mechanisms for scavenging oxidative stress-inducing molecules. The antioxidative enzyme 2-cysteine peroxiredoxin (2-Cys Prx) removes peroxides and protects the photosynthetic membrane from oxidative damage. In this study, transgenic potato (Solanum tuberosum L. cv. Atlantic) expressing At2-Cys Prx under control of the oxidative stress-inducible SWPA2 promoter or enhanced CaMV 35S promoter (referred to as SP and EP plants, respectively) was generated using Agrobacterium-mediated transformation. The transgenic plants were tested for tolerance to stress. Following treatment with 3 µM methyl viologen (MV), leaf discs from SP and EP plants showed approximately 33 and 15% less damage than non-transformed (NT) plants. When 300 µM MV was sprayed onto whole plants, the photosynthetic activity of SP plants decreased by 25%, whereas that of NT plants decreased by 60%. In addition, SP plants showed enhanced tolerance to high temperature at 42 °C. After treatment at high temperature, the photosynthetic activity of SP plants decreased by about 7% compared to plants grown at 25 °C, whereas it declined by 31% in NT plants. These results indicate that transgenic potato can efficiently regulate oxidative stress from various environmental stresses via overexpression of At2-Cys Prx under control of the stress-inducible SWPA2 promoter.