Evaluation of the Antiviral Efficacy of Subcutaneous Nafamostat Formulated with Glycyrrhizic Acid against SARS-CoV-2 in a Murine Model.

The ongoing COVID-19 pandemic highlights the urgent need for effective antiviral agents and vaccines. Drug repositioning, which involves modifying existing drugs, offers a promising approach for expediting the development of novel therapeutics. In this study, we developed a new drug, MDB-MDB-601a-NM, by modifying the existing drug nafamostat (NM) with the incorporation of glycyrrhizic acid (GA). We assessed the pharmacokinetic profiles of MDB-601a-NM and nafamostat in Sprague-Dawley rats, revealing rapid clearance of nafamostat and sustained drug concentration of MDB-601a-NM after subcutaneous administration. Single-dose toxicity studies showed potential toxicity and persistent swelling at the injection site with high-dose administration of MDB-601a-NM. Furthermore, we evaluated the efficacy of MDB-601a-NM in protecting against SARS-CoV-2 infection using the K18 hACE-2 transgenic mouse model. Mice treated with 60 mg/kg and 100 mg/kg of MDB-601a-NM exhibited improved protectivity in terms of weight loss and survival rates compared to the nafamostat-treated group. Histopathological analysis revealed dose-dependent improvements in histopathological changes and enhanced inhibitory efficacy in MDB-601a-NM-treated groups. Notably, no viral replication was detected in the brain tissue when mice were treated with 60 mg/kg and 100 mg/kg of MDB-601a-NM. Our developed MDB-601a-NM, a modified Nafamostat with glycyrrhizic acid, shows improved protectivity against SARS-CoV-2 infection. Its sustained drug concentration after subcutaneous administration and dose-dependent improvements makes it a promising therapeutic option.

Influence of Supplementary Vitamin D on Bone Mineral Density When Used in Combination with Selective Estrogen Receptor Modulators.

OBJECTIVES: Vitamin D is regarded as one of the major nutrients that significantly influence bone metabolism. This study aims to look at the effect of supplementary vitamin D on bone mineral density (BMD) in female osteoporosis patients. METHODS: The retrospective hospital record review was performed on 282 patients who were diagnosed with osteoporosis and treated with selective estrogen receptor modulators (SERMs) between January 2015 and December 2016. Of these patients, 151 were treated with SERMs only while 131 were treated using both SERMs and vitamin D supplements. The BMD and any occurrence of osteoporotic fracture episode were investigated after one year. The result of two groups was compared to find the significance of vitamin D. RESULTS: Overall, improvement in BMD score was observed in 76% of the patients. The BMD of the SERMs only group improved by 3% in spine and 1% in the hip while that of the SERMs with vitamin D group improved by 6% and 1% respectively. Statistical significance was noticed in the spine only. One distal radius fracture and one single level vertebral fracture occurred in patients of SERMs group while two distal radius fractures occurred in SERMs with vitamin D group. There was no occurrence of around hip fracture in both groups. CONCLUSION: The result of the current study suggests that additional vitamin D may have some additive effect on improving BMD of the spine. Further study with the larger study population and the extended study period is recommended.

Effects of cisplatin on photosensitizer-mediated photodynamic therapy in breast tumor-bearing nude mice.

OBJECTIVE: This study aimed to evaluate the potential effects of cisplatin on photodynamic therapy (PDT) in breast cancer using a breast tumor-bearing mouse model. METHODS: In this study, breast tumor (experimental mammary tumour-6 cell)-bearing nude mice were used as experimental animals. Photolon® (photosensitizer, 2.5 mg/kg body weight [BW]) was injected intraperitoneally; after 2 hours, the tumors were irradiated (660 nm, 80 J/cm2) using a diode laser tool. Cisplatin (3 mg/kg BW) was injected intraperitoneally 1 hour before the Photolon® injection. RESULTS: Tumor volume increased over time in the control group and was not different from that in the cisplatin group. In the PDT group, the tumor volume increased on day 3, but not on day 7. In the cisplatin+PDT group, tumor volume increased on day 3 but decreased on day 7. There was no significant difference in the levels of thiobarbituric acid reactive substance (TBARS) in tumor tissues between the control and cisplatin groups. The levels of TBARS in the cisplatin+PDT group were higher (47%) than those in the PDT group. Analysis of tumor tissue transcriptomes showed that the expression of genes related to the inflammatory response including CL and XCL genes increased, while that of Fn1 decreased in the cisplatin+PDT group compared with the PDT group. CONCLUSION: These results suggest that cisplatin enhances the therapeutic effect of PDT in a breast tumor-bearing mouse model. However, further clinical studies involving patients with breast cancer is needed.

Uterine arteriovenous malformation with repeated vaginal bleeding after dilatation and curettage.

Uterine arteriovenous vascular malformation (UAVM) is a disease that causes excessive bleeding. The symptoms do not subside without proper treatment and this can lead to life-threatening situations. The correct diagnosis of UAVM can be complicated if the patient's uterus did not completely discharge everything during abortion (in broader terms, retaining remnants of the products of conception). In this case, Doppler ultrasonography and computed tomography angiography with 3-dimensional rendering were used to analyze the cause of bleeding and provide proper treatment of this patient. Then, uterine artery embolization, dilatation, and curettage were performed safely and successfully. The patient no longer had symptoms of vaginal spotting during the planned follow up care. UAVM is uncommon; however, if reproductive-age women show repeated abnormal vaginal bleeding after dilatation and curettage, a diagnosis of UAVM must be considered based on the medical history and examination.

Characterization of gene expression profiles and functional analysis of peptidoglycan recognition protein 2 from rock bream (Oplegnathus fasciatus).

Peptidoglycan recognition protein 2 (PGRP2) is a Zn2+-dependent peptidase that plays important roles in binding to microbial components of the cell membrane, inducing phagocytosis and antimicrobial activity. Rock bream (Oplegnathus fasciatus) PGRP2 (RbPGRP2) was identified in the intestine by next generation sequencing (NGS) analysis. The open reading frame (ORF) the RbPGRP2 cDNA (470 amino acid residues) contains a peptidoglycan recognition protein domain (residues 300 to 446). Alignment analysis revealed that RbPGRP2 shares 37.6-53.5% overall sequence identity with the PGRP2s of other species. Phylogenetic analysis revealed that RbPGRP2 clustered together with PGRP2s from teleosts. In healthy rock bream, RbPGRP2 was found to be ubiquitously expressed in all of the examined tissues, especially in the liver. RbPGRP2 expression was significantly upregulated in all of the examined tissues of rock bream after infection with Edwardsiella piscicida, Streptococcus iniae and red sea bream iridovirus (RSIV) compared with the control. Purified rRbPGRP2 interactions with bacteria and inhibited the growth of bacteria in the presence of Zn2+. These results indicate that RbPGRP2 plays an important role in the innate immune response against bacterial infection.

Piscidin: Antimicrobial peptide of rock bream, Oplegnathus fasciatus.

The piscidin family consists of antimicrobial peptides (AMPs) that are mainly found in fish and are crucial effectors of fish innate immune responses. The piscidin family typically has broad-spectrum antimicrobial activity and can modulate immune responses. In this study, we cloned rock bream piscidin (Rbpisc) and investigated its gene expression and biological activity (including antimicrobial and cytotoxic activities). The coding region of Rbpisc consisted of 213 base pairs (bp) encoding 70 amino acid residues. The tertiary structure predicted for Rbpisc includes an amphipathic helix-loop-helix structure. The Rbpisc gene was highly expressed in the gills of healthy fish. The gene expression of Rbpisc increased in the gills after pathogen infection, while the expression was down-regulated in other tissues. A synthetic peptide based on the AMP 12 domain amino acid sequence of Rbpisc appeared to have broad-spectrum antimicrobial activity against various bacteria. However, the synthetic peptide exhibited weak haemolytic activity against fish erythrocytes. These results suggest that Rbpisc might play an important role in the innate immune responses of rock bream.

In situ field application of electrokinetic remediation for an As-, Cu-, and Pb-contaminated rice paddy site using parallel electrode configuration.

The applicability of an in situ electrokinetic process with a parallel electrode configuration was evaluated to treat an As-, Cu-, and Pb-contaminated paddy rice field in full scale (width, 17 m; length, 12.2 m; depth, 1.6 m). A constant voltage of 100 V was supplied and electrodes were spaced 2 m apart. Most As, Cu, and Pb were bound to Fe oxide and the major clay minerals in the test site were kaolinite and muscovite. The electrokinetic system removed 48.7, 48.9, and 54.5 % of As, Cu, and Pb, respectively, from the soil during 24 weeks. The removal of metals in the first layer (0-0.4 m) was higher than that in the other three layers because it was not influenced by groundwater fluctuation. Fractionation analysis showed that As and Pb bound to amorphous Fe and Al oxides decreased mainly, and energy consumption was 1.2 kWh/m(3). The standard deviation of metal concentration in the soil was much higher compared to the hexagonal electrode configuration because of a smaller electrical active area; however, the electrode configuration removed similar amounts of metals compared to the hexagonal system. From these results, it was concluded that the electrokinetic process could be effective at remediating As-, Cu-, and Pb-contaminated paddy rice field in situ.

In situ electrokinetic remediation of As-, Cu-, and Pb-contaminated paddy soil using hexagonal electrode configuration: a full scale study.

We investigated the in situ applicability of the electrokinetic process with a hexagonal electrode configuration in order to remediate arsenic (As)-, copper (Cu)-, and lead (Pb)-contaminated paddy rice field soil at a field scale (width 17 m, length 12.2 m, and depth 1.6 m). An iron electrode was used in order to prevent the severe acidification of the soil near the anode. We selected ethylenediaminetetraacetic acid (EDTA) as a pursing electrolyte to enhance the extraction of Cu and Pb. The system removed 44.4% of the As, 40.3% of the Cu, and 46.6% of the Pb after 24 weeks of operation. Fractionation analysis showed that the As bound to amorphous ion (Fe) and aluminum (Al) oxyhydroxides was changed into a form of As specifically bound. In the case of Cu and Pb, the fraction bound to Fe-Mn oxyhydroxide primarily decreased. The EDTA formed negatively charged complexes with Cu and Pb, and those complexes were transported toward the anode. The energy consumption was very low compared to that on a small scale because there was less energy consumption due to Joule heating. These results show that the in situ electrokinetic process could be applied in order to remediate paddy rice fields contaminated with multiple metals.

Field application of electrokinetic remediation for multi-metal contaminated paddy soil using two-dimensional electrode configuration.

In this study, we evaluated the feasibility of in situ electrokinetic remediation for arsenic (As)-, copper (Cu)-, and lead (Pb)-contaminated soil, in a pilot-scale field application with two-dimensional electrode configurations. Square and hexagonal configurations with different electrode spacing, 1 m and 2 m, were investigated under a constant 100 V. A square configuration with electrode spacing of 2 m removed 61.5 % of As, 11.4 % of Cu, and 0.9 % of Pb, respectively, and a hexagonal configuration with the same spacing showed a higher removal efficiency in top (59 % of As, 0-0.5 m) and middle (53 % of As, 0.5-1.0 m) layers, but much lower removal efficiency in the bottom layer (1-1.5 m), which was thought to be due to groundwater flow through periodic rise and fall of tides. Fractionation analysis showed that As bound to Fe-Mn oxyhydroxide was the main form of As removed by the electrokinetic process. The two-dimensional configuration wasted less electrical energy by Joule heating, and required fewer electrode installations, compared to the one-dimensional electrode configuration.

The association between reflux esophagitis and psychosocial stress.

BACKGROUND: The prevalence of reflux esophagitis is increasing in Korea. Reflux esophagitis aggravates the stress and fatigue level of daily life, but less is known about the association with stress and fatigue, which could be bi-directional. AIM: To evaluate the impact of reflux esophagitis on stress and fatigue and to compare the stress level of people with reflux esophagitis with that of controls with peptic ulcer disease and healthy controls. METHODS: Among a total of 9,033 subjects who underwent a comprehensive medical check-up including upper endoscopy, 6,834 subjects (75.7 %) were enrolled. Stress and fatigue scores were measured by a validated Korean version of the Brief Encounter Psychosocial Instrument and the Fatigue Severity Scale. RESULTS: Among 6,834 subjects, 13.2 % were in the high-stress group, and reflux esophagitis was found in 6.0 %. After adjustment for confounders, reflux esophagitis was significantly associated with high stress (odds ratio 1.94, 95 % confidence interval 1.25-3.02). Subjects with reflux esophagitis had significantly higher BEPSI-K scores compared with healthy controls (p = 0.027); and however, there was no significant difference in BEPSI-K scores between reflux esophagitis group and peptic ulcer disease controls. Fatigue severity scale was highly correlated with BEPSI-K (p < 0.001); however, there was no significant difference in fatigue severity scale level between the reflux esophagitis group and controls. The severity of reflux esophagitis was significantly correlated with BEPSI-K score (p = 0.008). CONCLUSIONS: Reflux esophagitis is significantly associated with psychosocial stress, and the severity of reflux esophagitis correlates with the degree of stress.

Associations between single nucleotide polymorphisms of MMP2, VEGF, and HIF1A genes and the risk of developing colorectal cancer.

UNLABELLED: The aim of this study was to investigate the association between the risk for colorectal cancer and single nucleotide polymorphisms (SNP) of matrix metalloproteinase-2 (MMP2) -1306C/T, vascular endothelial growth factor (VEGF) 936C/T and hypoxia inducible factor-1α (HIF1A) 1772C/T. PATIENTS AND METHODS: A total 50 colorectal cancer patients (46% women, mean age 68 ± 11 years) were enrolled. Healthy controls without evidence of cancer history or family cancer predispositions were frequency-matched to the cases by sex and age (±5 years). Genotyping was performed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method and the genotype distribution and risk estimate were analyzed. The correlation between the genotypes and clinicopathological parameters (Dukes stage, phenotype, location, differentiation and size) among colorectal cancer patients were investigated. RESULTS: There was a significant association between colorectal cancer and T allele-bearing genotype distribution of HIF1A 1772C/T polymorphism (Odds ratio, OR=3.63, 95% confidence interval, CI = 1.08-12.18, p = 0.03 for CT and TT genotypes relative to CC genotype). In addition, when stratified by age, the association remained in patients older than 60 years old (OR = 13.60, 95% CI = 1.63-113.24, p = 0.01). However, there was no association between the genotypes of the MMP2, VEGF and HIF1A SNP and clinicopathological parameters of colorectal cancer. CONCLUSION: There is a significant association between the HIF1A 1772C/T SNP and the risk of developing colorectal cancer, especially in individuals older than 60 years.

[Analysis of risk factors for low bone mineral density in patients with inflammatory bowel disease].

BACKGROUND/AIMS: Several clinical risk factors for low bone mineral density (BMD) in the patients with inflammatory bowel disease (IBD) have been suggested. However, its prevalence and pathophysiology in Korean population have not been fully studied. The aim of this study was to investigate the prevalence and risk factors for low BMD in Korean IBD patient. METHODS: BMD of the lumbar spine and femur was evaluated using dual-energy X-ray absorptiometry in 30 patients with IBD. Biochemical parameters of bone metabolism, such as serum calcium, phosphorus, osteocalcin, and deoxypyridinoline were measured. The associations between low BMD and clinical parameters such as disease duration, disease activity, drug history, body mass index (BMI), and others were evaluated retrospectively using medical records. RESULTS: Low BMD at the lumbar spine or femur was observed in 63.3% of the patients, and there was no significant difference between the patients with Crohns disease and ulcerative colitis. Clinical and biochemical parameters were irrelevant to BMD. In the patients without glucocorticoid treatment prior to BMD measurement, already 50.0% of patients had low BMD. CONCLUSIONS: Low BMD is a common feature in Korean IBD patients, even those who do not use glucocorticoid. The multiple factors may be involved in the pathogenesis of low BMD. Therefore, BMD should be examined in all IBD patients, irrespective of glucocorticoid treatment.

[Comparison of clinical characteristics and outcomes between geriatric and non-geriatric patients in peptic ulcer bleeding].

BACKGROUND/AIMS: In geriatric patients with peptic ulcer, the use of NSAID and prevalence of chronic illness have been increased, but the Helicobacter pylori (H. pylori) infected portion decreased. The aim of this study was to evaluate the clinical characteristics and outcomes of geriatric patients (aged 65 or older) with peptic ulcer bleeding and compare with non-geriatric patients (less than 65 years old). METHODS: We conducted a retrospective study of 88 patients with peptic ulcer bleeding treated with therapeutic endoscopy from January 2006 to December 2006. We compared the clinical characteristics and outcomes of geriatric patients (n=34, 38.6%) with those of non-geriatric patients (n=54, 61.4%). RESULTS: Hypertension (52.9% vs. 24.1%), cardiovascular disease (35.3% vs. 13.0%), and chronic obstructive pulmonary disease (20.6% vs. 3.7%) were more prevalent in the geriatric group, compared with the non-geriatric group (p<0.05). The geriatric group had taken more ulcerogenic drugs than the non-geriatric group (64.7% vs. 33.3%, p<0.05); aspirin plus clopidogrel (23.6% vs. 13.0%) and aspirin (20.6% vs. 11.0%). Sixteen (21.1%) of the 76 cases had H. pylori-negative ulcer. Between the two groups, there was no difference in the prevalence of H. pylori-negative ulcer (25.9% vs. 18.4%, p>0.05). The amount of transfusion length of ICU stay, rebleeding rate, operation rate and mortality were not different between the two groups. The length of hospital stay in the geriatric patients was significantly longer than the non-geriatric group (12.3+/-10.6 vs. 7.2+/-5.9 days, p<0.05). In multiple regression analysis, old age was a significant risk factor for longer hospital stay (p<0.05). CONCLUSIONS: The geriatric patients with bleeding peptic ulcer had longer hospital stay than the non-geriatric patients in our study. The important emerging etiologies such as ulcerogenic drug and associated chronic illness should be checked and treated in these patients.

[Efficacy of levofloxacin-based triple therapy as second-line Helicobacter pylori eradication].

BACKGROUND/AIMS: Bismuth-based quadruple therapy for second-line eradication treatment achieves the eradication rate ranging from 70% to 81% due to antimicrobial resistance and poor compliance. The aim of this study was to compare the eradication rate of levofloxacin-based triple therapy with that of bismuth-based quadruple therapy in second-line Helicobacter pylori (H. pylori) eradication therapy. METHODS: Seventy-six outpatients with persistent H. pylori infection after first-line triple therapy were enrolled in this prospective randomized trial. The subjects were randomized to receive levofloxacin 300 mg, amoxicillin 1 g, and pantoprazole 20 mg, given twice daily for 7 days (LAP group), or metronidazole 500 mg twice, tetracycline 500 mg four times, and pantoprazole 20 mg twice, bismuth subcitrate 600 mg twice daily for 7 days (MTPB group). Eradication was confirmed with (13)C-urea breath test or rapid urease test 4 weeks after the cessation of therapy. RESULTS: Among Seventy-six patients initially included, eleven were lost during follow-up. The eradication rates, expressed as intention to treat (ITT) and per protocol (PP) analyses, were 51.6% and 53.3% in the LAP group, and 48.9% and 62.9% in the MTPB group, respectively. There was no significant difference in H. pylori eradication rates between the two groups (p=0.815 by ITT, p=0.437 by PP). LAP regimen was better tolerated than MTPB regimen with lower incidence of side effects (10.0% versus 31.4%, p=0.03). CONCLUSIONS: H. pylori eradication rates of levofloxacin-based triple therapy and bismuth-based quadruple therapy were not significantly different in second-line H. pylori eradication therapy, and low incidence of side effects was observed in levofloxacin-based triple therapy.

[Role of anti-secretory treatment in addition to Helicobacter pylori eradication triple therapy in the treatment of peptic ulcer].

BACKGROUND/AIMS: It is not clear whether the anti-secretory therapy should be continued for symptomatic relief and ulcer healing before or after the eradication of H. pylori in patients with peptic ulcer disease. The aim of this study was to evaluate the effectiveness of additional anti-secretory therapy before or after H. pylori eradication in peptic ulcer disease. METHODS: Thirty eight patients with H. pylori-positive active peptic ulcer were included. Patients were randomly allocated into 3 groups; standard 1-week triple therapy followed by omeprazole (20 mg, qd) for 3 weeks (group A), standard 1-week triple therapy only (group B), and omeprazole (20 mg, qd) for 3 weeks followed by 1-week triple therapy (group C). Endoscopy with the rapid urease test and histology for H. pylori was performed 4-8 weeks after the completion of treatment. The symptom was scored by a visual analog scale. RESULTS: Of the 38 patients, 10 were excluded from the per-protocol analysis of this study. The H. pylori eradication rates were 87.5% (group A), 80.0% (group B) and 90.0% (group C) respectively. The peptic ulcer healing rates were 100% in group A, 70.0% in group B, and 90.0% in group C. There was no difference in H. pylori eradication rates and ulcer healing rates among three groups (p>0.05). Symptom score differences between pre-treatment and post-treatment group were not significantly different (p>0.05). CONCLUSIONS: The standard one week triple therapy with or without 3-weeks anti-secretory treatment with omeprazole before or after the therapy does not affect H. pylori eradication rates, peptic ulcer healing rates, and symptom score improvement.

Primary esophageal adenoid cystic carcinoma.

Adenoid cystic carcinoma (ACC) is common in the salivary glands but rare in the esophagus. Routine esophagogastroscopy performed in a 54-year-old woman as part of a medical check-up revealed a submucosal tumor (1.5x1.0 cm) at the mid-esophagus. Endoscopic ultrasonography revealed a lesion with mixed echogenicity in the submucosal layer. The submucosal mass was removed by incisional endoscopic enucleation, and pathological analysis revealed epithelial cells with small hyperchromatic angular nuclei in tubular and cribriform patterns. The lesion was pathologically confirmed as an ACC of the esophagus.